MICROBIAL CONSORTIA PRODUCING DIPICOLINIC ACID AND METHODS FOR SELECTING MICROBES FOR CO-FORMULATION WITH CARRIERS

Abstract

Methods for selecting a microbe for co-formulation with a carrier are provided. In some examples, the methods include identifying a microbe that comprises one or more dipicolinic acid (DPA) synthase genes, a microbe that expresses one or more DPA synthase proteins, and/or a microbe that produces DPA; and selecting the microbe for co-formulation with a carrier. The methods optionally include co-formulating the selected microbe with the carrier. In some examples, the methods include detecting one or more DPA synthase genes or one or more DpaA and/or DpaB proteins in a microbe. In other examples, the methods include detecting DPA in a microbe or medium containing a microbe, for example, utilizing a fluorescence assay. Microbial compositions including one or more microbes that comprise one or more DPA synthase genes, express one or more DPA synthase proteins and/or produce DPA are also provided.

Description

CROSS REFERENCE TO RELATED APPLICATION

[0001] This claims the benefit of U.S. Provisional Application No. 62/681,469, filed Jun. 6, 2018, which is incorporated herein by reference in its entirety.

FIELD

[0002] This disclosure relates to microbes producing dipicolinic acid and methods of identifying microbes with improved viability, methods of co-formulating microbes with carriers, and compositions including the microbes and/or co-formulations.

BACKGROUND

[0003] Microbe-based plant biostimulants offer sustainable agriculture practices that protect the health of the ecosystem. Moreover, supplementation of the plant and soil microbiome with beneficial microorganisms has potential in promoting plant growth and plant fitness, increasing productivity, improving soil fertility, and reducing chemical inputs, resulting in more sustainable agricultural practices. In current agricultural practices, microbial biostimulants can be co-applied and/or co-formulated with numerous wet or dry carriers.

SUMMARY

[0004] Microbial inoculants can be susceptible to the chemistry of the carrier(s) used. Moreover, storage conditions and length of storage before application can also affect microbes. These factors can negatively impact their viability and ultimately limit their efficacy in the field. Disclosed herein are compositions and methods that result in improved microbe survival and/or improved co-formulation of microbes with carriers or seeds. In some embodiments, the methods include selecting one or more microbes with extended viability or survival either alone and/or in co-formulation with one or more carriers or seeds.

[0005] In some embodiments, disclosed herein are microbes that produce dipicolinic acid (DPA) and compositions including such microbes. In one example, the composition includes Bacillus amyloliquefaciens, Bacillus firmus, Bacillus flexus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus koreensis, Bacillus drentensis, Bacillus subtilis, Clostridium bifermentans, Clostridium beijerinckii, Clostridium pasteurianum, Lactobacillus paracasei, Fontibacillus sp. (panacisegetis), Oceanobacillus oncorhynchi, Paenibacillus lautus, Paenibacillus azoreducens, Paenibacillus chibensis, Paenibacillus cookii, Paenibacillus sp. (chitinolyticus), Paenibacillus sp. (P1XP2), Pseudomonas sp., and Streptomyces griseus (in some examples, referred to herein as the "DFC" consortium). In one embodiment, the composition includes cells of microbial species deposited with the American Type Culture Collection (ATCC, Manassas, Va.) on May 16, 2019 and assigned deposit number PTA-125924. In other embodiments the disclosed microbial consortia or compositions include, consist essentially of, or consist of two or more (such as 5 or more, 10 or more, 15 or more, 20 or more, or all) microbes having 16S rDNA sequences with at least 95% identity (such as at least 96%, 97%, 98%, 99% identity, or more) with SEQ ID NOs: 3-25.

[0006] Also disclosed are compositions including the disclosed microbes or consortia (for example, the DFC consortium) and one or more carriers (such as a dry carrier or a liquid carrier) or one or more seeds. In some examples, the carrier includes a liquid or dry fertilizer, a soil-derived substance, an organic substance, an inert material, a dust control chemical, or a mixture of two or more thereof.

[0007] In some embodiments, the methods include selecting a microbe for co-formulation with a carrier or seed, including identifying a microbe that comprises one or more dipicolinic acid (DPA) synthase genes, a microbe that expresses one or more DPA synthase proteins, and/or a microbe that produces detectable amounts of DPA; and selecting the microbe for co-formulation with a carrier. In some embodiments, the methods also include co-formulating the selected microbe with the carrier or seed. In some examples, the selected microbes include one or more of those included in Tables 25 or 26, including, but not limited to all of those listed in Table 26.

[0008] In some examples, the methods include detecting one or more DPA synthase genes (such as a DPA synthase subunit A (DpaA) gene and/or or a DPA synthase subunit B (DpaB) gene) or one or more DpaA and/or DpaB proteins in a microbe. DpaA genes include nucleic acids that encode a DpaA protein with at least 20% (for example, at least 60%) sequence identity to any one of the amino acid sequences in FIG. 1 (e.g., SEQ ID NOs: 26-41). DpaB genes include nucleic acids that encode a DpaB protein with at least 20% (for example, at least 60%) sequence identity to any one of the amino acid sequences in FIG. 2 (e.g., SEQ ID NOs: 42-56). DpaA proteins include DpaA proteins with at least 20% (such as at least 60%) sequence identity to any one of the amino acid sequences in FIG. 1 (e.g., SEQ ID NOs: 26-41). DpaB proteins include DpaB proteins with at least 20% (such as at least 60%) sequence identity to any one of the amino acid sequences in FIG. 2 (e.g., SEQ ID NOs: 42-56). In further examples, the methods include detecting one or more Isf genes or proteins in a microbe. Isf genes include nucleic acids that encode an Isf protein with at least 20% (for example, at least 60%) sequence identity to any one of the amino acid sequences in FIG. 3 (e.g., SEQ ID NOs: 57-67). Isf proteins include Isf proteins with at least 20% (such as at least 60%) sequence identity to any one of the amino acid sequences in FIG. 3 (e.g., SEQ ID NOs: 57-67). In other examples, the methods include detecting DPA in a microbe or medium containing a microbe, for example, utilizing a fluorescence assay.

[0009] In some embodiments, the method includes co-formulating one or more selected microbes with a carrier by contacting the selected microbes (including, but not limited to the microbial consortia disclosed herein) in liquid or dry form with one or more liquid or dry carriers. In some examples, the carrier includes a liquid or dry fertilizer, a soil-derived substance, an organic substance, an inert material, a dust control chemical, or a mixture of two or more thereof. In other examples, the methods include treating seeds with the one or more selected microbes (including, but not limited to the microbial consortia disclosed herein). In some examples, the methods further include co-formulating the one or more selected microbes and one or more microbes that do not comprise one or more DPA synthase genes, do not express one or more DPA synthase proteins, and/or a microbe that does not produce detectable amounts of DPA with the carrier or seed.

[0010] The foregoing and other features of the disclosure will become more apparent from the following detailed description, which proceeds with reference to the accompanying figures.

BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIG. 1 is an alignment of DpaA protein sequences from the indicated bacteria. Fourteen DpaA sequences from 13 strains (SEQ ID NOs: 26-39) were aligned using Clustal Omega (clustal.org/omega) with default settings. A consensus sequence was then generated ("Consensus60"; SEQ ID NO: 40) using a minimum sequence identity threshold of 60%. PRK08306 (SEQ ID NO: 41) is the consensus sequence of the DpaA superfamily retrieved from the NCBI CDD Conserved Domain Family database (ncbi.nih.gov/Structure/cdd/cddsrv.cgi).

[0012] FIG. 2 is an alignment of DpaB protein sequences from the indicated bacteria. Thirteen DpaB sequences from 13 strains (SEQ ID NOs: 42-54) were aligned using Clustal Omega (clustal.org/omega) with default settings. A consensus sequence was then generated ("Consensus60"; SEQ ID NO: 55) using a minimum sequence identity threshold of 60%. PRK08305 (SEQ ID NO: 56) is the consensus sequence of the DpaB superfamily retrieved from the NCBI CDD Conserved Domain Family database (ncbi.nih.gov/Structure/cdd/cddsrv.cgi).

[0013] FIG. 3 shows an alignment of 10 Isf protein sequences from five bacteria (SEQ ID NOs: 57-66) and a consensus sequence (SEQ ID NO: 67).

[0014] FIG. 4 is a graph summarizing survival of bacteria in combination with the indicated carriers.

[0015] FIG. 5 is graph showing 32 day cucumber shoot dry weight in plants treated with perlite or perlite impregnated with a microbial consortium (AMC1).

[0016] FIG. 6 is a graph showing 32 day cucumber shoot dry weight in plants treated with perlite, perlite impregnated with DFC microbial consortium, bentonite, and bentonite impregnated with DFC microbial consortium.

[0017] FIG. 7 is a graph showing 32 day cucumber shoot dry weight in untreated plants, plants treated with liquid DFC consortium, and plants treated with perlite impregnated with DFC microbial consortium.

[0018] FIG. 8 is a graph showing survival of bacteria from DFC consortium in combination with corn and soybean seeds, along with insecticide/fungicide treatments.

SEQUENCE LISTING

[0019] Any nucleic acid and amino acid sequences listed herein or in the accompanying Sequence Listing are shown using standard letter abbreviations for nucleotides and amino acids, as defined in 37 C.F.R. .sctn. 1.822. In at least some cases, only one strand of each nucleic acid sequence is shown, but the complementary strand is understood as included by any reference to the displayed strand.

[0020] SEQ ID NO: 1 is a consensus 16S rDNA nucleic acid sequence from Streptomyces pratensis.

[0021] SEQ ID NO: 2 is a consensus 16S rDNA nucleic acid sequence from Streptomyces venezuelae.

[0022] SEQ ID NO: 3 is a 16S rDNA nucleic acid sequence from Bacillus firmus.

[0023] SEQ ID NO: 4 is a consensus 16S rDNA nucleic acid sequence from Paenibacillus azoreducens.

[0024] SEQ ID NO: 5 is a 16S rDNA nucleic acid sequence from Bacillus amyloliquefaciens.

[0025] SEQ ID NO: 6 is a 16S rDNA nucleic acid sequence from Bacillus flexus.

[0026] SEQ ID NO: 7 is a 16S rDNA nucleic acid sequence from Bacillus licheniformis.

[0027] SEQ ID NO: 8 is a 16S rDNA nucleic acid sequence from Bacillus megaterium.

[0028] SEQ ID NO: 9 is a 16S rDNA nucleic acid sequence from Bacillus pumilus.

[0029] SEQ ID NO: 10 is a 16S rDNA nucleic acid sequence from Bacillus koreensis.

[0030] SEQ ID NO: 11 is a 16S rDNA nucleic acid sequence from Bacillus drentensis.

[0031] SEQ ID NO: 12 is a 16S rDNA nucleic acid sequence from Bacillus subtilis.

[0032] SEQ ID NO: 13 is a 16S rDNA nucleic acid sequence from Clostridium bifermentans.

[0033] SEQ ID NO: 14 is a 16S rDNA nucleic acid sequence from Clostridium beijerinckii.

[0034] SEQ ID NO: 15 is a 16S rDNA nucleic acid sequence from Clostridium pasteurianum.

[0035] SEQ ID NO: 16 is a 16S rDNA nucleic acid sequence from Lactobacillus paracasei.

[0036] SEQ ID NO: 17 is a partial 16S rDNA nucleic acid sequence from Fontibacillus sp. (panacisegetis).

[0037] SEQ ID NO: 18 is a 16S rDNA nucleic acid sequence from Oceanobacillus oncorhynchi.

[0038] SEQ ID NO: 19 is a 16S rDNA nucleic acid sequence from Paenibacillus lautus.

[0039] SEQ ID NO: 20 is a 16S rDNA nucleic acid sequence from Paenibacillus chibensis.

[0040] SEQ ID NO: 21 is a 16S rDNA nucleic acid sequence from Paenibacillus cookii.

[0041] SEQ ID NO: 22 is a 16S rDNA nucleic acid sequence from Paenibacillus sp. (chitinolyticus).

[0042] SEQ ID NO: 23 is a partial 16S rDNA nucleic acid sequence from Paenibacillus sp. (P1XP2).

[0043] SEQ ID NO: 24 is a 16S rDNA nucleic acid sequence from Pseudomonas sp.

[0044] SEQ ID NO: 25 is a 16S rDNA nucleic acid sequence from Streptomyces griseus.

[0045] SEQ ID NOs: 26-39 are DpaA amino acid sequences.

[0046] SEQ ID NO: 40-41 are DpaA consensus amino acid sequences.

[0047] SEQ ID NOs: 42-54 are DpaB amino acid sequences.

[0048] SEQ ID NO: 55-56 are DpaB consensus amino acid sequences.

[0049] SEQ ID NOs: 57-66 are Isf amino acid sequences.

[0050] SEQ ID NO: 67 is an Isf consensus amino acid sequence.

DETAILED DESCRIPTION

[0051] Microbes that do not form spores are often more susceptible to deleterious factors occurring during processing and field application than microbes that form spores. The selection process for identifying microbes that form spores from large microbial inventories can be tedious and time consuming. This usually involves wet-lab testing for survivability (for example, in co-formulations with carriers of choice), with limited guarantee of survivability and final product extended shelf-life.

[0052] Disclosed herein is a novel strategy and method for selecting microbes with high confidence of extended shelf-life as either standalone biostimulant formulations and/or in co-formulation with wet or dry carriers or seeds, thus significantly accelerating lead time to new product testing in field trials. As described herein, spore forming bacteria with identified DPA genes or proteins and/or producing DPA outperform strains with no identified DPA genes and no detectable DPA production in terms of survivability in co-formulation with carriers over time. Finally, a consortium of microbes that includes DPA-producing strains is described.

I. Terms

[0053] Unless otherwise noted, technical terms are used according to conventional usage. Definitions of common terms in molecular biology may be found in Krebs et al., Lewin's Genes XI, published by Jones and Bartlett Learning, 2012 (ISBN 1449659853); Kendrew et al. (eds.), The Encyclopedia of Molecular Biology, published by Blackwell Publishers, 1994 (ISBN 0632021829); Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by Wiley, John & Sons, Inc., 2011 (ISBN 8126531789); and George P. Redei, Encyclopedic Dictionary of Genetics, Genomics, and Proteomics, 2nd Edition, 2003 (ISBN: 0-471-26821-6).

[0054] The following explanations of terms and methods are provided to better describe the present disclosure and to guide those of ordinary skill in the art to practice the present disclosure. The singular forms "a," "an," and "the" refer to one or more than one, unless the context clearly dictates otherwise. For example, the term "comprising a cell" includes single or plural cells and is considered equivalent to the phrase "comprising at least one cell." As used herein, "comprises" means "includes." Thus, "comprising A or B," means "including A, B, or A and B," without excluding additional elements. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety for all purposes. In case of conflict, the present specification, including explanations of terms, will control.

[0055] Although methods and materials similar or equivalent to those described herein can be used to practice or test the disclosed technology, suitable methods and materials are described below. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0056] To facilitate review of the various embodiments of this disclosure, the following explanations of specific terms are provided:

[0057] Carrier: A substance that can be used as a delivery vehicle (for example, in co-formulation or as inoculant) for microbes, such as the microbes or microbial consortia described herein (also referred to herein as "agro-carriers"). The carrier may be liquid or solid (dry). Exemplary carriers include liquid or dry fertilizers, soil-derived substances (for example, charcoal, clays, turf) organic substances (for example, sawdust, wheat/soy/oat bran, composts,) and inert materials (for example, perlite, vermiculite, bentonite, Azomite.RTM., kaolin, silicates, talc). In some examples, seeds may also be referred to as carriers.

[0058] Contacting: Placement in direct physical association, including in either solid and/or liquid form. For example, contacting can occur with one or more microbes (such as the microbes in a microbial consortium) and a carrier or a seed. Contacting can also occur with one or more microbes, microbe/carrier co-formulation, or microbe/seed co-formulation and soil, plants, and/or plant parts (such as foliage, stem, seedling, roots, and/or seeds).

[0059] Culturing: Intentional growth of one or more organisms or cells in the presence of assimilable sources of carbon, nitrogen and mineral salts. In an example, such growth can take place in a solid or semi-solid nutritive medium, or in a liquid medium in which the nutrients are dissolved or suspended. In a further example, the culturing may take place on a surface or by submerged culture. The nutritive medium can be composed of complex nutrients or can be chemically defined.

[0060] Dipicolinic acid (pyridine-2,6-dicarboxylic acid; DPA): A compound with the structure

##STR00001##

[0061] In most microbes, DPA is produced by conversion of dihydrodipicolinate to DPA by the enzyme dipicolinate synthase. DPA synthase has two subunits, subunit A (DpaA or spoVFA) and subunit B (DpaB or spoVFB). Exemplary DpaA and DpaB amino acid sequences are provided herein (FIGS. 1 and 2)

[0062] Some bacteria (e.g., some Clostridium) are able to synthesize DPA, despite lacking identifiable DpaA and DpaB genes. Without being bound by theory, these bacteria are proposed to utilize a structurally related protein, electron transfer flavoprotein (etfA), which is a flavin mononucleotide (FMN) oxidoreductase. EtfA is thought to catalyze the final step in the biosynthesis pathway by converting dihydrodipicolinate to dipicolinic acid (Orsburn et al., Mol. Microbiol. 75:178-186, 2010). Alternatively, some bacteria may utilize and iron-sulfur flavoprotein (Isf) in production of DPA.

[0063] Heterologous: Originating from a different genetic sources or species. For example, a nucleic acid that is heterologous to a cell originates from an organism or species other than the cell in which it is expressed. Methods for introducing a heterologous nucleic acid into bacterial cells include for example transformation with a nucleic acid, including electroporation, lipofection, and particle gun acceleration.

[0064] In another example of use of the term heterologous, a nucleic acid operably linked to a heterologous promoter is from an organism, species, or gene other than that of the promoter. In other examples of the use of the term heterologous, a nucleic acid encoding a polypeptide or portion thereof is operably linked to a heterologous nucleic acid encoding a second polypeptide or portion thereof, for example to form a non-naturally occurring fusion protein.

[0065] Isolated: An "isolated" biological component (such as a nucleic acid, protein or organism) has been substantially separated or purified away from other biological components (such as other cells, cell debris, or other proteins or nucleic acids). Biological components that have been "isolated" include those components purified by standard purification methods. The term also embraces recombinant nucleic acids, proteins, or microbes, as well as chemically synthesized nucleic acids or peptides. The term "isolated" (or "enriched" or "purified") does not require absolute purity, and can include microbes or molecules that are at least 50% isolated, such as at least 75%, 80%, 90%, 95%, 98%, 99% or even 100% isolated.

[0066] Microbe: A microorganism, including but not limited to bacteria, archaebacteria, fungi, and algae (such as microalgae). In some examples, microbes are single-cellular organisms (for example, bacteria, cyanobacteria, some fungi, or some algae). In other examples, the term microbes includes multi-cellular organisms, such as certain fungi or algae (for example, multicellular filamentous fungi or multicellular algae).

[0067] Microbial composition: A composition (which can be solid, liquid, or at least partially both) that includes cells of at least one type (or species) of microbe (or a population of cells of at least one type of microbe). In some examples, a microbial composition comprises cells of one or more types (species) of microbes (or one or more populations of microbes) in a liquid (such as a storage, culture, or fermentation medium or a liquid fertilizer), for example, as a suspension in the liquid. In other examples, a microbial composition includes cells of one or more types (species) of microbes (or one or more populations of microbes) on the surface of or embedded in a solid or gelatinous medium (including but not limited to a culture plate), or a slurry or paste. In other examples, a microbial composition includes cells of one or more types (or species) of microbes (or one or more populations of microbes) in association with a dry material or seed, such as on the surface of or impregnated in a dry material or seed.

[0068] Microbial consortium: A mixture, association, or assemblage of cells of two or more microbial species, which in some instances are in physical contact with one another. The microbes in a consortium may affect one another by direct physical contact or through biochemical interactions, or both. For example, microbes in a consortium may exchange nutrients, metabolites, or gases with one another. Thus, in some examples, at least some of the microbes in a consortium are metabolically interdependent. Such interdependent interactions may change in character and extent through time and with changing culture conditions.

[0069] Transduced and Transformed: A virus or vector "transduces" a cell when it transfers nucleic acid into the cell. A cell is "transformed" by a nucleic acid transduced into the cell when the DNA becomes replicated by the cell, either by incorporation of the nucleic acid into the cellular genome, or by episomal replication. As used herein, the term transformation encompasses all techniques by which a nucleic acid molecule might be introduced into such a cell, including bacterial conjugation, transfection with viral vectors, transformation with plasmid vectors, and introduction of naked DNA by electroporation, lipofection, and particle gun acceleration.

[0070] Vector: A nucleic acid molecule that can be introduced into a host cell, thereby producing a transformed or transduced host cell. Recombinant DNA vectors are vectors including recombinant DNA. A vector can include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication. A vector can also include one or more selectable marker genes, a cloning site for introduction of heterologous nucleic acids, a promoter (for example for expression of an operably linked nucleic acid), and/or other genetic elements known in the art. Vectors include plasmid vectors, including plasmids for expression in gram negative and/or gram positive bacterial cells. Exemplary vectors include those for use in E. coli.

[0071] Viability: Ability of a cell (such as a microbial cell) to grow or reproduce under appropriate conditions for growth or reproduction. In some examples, "survival" or "survivability" refers to the viability of a cell (such as a microbial) cell after a period of storage in a liquid or dry state, alone, in a mixture with other microbial cells, and/or when co-formulated with a carrier or seed.

II. Methods of Identifying Microbes with Viability in Co-Formulations

[0072] Disclosed herein are methods of identifying microbes that remain viable or survive when co-formulated with a liquid or solid carrier. The microbes may be individually co-formulated with a carrier or seed (e.g., a single strain or species of microbes is co-formulated with a carrier or seed) or may be part of a consortium or mixture of microbes (e.g., two or more strains or species of microbes) that is co-formulated with a carrier or seed. In other embodiments, the methods include identifying microbes that remain viable or survive for an extended period of time in a consortium (as a standalone consortium or co-formulated with a carrier or seed).

[0073] In some examples, microbes identified with the methods disclosed herein, for example, microbes that include one or more DPA synthase genes, express one or more DPA proteins, and/or produce detectable amounts of DPA have improved viability (alone or in a co-formulation) than microbes that do not include one or more DPA synthase genes, do not express one or more DPA synthase proteins, and/or do not produce detectable amounts of DPA. In some examples, the microbes identified with the methods disclosed herein have at least 10% increased viability (for example, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 1.5-fold, at least 2-fold, at least 5-fold, or more increased viability) compared to a microbe that does not include one or more DPA synthase genes, does not express one or more DPA synthase proteins, and/or does not produce detectable amounts of DPA. Increased viability may include a greater number of viable cells after a set period of time and/or a viability for a longer period of time.

[0074] In some embodiments, the methods disclosed herein include identifying microbes that include in their genome one or more genes encoding a DPA synthase, express one or more DPA synthase proteins, and/or produce detectable amounts of DPA. Such microbes are identified as microbes that can remain viable or survive individually or when co-formulated with a liquid or solid carrier (for example, compared to one or more microbes that do not include genes encoding a DPA synthase, do not express one or more DPA synthase proteins, and/or do not produce detectable amounts of DPA). The identified microbes may further be selected for downstream use, such as for co-formulation with a liquid or solid carrier or seed. In some examples, the microbes remain viable or survive (either individually or when co-formulated with a carrier or seed) for at least 1 day, at least 3 days, at least 5 days, at least 7 days, at least 10 days, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 8 months, at least 10 months, at least 1 year, at least 2 years, or more (for example, at least 1-28 days, at least 5-21 days, at least 2-6 weeks, at least 4-8 weeks, at least 2-6 months, at least 3-9 months, at least 4-10 months, at least 6 months to 1 year, at least 1-2 years, or more).

[0075] Methods of determining viability or survival of a microbe include detecting growth of the microbe in culture. In some examples, a preparation containing microbial cells (in a liquid or dry state, or in co-formulation with a carrier or seed) is inoculated in a liquid medium, incubated under conditions suitable for microbial growth, and presence and/or amount of microbes after a defined period of time are measured. In other examples a preparation containing microbial cells (in a liquid or dry state, or in co-formulation with a carrier or seed) is streaked on a plate containing solid or semi-solid medium, incubated under conditions suitable for microbial growth, and presence and/or amount of microbes (such as presence, size, and/or number of colonies) are measured. In some examples, the microbes are identified, for example, using PCR methods. Exemplary methods for determining microbial cell viability and identity are provided in Example 1.

[0076] In further embodiments, the methods include selecting one or more microbes that include one or more DPA synthase genes, express one or more DPA synthase proteins, and/or produce detectable amounts of DPA and optionally co-formulating the one or more selected microbes with one or more carriers or seeds. In some examples, the methods include preparing a co-formulation of one or more of the selected microbes with one or more carriers or seed. The methods include contacting the one or more microbes with the one or more carriers or seeds, for example, in a solid (dry) or liquid form. In some example, the carrier(s) or seed(s) are contacted with a mixture of microbes. The mixture includes microbes that express DPA synthase or produce DPA (such as microbes selected or produced using the methods described herein, including, but not limited to DFC) and may also include one or more microbes that do not express DPA synthase or produce DPA. In some examples, the carrier is contacted with a liquid that includes about 10.sup.3-10.sup.9 cells/mL or more (e.g., about 1.times.10.sup.3 cells/mL, about 5.times.10.sup.3 cells/mL, about 1.times.10.sup.4 cells/mL, about 5.times.10.sup.4 cells/mL, about 1.times.10.sup.5 cells/mL, about 5.times.10.sup.5 cells/mL, about 1.times.10.sup.6 cells/mL, about 5.times.10.sup.6 cells/mL, about 1.times.10.sup.7 cells/mL, about 5.times.10.sup.7 cells/mL, about 1.times.10.sup.8 cells/mL, about 5.times.10.sup.8 cells/mL, about 1.times.10.sup.9 cells/mL, about 5.times.10.sup.9 cells/mL, or more) of each microbe.

[0077] In some embodiments, a liquid including one or more of the selected microbes (and optionally one or more additional microbes) is placed in contact with one or more dry carriers or seeds. In some examples, the liquid including the microbes is a fresh or frozen bacterial culture or a mixture of fresh or frozen bacterial cultures. In other examples, the liquid including the microbes is a liquid to which freeze-dried microbes have been added. The liquid including the one or more microbes is allowed to soak into the dry carrier or seed. In some examples, an amount of liquid including the one or more microbes is used so that the dry carrier or seed is saturated, for example to provide relatively even distribution of the microbes throughout the carrier or seed. However, non-saturating amounts of liquid may also be used. In non-limiting examples, the amount is about 35 .mu.L/g to 6 mL/g. In some examples, the microbe-impregnated carrier or seed is dried (such as at room temperature or at about 30-35.degree. C.) and stored at ambient temperature (for example, in a closed or air-tight container).

[0078] In other embodiments, a liquid including one or more of the selected microbes (and optionally one or more additional microbes) is mixed with one or more liquid carriers. In some examples, the liquid including the microbes is a fresh or frozen bacterial culture or a mixture of fresh or frozen bacterial cultures. In other examples, the liquid including the microbes is a liquid to which freeze-dried microbes have been added. The liquid including the microbes can be mixed with the liquid carrier at any selected amount, for example, from 0.1%-90% (v/v), such as 0.5-1%, 1-5%, 2-10%, 3-6%, 4-8%, 5-15%, 8-20%, 10-25%, 20-40%, 30-50%, 40-60%, 50-75%, or 70-90% (v/v). In some examples, the microbes are mixed with the liquid carrier at about 0.1%, about 0.2%, about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% (v/v). In one non-limiting example, the mixture of microbes is added to the liquid carrier at 0.5% (v/v) or a ratio of 1:180. In another non-limiting example, the mixture of microbes is added to a concentrated liquid carrier (such as a 10.times. concentrated liquid carrier) at 90% (v/v) to produce a 1.times. concentration of the liquid carrier. The amount of microbial cells in the mixture can be adjusted to achieve a desired final concentration of microbial cells, depending on the dilution factor that will be used. The mixture of microbes is stored at ambient temperature (for example, in a closed or air-tight container).

[0079] In some embodiments, a dry preparation of microbes (such as freeze-dried microbes) is used in the co-formulation with a dry carrier or seed. In some examples, freeze-dried microbes are mixed with a dry carrier or seed (such as about 40 mg microbes/kg carrier or seed to about 1 g microbes/kg carrier or seed). In some examples, of this embodiment, the freeze-dried microbes are added to a liquid that is then contacted with the dry carrier or seed. In other examples, the freeze-dried microbes are added to a liquid and then contacted with the dry carrier or seed as described above.

[0080] A. Detecting DPA Synthase Nucleic Acids

[0081] In some embodiments, the methods include identifying presence of one or more DPA synthase nucleic acid molecules (such as DNA, cDNA, or mRNA) in a microbe or population of microbes. In some examples, the methods include detecting one or more DPA synthase genes (such as DpaA and/or DpaB) in the genome of a microbe. In some examples, a microbe includes both DpaA and DpaB genes. Exemplary DPA synthase genes include B. subtilis DpaA (GenBank Accession No. NC_000964.3, 1744367-1745260, incorporated herein by reference as present in GenBank on Jun. 3, 2018) and DpaB (GenBank Accession No. NC_000964.3, 1745236-1745865, incorporated herein by reference as present in GenBank on Jun. 3, 2018). In some examples, a DpaA gene encodes a protein shown in FIG. 1 or a protein with at least 20% sequence identity (such as at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more) with a protein shown in FIG. 1 (e.g., SEQ ID NOs: 26-39). In some examples, a DpaB gene encodes a protein shown in FIG. 2 or a protein with at least 20% sequence identity (such as at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more) with a protein shown in FIG. 2 (e.g., SEQ ID NOs: 42-54. In some examples, a DpaA or DpaB protein has one or more conserved regions identified in the "Consensus60" sequences in FIGS. 1 and 2, respectively (SEQ ID NOs: 40 and 55, respectively).

[0082] In other embodiments, the methods include identifying presence of one or more nucleic acids (such as DNA, mRNA, or cDNA) that are involved in an alternative pathway for DPA synthesis. In some examples, the method includes identifying presence or expression of one or more nucleic acids encoding an electron transfer flavoprotein (such as EtfA) or an iron-sulfur flavoprotein (such as Isf). The electron transfer flavoprotein is a heterodimer consisting of an alpha and a beta subunit, and are part of the adenine nucleotide alpha hydrolase superfamily. Exemplary bacterial EtfA nucleic acid sequences include GenBank Accession Nos. CP000312.1 (2508382-2509389), NC_004578.1 (2407768-2408697), NC_009089.1 (977905-978927), NC_019382.1 (1357738-1356809, complement), NC_003030.1 (2833696-2833268, complement), NC_002971.4 (1062557-1061613, complement), and NC_003063.2 (650447-651376), each of which is incorporated herein by reference as present in GenBank on Jun. 3, 2018. Exemplary bacterial EtfA amino acid sequences include ABG86939, NP_792007.1, YP_001087282.1, YP_006967336.1, NP_349315.1, NP_820116.1, and NP_357016.2, each of which is incorporated herein by reference as present in GenBank on Jun. 3, 2018). Exemplary Isf nucleic acid and protein sequences include GenBank Accession Nos. CP016318 (3060700-3061308) and ARE63607, respectively (incorporated herein by reference as present in GenBank on Jun. 3, 2018) and those shown in FIG. 3 (e.g., SEQ ID NOs: 57-66).

[0083] In some examples, DPA synthase nucleic acids (or EtfA or Isf nucleic acids) can be identified by sequence analysis of a microbe (for example, whole genome sequencing and/or sequencing using DPA synthase-specific oligonucleotides). In some examples, the sequence analysis is performed using sequences present in one or more databases, including GenBank (ncbi.nlm.nih.gov/nucleotide/), ENSEMBL (ensembl.org/index.html), IMG (img.jgi.doe.gov), MicrobesOnline (microbesonline.org), SEED (theseed.org), or GOLD (gold.jgi-psf.gov). Exemplary methods for identifying DPA synthase genes are provided in Example 1, below. Similar methods can be used for identifying EtfA or Isf genes.

[0084] In some examples, nucleic acids from a microbe or population of microbes are isolated, amplified, or both, prior to detection. In some examples, amplification and detection of expression occur simultaneously or nearly simultaneously. In some examples, nucleic acid expression can be detected by PCR (for example, PCR, real-time PCR, RT-PCR or quantitative RT-PCR). For example, nucleic acids can be isolated and amplified by employing commercially available kits. In an example, the nucleic acids can be incubated with primers that permit the amplification of DpaA and/or DpaB (or EtfA or Isf) nucleic acids, under conditions sufficient to permit amplification of such products. The resulting amplicons can be detected.

[0085] In another example, nucleic acids from a microbe or population of microbes are incubated with probes that can bind to DpaA and/or DpaB (or EtfA or Isf) nucleic acid molecules (such as cDNA, genomic DNA, or RNA (such as mRNA)) under high stringency conditions. The resulting hybridization can then be detected. In other examples, a microbe or population of microbes is screened by applying isolated nucleic acid molecules obtained from the microbe(s) to an array. In one example, the array includes oligonucleotides complementary to DpaA and/or DpaB (or EtfA or Isf) nucleic acids. In an example, the microbial nucleic acid molecules are incubated with an array including oligonucleotides complementary to DpaA and/or DpaB (or EtfA or Isf) for a time sufficient to allow hybridization between the isolated nucleic acid molecules and oligonucleotide probes, thereby forming isolated nucleic acid molecule:oligonucleotide complexes. The isolated nucleic acid molecule:oligonucleotide complexes are then analyzed to determine if the nucleic acids are present in the sample.

[0086] B. Detecting DPA Synthase Proteins

[0087] As an alternative, or in addition to detecting DPA synthase nucleic acids, proteins can be detected using methods such as immunoassays (such as Western blot, immunohistochemistry, flow cytometry, or ELISA) or mass spectrometry. In some examples, a DpaA protein includes a protein with at least 20% sequence identity (such as at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more) with a protein shown in FIG. 1 (e.g., SEQ ID NOs: 26-39). In some examples, a DpaB protein includes a protein with at least 20% sequence identity (such as at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more) with a protein shown in FIG. 2 (e.g., SEQ ID NOs: 42-54). In some examples, a DpaA or DpaB protein has one or more conserved regions identified in the "Consensus60" sequences in FIGS. 1 and 2, respectively (SEQ ID NOs: 40 and 55, respectively). In other examples, an Isf protein includes a protein with at least 20% sequence identity (such as at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or more) with a protein shown in FIG. 3 (e.g., SEQ ID NOs: 57-66).

[0088] In some examples, proteins are purified before detection. In one example, DpaA and/or DpaB (or Isf) proteins can be detected by incubating a microbial sample with an antibody that specifically binds to DpaA and/or DpaB (or Isf). The antibody ("primary antibody") can include a detectable label. For example, the primary antibody can be directly labeled, or the sample can be subsequently incubated with a secondary antibody that is labeled (for example with a fluorescent label). The label can then be detected, for example by microscopy, ELISA, flow cytometry, or spectrophotometry. In another example, the sample is analyzed by Western blotting for detecting expression of DpaA and/or DpaB proteins. Antibodies for DpaA, DpaB, or Isf can be generated by one of ordinary skill in the art, for example, using the amino acid sequences in FIGS. 1-3.

[0089] Suitable labels for the antibody or secondary antibody include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, magnetic agents and radioactive materials. Non-limiting examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase. Non-limiting examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin. Non-limiting examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin. A non-limiting exemplary luminescent material is luminol; a non-limiting exemplary magnetic agent is gadolinium, and non-limiting exemplary radioactive labels include .sup.125I, .sup.131I, .sup.35S or .sup.3H.

[0090] C. Detecting DPA

[0091] In some embodiments, the methods include identifying a microbe or population of microbes that produces DPA (such as detectable levels of DPA). In some examples, the methods include detecting at least 1 nM DPA (such as at least 2 nM, at least 5 nM, at least 10 nM, at least 25 nM, at least 50 nM, at least 100 nM, at least 200 nM, at least 500 nM DPA, or more). In one example, the methods include detecting DPA using a terbium-DPA fluorescence assay (see, e.g., Rosen, Anal. Chem. 69:1082-1085, 1997; Pellegrino et al., Anal. Chem. 70:1755-1760, 1998; Ammann et al., Int. J. Microbiol. 2011:435281, 2011). Briefly, contacting DPA with terbium(III) forms a complex that has increased fluorescence compared to terbium(III), allowing detection and/or quantitation of DPA in a sample. An exemplary Terbium-DPA assay is described in Example 1.

III. Microbes and Co-Formulations

[0092] Disclosed herein are microbes that include one or more DPA synthase genes in their genome, express one or more DPA synthase proteins, and/or produce DPA. In some examples, the microbes are modified to include one or more DPA synthase genes in their genome, express one or more DPA synthase proteins, and/or produce DPA. Also disclosed are co-formulations of the microbes with one or more carriers or seeds.

[0093] A. Microbes

[0094] Microbes that possess one or more DPA synthase genes, express one or more DPA synthase proteins, and/or produce DPA include, but are not limited to, Bacillus amyloliquefaciens, Bacillus flexus, Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis, Bacillus sp. (closely related to B. kochii, B. pocheonensis, and Bacillus sp. (strain R-27341)), Clostridium beijerinckii, Oceanobacillus oncorhynchi, Paenibacillus chibensis, Paenibacillus cookii, Paenibacillus lautus, Virgibacillus halophilus, Paenibacillus azoreducens, and Bacillus firmus. In some examples, these bacteria include those described in PCT Publication No. WO 2018/045004 (incorporated herein by reference in its entirety). Additional microbes include those listed in Tables 25 and 26. In some examples, these microbes also have sporulation ability; however, sporulation ability and presence of identifiable DPA synthase genes or DPA production are not completely concordant (see, e.g., Table 6).

[0095] In additional embodiments, disclosed are compositions including microbes that possess one or more DPA synthase genes, express one or more DPA synthase proteins, and/or produce DPA, including those referred to herein as Dry Formulation Consortium (DFC). The microbes in DFC include, but are not limited to Bacillus amyloliquefaciens, Bacillus firmus, Bacillus flexus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus koreensis, Bacillus drentensis, Bacillus subtilis, Clostridium bifermentans, Clostridium beijerinckii, Clostridium pasteurianum, Lactobacillus paracasei, Fontibacillus sp. (panacisegetis), Oceanobacillus oncorhynchi, Paenibacillus lautus, Paenibacillus azoreducens, Paenibacillus chibensis, Paenibacillus cookii, Paenibacillus sp. (chitinolyticus), Paenibacillus sp. (P1XP2), Pseudomonas sp., and Streptomyces griseus. In one embodiment, the composition includes cells of microbial species deposited with the American Type Culture Collection (ATCC, Manassas, Va.) on May 16, 2019 and assigned deposit number PTA-125924.

[0096] One of ordinary skill in the art will recognize that identification of microbes, particularly at the species or strain level, is not always possible. In some examples, the microbes in the compositions described herein were analyzed by 16S rDNA sequencing and whole genome sequencing followed by comparison to sequences in public databases. However, due to limitations of information in sequence databases (including little or no information for some species or strains and/or changes in nomenclature over time) it can be challenging to provide definitive species or strain identifications. Thus, in some embodiments, the microbial species included in the disclosed compositions are identified by their sequence identity to the 16S rDNA sequences provided herein (SEQ ID NOs: 3-25). In some examples, the disclosed microbial consortia or compositions include, consist essentially of, or consist of two or more (such as 5 or more, 10 or more, 15 or more, 20 or more, or all) of the microbes having 16S rDNA sequences with at least 95% identity (such as at least 96%, 97%, 98%, 99%, or more) to SEQ ID NOs: 3-25.

[0097] Microbes that possess one or more DPA synthase genes, express one or more DPA synthase proteins, and/or produce detectable amounts of DPA also include microbes that do not naturally have one or more DPA synthase genes, express one or more DPA synthase proteins, and/or produce DPA, but are modified to do so. In some examples, a microbe that does not naturally have one or more DPA synthase genes, express one or more DPA synthase proteins, and/or produce DPA is modified to express one or more heterologous DPA synthase genes, such as DpaA and/or DpaB or one or more Isf genes. Exemplary DpaA and DpaB genes and proteins and Isf genes and proteins are described in Section II and FIGS. 1-3, including SEQ ID NOs: 26-67).

[0098] Bacteria that may be modified to express one or more heterologous DPA synthase genes include, but are not limited to, Azotobacter (such as Azotobacter vinelandii), Clostridium (such as Clostridium pasteurianum), Streptomyces (such as Streptomyces griseus, Streptomyces venezuelae, Streptomyces pratensis), Sporolactobacillus spp. (e.g., Sporolactobacillus dextrus). Sporosarcina spp. (e.g., Sporosarcina halophila), Desulfotomaculum spp. (e.g., Desulfotomaculum guttoideum). Nocardiopsis Spp. (e.g., Nocardiopsis sinuspersici). Promicromonospora spp. (e.g., Promicromonospora enterophila, Promicromonospora Brevibacillus spp. (e.g., Brevibacillus centrosporus), Rummeliibacillus spp. (e.g., Rummeliibacillus pycnus), Lysinibacillus spp., Terribacillus spp. (e.g., Terribacillus shanxiensis), Micromonospora spp. (e.g., Micromonospora fulva, Micromonospora palomenea). Saccharopolyspora spp. (e.g., Saccharopolyspora spinose, Saccharopolyspora indica), and Fontibacillus spp. (e.g., Fontibacillus panacisegetis). In some examples, these bacteria include those described in PCT Publication No. WO 2018/045004 (incorporated herein by reference in its entirety).

[0099] In some examples, the heterologous DpaA and/or DpaB gene is placed under control of a promoter. In some examples, the promoter is a constitutive promoter, while in other examples, the promoter is inducible (for example, an inducible T7 promoter). In additional examples, the promoter is an arabinose-inducible promoter (for example, the pBAD system), a lac promoter (direct IPTG/lactose induction), a trc promoter (direct IPTG/lactose induction), a tetracycline-inducible promoter, or a pho promoter (phosphate deprivation induced). The heterologous DpaA and/or DpaB gene may be included in a vector, for example operatively linked to a promoter. Similar methods can be used for EtfA or Isf genes.

[0100] Multiple genes (such as two or more DPA synthase and/or Isf genes) can be expressed simultaneously in bacteria. To ensure adequate and coordinate production of multiple enzymes from a single pathway, each nucleic acid encoding a heterologous gene is optionally placed under control of a single type of promoter, such as the inducible T7 promoter. One example is the Duet.TM. vectors (Novozymes), which are designed with compatible replicons and drug resistance genes for effective propagation and maintenance of four plasmids in a single cell. This allows for the coexpression of up to eight different proteins. In other examples, the vector is a pET vector, such as a pET21 or pET28 vector. pET and pET-based vectors are commercially available, for example from Novagen (San Diego, Calif.), or Clontech (Mountain View, Calif.).

[0101] In one example, the vector is pET21a or pET28a. In some examples, the pET vector includes a resistance marker (e.g. ampicillin or kanamycin resistance) and a T7 promoter. The multiple cloning site has been manipulated such that more than one gene (such as 2, 3, 4, or more) can be expressed from a single vector. In some examples, the genes are expressed as a multicistronic product (for example, a bi-cistronic, tri-cistronic, etc. product), with a single mRNA and multiple polypeptides produced. In other examples, the genes are expressed as multiple monocistronic products, with an individual mRNA and polypeptide produced for each gene. Appropriate vectors can be selected depending on the gene(s) to be expressed and the host cell being transformed.

[0102] In some examples, a plasmid is introduced extrachromosomally and replicated within the host microbe. In other examples, after introduction of the plasmid, a double homologous recombination event occurs and the one or more genes are inserted into the genome.

[0103] Transformation of a bacterial cell with recombinant DNA can be carried out. Where the host is bacterial, competent cells which are capable of DNA uptake can be prepared from cells harvested after exponential growth phase and subsequently treated by the CaCl.sub.2 method using procedures well known in the art. Alternatively, MgCl.sub.2 or RbCl can be used. Bacteria can also be transformed by electroporation, conjugation, or transduction.

[0104] B. Carriers and Seed Treatments

[0105] Disclosed herein are methods for co-formulating one or more microbes with one or more carriers and compositions including one or more microbes and one or more carriers. The carrier may be liquid or solid (dry). Carriers include liquid or dry fertilizers (such as fertilizers including urea, potash, ammonium phosphate, and/or ammonium nitrate), soil-derived substances (for example, clay, peat, coal, inorganic soil) organic substances (for example, charcoal, sawdust, wheat/soy/oat bran, compost, coco coir), and/or inert materials (for example, perlite, vermiculite, bentonite, Azomite.RTM., kaolin, silicates, pumice, talc). Exemplary carriers include Azomite.RTM., perlite, biochar, dry fertilizers (such as urea, MOP, or MAP), liquid fertilizer (such as UAN), and dust control chemicals (such as those available from ArrMaz, FL, USA). Additional exemplary carriers include montmorillonite, attapulgite, hydrous aluminosilicate (Agsorb Products Group, IL, USA), akadama (Eastern Leaf Inc, CA, USA), Seramis Clay granules (Greens hydroponics, UK), Aquasmart.TM. Pro (Aquasmart, TX, USA), Pyro-Gro (Green Air products, OR, USA), crushed lava, clay pebbles).

[0106] In some embodiments, co-formulations with carriers include the consortium of 22 microbes described in WO 2018/045004 (incorporated herein by reference in its entirety; referred to herein as AMC1) and one or more of the carriers described herein. In other embodiments, co-formulations with carriers include the consortium of 23 microbes disclosed herein (e.g., the microbes listed in Table 26 or ATCC deposit PTA-125924) and one or more carriers. Co-formulations also include one or more (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or more) microbes and one or more of the carriers described herein. In some examples, a co-formulation includes at least one microbe that includes or expresses one or more DPA synthase and/or Isf gene(s) (or produces DPA) and a carrier. In other examples, a co-formulation includes at least one microbe that includes or expresses DPA synthase gene(s) or produces DPA and at least one microbe that does not include or express DPA synthase gene(s) or does not produce DPA with a carrier. Methods of co-formulating a carrier and one or more microbes include contacting the one or more carriers with the one or more microbes. In some examples, the one or more microbes are in liquid form (e.g., are in a liquid medium) or are in a solid or dry form.

[0107] Also disclosed herein are methods for treating seeds with one or more microbes (e.g., co-formulating one or more microbes with one or more seeds) and compositions including one or more microbes and one or more seeds. In such embodiments, the seeds are the "carrier" for the microbes. In some embodiments, seed treatments include the consortium of 22 microbes described in WO 2018/045004 (incorporated herein by reference in its entirety) and one or more seeds. In other embodiments, seed treatments include the consortium of 23 microbes disclosed herein (e.g., the microbes listed in Table 26 or ATCC deposit PTA-125924) and one or more seeds. In other examples, a co-formulation includes at least one microbe that includes or expresses one or more DPA synthase and/or Isf gene(s) (or produces DPA) and a seed. In other examples, a co-formulation includes at least one microbe that includes or expresses DPA synthase gene(s) or produces DPA and at least one microbe that does not include or express DPA synthase gene(s) or does not produce DPA and a seed. Exemplary seeds that can be treated with the one or more microbes include, but are not limited to, corn seeds, sunflower seeds, canola seeds, wheat seeds, cucumber seeds, tomato seeds, rice seeds, and cotton seeds.

[0108] In some examples, microbe-treated seeds are prepared by applying microbes directly to seeds (e.g., contacting seed with one or more microbes). In other examples, microbe-treated seeds are prepared by applying the microbes as an overcoat to seeds that have been previously treated with an insecticide and/or fungicide (e.g., contacting insecticide and/or fungicide treated seed with one or more microbes). In yet further examples, microbe-treated seeds are prepared by mixing the microbes with an insecticide and/or fungicide (such as an insecticide/fungicide slurry) and applying the mixture to the seeds (e.g., contacting seed with a mixture of insecticide and/or fungicide and one or more microbes). Exemplary insecticides and fungicides that can be used in combination with the microbes include, but are not limited to, metalaxyl, trifloxystrobin, ipconazole, clothianidin, thiamethoxam, fludioxonil, mefenoxam, azoxystrobin, thiabendazole, pyraclostrobin, imidacloprid, fluxapyroxad, and/or sedexane. In some examples, the one or more microbes applied to the seed are in liquid form (e.g., are in a liquid medium) or are in a solid or dry form. Methods of preparing treated seeds include, but are not limited to those described in Seed Treatment: Oregon Pesticide Applicator Training Manual (Paulsrud et al., Urbana, Ill., 2001) and Example 13.

IV. Methods of Use

[0109] The disclosed microbial compositions, alone or in co-formulation with one or more liquid or dry carriers, can be used to treat soil, plants, or plant parts (such as roots, stems, foliage, seeds, or seedlings). In other examples, the disclosed microbial compositions can be used in the form of treated seeds.

[0110] In some examples, treatment with the disclosed compositions and/or carriers or seeds treated with the disclosed compositions improve plant growth, improve stress tolerance, and/or increase crop yield. In some embodiments the methods include contacting soil, plants (such as plant foliage, stems, roots, seedlings, or other plant parts), or seeds with a microbial composition or co-formulation disclosed herein. In other embodiments, the methods include planting seeds treated with the disclosed compositions. The methods may also include growing the treated plants, plant parts, or seeds and/or cultivating plants, plant parts, or seeds in the treated soil.

[0111] In some examples, the amount of the composition(s), alone or as a co-formulation of one or more microbes and carriers or seeds to be applied (for example, per acre or hectare) is calculated and the composition is diluted in water (or in some examples, liquid fertilizer) to an amount sufficient to spray or irrigate the area to be treated (if the composition is a liquid). The composition can be applied at the time of seed planting at a rate of 0.5-2 liters per acre (such as 0.5 L/acre, 1 L/acre, 1.5 L/acre, or 2 L/acre). The composition can also be applied to the soil (e.g., near the plant roots) or plant one or more times during growth, in the same or a different amount. In other examples, the composition can be mixed with diluted herbicides, insecticides, pesticides, or plant growth regulating chemicals. If the composition to be applied is a solid (such as a dry formulation), the solid can be applied directly to the soil, plants, or plant parts or can be suspended or dissolved in water (or other liquid) prior to use.

[0112] In some examples, treatment of soil, seeds, plants, or plant parts with a disclosed composition increases plant growth (such as overall plant size, amount of foliage, root number, root diameter, root length, production of tillers, fruit production, pollen production, and/or seed production) by at least about 5% (for example, at least about 10%, at least about 30%, at least about 50%, at least about 75%, at least about 100%, at least about 2-fold, at least about 3-fold, at least about 5-fold, at least about 10-fold, or more). In other examples, the disclosed methods result in increased crop production by about 10-75% (such as about 20-60% or about 30-50%) compared to untreated crops. Other measures of crop performance include quality of fruit, yield, starch or solids content, sugar content or brix, shelf-life of fruit or harvestable product, production of marketable yield or target size, quality of fruit or product, grass tillering and resistance to foot traffic in turf, pollination and fruit set, bloom, flower number, flower lifespan, bloom quality, rooting and root mass, crop resistance to lodging, abiotic stress tolerance to heat, drought, cold and recovery after stress, adaptability to poor soils, level of photosynthesis and greening, and plant health. To determine efficacy of products, controls include the same agronomic practices without addition of microbes, performed in parallel.

[0113] The disclosed methods and compositions and/or co-formulations can be used in connection with any crop (for example, for direct crop treatment or for soil treatment prior to or after planting). Exemplary crops include, but are not limited to alfalfa, almond, banana, barley, broccoli, cabbage, cannabis, canola, carrots, citrus and orchard tree crops, corn, cotton, cucumber, flowers and ornamentals, garlic, grapes, hops, horticultural plants, leek, melon, oil palm, onion, peanuts and legumes, pineapple, poplar, pine and wood-bearing trees, potato, raspberry, rice, sesame, sorghum, soybean, squash, strawberry, sugarcane, sunflower, tomato, turf and forage grasses, watermelon, wheat, and eucalyptus.

EXAMPLES

[0114] The following examples are provided to illustrate certain particular features and/or embodiments. These examples should not be construed to limit the disclosure to the particular features or embodiments described.

Example 1

Materials and Methods

[0115] Isolation and Identification of Microbes: All microbes were derived from Agrinos microbial collection (AMC) and previously described in WO 2018/045004, except for four additional microbes described herein. These additional microbes were isolated as described below.

[0116] The bacteria Streptomyces pratensis and Streptomyces venezuelae were isolated from bulk soil (N38.degree. 38' 49.402'', W121.degree. 40'' 5.775''). Briefly, the soil sample was suspended in a sterile phosphate buffered saline-TWEEN.RTM. 80 solution before serial dilution and plating onto several types semi-solid media. S. pratensis and S. venezuelae were isolated from Azotobacter medium agar with mannitol (HIMEDIA #M372) plates after incubation for up to 3 days at 30.degree. C. The strains were repeatedly streaked onto semi-solid media MP (see below) until isogenic.

[0117] Bacillus firmus was isolated from a sample of HYT.RTM. A (Agrinos AS) which had been previously mixed with fertilizer (UAN32) at a ratio of 1:180. After three weeks incubation at room temperature, aliquots of the mixture were plated on several types of semi-solid media and incubated for up to 3 days at 30.degree. C. A B. firmus colony was collected from a Pikovskaya's medium agar plate (HIMEDIA #M520). The strain was repeatedly streaked onto semi-solid media MP (see below) until isogenic.

[0118] Paenibacillus azoreducens was isolated from a sample of HYT.RTM. A (Agrinos AS). P. azoreducens was isolated as a colony growing on 1-10 mM ammonium sulfate agar media (0.585 g/L NaCl, 0.075 g/L KCl, 0.147 g/L CaCl.sub.2, 0.049 g/L MgSO.sub.4, 1.32-0.132 g/L (NH.sub.4).sub.2SO.sub.4, 0.054 g/L KH.sub.2PO.sub.4 in HEPES buffer pH 7.5). The strain was repeatedly streaked onto semi-solid media MP (see below) until isogenic.

[0119] Taxonomic classification of newly described microbes: For all four newly described strains, whole-genome sequencing of biologically pure isolates was performed as described below. De novo genome assembly was performed with Hierarchical Genome Assembly Process (HGAP, Pacific Biosciences, Menlo Park, Calif. USA).

[0120] Taxonomic identifications were primarily made using 16S ribosomal RNA (rRNA) sequences. 16S rRNA sequences were first identified within the de novo genome assembly using RNAmmer (cbs.dtu.dk/services/RNAmmer/). 16S sequences were then classified using pairwise alignment with NCBI BLASTn, the Ribosomal Database Project (RDP) Nave Bayesian Classifier (Wang et al. Appl. Environ. Microbiol. 73:5261-5267, 2007), and Greengenes de novo phylogenetic tree construction and rank mapping (DeSantis et al. Appl. Environ. Microbiol. 72:5069-502, 2006). Species assignments were then made using a consensus of the three methods.

[0121] Based on the above, microbial identifications were made as follows:

[0122] Streptomyces pratensis. Using 16S sequences, a whole genome taxonomic classification was also performed. Protein coding sequences were identified using Prodigal (Hyatt et al. BMC Bioinformatics, 11:119, 2010). This classification utilized a set of 49 conserved Clusters of Orthologous Groups (COG) families (Tatsuov et al. Science 278:631-637, 1997) to find the matching corresponding set of sequences for a specific genome. The sequences from the selected genome were then inserted into the reference alignments, the closest neighbors were extracted and concatenated, and a tree was rendered from them using FastTree2 (an approximate maximum likelihood method; Price et al. PLoS One 5:e9490, 2010). This rigorous classification method selected Streptomyces pratensis as the most appropriate reference species. The reference genome for Streptomyces pratensis ATCC 33331 was downloaded from NCBI RefSeq and aligned against the obtained whole genome sequence using MUMmer mummer.sourceforge.net/). The alignment revealed broad global agreement and confirmed that the two are very closely related on a genome-wide scale. A consensus 16S sequence is provided as SEQ ID NO: 1.

[0123] Streptomyces venezuelae. The results of all analyses strongly supported the identification of this isolate as S. venezuelae. A consensus 16S sequence is provided as SEQ ID NO: 2.

[0124] Bacillus firmus. The results of all analyses strongly supported the identification of this isolate as B. firmus. A consensus 16S sequence is provided as SEQ ID NO: 3.

[0125] Paenibacillus azoreducens. The results of all analyses strongly supported the identification of this isolate as P. azoreducens. A consensus 16S sequence is provided as SEQ ID NO: 4.

[0126] Identification of Microbial Metabolic Activity Potential: All potential microbial metabolic activities were assessed using laboratory assays as described in WO 2018/045004, incorporated herein by reference in its entirety. In order to determine whether the microbe reduces sulfur-containing compounds to sulfides during the process of metabolism, bioMerieux's API.RTM. identification products were used according to the manufacturer's recommendations (bioMerieux, Inc., Durham, N.C. USA). The results of the key metabolic activity profiling for newly identified microbes are shown in Table 1.

TABLE-US-00001 TABLE 1 Metabolic activities of new microbial isolates Other Plant N Salt Mineral Cellulolytic/ Beneficial Sulfur Iron metab./ Microbe metab. tolerant solubil. Chitinolytic Activity metab. Siderophore Streptomyces N, D .gtoreq.2.5% Chitin + M + IAA Siderophore venezuelae Cellulose biosynthesis and transport Streptomyces N .gtoreq.2.5% Chitin + M + IAA Siderophore pratensis Cellulose biosynthesis and transport Bacillus N <5% Cellulose M + IAA Siderophore firmus transport Paenibacillus N <2.5% P, Ca IAA H2S Siderophore azoreducens prod. transport D: denitrification, N: Nitrogen fixation, P: phosphate, Ca: calcium, IAA: Indole-3-acetic acid production, M: Malic acid assimilation, H2S: production of hydrogen sulfide

[0127] Evaluation of di picolinic acid (DPA) production in bacteria: Evaluation of DPA production in bacterial strains of interest was performed with a terbium-DPA fluorescence assay, essentially as described by Rosen (Anal. Chem. 69:1082-1085, 1997); Pellegrino et al. (Anal. Chem. 70:1755-1760, 1998); and Ammann et al. (Int. J. Microbiol. 2011:435281, 2011). Briefly, each isogenic strain was grown on agar media (see Table 2) either aerobically or anaerobically. Aerobic strains were grown at 30.degree. C. for up to 3 days, while anaerobic strains were grown at 35.degree. C. in BD GasPak EZ container systems (Becton, Dickinson and Company, Franklin Lakes, N.J. USA) with Pack-Anaero anaerobic gas generating sachets (Mitsubishi Gas Chemical Company, Inc., Tokyo, Japan) for up to 3 days. For the DPA assay, approximately 5 .mu.L of bacteria was taken from a colony growing on a plate and resuspended in 10 mL of sodium acetate buffer (0.2 M, pH 5). The suspension was autoclaved for 15 min at 121.degree. C., 15 psig and cooled at room temperature for about 30 min. Equal volumes of the autoclaved suspension and a 30 .mu.M Terbium(III) chloride hexahydrate (Sigma-Aldrich, Saint Louis, Mo. USA) solution were subsequently mixed. Fluorescence was then measured (272 nm excitation, 545 nm emission) using a the Cytation 5 Imaging Reader (BioTek, Winooski, Vt. USA).

[0128] Culturing of bacteria used in this study on semi-solid media: Isogenic bacterial strains stored at -80.degree. C. as master cell banks were grown on agar media (Table 2) until formation of distinct colonies was observed. Briefly, aerobic strains were grown at 30.degree. C. for up to 3 days, while anaerobic strains were grown at 35.degree. C. in BD GasPak EZ container systems (Becton, Dickinson and Company, Franklin Lakes, N.J. USA) with Pack-Anaero anaerobic gas generating sachets (Mitsubishi Gas Chemical Company, Inc., Tokyo, Japan) for up to 3 days.

TABLE-US-00002 TABLE 2 Agar media used to culture microbes Genus Agar-based medium* Bacillus spp. MP, NA (amyloliquefaciens; flexus; licheniformis; subtilis), BHI (subtilis; licheniformis; flexus), RCM (firmus; sp.(pocheonensis)), YPD (megaterium; amyloliquefaciens), RhX (amyloliquefaciens) Lactobacillus spp. RCM, MP, MRS Virgibacillus spp. YPD, MP Paenibacillus spp. BHI, MP, NA (cookii, lautus), RCM (azoreducens), R2A (chibensis) Clostridium spp. RCM (pasteurianum; beijerinckii), AMAS (pasteurianum), NA (beijerinckii), MP (pasteurianum; beijerinckii), MP Oceanobacillus spp. BHI, RhX, NA, MP Acetobacter spp. RCM, YPD, MP Pseudomonas spp. MP, YPD, BHI (putida), RCM (sp. (fluorescens)) Streptomyces spp. MP, BHI, NA (pratensis), YPD (griseus) Azotobacter spp. MP, AMAS, RhX *NA: nutrient agar (BD #213000); YPD: yeast peptone dextrose (BD #242720); BHI: Brain Heart infusion agar (Teknova, CA, USA); RhX: 111 Rhizobium .times. medium (ATCC); AMAS: azotobacter medium agar (HIMEDIA #M372); RCM: reinforce Clostridium medium (BD#218081); MRS: Lactobacilli MRS (BD# 288210); R2A: R2A agar (HIMEDIA #SMEB962), MP: Molasses medium agar (2% w/v molasses, 0.15 g/L MgSO.sub.4, 0.1 g/L CaCl.sub.2, 0.12 g/L FeSO.sub.4, 1 g/L K.sub.2SO.sub.4, 5 g/L Yeast extract, 10 g/L peptone, 5 g/L NaCl, 0.1 g/L NaMoO.sub.4, 0.01 g/L MnCl.sub.2, 0.03 g/L KH.sub.2PO.sub.4, 0.03 g/L Na.sub.2HPO.sub.4 and 15 g/L Agar)

[0129] Culturing of individual bacteria strains used in this study in liquid media: Selected colonies from agar-grown strains were inoculated in appropriate sterilized liquid media (Table 3) and incubated for up to 3 days. Aerobic strains were cultured at 30.degree. C. with shaking (125-175 rpm) while anaerobic strains were cultured under N2 gas in sealed serum bottles or Hungate tubes at 35.degree. C. with no agitation. When needed, microbial consortia were produced by mixing equal volumes of individually grown strains. A typical result illustrating the number of cells per mL per strain (see below) is summarized in Table 4. Microbial content was determined by Droplet Digital PCR (ddPCR) using Supermix For Probes (Bio-Rad Laboratories, Hercules, Calif.), as described in WO 2018/045004.

TABLE-US-00003 TABLE 3 Liquid media used to culture microbes Genus Liquid medium* Bacillus spp. BHI (flexus; licheniformis; sp.(pocheonensis); subtilis), YPDS (megaterium; amyloliquefaciens), NB (firmus) Lactobacillus spp. RCM Virgibacillus spp. BHIS Paenibacillus spp. BHI (chibensis; cookii; lautus), NB (azoreducens) Clostridium spp. RCM Oceanobacillus spp. BHIS Acetobacter spp. YPD Pseudomonas spp. YPDS 0.5% Streptomyces spp. YEME Azotobacter spp. MP *YPD: yeast peptone dextrose (BD #242720); YPDS: yeast peptone dextrose (BD #242720) supplemented with 0.5 g/L NaCl; BHI: Brain Heart infusion (Teknova, CA, USA); RCM: reinforce clostridium medium (BD#218081); ); BHIS: Brain Heart infusion (Teknova, CA, USA) supplemented with 45 g/L NaCl; RCM: reinforce clostridium medium (BD#218081); MP: Molasses medium (2% w/v molasses, 0.15 g/L MgSO.sub.4, 0.1 g/L CaCl.sub.2, 0.12 g/L FeSO.sub.4, 1 g/L K.sub.2SO.sub.4, 5 g/L Yeast extract, 10 g/L peptone, 5 g/L NaCl, 0.1 g/L NaMoO.sub.4, 0.01 g/L MnCl.sub.2, 0.03 g/L KH.sub.2PO.sub.4 and 0.03 g/L Na.sub.2HPO.sub.4); YEME: Yeast extract-malt extract medium (3 g/L yeast extract, 5 g/L bacto-peptone; 3 g/L malt extract; 10 g/L glucose; 340 g/L sucrose 5 mM MgCl.sub.2); NB: nutrient broth (BD #234000)

TABLE-US-00004 TABLE 4 Typical number of bacteria cells per mL of final liquid formulation by mixing individually cultured strains. Number of bacteria cells per mL of final Microorganisms liquid formulation (mix of individuals) Bacillus megaterium 2.53E+07 Lactobacillus paracasei/casei 7.66E+07 Clostridium beijerinckii 1.97E+07 Acetobacter pasteurianus 4.58E+07 Lactobacillus buchneri 1.77E+07 Bacillus subtilis 5.26E+07 Paenibacillus cookii 9.14E+07 Lactobacillus vini 9.06E+07 Bacillus licheniformis 5.00E+08 Paenibacillus lautus 5.16E+07 Oceanobacillus oncorhynchi 2.12E+07 Bacillus amyloliquefaciens 8.30E+07 Bacillus sp. 1.26E+08 Pseudomonas putida 1.02E+08 Pseudomonas sp. 1.96E+08 Streptomyces griseus 3.76E+07 Paenibacillus chibensis 9.34E+07 Bacillus flexus 5.88E+07 Clostridium pasteurianum 3.29E+07 Azotobacter vinelandii 6.02E+07 Virgibacillus halophilus 1.95E+07 Lactobacillus delbrueckii 3.36E+07

[0130] Production of co-cultivated microbial consortia by fermentation: Both aerobic and/or anaerobic bacteria were cultured in medium containing 2% molasses supplemented with essential elements such as phosphates, sodium, potassium and chlorides (in the form of commercially available Phosphate Buffered Saline) as well as amino acids, nitrogen and peptides/proteins in the form of food grade Whey powder (0.1% w/v) and non-GMO soybean extract produced enzymatically (0.25% w/v; Ferti-Nitro Plus Plant N; Ferti-Organic, Brownsville, Tex. USA). Sodium chloride concentrations ranged from 0-4% w/v. Strains from Table 4 described above (referred to herein as AMC1) were inoculated into 2 L DASGIP bioreactors (Eppendorf North America Hauppauge, N.Y.) with a 1.5 liter working volume at a final inoculation of OD.sub.600 for each strain ranging between 6.67E-05 to 6.67E-04. Ammonium hydroxide and phosphoric acid were used as base and acid solutions respectively to maintain pH between pH 5.5 and 6.9. Temperature was controlled between 28.degree. C. and 35.degree. C. Anaerobic fermentations were continuously sparged with N2 gas to maintain an anaerobic environment while sparged air was used in aerobic fermentations as a source of oxygen for the microbes during the length of fermentation (typically up to 3 days). For some experiments, after fermentation batches containing different strains were pooled to generate one complete bacterial mixture of 22 strains. A typical result illustrating the number of cells per mL per strain (see below) is summarized in Table 5. Microbial content in fermentates was determined by Droplet Digital PCR (ddPCR) using Supermix For Probes (Bio-Rad Laboratories, Hercules, Calif.), as described describes in WO 2018/045004.

TABLE-US-00005 TABLE 5 Typical number of bacteria cells per mL of final liquid formulation through co-cultivation Number of bacteria cells per mL of final Microorganisms liquid formulation (co-culture) Bacillus megaterium 1.40E+04 Lactobacillus paracasei/casei 9.60E+05 Clostridium beijerinckii 1.60E+07 Acetobacter pasteurianus 1.20E+07 Lactobacillus buchneri 1.00E+05 Bacillus subtilis 1.30E+06 Paenibacillus cookii 6.60E+03 Lactobacillus vini 8.20E+04 Bacillus licheniformis 9.30E+05 Paenibacillus lautus 2.00E+09 Oceanobacillus oncorhynchi 2.00E+09 Bacillus amyloliquefaciens 6.00E+05 Bacillus sp. 3.20E+05 Pseudomonas putida 2.30E+07 Pseudomonas sp. 1.20E+07 Streptomyces griseus 7.50E+06 Paenibacillus chibensis 5.80E+03 Bacillus flexus 6.00E+06 Clostridium pasteurianum 4.80E+06 Azotobacter vinelandii 1.30E+07 Virgibacillus halophilus 6.10E+06 Lactobacillus delbrueckii 2.00E+06

[0131] Production of freeze-dried microbial consortium: In some experiments, the consortium of microbes produced was freeze-dried prior to experimentation in co-formulation with agro-carriers. Consortia were produced either by pooling an equal volume of individually cultured bacteria or using co-cultured fermentates (described above). Freeze-drying was performed essentially as described in WO 2018/045004. Briefly, freeze-dried microbial formulations were produced by mixing the liquid microbial consortia with mannitol/lyoprotectant solution (OPS Diagnostics Lebanon, N.J., USA) as per manufacturer's recommendation and the microbial suspension was aliquoted into lyophilization vials (OPS Diagnostics, Lebanon, N.J., USA). After 60 minutes at -80.degree. C., the mixtures were placed in the FreeZone 6 freeze dry system (Labconco, Kansas City, Mo.), vacuum was applied, and the water in the samples was allowed to sublimate. Samples were stored at 4.degree. C. until needed. In some experiments, the lyoprotectant solution was prepared by adding the following chemicals to microbial cultures for a final concentration of 0.75 g/L Tryptic Soy Broth (Becton, Dickinson and Company, USA), 10 g/L sucrose (Sigma Aldrich, USA) and 5 g/L skim milk (Carnation, Nestle S.A, CH).

[0132] Co-formulation of microbes with agro-carriers: Liquid microbial consortia (produced from co-culture or individually grown and then pooled) or individual bacteria strains produced as described above, were impregnated onto agro-carriers such as perlite, Azomite.RTM. (Azomite, UT, USA), pumice, Monobasic Ammonium phosphate fertilizer (MAP; Mosaic, MN, USA), Muriate of potash fertilizer (MOP; Mosaic, MN, USA), and Biochar (Cool Terra.RTM.; Cool Planet Energy system, CO, USA). Depending on the carrier's water retention characteristics, different volumes of microbial consortia were used so as to saturate the carrier from as low as 35 .mu.L for per gram up to 6 mL per gram. The microbe/agro-carrier mixture was then dried overnight at 30.degree. C.-35.degree. C. before storing in air tight containers for further shelf life microbial survivability studies and plant assays.

[0133] In the case of co-formulation with liquid agro-chemicals such as urea and ammonium nitrate in water (UAN.sub.32; TGI, CA, USA) or fertilizer dust control agents (DUSTROL; ArrMaz FL, USA), liquid microbial consortia, produced as detailed above, were mixed at various ratios (described in examples below) prior to storage and microbial survivability analysis. All work was performed under sanitary conditions to minimize contamination.

[0134] In some instances, freeze-dried bacterial consortia were used, in order to minimize the effects of the culture broth on the carrier's chemistry. Details are described in examples as appropriate.

[0135] Analysis of Bacteria Genomes for DPA Synthase Production

[0136] Microbial genomic DNA extraction: Bacterial cells of different species were grown and harvested from optimized liquid broth and culture conditions. PowerSoil DNA isolation kit (MO BIO Laboratories, Inc, Carlsbad, Calif. USA) was used for small scale genomic DNA extractions. For large scale genomic DNA extractions, the GenElute Bacterial Genomic DNA kit (Sigma-Aldrich, St. Louis, Mo. USA) or Qiagen Genomic DNA Buffer Set and Genomic-tip 500/G (Qiagen, Hilden, Germany) were used following the methods recommended by the manufacturer. The resulting genomic DNA was subsequently precipitated with equal volume of isopropanol, washed with 70% ethanol, air-dried, and resuspended in TE buffer.

[0137] Whole genome sequencing (WGS): Whole Genome Sequencing of biologically pure isolates was performed using PacBio RSII system (Pacific Biosciences, Menlo Park, Calif. USA) following the manufacturer's recommended method for sequence library preparation and sequencing. An average of 73,000 reads of 24 kb in length on average were generated from the microbial isolates, followed by de novo genome assembly with Hierarchical Genome Assembly Process (HGAP, Pacific Biosciences, Menlo Park, Calif. USA).

[0138] Identification of DPA synthase coding sequence in selected bacteria strains: Initial bioinformatic analyses included select members of the class Bacilli; Bacillus (Bacillus megaterium, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus sp., and Bacillus flexus), as well as Lactobacillus (Lactobacillus delbrueckii). First, using previously acquired whole-genome sequences, a bacterial pan-genome analysis was performed using BPGA-Version-1.3 (iicb.res.in/bpga/index.html) to estimate the genomic diversity and to determine the core (conserved), accessory (dispensable), and unique (strain-specific) gene pool. A search was then performed for any dipicolinate synthase subunit A (DpaA) in the accessory gene set. This was followed by a complete search of the strains in Table 1 for DpaA and DpaB using a custom Bash script (Free Software Foundation, 2007).

[0139] Briefly, protein coding genes were annotated in two stages. Prodigal (PROkaryotic DYnamic programming Gene-finding ALgorithm (Hyatt et al., BMC Bioinformatics 11:119, 2010)) was used to identify the coordinates of candidate genes but does not describe the putative gene product. These candidate genes were then compared to large databases in a hierarchical manner, starting with a smaller trustworthy database, moving to medium-sized but domain-specific databases, and finally to curated models of protein families. By default, an e-value threshold of 10.sup.-6 was used with the following series of databases:

[0140] 1. All bacterial proteins in UniProt that have real protein or transcript evidence and are not a fragment. BLAST+ is used for the search.

[0141] 2. All proteins from finished bacterial genomes in RefSeq for a specified genus. BLAST+ is used for the search.

[0142] 3. A series of hidden Markov model profile databases, including Pfam (Punta et al., 2012) and TIGRFAMs (Haft et al., 2013). This is performed using hmmscan from the HMMER 3.1 package (Eddy, 2011).

[0143] 4. If no matches can be found, label as `hypothetical protein`.

[0144] These data were then tabulated along with the capacity for sporulation and dry-formulation survivability (see Table 6) to establish correlations between DPA synthase and viability. Alignments of DpaA and DpaB genes were then performed using Clustal Omega (www.ebi.ac.uk/Tools/msa/clustalo/) and amino acid composition and frequency statistics calculated using Seaview 4 (Gouy et al. Mol. Biol. Evol. 27:221-224, 2010). DpaA and DpaB amino acid alignments were then used to calculate consensus sequences and probability matrices in R using packages Biostrings and seqinr (Pages et al., 2016; Charif and Lobry, Structural Approaches to Sequence Evolution, pp. 207-232, 2007; R Core Team, 2016).

TABLE-US-00006 TABLE 6 Summary of strains capable of sporulating and producing DPA Sporulation Presence of DPA Bacteria strains ability synthase genes DPA production A. pasteurianus N NF ND A. vinelandii Y NF ND B. amyloliquefaciens Y Y Y B. flexus Y Y Y B. licheniformis Y Y Y B. megaterium Y Y Y B. subtilis Y Y Y Bacillus sp. Y Y Y C. beijerinckii Y NF Y C. pasteurianum Y NF ND L. buchneri N NF ND L. casei/paracasei N NF ND L. delbrueckii N NF ND L. vini N NF ND O. oncorhynchi Y Y ND P. chibensis Y Y Y P. cookii Y Y Y P. lautus Y Y Y P. putida N NF ND Pseudomonas sp. N NF ND S. griseus Y NF ND V. halophilus Y Y ND S. venezuelae Y NF ND S. pratensis Y NF ND P. azoreducens Y Y Y B. firmus Y Y Y Y: yes; N: no; NF: not found; ND: not detected

[0145] Bacteria in Consortia Survivability Assay

[0146] Liquid co-formulation sample preparation: For each shelf-life timepoint, 1 mL of the co-formulation (agrochemical/bacterial consortium) was serially diluted from 10.sup.-1 to 10.sup.-5 in sterile peptone water.

[0147] Dry formulation sample preparation: For each shelf-life timepoint, 0.03-1 g of dry formulation (agro-carrier/bacterial consortium) was suspended in up to 3 mL of culture broth such as peptone water or other appropriate medium and incubated up to 1 hr at room temperature. In some instances, gentle sonication (35 khz) was used to release bacteria from dry matrices using a water bath sonicator (VWR ultrasonic cleaner). The suspension was subsequently serially diluted from 10.sup.-1 to 10.sup.-5 in sterile peptone water or other type of culture broth. In other cases, dry material was added directly to liquid media for growth.

[0148] Survivability assay: The bacterial strains which underwent treatment(s) were given the opportunity to multiply. In order to maximize the growth potential of each strain in the consortium, several different agar media were used such as chA (semi-dry chitin, 5 g/L; K.sub.2HPO.sub.4, 0.7 g/L; KH.sub.2PO.sub.4, 0.3 g/L; MgSO.sub.4.5H.sub.2O, 0.5 g/L; FeSO.sub.4.7H.sub.2O, 0.01 g/L; ZnSO.sub.4, 0.001 g/L; MnCl.sub.2, 0.001 g/L and agar, 15 g/L), RCM, NA, MP, YPD, RhX, AMAS, and/or BHIS. In some instances, each serial dilution produced above was spread plated onto one or more agar media in duplicate. In other cases, dry material produced above was suspended into one or more liquid media in sextuplet. For plate cultures, plates were incubated in static incubators, one set aerobically at 30.degree. C. and the other set anaerobically at 35.degree. C. for 3 days. For liquid cultures, tubes were incubated either shaking aerobically at 30.degree. C. or static anaerobically at 35.degree. C. for 7 days. Bacteria which had survived the treatment(s) grew either by forming colonies or multiplied in liquid cultures, and this growth was then be sampled and identified using droplet digital PCR (ddPCR).

[0149] Genomic DNA was extracted from harvested cells using the DNeasy PowerLyzer PowerSoil kit (Qiagen, Inc., Germantown, Md. USA) per the manufacturer's recommendations. DNA was then quantified using the Quantas Fluorometer and the QuantiFluor dsDNA (Promega Corporation, Madison, Wis. USA) and processed for identification and quantification using strain specific probes in conjunction with ddPCR (Dreo et al., Anal. Bioanal. Chem. 406:6513-6528, 2014; Yin, et al., Journal of Microbiological Methods 65:21-31, 2006). Briefly, ddPCR reactions were prepared by combining DNA sample, primers, and probes (designed using unique sequences from the 16S genes and/or unique coding gene sequences identified from WGS genome assemblies as previously described in WO 2018/045004, incorporated herein by reference) with Bio-Rad's ddPCR Supermix for Probes per the manufacturer's recommendations. Droplets were then generated using either the QX200.TM. droplet generator or the AutoDG.TM. Instrument per the manufacturer's recommendations. Polymerase chain reaction (PCR) was carried out using the Eppendorf Mastercycler.RTM. nexus gradient using the recommended thermal cycling conditions from Bio-Rad's ddPCR Supermix for Probes. Following the PCR protocol, reactions were read using the QX200 droplet reader. Finally, concentrations were analyzed with QuantaSoft.TM. software.

[0150] In the case of single strain survivability identification after co-formulation with liquid or dry agro-carrier, simple plating was performed using agar medium best suited of the given strains, as described in Table 2.

Example 2

Identification of DPA Synthase Coding Sequence in Selected Bacterial Strains

[0151] Dipicolinate synthase subunit A (DpaA): An amino acid alignment of DpaA from the bacterial strains revealed that it is divergent, with only 28.4% of amino acids conserved. Results are summarized in Tables 7-9 and FIG. 1. Oceanobacillus oncorhynchi appeared more divergent than most other strains. By calculating the percent identity with the denominator defined as aligned positions, O. oncorhynchi had a 58% percent identity with the consensus sequence using BLOSUM62. In addition, these amino acid changes appeared to lie in conserved regions for most of the strains analyzed. The amino acid alignment of DpaA also revealed that Virgibacillus halophilus had two copies of the gene, likely due to a duplication of the gene. The first copy was highly divergent (51% identity with the consensus sequence, same method as above), and the second copy appeared to have been truncated by 29 amino acids, in addition to significant amino acid changes. In addition, while all other instances of DpaA were located sequentially with DpaB as part of an operon, in V. halophilus the two genes are separated by 539 other genes.

TABLE-US-00007 TABLE 7 Number of DpaA gene copies Strains with DPA genes Number of DpaA gene copies Bacillus megaterium 1 Bacillus subtilis 1 Paenibacillus cookii 1 Bacillus licheniformis 1 Paenibacillus lautus 1 Oceanobacillus oncorhynchi 1 Bacillus amyloliquefaciens 1 Bacillus sp. 1 Paenibacillus chibensis 1 Bacillus flexus 1 Bacillus firmus 1 Virgibacillus halophilus 2 Paenibacillus azoreducens 1

TABLE-US-00008 TABLE 8 Sequence diversity of DpaA genes Selected sites: 303 100.0% Complete (no gaps, no X): 262 86.5% Variable: 208 68.6% Informative: 176 58.1% Gaps or X: 41 13.5% Identical: 54 17.8% Conserved, not identical: 32 10.6% Total conserved: 86 28.4%

TABLE-US-00009 TABLE 9 Amino acid composition of DpaA genes Amino Acid Composition (all sites) Group 1: 30.50% E: 5.7 D: 6 Q: 3.4 N: 3.1 H: 2.3 R: 4.3 K: 5.6 Group 2: 29.90% I: 9.3 L: 10.2 M: 2.8 V: 7.6 Group 3: 33.50% A: 9 P: 3.7 S: 5.8 G: 8.3 T: 6.7 Group 4: 4.90% F: 3.2 Y: 1.6 Group 5: 1.30% W: 0.1 C: 1.2

[0152] Dipicolinate synthase subunit B (DpaB): An amino acid alignment of DpaB from the bacterial strains revealed that it is more conserved that DpaA, with 45.5% of the amino acids conserved. Results are summarized in Tables 10-12 and FIG. 2. No duplications or major truncations were detected. As mentioned above, all copies of DpaB were located sequentially with DpaA as part of an operon, except for V. halophilus whose DpaB lies 539 genes upstream from DpaA.

TABLE-US-00010 TABLE 10 Number of DpaB gene conies Strains with DPA genes Number of DpaB gene copies Bacillus megaterium 1 Bacillus subtilis 1 Paenibacillus cookii 1 Bacillus licheniformis 1 Paenibacillus lautus 1 Oceanobacillus oncorhynchi 1 Bacillus amyloliquefaciens 1 Bacillus sp. 1 Paenibacillus chibensis 1 Bacillus flexus 1 Bacillus firmus 1 Virgibacillus halophilus 1 Paenibacillus azoreducens 1

TABLE-US-00011 TABLE 11 DpaB sequence diversity Selected sites: 202 100.0% Complete (no gaps, no X): 196 97.0% Variable: 133 65.8% Informative: 104 51.5% Gaps or X: 6 3.0% Identical: 63 31.2% Conserved, not identical: 29 14.4% Total conserved: 92 45.5%

TABLE-US-00012 TABLE 12 Amino acid composition of DpaB genes Amino Acid Composition (all sites) Group 1: 30.30% E: 4.8 D: 4.7 Q: 3.8 N: 6.2 H: 1.4 R: 3.2 K: 6.3 Group 2: 29.00% I: 6.5 L: 9.8 M: 4.3 V: 8.4 Group 3: 34.30% A: 8.3 P: 6.6 S: 5.2 G: 7 T: 7.1 Group 4: 4.70% F: 3 Y: 1.7 Group 5: 1.70% W: 0.6 C: 1.1

[0153] Correlation between production of DPA (in assays described above) and presence of DPA genes: As noted in Table 6, Clostridium beijerinckii was detected as a DPA-producing strain via the Terbium-DPA fluorescence assay, but neither DpaA or DpaB were identifiable in its genome. We found that C. beijerinckii possesses an iron-sulfur flavoprotein (Isf) that is structurally related to EtfA, which has been previously implicated in DPA production (Orsburn et al., Mol. Microbiol. 75:178-186, 2010). Six copies of the iron-sulfur flavoprotein were detected in the genome of C. beijerinckii. These sequences were then aligned with isf sequences from four diverse bacteria: Archaeoglobus fulgidus, Methanocaldococcus jannaschii, Methanosarcina thermophila, and Peptoclostridium difficile (FIG. 3). This Isf protein is absent in Clostridium pasteurianum, and production of DPA was not detected in C. pasteurianum. Additional support for the role of Isf in DPA production is provided in Example 12.

Example 3

Survival of Selected Single Strains on a Carrier

[0154] An evaluation of bacterial survivability in Azomite.RTM. impregnated with individual strains was performed. Four strains were selected based on their ability to produce DPA in lab assays and/or on the results of DPA gene identification. The impregnated material was stored dry for 5 days before evaluating bacteria survivability. Results are as indicated in Table 13. 100% of strains with detectable production of DPA (2/2) remained viable for 5 days. For strains with undetectable DPA production, 50% (1/2 strains) remained viable for 5 days.

TABLE-US-00013 TABLE 13 Survival of individual bacteria in co-formulation with dry Azomite .RTM. vs. DPA production DPA 5-day Microbe production viability Streptomyces venezuelae 0 1 Streptomyces pratensis 0 0 Bacillus firmus 1 1 Paenibacillus azoreducens 1 1 1 = detected and 0 = not detected or below detection limit. The DPA production column represents those strains that tested positive for DPA production via the Terbium-DPA fluorescence assay.

[0155] In a second experiment, the number of strains was expanded. An evaluation of bacteria survivability in Azomite.RTM. following impregnation was performed over a 1-month period. Briefly, Azomite.RTM. was impregnated with individual isogenic strains, dried, and samples were taken on days 3, 7, 14, 21, and 28 for bacteria viability analysis. Results are as indicated in Table 14 and show that 100% (4/4) bacteria with identified DPA genes and producing DPA remained viable for 1 month. For strains with no identified DPA genes and undetectable DPA production, 40% (2/5 strains) remained viable for 1 month.

TABLE-US-00014 TABLE 14 Survival of individual bacteria in co-formulation with dry Azomite .RTM. vs. DPA gene identification and DPA production DPA DPA 3 7 14 21 28 Microbe Genes production days days days days days Bacillus subtilis 1 1 1 1 1 1 1 Paenibacillus putida 0 0 1 0 0 0 0 Pseudomonas sp. 0 0 1 1 1 1 1 Streptomyces griseus 0 0 1 1 1 1 1 Streptomyces 0 0 1 0 0 0 0 venezuelae Streptomyces pratensis 0 0 0 0 0 0 0 Bacillus firmus 1 1 1 1 1 1 1 Paenibacillus 1 1 1 1 1 1 1 azoreducens 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that tested positive for DPA production via the Terbium-DPA fluorescence assay.

[0156] A second agro-carrier (perlite) was then tested. A microbial survivability evaluation was performed over a 1-month period, as previously was described with Azomite.RTM.. The results are as indicated in Table 15 and show that 100% (4/4) of bacteria with identified DPA genes and producing DPA remained viable for 1 month. For strains with no detected DPA genes and undetectable DPA production, 60% (3/5 strains) remained viable for 1 month.

TABLE-US-00015 TABLE 15 Survival of individual bacteria in co-formulation with dry perlite vs. DPA gene identification and DPA production DPA DPA 3 7 14 21 28 Microbe Genes production days days days days days Bacillus subtilis 1 1 1 1 1 1 1 Paenibacillus putida 0 0 1 1 1 1 0 Pseudomonas sp. 0 0 1 1 1 1 0 Streptomyces griseus 0 0 1 1 1 1 0 Streptomyces 0 0 0 0 0 0 0 venezuelae Streptomyces pratensis 0 0 0 0 0 0 0 Bacillus firmus 1 1 1 1 1 1 1 Paenibacillus 1 1 1 1 1 1 1 azoreducens 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that tested positive for DPA production via the Terbium-DPA fluorescence assay.

Example 4

Survival of Microbial Consortia on a Carrier

[0157] In the following experiment, 22 bacteria were grown individually and subsequently mixed to produce a consortium, as described above. Dry Azomite.RTM. in a pelletized form was impregnated with 0.23-0.4 mL of consortium (to preserve the integrity of the pellet), dried and stored for two weeks before analyzing which bacteria survived using the assay for assaying microbial viability from a consortium (Example 1). Two independent trials were conducted, and the results are summarized in Table 16. We observed that 83% and 92% of bacteria strains with identified DPA genes and/or producing DPA remained viable 2 weeks after impregnation on Azomite.RTM.. The strain which did not appear to show consistent viability in the two trials was O. oncorhynchi. As mentioned above, O. oncorhynchi DPA synthase subunit A appears more divergent than most other examined strains, with a 58% percent identity with the consensus sequence using BLOSUM62. For strains with no identified DPA genes and no detectable DPA production, the results varied between 60% and 70% of bacteria remaining viable within the same period. Reproducibility between the two trials was low and may be attributed to different metabolic states of this set of microbes at the time of impregnation.

TABLE-US-00016 TABLE 16 Survival of individual bacteria in co-formulation with dry Azomite .RTM. DPA DPA Trial 1 Trial 2 Bacteria genus/species Genes production 2 weeks 2 weeks Bacillus 1 1 1 1 Bacteria with amyloliquefaciens identified Bacillus flexus 1 1 1 1 DPA genes Bacillus licheniformis 1 1 1 1 and/or Bacillus megaterium 1 1 1 1 producing Bacillus sp. 1 1 1 1 DPA Bacillus subtilis 1 1 1 1 Paenibacillus chibensis 1 1 1 1 Paenibacillus cookii 1 1 1 1 Oceanobacillus 1 0 1 0 oncorhynchi Paenibacillus lautus 1 0 1 1 Virgibacillus halophilus 1 0 0 0 Clostridium beijerinckii 0 1 1 1 Acetobacter pasteurianus 0 0 0 1 Bacteria with Azotobacter vinelandii 0 0 1 1 no identified Clostridium pasteurianum 0 0 1 1 DPA genes Lactobacillus buchneri 0 0 1 1 and no Lactobacillus delbrueckii 0 0 0 0 detectable Lactobacillus 0 0 1 0 DPA paracasei/casei production Lactobacillus vini 0 0 0 0 Pseudomonas putida 0 0 1 0 Pseudomonas sp. 0 0 1 1 Streptomyces griseus 0 0 1 1 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay.

[0158] Similar to the experiment with Azomite.RTM. described above, the 22 bacteria were grown individually and subsequently mixed to produce a consortium. Perlite was impregnated with 2-6 mL of consortium, dried and stored for 2 weeks before analyzing which bacteria survived using the assay for assaying microbial viability from a consortium, (Example 1). Two independent trials were conducted, and the results are summarized in Table 17. We observed that 58% and 100% of bacteria strains with identified DPA genes and/or producing DPA remain viable two weeks after impregnation on perlite. For strains with no identified DPA genes and no detectable DPA production, the results varied between 60% and 80% of bacteria remaining viable within the same period. Compared with the Azomite.RTM. experiment, the bacteria with identified DPA genes and/or producing DPA showed good reproducibility even using a different impregnation substrate. They also outperformed those with no identified DPA genes or with no detectable DPA production in our assay.

TABLE-US-00017 TABLE 17 Survival of individual bacteria from consortium in co-formulation with perlite Bacteria DPA DPA Trial 1 Trial 2 genus/species Genes production 2 weeks 2 weeks Bacillus 1 1 1 1 Bacteria with amyloliquefaciens identified DPA Bacillus flexus 1 1 0 1 genes and/or Bacillus licheniformis 1 1 1 1 producing DPA Bacillus megaterium 1 1 1 1 Bacillus sp. 1 1 1 1 Bacillus subtilis 1 1 1 1 Paenibacillus chibensis 1 1 1 1 Paenibacillus cookii 1 1 1 1 Oceanobacillus 1 0 0 1 oncorhynchi Paenibacillus lautus 1 0 1 1 Virgibacillus halophilus 1 0 0 1 Clostridium beijerinckii 0 1 1 1 Acetobacter 0 0 1 0 Bacteria with no pasteurianus identified DPA Azotobacter vinelandii 0 0 0 0 genes and no Clostridium 0 0 1 1 detectable DPA pasteurianum production Lactobacillus buchneri 0 0 1 1 Lactobacillus 0 0 0 0 delbrueckii Lactobacillus 0 0 1 1 paracasei/casei Lactobacillus vini 0 0 1 0 Pseudomonas putida 0 0 1 1 Pseudomonas sp. 0 0 1 1 Streptomyces griseus 0 0 1 1 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay.

Example 4

Survival of Microbial Consortia Co-Formulated with Liquid Dust Control Agents

[0159] In the experiments below, bacterial strains of interest were grown on semi-solid agar plates using media as described above (see Table 2). Dust control chemical/bacteria co-formulations were generated by suspending a 1 .mu.L loopful scraped from an agar plate in 200 .mu.L of sterile peptone water, and then mixing with dust control chemicals at a rate of 5% v/v. Survivability of each strain in co-formulation was then determined by inoculating fresh media (Table 3) with the bacteria/agrochemical mixture and scoring for signs of growth.

[0160] Survivability of bacteria co-formulated with MDC-200: An evaluation of bacterial viability following co-formulation with the dust control agent MDC-200 (ArrMaz, FL, USA) was performed over a 1-month period. Results are as indicated in Table 18. We observed that 92% (11/12) of bacteria with identified DPA genes and/or producing DPA remained viable for 1 month. For strains with no identified DPA genes and undetectable DPA production, 0% (0/10 strains) remained viable for 1 month.

[0161] Survivability of bacteria co-formulated with DUSTROL.RTM. 3275: An evaluation of bacterial viability following co-formulation with DUSTROL.RTM. 3275 (ArrMaz, FL, USA) was performed over a 1-month period. Results are as indicated in Table 18. We observed that 100% (12/12) of bacteria with identified DPA genes and/or producing DPA remained viable for 1 month. For strains with no identified DPA genes and undetectable DPA production, 0% (0/10 strains) remained viable for 1 month.

[0162] Survivability of bacteria co-formulated with DUSTROL.RTM. 3133: An evaluation of bacterial viability following co-formulation with DUSTROL.RTM. 3133 (ArrMaz, FL, USA) was performed over a 1-month period. Results are as indicated in Table 18. We observed that 100% (12/12) of bacteria with identified DPA genes and/or producing DPA remained viable for 1 month. For strains with no identified DPA genes and undetectable DPA production, 0% (0/10 strains) remained viable for 1 month.

[0163] Survivability of bacteria co-formulated with DUSTROL.RTM. 3139: An evaluation of bacterial viability following co-formulation with DUSTROL.RTM. 3139 (ArrMaz, FL, USA) was performed over a 1-month period. Results are as indicated in Table 18. We observed that 92% (11/12) of bacteria with identified DPA genes and/or producing DPA remained viable for 1 month. For strains with no identified DPA genes and undetectable DPA production, 0% (0/10 strains) remained viable for 1 month.

[0164] Survivability of bacteria co-formulated with DUSTROL.RTM. 3001: An evaluation of bacterial viability following co-formulation with DUSTROL.RTM. 3001 (ArrMaz, FL, USA) was performed over a 1-month period. Results are as indicated in Table 18. We observed that 33% (4/12) of bacteria with identified DPA genes and/or producing DPA remained viable for 1 month. Strains with no identified DPA genes and undetectable DPA production, 0% (0/10 strains) remained viable for 1 month. The reduced survival in this experiment may be due to incompatibility of microbes (whether or not they produce DPA) with one or more components of DUSTROL.RTM. 3001.

[0165] Survivability of bacteria co-formulated with DUSTROL.RTM. 3010: An evaluation of bacterial viability following co-formulation with DUSTROL.RTM. 3010 (ArrMaz, FL, USA) was performed over a 1-month period. Results are as indicated in Table 18. We observed that 75% (9/12) of bacteria with identified DPA genes and/or producing DPA remained viable for 1 month. For strains with no identified DPA genes and undetectable DPA production, 20% (2/10 strains) remained viable for 1 month.

TABLE-US-00018 TABLE 18 One-month survival of individual bacteria from consortium in co-formulation with liquid dust control chemicals Bacteria DPA DPA MDC- DUSTROL .RTM. DUSTROL .RTM. DUSTROL .RTM. DUSTROL .RTM. DUSTROL .RTM. Genus/Species genes production 200 3275 3133 3139 3001 3010 Bacillus 1 1 1 1 1 1 1 1 Bacteria with amyloliquefaciens identified Bacillus flexus 1 1 1 1 1 1 0 1 DPA genes Bacillus 1 1 1 1 1 1 0 1 and/or DPA licheniformis production Bacillus megaterium 1 1 1 1 1 1 0 1 Bacillus sp. 1 1 1 1 1 1 0 1 Bacillus subtilis 1 1 1 1 1 1 1 1 Paenibacillus 1 1 1 1 1 1 1 1 chibensis Paenibacillus cookii 1 1 1 1 1 1 0 0 Oceanobacillus 1 0 1 1 1 1 0 1 oncorhynchi Paenibacillus lautus 1 0 1 1 1 1 1 0 Virgibacillus 1 0 0 1 1 0 0 0 halophilus Clostridium 0 1 1 1 1 1 0 1 beijerinckii Acetobacter 0 0 0 0 0 0 0 1 Bacteria with pasteurianus no identified Azotobacter 0 0 0 0 0 0 0 0 DPA genes vinelandii and no Clostridium 0 0 0 0 0 0 0 0 detectable pasteurianum DPA Lactobacillus 0 0 0 0 0 0 0 0 production buchneri Lactobacillus 0 0 0 0 0 0 0 0 delbrueckii Lactobacillus casei 0 0 0 0 0 0 0 0 Lactobacillus vini 0 0 0 0 0 0 0 0 Pseudomonas putida 0 0 0 0 0 0 0 0 Pseudomonas sp. 0 0 0 0 0 0 0 0 Streptomyces 0 0 0 0 0 0 0 1 griseus 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay. The next six columns (MDC-200, DUSTROL .RTM. 3275, 3133, 3139, 3001, and 3010) represent 1-month viability in each co-formulation. The final column summarizes survivability for the group of strains that either A) possesses known DPA genes, produces DPA, or both and B) neither possesses known DPA genes, nor produces DPA.

Example 5

Survival of Microbial Consortia Co-Formulated with Liquid UAN Fertilizer

[0166] An evaluation of bacterial viability following co-formulation with liquid UAN fertilizer was performed at various timepoints over a 1-month period. Briefly, each isogenic strain was grown in liquid medium (as described above) either aerobically or anaerobically. Cultures were then mixed in equal ratios, and the mixture was then combined with UAN32 at a ratio of 1:180 (microbes:UAN). Microbial survivability was assessed on days 0, 7, 14, and 28 and is summarized in Table 19. For the Day 0 timepoint (.about.1 hour after co-formulation), 83% (10/12) of bacterial strains with identified DPA genes and/or producing DPA remained viable. For bacteria with no detectable DPA production, only 20% (2/10 strains) remained viable for the Day 0 timepoint. By the Day 7 timepoint, 67% (8/12) of bacteria strains with identified DPA genes and/or producing DPA remained viable remained viable. Following Day 7, there was no change in viability for this set of strains. For bacteria with no detectable DPA production, only 10% (1/10 strains) remained viable for the Day 7 timepoint.

TABLE-US-00019 TABLE 19 Bacterial survivability over a one-month period in co-formulation with UAN fertilizer DPA DPA UAN - UAN - UAN - UAN - Genus/Species genes production 0 Day 7 Day 14 Day 30 Day Bacillus 1 1 1 0 0 0 Bacteria with amyloliquefaciens identified Bacillus flexus 1 1 1 1 1 1 DPA genes Bacillus 1 1 1 1 1 1 and/or licheniformis producing DPA Bacillus megaterium 1 1 1 1 1 1 Bacillus sp. 1 1 1 1 1 1 Bacillus subtilis 1 1 1 1 1 1 Paenibacillus 1 1 1 1 1 1 chibensis Paenibacillus cookii 1 1 1 1 1 1 Oceanobacillus 1 0 0 0 0 0 oncorhynchi Paenibacillus lautus 1 0 1 1 1 1 Virgibacillus 1 0 0 0 0 0 halophilus Clostridium 0 1 1 0 0 0 beijerinckii Acetobacter 0 0* 0* 0* 0* 1 Bacteria pasteurianus with no Azotobacter 0 0 0 0 0 0 identified vinelandii DPA genes Clostridium 0 0 1 1 1 1 and no pasteurianum detectable Lactobacillus 0 0 0 0 0 0 DPA production buchneri Lactobacillus 0 0 0 0 0 0 delbrueckii Lactobacillus casei 0 0 0 0 0 0 Lactobacillus vini 0 0 0 0 0 0 Pseudomonas putida 0 0 0 0 0 0 Pseudomonas sp. 0 0 0 0 0 0 Streptomyces 0 0 1 0 0 0 griseus 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay. The next 4 columns represent timepoints where the co-formulation was tested for viability using the assay described above. *On Day 30, Acetobacter pasteurianus (a strain unable to produce DPA) was detected as viable for the first time, indicating false negatives for the 0, 7, and 14-day timepoints.

Example 6

Survival of Microbial Consortia Co-Formulated with Biochar

[0167] In this experiment, the 22 bacteria shown in Table 4 were grown individually and subsequently mixed to produce a consortium, as described above. One gram of Biochar (Cool Terra.RTM.; Cool Planet Energy system, CO, USA) was impregnated with .about.0.5 mL of consortium, dried, and stored for 7 days before analyzing which bacteria survived using the survivability assay previously described. The results are summarized in Table 20. 66.7% (7/12 strains) of the bacteria with identified DPA genes and/or producing DPA remained viable 7 days after impregnation on biochar. For strains with no identified DPA genes and no detectable DPA production, 30% (3/10 strains) remained viable within the same time frame.

TABLE-US-00020 TABLE 20 Survival of bacteria from consortium in biochar after 7 days Bacteria DPA DPA genus/species Genes production 7 days Bacillus 1 1 1 Bacteria with amyloliquefaciens identified DPA Bacillus flexus 1 1 1 genes and/or Bacillus licheniformis 1 1 1 producing DPA Bacillus megaterium 1 1 1 Bacillus sp. 1 1 1 Bacillus subtilis 1 1 1 Paenibacillus chibensis 1 1 0 Paenibacillus cookii 1 1 0 Oceanobacillus 1 0 0 oncorhynchi Paenibacillus lautus 1 0 1 Virgibacillus halophilus 1 0 0 Clostridium beijerinckii 0 1 1 Acetobacter pasteurianus 0 0 0 Bacteria with Azotobacter vinelandii 0 0 0 no identified Clostridium 0 0 1 DPA genes and pasteurianum no detectable Lactobacillus buchneri 0 0 1 DPA production Lactobacillus delbrueckii 0 0 0 Lactobacillus 0 0 1 paracasei/casei Lactobacillus vini 0 0 0 Pseudomonas putida 0 0 0 Pseudomonas sp. 0 0 0 Streptomyces griseus 0 0 0 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay.

Example 7

Survival of Microbial Consortia Co-Formulated with Dry Fertilizer Granules

[0168] In this experiment, the 22 bacteria shown in Table 4 were grown individually and subsequently mixed to produce a consortium and impregnated on MOP (0-0-60) dry fertilizer at a rate of 35 .mu.L/g, as described above. The fertilizer was then stored at room temperature and periodically sampled to assess microbial survivability as previously described. The results are summarized in Table 21. 66.7% (8/12 strains) of the bacteria with identified DPA genes and/or producing DPA remained viable up to 7 days. For strains with no identified DPA genes and no detectable DPA production, 30% (3/10 strains) remained viable within the same time frame.

TABLE-US-00021 TABLE 21 Survival of bacteria from consortium in MOP after 7 days Bacteria DPA DPA Survival genus/species genes production on day 7 Bacillus amyloliquefaciens 1 1 1 Bacteria with Bacillus flexus 1 1 1 identified Bacillus licheniformis 1 1 1 DPA genes and/ Bacillus megaterium 1 1 1 or producing DPA Bacillus sp. 1 1 0 Bacillus subtilis 1 1 1 Paenibacillus chibensis 1 1 0 Paenibacillus cookii 1 1 1 Oceanobacillus 1 0 1 oncorhynchi Paenibacillus lautus 1 0 1 Virgibacillus halophilus 1 0 0 Clostridium beijerinckii 0 1 0 Acetobacter pasteurianus 0 0 1 Bacteria with Azotobacter vinelandii 0 0 0 no identified Clostridium pasteurianum 0 0 1 DPA genes and Lactobacillus buchneri 0 0 0 no detectable Lactobacillus delbrueckii 0 0 0 DPA production Lactobacillus 0 0 0 paracasei/casei Lactobacillus vini 0 0 0 Pseudomonas putida 0 0 0 Pseudomonas sp. 0 0 1 Streptomyces griseus 0 0 1 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay.

[0169] The 22 bacteria shown in Table 4 were also grown individually and subsequently mixed to produce a consortium and impregnated on MAP (11-52-0) at a rate of 35 .mu.L/g, as described above. The fertilizer was then stored at room temperature and periodically sampled to assess microbial survivability as previously described. The results are summarized in Table 22. In this case, 58.3% (7/12 strains) of the bacteria with identified DPA genes and/or producing DPA remained viable up to 7 days. For strains with no identified DPA genes and no detectable DPA production, 40% (4/10 strains) remained viable within the same time frame.

TABLE-US-00022 TABLE 22 Survival of bacteria from consortium in MAP after 7 days Bacteria DPA DPA Survival genus/species genes production on day 7 Bacillus amyloliquefaciens 1 1 1 Bacteria with Bacillus flexus 1 1 0 identified DPA Bacillus licheniformis 1 1 1 genes and/or Bacillus megaterium 1 1 1 producing DPA Bacillus sp. 1 1 0 Bacillus subtilis 1 1 0 Paenibacillus chibensis 1 1 0 Paenibacillus cookii 1 1 1 Oceanobacillus 1 0 1 oncorhynchi Paenibacillus lautus 1 0 1 Virgibacillus halophilus 1 0 0 Clostridium beijerinckii 0 1 1 Acetobacter pasteurianus 0 0 1 Bacteria with no Azotobacter vinelandii 0 0 0 identified DPA Clostridium pasteurianum 0 0 1 genes and no Lactobacillus buchneri 0 0 0 detectable DPA Lactobacillus delbrueckii 0 0 0 production Lactobacillus 0 0 0 paracasei/casei Lactobacillus vini 0 0 0 Pseudomonas putida 0 0 1 Pseudomonas sp. 0 0 1 Streptomyces griseus 0 0 0 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay.

Example 9

Survival of Microbes from Consortium Compared to DPA Expression/Production

[0170] In this experiment, the 22 bacteria shown in Table 4 were grown individually and subsequently mixed to produce a consortium, as described above. The consortium was then stored at room temperature and periodically sampled to assess microbial survivability, as previously described. The results are summarized in Table 23.

[0171] On week 2, 83% (10/12 strains) of the bacteria with identified DPA genes and/or producing DPA remained viable. For strains with no identified DPA genes and no detectable DPA production, 60% (6/10 strains) remained viable within the same time frame. After 7.5 weeks, that 58% (7/12 strains) of the bacteria with identified DPA genes and/or producing DPA remained viable. For strains with no identified DPA genes and no detectable DPA production, 50% (5/10 strains) remained viable within the period.

TABLE-US-00023 TABLE 23 Survival of bacteria from consortium in spent medium Bacteria DPA DPA 2 7.5 genus/species genes production weeks weeks Bacillus 1 1 1 1 Bacteria with amyloliquefaciens identified DPA Bacillus flexus 1 1 1 0 genes and/or Bacillus 1 1 1 1 producing DPA licheniformis Bacillus megaterium 1 1 1 1 Bacillus sp. 1 1 1 1 Bacillus subtilis 1 1 1 1 Paenibacillus 1 1 1 1 chibensis Paenibacillus cookii 1 1 1 0 Oceanobacillus 1 0 0 0 oncorhynchi Paenibacillus lautus 1 0 1 0 Virgibacillus 1 0 0 0 halophilus Clostridium 0 1 1 1 beijerinckii Acetobacter 0 0 1 1 Bacteria with no pasteurianus identified DPA Azotobacter 0 0 0 0 genes and no vinelandii detectable DPA Clostridium 0 0 1 1 production pasteurianum Lactobacillus 0 0 1 1 buchneri Lactobacillus 0 0 1 0 delbrueckii Lactobacillus 0 0 1 1 paracasei/casei Lactobacillus vini 0 0 1 1 Pseudomonas putida 0 0 0 0 Pseudomonas sp. 0 0 0 0 Streptomyces 0 0 0 0 griseus 1 = detected and 0 = not detected or below detection limit. The DPA genes column represents those strains that possess both DpaA and DpaB genes. The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay.

Example 10

Summary of Microbial Survival

[0172] In all instances tested, spore forming bacteria with identified DPA genes and/or producing DPA outperformed strains with no identified DPA genes and no detectable DPA production in terms of survivability in co-formulation with agro-carriers over time, as either liquid or dry formulations (Table 24 and FIG. 4). Therefore in the rational design of microbial consortia with selected plant/soil beneficial traits for co-formulation with agro-carriers (wet and/or dry) or as seed treatments, the selection of spore forming and DPA-producing microbes can assist in providing that desired microbial functionalities (such as but not limited to nitrogen metabolism, sulfur metabolism, salt tolerance, mineral salt solubilization, cellulose degradation, chitin degradation, phytohormone production, iron metabolism, dephosphorylation of organic matter) are retained in the surviving microbes. In some instances, this can ensure sufficient redundancy in microbial functionalities present in the rationally designed consortium in order to comply with target crops, soils, and application practice needs as well as geographical and regulatory challenges.

TABLE-US-00024 TABLE 24 Survival of microbial strains in co-formulation with agro-chemicals/carriers Bacteria Bacteria with no identified with identified DPA genes and no Carrier DPA genes and/or detectable DPA (shelf-life tested) producing DPA production MDC-200 (1 mo) 92% 0% DUSTROL .RTM. 3275 (1 mo) 100% 0% DUSTROL .RTM. 3133 (1 mo) 100% 0% DUSTROL .RTM. 3139 (1 mo) 92% 0% DUSTROL .RTM. 3001 (1 mo) 33% 0% DUSTROL .RTM. 3010 (1 mo) 75% 20% UAN (1 mo) 67% 0% Perlite (2 weeks) 75-100% 60-80% .sup. Azomite .RTM. (2 weeks) 83.3%.sup. 60% Biochar (1 week) 66.7%.sup. 30% MOP (1 week) 66.7%.sup. 30% MAP (1 week) 58.3%.sup. 40% Aged liquid/Spent medium 58% 50% (7.5 weeks)

Example 11

Evaluation of Plant Growth Promoting Activity

[0173] Cucumber seeds purchased from The Seed Kingdom (Lubbock, Tex.) were pre-germinated for 4 days at 22-24.degree. C. in rolled germination paper (Anchor Paper, Saint Paul, Minn.) impregnated with a dilute mixture of liquid fertilizer (25 ppm NPK in water). Potting medium (Sunshine Mix) was pre-treated with a Hoagland solution (Hoagland, Calif. Agric. Exp. Stn. Bull. 347:36-39, 1938), modified to contain P, 30.97 ppm; K, 39.1 ppm; Ca, 40.0 ppm; Mg, 14.59 ppm; S, 20.143 ppm; Fe, 1.010 ppm; Cu, 0.019 ppm; Co, 0.012 ppm; B, 2.44 ppm; Mn, 0.494 ppm; Mo, 0.001 ppm and Zn, 0.056 ppm. A rate of 1 L per pound of potting medium was used. To each pot, 1 mL of a 10% w/v urea solution was added before pre-germinated cucumber seedlings with similar length were transplanted. For each treatment (including control) 17-18 plants were randomized in flats in defined growth conditions, controlling for temperature (16-24.degree. C.) and 12 hours photoperiod. The control pots contained 2 g of untreated perlite. The experimental pots were treated with perlite impregnated with 2 mL of the 22 bacteria shown in Table 4. The impregnated perlite had been stored dry for two weeks prior to use in this assay. The flats were watered 3 times a week with modified Hoagland solution. After 28 to 32 days, shoots were dried, and weights were recorded for each plant. The data were analyzed by One-way ANOVA (Analysis Of Variance) and with post-hoc Tukey test to compare samples within the experiment. Shoot dry weight was increased in plants treated with the microbe-impregnated perlite, though it did not reach statistical significance (FIG. 5). As described in Example 4 (Table 17) mainly microbes with identified DPA genes and/or with detectable DPA production survived on impregnated perlite, a subset of the starting consortium.

Example 12

DPA Producing Consortium

[0174] To identify suitable strains for use in designing an additional DPA producing consortium, an in-house microbial collection was screened. Strains derived from genera that are known to sporulate were revived from bacterial glycerol stocks on appropriate media and allowed to grow for up to three days. Single colonies were then selected and passaged at least three times to verify purity. Strains were tested for DPA production using the terbium-DPA fluorescence assay as described in Example 1. Strains shown to produce DPA were selected for whole-genome sequencing. Whole-genome sequencing of biologically pure isolates was performed using Illumina NovaSeq 6000 sequencing system (Illumina, Inc., San Diego, Calif. USA) following the manufacturer's recommended method for sequence library preparation and sequencing. An average of 5,133,928 reads of 87 bp in length on average were generated from the microbial isolates. De novo genome assembly was performed using SPAdes 3.12.0 (Nurk et al., J. Comput. Biol. 20:714-737, 2013). Assembled contigs were then annotated with PROKKA (Seemann, Bioinformatics 30(14):2068-2069, 2014). Annotated genomes were then assessed for the presence of Dipicolinate Synthase subunit A (DpaA) and Dipicolinate Synthase subunit B (DpaB) and Iron-sulfur Flavoprotein (Isf, Table 25). In most cases, strains that produced DPA possessed both DpaA and DpaB genes and lacked Isf genes. All strains that produced DPA and lacked DpaA and DpaB possessed one or more copies of Isf genes. In three cases strains possessed all three (DpaA, DpaB, and Isf) genes. Therefore, detection of DPA production via the terbium-DPA fluorescence assay was perfectly correlated with the presence of DpaA, DpaB, and/or Isf genes.

TABLE-US-00025 TABLE 25 Whole-genome taxonomic classification, DPA-production, and gene copy number for DpaA, DpaB, and Isf Whole DPA- Gene Copy Number Genome ID production DpaA DpaB Isf Bacillus megaterium 1 1 1 0 Bacillus koreensis 1 1 1 0 Bacillus clausii 1 1 1 0 Clostridium beijerinckii 1 0 0 6 Bacillus subtilis 1 1 1 0 Clostridium sp. (carboxidivorans) 1 0 0 1 Paenibacillus cookii 1 1 1 0 Clostridium sp. (scatologenes) 1 0 0 1 Bacillus licheniformis 1 1 1 0 Paenibacillus lautus 1 1 1 0 Brevibacillus sp. (choshinensis) 1 1 1 0 Bacillus amyloliquefaciens 1 1 1 0 Bacillus sp. (pocheonensis) 1 1 1 0 Lysinibacillus sp. (fusiformis) 1 1 1 0 Lysinibacillus sp. (chungkukjangi) 1 1 1 0 Bacillus pumilus 1 1 1 0 Rummeliibacillus sp. (pycnus) 1 1 1 0 Bacillus cereus 1 1 1 1 Paenibacillus chibensis 1 1 1 0 Bacillus flexus 1 1 1 0 Brevibacillus parabrevis 1 1 1 0 Bacillus aryabhattai 1 1 1 0 Paenibacillus sp. (amylolyticus) 1 1 1 0 Clostridium beijerinckii 1 0 0 11 Bacillus subtilis 1 1 1 0 Paenibacillus sp. (chitinolyticus) 1 1 1 0 Bacillus sp. (wudalianchiensis) 1 1 1 0 Paenibacillus larvae 1 1 1 0 Bacillus licheniformis 1 3 3 0 Paenibacillus sp. (agaridevorans) 1 1 1 0 Clostridium sp. (tyrobutyricum) 1 0 0 1 Paenibacillus sp. (alvei) 1 1 1 0 Bacillus sp. (selenatarsenatis) 1 1 1 0 Paenibacillus sp. (P1XP2) 1 1 1 0 Fontibacillus sp. (panacisegetis) 1 1 1 0 Bacillus firmus 1 1 1 0 Bacillus subtilis 1 1 1 0 Lysinibacillus sp. (chungkukjangi) 1 1 1 0 Paenibacillus sp. (ehimensis) 1 1 1 0 Paenibacillus lactis 1 1 1 0 Bacillus farraginis 1 1 1 0 Paenibacillus sp. (P1XP2) 1 1 1 0 Brevibacillus massiliensis 1 1 1 0 Paenibacillus azoreducens 1 1 1 0 Bacillus litoralis 1 1 1 0 Bacillus endophyticus 1 1 1 0 Paenibacillus xylanexedens 1 1 1 0 Bacillus frigoritolerans 1 1 1 0 Bacillus velezensis 1 1 1 0 Clostridium aerotolerans 1 1 1 1 Sphingomonas koreensis 1 0 0 2 Clostridium beijerinckii 1 0 0 10 Clostridium aerotolerans 1 1 1 2 Brevibacillus sp. (brevis) 1 2 2 0 Bacillus sp. (cereus) 1 1 1 0 Bacillus badius 1 1 1 0 Bacillus aryabhattai 1 1 1 0 Bacillus anthracis 1 1 1 0 Bacillus flexus 1 1 1 0 Bacillus drentensis 1 1 1 0 Bacillus sp. (cuccumis) 1 1 1 0 Paenibacillus peoriae 1 1 1 0 Bacillus sp. (cereus) 1 1 1 0 Bacillus aryabhattai 1 1 1 0 Bacillus sp. (solani) 1 1 1 0 Terribacillus sp. (aidingensis) 1 1 1 0 Bacillus sp. (circulans) 1 1 1 0 Brevibacterium frigoritolerans 1 1 1 0 The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay, 1 = detected and 0 = not detected or below detection limit.

[0175] For strains that tested positive for DPA production, potential microbial metabolic activities (such as but not limited to nitrogen metabolism, sulfur metabolism, salt tolerance, mineral salt solubilization, cellulose degradation, chitin degradation, phytohormone production, iron metabolism, dephosphorylation of organic matter) were assessed using laboratory assays as described in WO 2018/045004 and Example 1. Strains possessing the greatest number of metabolic activities were selected for co-fermentation in microbial consortia along with 5 strains that lack DPA production which were selected for their metabolic activities (see Example 1). Those strains that proved to be amenable to co-fermentation in consortium (see Table 26), also referred to herein as Dry Formulation Consortium (DFC), were selected for co-formulation with a carrier such as bentonite, perlite and/or urea.

[0176] All consortia included in this example were fermented as follows. Both aerobic and anaerobic bacteria strains were cultured on medium containing 2-10% sugar source. Amino acids, nitrogen and peptides were provided in the form of one or more of the following: food grade whey powder (0.1-0.5% w/v), yeast extract (0.1-0.5% w/v), non-GMO soybean extract produced enzymatically (0.1-0.5% w/v; Ferti-Nitro Plus Plant N; Ferti-Organic, Brownsville, Tex. USA), spirulina (0.1-0.5% w/v), and/or peptone (0.1-0.5% w/v). When needed, additional vitamins and micronutrients were provided by kelp extract (0.1-0.5% w/v), purified B-vitamins (Sigma), and/or Wolfe's trace metal solution. When needed, additional salts were added as phosphate buffered saline solution and/or sodium chloride addition (0-4% w/v). Strains from AMC1 (described above) were inoculated into 2 L DASGIP bioreactors (Eppendorf North America Hauppauge, N.Y.) with a 1.5 liter working volume. The pH during fermentation was maintained between 5.0 and 7.0. Aeration conditions during fermentation were controlled by varying agitation, gas composition (air and/or nitrogen gas) and gas flow rates to obtain target oxygen transfer rate (estimated by using k.sub.La) and ranged from having a k.sub.La (per hour) of 0 to 110. Temperature was controlled between 28.degree. C. and 35.degree. C.

TABLE-US-00026 TABLE 26 Co-cultivated consortium ("DFC") designed for co-formulation with carriers or as seed treatment Taxonomy DPA-production Bacillus amyloliquefaciens 1 Bacillus firmus 1 Bacillus flexus 1 Bacillus licheniformis 1 Bacillus megaterium 1 Bacillus pumilus 1 Bacillus koreensis 1 Bacillus drentensis 1 Bacillus subtilis 1 Clostridium bifermentans 1 Clostridium beijerinckii 1 Clostridium pasteurianum 0 Lactobacillus paracasei 0 Fontibacillus sp. (panacisegetis) 1 Oceanobacillus oncorhynchi 0 Paenibacillus lautus 1 Paenibacillus azoreducens 1 Paenibacillus chibensis 1 Paenibacillus cookii 1 Paenibacillus sp. (chitinolyticus) 1 Paenibacillus sp. (P1XP2) 1 Pseudomonas sp. 0 Streptomyces griseus 0 The DPA production column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay, 1 = detected and 0 = not detected or below detection limit.

[0177] To produce co-formulations, DFC fermentate was applied to bentonite or perlite, and then dried for approximately 24 hours. In some cases, fermentate was concentrated via centrifugation prior to application. Co-formulations were then tested for viability. In addition, certain co-formulations were tested in the plant growth room trials to verify plant beneficial characteristics.

[0178] Viability of DFC and AMC1 with carriers: In preparation for field trials that included DFC liquid, AMC1 liquid, DFC-impregnated perlite, AMC1-impregnated perlite, DFC-impregnated bentonite, and AMC1-impregnated bentonite, DFC and AMC1 were each co-fermented and perlite or bentonite carriers were impregnated with 2 mL/g or 1 mL/g respectively. The impregnated perlite and bentonite were stored dry for one to three weeks, at which point the viability assay was performed to determine the number of strains that were viable at the time of application. In addition, liquid was stored for one week, at which point the viability assay was performed to determine the number of strains that were viable at the time of application. In liquid, 16 out of 23 DFC strains were viable after one week versus 14 out of 22 AMC1 strains (Table 27).

TABLE-US-00027 TABLE 27 Viability of DFC and AMC1 consortia in liquid following aging DFC Liquid AMC1 Liquid T0 - At Application Bacillus amyloliquefaciens 1 1 Bacillus flexus 0 1 Bacillus licheniformis 1 1 Bacillus megaterium 1 1 Bacillus subtilis 0 0 Paenibacillus chibensis 1 1 Paenibacillus cookii 1 1 Oceanobacillus oncorhynchi 0 1 Paenibacillus lautus 1 1 Clostridium beijerinckii 1 1 Clostridium pasteurianum 1 1 Lactobacillus paracasei 1 1 Pseudomonas sp. 0 0 Streptomyces griseus 1 0 Bacillus koreensis 0 Bacillus pumilus 1 Paenibacillus sp. 1 (chitinolyticus) Paenibacillus sp. (P1XP2) 1 Fontibacillus sp. 1 (panacisegetis) Bacillus firmus 0 Clostridium bifermentans 1 Paenibacillus azoreducens 1 Bacillus drentensis 0 Bacillus sp. 0 Virgibacillus halophilus 0 Acetobacter pasteurianus 1 Azotobacter vinelandii 0 Lactobacillus buchneri 1 Lactobacillus delbrueckii 0 Lactobacillus vini 1 Pseudomonas putida 0 Total # Viable: 16 14 1 = detected and 0 = not detected or below detection limit

[0179] On impregnated perlite, 17 out of 23 DFC strains versus 14 out of 22 AMC1 strains were viable after up to three weeks (Table 28).

TABLE-US-00028 TABLE 28 Viability of DFC and AMC1 consortia impregnated on perlite following aging. DFC Perlite T0 - At Application AMC1 Perlite Bacillus amyloliquefaciens 1 1 Bacillus flexus 1 1 Bacillus licheniformis 1 1 Bacillus megaterium 1 1 Bacillus subtilis 1 1 Paenibacillus chibensis 1 1 Paenibacillus cookii 1 1 Oceanobacillus oncorhynchi 0 0 Paenibacillus lautus 1 1 Clostridium beijerinckii 1 1 Clostridium pasteurianum 1 1 Lactobacillus paracasei 0 1 Pseudomonas sp. 0 0 Streptomyces griseus 0 0 Bacillus koreensis 1 Bacillus pumilus 1 Paenibacillus sp. 1 (chitinolyticus) Paenibacillus sp. (P1XP2) 1 Fontibacillus sp. 1 (panacisegetis) Bacillus firmus 1 Clostridium bifermentans 0 Paenibacillus azoreducens 1 Bacillus drentensis 0 Bacillus sp. 1 Virgibacillus halophilus 0 Acetobacter pasteurianus 1 Azotobacter vinelandii 0 Lactobacillus buchneri 1 Lactobacillus delbrueckii 0 Lactobacillus vini 0 Pseudomonas putida 0 Total # Viable: 17 14 1 = detected and 0 = not detected or below detection limit.

[0180] On impregnated bentonite, 17 out of 23 DFC strains versus 14 out of 22 AMC1 strains were viable after up to three weeks (Table 29). This illustrates how DPA-producing strains maintain improved viability when stored in liquid, as well as when dried on a carrier such as perlite or bentonite.

TABLE-US-00029 TABLE 29 Viability of DFC and AMC1 consortia impregnated on bentonite following aging DFC Bentonite T0 - At Application AMC1 Bentonite Bacillus amyloliquefaciens 1 1 Bacillus flexus 1 1 Bacillus licheniformis 1 1 Bacillus megaterium 1 1 Bacillus subtilis 1 1 Paenibacillus chibensis 1 1 Paenibacillus cookii 1 1 Oceanobacillus oncorhynchi 0 0 Paenibacillus lautus 1 1 Clostridium beijerinckii 1 1 Clostridium pasteurianum 1 1 Lactobacillus paracasei 1 1 Pseudomonas sp. 0 0 Streptomyces griseus 0 0 Bacillus koreensis 0 Bacillus pumilus 1 Paenibacillus sp. 1 (chitinolyticus) Paenibacillus sp. (P1XP2) 1 Fontibacillus sp. 1 (panacisegetis) Bacillus firmus 1 Clostridium bifermentans 0 Paenibacillus azoreducens 1 Bacillus drentensis 0 Bacillus sp. 1 Virgibacillus halophilus 0 Acetobacter pasteurianus 0 Azotobacter vinelandii 0 Lactobacillus buchneri 1 Lactobacillus delbrueckii 0 Lactobacillus vini 1 Pseudomonas putida 0 Total # Viable: 17 14 1 = detected and 0 = not detected or below detection limit

[0181] Following 3 months of aging, DFC-impregnated perlite showed 15 strains as viable, whereas AMC1-impregnated perlite showed only 11 strains as viable and only one of which was not DPA producing (Tables 30 and 31).

TABLE-US-00030 TABLE 30 Viability of AMC1 impregnated on perlite and bentonite over time DPA Perlite - 3 months Bentonite - 3 months Bacillus amyloliquefaciens 1 1 1 Bacillus flexus 1 1 1 Bacillus licheniformis 1 1 1 Bacillus megaterium 1 1 1 Bacillus sp. 1 0 0 Bacillus subtilis 1 1 1 Paenibacillus chibensis 1 1 1 Paenibacillus cookii 1 1 1 Oceanobacillus oncorhynchi 1 1 1 Paenibacillus lautus 1 1 1 Virgibacillus halophilus 0 0 0 Clostridium beijerinckii 1 1 1 Acetobacter pasteurianus 0 0 0 Azotobacter vinelandii 0 0 0 Clostridium pasteurianum 0 1 1 Lactobacillus buchneri 0 0 1 Lactobacillus delbrueckii 0 0 0 Lactobacillus paracasei 0 0 1 Lactobacillus vini 0 0 0 Pseudomonas putida 0 0 0 Pseudomonas sp. 0 0 0 Streptomyces griseus 0 0 0 Total # Viable: 11 13 The DPA column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay. 1 = detected and 0 = not detected or below detection limit.

[0182] Following the same 3 months of aging, DFC-impregnated bentonite showed 19 strains as viable, whereas AMC1-impregnated perlite showed on 13 strains as viable and only three of which were not DPA producing (Tables 30 and 31).

TABLE-US-00031 TABLE 31 Viability of DFC impregnated on perlite and bentonite over time DPA Perlite - 3 months Bentonite - 3 months Bacillus amyloliquefaciens 1 1 1 Bacillus flexus 1 0 1 Bacillus licheniformis 1 1 1 Bacillus megaterium 1 1 1 Bacillus subtilis 1 1 1 Paenibacillus chibensis 1 1 1 Paenibacillus cookii 1 1 1 Oceanobacillus oncorhynchi 1 1 1 Paenibacillus lautus 1 1 1 Clostridium beijerinckii 1 1 1 Clostridium pasteurianum 0 1 1 Lactobacillus paracasei 0 0 1 Pseudomonas sp. 0 0 0 Streptomyces griseus 0 0 0 Bacillus koreensis 1 0 1 Bacillus pumilus 1 1 1 Paenibacillus sp. (chitinolyticus) 1 1 1 Paenibacillus sp. (P1XP2) 1 1 1 Fontibacillus sp. (panacisegetis) 1 1 1 Bacillus firmus 1 0 1 Clostridium bifermentans 1 0 0 Paenibacillus azoreducens 1 1 1 Bacillus drentensis 1 0 0 Total # viable: 15 19 The DPA column represents those strains that test positive for DPA production via the Terbium-DPA fluorescence assay. 1 = detected and 0 = not detected or below detection limit.

[0183] Evaluation of DFC Plant Beneficial Activity: Cucumber seeds purchased from The Seed Kingdom (Lubbock, Tex.) were pre-germinated for 4 days at 22.degree. C. in rolled germination paper (Anchor Paper, Saint Paul, Minn.) impregnated with a dilute mixture of liquid fertilizer (25 ppm NPK in water). At the time of seed preparation, the potting medium (Sunshine Mix) was prepared with a pre-treatment of 67.60 kg Tricalcium phosphate (TCP) ha.sup.-1 and a modified Hoagland solution (Hoagland, Calif. Agric. Exp. Stn. Bull. 347:36-39, 1938). This Hoagland solution, NK+ was modified to contain N, 56.03 ppm; P, 0 ppm; K, 39.1 ppm; Ca, 40.0 ppm; Mg, 14.59 ppm; S, 20.143 ppm; Fe, 1.010 ppm; Cu, 0.019 ppm; Co, 0.012 ppm; B, 2.44 ppm; Mn, 0.494 ppm; Mo, 0.001 ppm and Zn, 0.056 ppm, which was applied a rate of 1 L per pound of potting medium. The control treatments also contained 10 g of untreated perlite or 20 g of untreated bentonite per pound of potting medium. In comparison, the experimental pots had the same amount of perlite or bentonite as the control pots however those carriers were impregnated with 2 mL/g of DFC or 1 mL/g of DFC, respectively. The impregnated perlite and bentonite had been stored dry for two weeks prior to use in this assay, at which point the viability assay was performed to determine the number of strains from DFC that were viable at the time of application. The large majority of strains (15-18 out of 23 strains viable) survived the co-formulation and aging process.

[0184] Following the pre-germination of the cucumber seeds, similar length seedling were selected and one was transplanted into each pot. For each treatment (including control) 18 plants were randomized in flats in defined growth conditions, controlling for temperature (16-24.degree. C.) and 12 hours photoperiod. The flats were watered for the first time three days after transplanting with a modified Hoagland solution, PK+ which contains N, 0 ppm; P, 14.49 ppm; K, 19.55 ppm; Ca, 20.0 ppm; Mg, 14.59 ppm; S, 20.143 ppm; Fe, 1.010 ppm; Cu, 0.019 ppm; Co, 0.012 ppm; B, 2.44 ppm; Mn, 0.494 ppm; Mo, 0.001 ppm and Zn, 0.056 ppm. The flats were then watered 3 times a week with NK+ Hoagland solution. After 32 days, shoots were harvested, and dried weights were recorded for each plant. The data were analyzed by One-way ANOVA (Analysis Of Variance) and with a post-hoc Tukey test to compare samples within the experiment. These trials were performed twice on two separate occasions.

[0185] The results of the initial trial showed a significant increase in shoot weight for both DFC impregnated perlite and DFC impregnated bentonite when compared to controls (FIG. 6). The results of the second trial also showed a significant increase in shoot weight for both DFC liquid treatment and DFC impregnated perlite, and while the DFC impregnated perlite performed better that the DFC liquid treatment, they were not significantly different (FIG. 7). Thus, co-formulation with a carrier did not impact efficacy when compared to fresh liquid product.

Example 13

Microbe Seed Treatment

[0186] Microbe seed treatments were applied to corn and soybean seed using a batch treater SGS (SGS North America, Brookings, S. Dak. USA). One kg of seed was treated for each treatment. In separate seed treatments, microbes were either applied directly to untreated seed, applied as an overcoat to seed previously treated with an insecticide/fungicide package, or mixed with the insecticide/fungicide slurry prior to seed application. For corn, insecticide/fungicide treatment consisted of either an Acceleron mix containing metalaxyl, trifloxystrobin, ipconazole, and clothianidin, or a CruiserMaxx mix containing Cruiser (thiamethoxam), fludioxonil, mefenoxam, azoxystrobin, and thiabendazole. For soybean, insecticide/fungicide treatment consisted of either an Acceleron mix containing metalaxyl, pyraclostrobin, imidacloprid, and fluxapyroxad, or a CrusierMaxx mix containing Cruiser (thiamethoxam), mefenoxam, fludioxonil, and sedexane.

[0187] Following seed treatment, viability was tested (as in Example 1) within 48 hours and three weeks thereafter. The overwhelming majority of strains (19-20 out of 23 strains viable) survived the initial co-formulation process. After three weeks, only a slight reduction in viability was observed (17-18 out of 23 strains viable; FIG. 8). This illustrated that selecting DPA producing strains was effective not only for dry fertilizers such as bentonite and perlite, but also effective as a seed treatment. In addition, seeds were tested for germination to determine the impact of seed treatment on germination potential. A Cold Vigor test was performed, where 4 replications of 100 seeds were tested for germination. Each 100 seed replicate was planted on moistened crepe cellulose paper and chilled overnight at 10.degree. C. The seeds were then covered with one inch of non-sterile sand wet to 70% water holding capacity and returned to 10.degree. C. for seven days without light. The seeds were then moved into 25.degree. C. for four days. Seedlings that emerged through the sand were evaluated. Results were reported as a percentage that represents the number of seedlings categorized as normal according to AOSA rules. Scores of 82% or higher are considered to be the minimum acceptable for marketing a corn seed lot according to Iowa State Seed Lab and SGS. All treatments had a germination percentage of at least 83.8% or greater (Table 32). This illustrates that in addition to performing well in the form of a seed treatment, DPA-producing strains did not negatively impact germination of said seeds.

TABLE-US-00032 TABLE 32 Average germination rate of DFC treated corn and soybean seeds using the cold vigor germination test Corn Soybean Untreated DFC Untreated DFC No Fungicide Treatment 90.30% 87.50% 87.50% 83.80% Acceleron Treated 89.30% 87.90% 86.00% 84.90% CruiserMaxx Treated 88.00% 86.00% 85.00% 88.65%

[0188] In view of the many possible embodiments to which the principles of the disclosure may be applied, it should be recognized that the illustrated embodiments are only examples and should not be taken as limiting the scope of the invention. Rather, the scope of the invention is defined by the following claims. We therefore claim as our invention all that comes within the scope and spirit of these claims.

Sequence CWU 1

1

6711524DNAStreptomyces pratensis 1catggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcttaac acatgcaagt 60cgaacgatga agcccttcgg ggtggattag tggcgaacgg gtgagtaaca cgtgggcaat 120ctgcccttca ctctgggaca agccctggaa acggggtcta ataccggata acactctgtc 180cctcatgggg cggggttaaa agctccggcg gtgaaggatg agcccgcggc ctatcagctt 240gttggtgggg taatggccta ccaaggcgac gacgggtagc cggcctgaga gggcgaccgg 300ccacactggg actgagacac ggcccagact cctacgggag gcagcagtgg ggaatattgc 360acaatgggcg aaagcctgat gcagcgacgc cgcgtgaggg atgacggcct tcgggttgta 420aacctctttc agcagggaag aagcgcaagt gacggtacct gcagaagaag caccggctaa 480ctacgtgcca gcagccgcgg taatacgtag ggtgcgagcg ttgtccggaa ttattgggcg 540taaagagctc gtaggcggct tgtcacgtcg gatgtgaaag ctcggggctt aaccccgagt 600ctgcattcga tacgggctag ctagagtgtg gtaggggaga tcggaattcc tggtgtagcg 660gtgaaatgcg cagatatcag gaggaacacc ggtggcgaag gcggatctct gggccattac 720tgacgctgag gagcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780cgtaaacgtt gggaactagg tgttggcgac attccacgtc gtcggtgccg cagctaacgc 840attaagttcc ccgcctgggg agtacggccg caaggctaaa actcaaagga attgacgggg 900gcccgcacaa gcagcggagc atgtggctta attcgacgca acgcgaagaa ccttaccaag 960gcttgacata taccggaaag catcagagat ggtgcccccc ttgtggtcgg tatacaggtg 1020gtgcatggct gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080acccttgttc tgtgttgcca gcatgccctt cggggtgatg gggactcaca ggagactgcc 1140ggggtcaact cggaggaagg tggggacgac gtcaagtcat catgcccctt atgtcttggg 1200ctgcacacgt gctacaatgg ccggtacaat gagctgcgat gccgcgaggc ggagcgaatc 1260tcaaaaagcc ggtctcagtt cggattgggg tctgcaactc gaccccatga agtcggagtt 1320gctagtaatc gcagatcagc attgctgcgg tgaatacgtt cccgggcctt gtacacaccg 1380cccgtcacgt cacgaaagtc ggtaacaccc gaagccggtg gcccaacccc ttgtgggagg 1440gagctgtcga aggtgggact ggcgattggg acgaagtcgt aacaaggtag ccgtaccgga 1500aggtgcggct ggatcacctc cttt 152421524DNAStreptomyces venezuelae 2cacggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcttaac acatgcaagt 60cgaacgatga agcccttcgg ggtggattag tggcgaacgg gtgagtaaca cgtgggcaat 120ctgcccttca ctctgggaca agccctggaa acggggtcta ataccggata acaccggctc 180ctgcatgggg gctggttaaa agctccggcg gtgaaggatg agcccgcggc ctatcagctt 240gttggtgggg taatggccta ccaaggcgac gacgggtagc cggcctgaga gggcgaccgg 300ccacactggg actgagacac ggcccagact cctacgggag gcagcagtgg ggaatattgc 360acaatgggcg aaagcctgat gcagcgacgc cgcgtgaggg atgacggcct tcgggttgta 420aacctctttc agcagggaag aagcgaaagt gacggtacct gcagaagaag cgccggctaa 480ctacgtgcca gcagccgcgg taatacgtag ggcgcaagcg ttgtccggaa ttattgggcg 540taaagagctc gtaggcggct tgtcacgtcg ggtgtgaaag cccggggctt aaccccgggt 600ctgcatccga tacgggcagg ctagagtgtg gtaggggaga tcggaattcc tggtgtagcg 660gtgaaatgcg cagatatcag gaggaacacc ggtggcgaag gcggatctct gggccattac 720tgacgctgag gagcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780cgtaaacgtt gggaactagg tgttggcgac attccacgtc gtcggtgccg cagctaacgc 840attaagttcc ccgcctgggg agtacggccg caaggctaaa actcaaagga attgacgggg 900gcccgcacaa gcagcggagc atgtggctta attcgacgca acgcgaagaa ccttaccaag 960gcttgacata taccggaaag cattagagat agtgcccccc ttgtggtcgg tatacaggtg 1020gtgcatggct gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080acccttgtcc tgtgttgcca gcatgccctt cggggtgatg gggactcaca ggagaccgcc 1140ggggtcaact cggaggaagg tggggacgac gtcaagtcat catgcccctt atgtcttggg 1200ctgcacacgt gctacaatgg ccggtacaaa gagctgcgat gccgtgaggc ggagcgaatc 1260tcaaaaagcc ggtctcagtt cggattgggg tctgcaactc gaccccatga agtcggagtt 1320gctagtaatc gcagatcagc attgctgcgg tgaatacgtt cccgggcctt gtacacaccg 1380cccgtcacgt cacgaaagtc ggtaacaccc gaagccggtg gcccaacccc ttgtgggagg 1440gagctgtcga aggtgggact ggcgattggg acgaagtcgt aacaaggtag ccgtaccgga 1500aggtgcggct ggatcacctc cttt 152431548DNABacillus firmus 3tatggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcggaca gatgggagct tgctccctga agtcagcggc ggacgggtga gtaacacgtg 120ggcaacctgc ctgtaagact gggataactc cgggaaaccg gggctaatac cggataattc 180tttccctcac atgagggaaa gctgaaagat ggtttcggct atcacttaca gatgggcccg 240cggcgcatta gctagttggt gaggtaacgg ctcaccaagg caacgatgcg tagccgacct 300gagagggtga tcggccacac tgggactgag acacggccca gactcctacg ggaggcagca 360gtagggaatc ttccgcaatg gacgaaagtc tgacggagca acgccgcgtg agtgatgaag 420gttttcggat cgtaaaactc tgttgttagg gaagaacaag taccggagta actgccggta 480ccttgacggt acctaaccag aaagccacgg ctaactacgt gccagcagcc gcggtaatac 540gtaggtggca agcgttgtcc ggaattattg ggcgtaaagc gcgcgcaggc ggttccttaa 600gtctgatgtg aaagcccccg gctcaaccgg ggagggtcat tggaaactgg ggaacttgag 660tgcagaagag aagagtggaa ttccacgtgt agcggtgaaa tgcgtagaga tgtggaggaa 720caccagtggc gaaggcgact ctttggtctg taactgacgc tgaggcgcga aagcgtgggg 780agcaaacagg attagatacc ctggtagtcc acgccgtaaa cgatgagtgc taagtgttag 840agggtttccg ccctttagtg ctgcagcaaa cgcattaagc actccgcctg gggagtacgg 900ccgcaaggct gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt 960ttaattcgaa gcaacgcgaa gaaccttacc aggtcttgac atctcctgac aaccctagag 1020atagggcgtt ccccttcggg ggacaggatg acaggtggtg catggttgtc gtcagctcgt 1080gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta gttgccagca 1140ttcagttggg cactctaagg tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt 1200caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggat ggtacaaagg 1260gctgcgagac cgcgaggtta agcgaatccc ataaaaccat tctcagttcg gattgcaggc 1320tgcaactcgc ctgcatgaag ccggaatcgc tagtaatcgc ggatcagcat gccgcggtga 1380atacgttccc gggccttgta cacaccgccc gtcacaccac gagagtttgt aacacccgaa 1440gtcggtgggg taaccttttg gagccagccg cctaaggtgg gacagatgat tggggtgaag 1500tcgtaacaag gtagccgtat cggaaggtgc ggctggatca cctccttt 154841552DNAPaenibacillus azoreducens 4cttggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcggatt tgatgaggag cttgctcctc tgatggttag cggcggacgg gtgagtaaca 120cgtaggcaac ctgcctgcaa gaccgggata actagcggaa acgttagcta ataccggata 180atttatcgct ttgcatgaag cgataatgaa agacggagca atctgtcact tgcagatggg 240cctgcggcgc attagctagt tggtgaggta acggctcacc aaggcgacga tgcgtagccg 300acctgagagg gtgaacggcc acactgggac tgagacacgg cccagactcc tacgggaggc 360agcagtaggg aatcttccgc aatgggcgaa agcctgacgg agcaacgccg cgtgagtgat 420gaaggttttc ggatcgtaaa gctctgttgc cagggaagaa cgaccgttag agtaactgct 480aacggagtga cggtacctga gaagaaagcc ccggctaact acgtgccagc agccgcggta 540atacgtaggg ggcaagcgtt gtccggaatt attgggcgta aagcgcgcgc aggcggtcgc 600ttaagtctgg tgtttaaggc caaggctcaa ccttggttcg cactggaaac tgggtgactt 660gagtgcagaa gaggagagtg gaattccacg tgtagcggtg aaatgcgtag agatgtggag 720gaacaccagt ggcgaaggcg actctctggg ctgtaactga cgctgaggcg cgaaagcgtg 780gggagcaaac aggattagat accctggtag tccacgccgt aaacgatgaa tgctaggtgt 840taggggtttc gatacccttg gtgccgaagt taacacatta agcattccgc ctggggagta 900cggtcgcaag actgaaactc aaaggaattg acggggaccc gcacaagcag tggagtatgt 960ggtttaattc gaagcaacgc gaagaacctt accaggtctt gacatccctc tgaccggact 1020agagatagtc ctttccttcg ggacagagga gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatttt agttgccagc 1140actttaaggt gggcactcta aaatgactgc cggtgacaaa ccggaggaag gcggggatga 1200cgtcaaatca tcatgcccct tatgacctgg gctacacacg tactacaatg gccagtacaa 1260cgggaagcga aatcgcgaga tggagccaat cctatcaaag ctggtctcag ttcggattgc 1320aggctgcaac tcgcctgcat gaagtcggaa ttgctagtaa tcgcggatca gcatgccgcg 1380gtgaatacgt tcccgggtct tgtacacacc gcccgtcaca ccacgagagt ttacaacacc 1440cgaagtcggt gaggtaaccg caaggagcca gccgccgaag gtggggtaga tgattggggt 1500gaagtcgtaa caaggtagcc gtatcggaag gtgcggctgg atcacctcct tt 155251548DNABacillus amyloliquefaciens 5atcggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcggaca gatgggagct tgctccctga tgttagcggc ggacgggtga gtaacacgtg 120ggtaacctgc ctgtaagact gggataactc cgggaaaccg gggctaatac cggatggttg 180tctgaaccgc atggttcaga cataaaaggt ggcttcggct accacttaca gatggacccg 240cggcgcatta gctagttggt gaggtaacgg ctcaccaagg cgacgatgcg tagccgacct 300gagagggtga tcggccacac tgggactgag acacggccca gactcctacg ggaggcagca 360gtagggaatc ttccgcaatg gacgaaagtc tgacggagca acgccgcgtg agtgatgaag 420gttttcggat cgtaaagctc tgttgttagg gaagaacaag tgccgttcaa atagggcggc 480accttgacgg tacctaacca gaaagccacg gctaactacg tgccagcagc cgcggtaata 540cgtaggtggc aagcgttgtc cggaattatt gggcgtaaag ggctcgcagg cggtttctta 600agtctgatgt gaaagccccc ggctcaaccg gggagggtca ttggaaactg gggaacttga 660gtgcagaaga ggagagtgga attccacgtg tagcggtgaa atgcgtagag atgtggagga 720acaccagtgg cgaaggcgac tctctggtct gtaactgacg ctgaggagcg aaagcgtggg 780gagcgaacag gattagatac cctggtagtc cacgccgtaa acgatgagtg ctaagtgtta 840gggggtttcc gccccttagt gctgcagcta acgcattaag cactccgcct ggggagtacg 900gtcgcaagac tgaaactcaa aggaattgac gggggcccgc acaagcggtg gagcatgtgg 960tttaattcga agcaacgcga agaaccttac caggtcttga catcctctga caatcctaga 1020gataggacgt ccccttcggg ggcagagtga caggtggtgc atggttgtcg tcagctcgtg 1080tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc ttgatcttag ttgccagcat 1140tcagttgggc actctaaggt gactgccggt gacaaaccgg aggaaggtgg ggatgacgtc 1200aaatcatcat gccccttatg acctgggcta cacacgtgct acaatggaca gaacaaaggg 1260cagcgaaacc gcgaggttaa gccaatccca caaatctgtt ctcagttcgg atcgcagtct 1320gcaactcgac tgcgtgaagc tggaatcgct agtaatcgcg gatcagcatg ccgcggtgaa 1380tacgttcccg ggccttgtac acaccgcccg tcacaccacg agagtttgta acacccgaag 1440tcggtgaggt aacctttatg gagccagccg ccgaaggtgg gacagatgat tggggtgaag 1500tcgtaacaag gtagccgtat cggaaggtgc ggctggatca cctccttt 154861550DNABacillus flexus 6tcggagagtt tgatcctggc tcaggatgaa cgctggcggc gtgcctaata catgcaagtc 60gagcgaactg attagaagct tgcttctatg acgttagcgg cggacgggtg agtaacacgt 120gggcaacctg cctgtaagac tgggataact ccgggaaacc ggagctaata ccggataaca 180ttttctcttg cataagagaa aattgaaaga tggtttcggc tatcacttac agatgggccc 240gcggtgcatt agctagttgg tgaggtaacg gctcaccaag gcaacgatgc atagccgacc 300tgagagggtg atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc 360agtagggaat cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa 420ggctttcggg tcgtaaaact ctgttgttag ggaagaacaa gtacaagagt aactgcttgt 480accttgacgg tacctaacca gaaagccacg gctaactacg tgccagcagc cgcggtaata 540cgtaggtggc aagcgttatc cggaattatt gggcgtaaag cgcgcgcagg cggtttctta 600agtctgatgt gaaagcccac ggctcaaccg tggagggtca ttggaaactg gggaacttga 660gtgcagaaga gaaaagcgga attccacgtg tagcggtgaa atgcgtagag atgtggagga 720acaccagtgg cgaaggcggc tttttggtct gtaactgacg ctgaggcgcg aaagcgtggg 780gagcaaacag gattagatac cctggtagtc cacgccgtaa acgatgagtg ctaagtgtta 840gagggtttcc gccctttagt gctgcagcta acgcattaag cactccgcct ggggagtacg 900gtcgcaagac tgaaactcaa aggaattgac gggggcccgc acaagcggtg gagcatgtgg 960tttaattcga agcaacgcga agaaccttac caggtcttga catcctctga caactctaga 1020gatagagcgt tccccttcgg gggacagagt gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc 1140atttagttgg gcactctaag gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg 1200tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatgga tggtacaaag 1260ggctgcaaga ccgcgaggtc aagccaatcc cataaaacca ttctcagttc ggattgtagg 1320ctgcaactcg cctacatgaa gctggaatcg ctagtaatcg cggatcagca tgccgcggtg 1380aatacgttcc cgggccttgt acacaccgcc cgtcacacca cgagagtttg taacacccga 1440agtcggtggg gtaaccttta tggagccagc cgcctaaggt gggacagatg attggggtga 1500agtcgtaaca aggtagccgt atcggaaggt gcggctggat cacctccttt 155071549DNABacillus licheniformis 7catggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcggacc gacgggagct tgctccctta ggtcagcggc ggacgggtga gtaacacgtg 120ggtaacctgc ctgtaagact gggataactc cgggaaaccg gggctaatac cggatgcttg 180attgaaccgc atggttccaa tcataaaagg tggcttttag ctaccactta cagatggacc 240cgcggcgcat tagctagttg gtgaggtaac ggctcaccaa ggcgacgatg cgtagccgac 300ctgagagggt gatcggccac actgggactg agacacggcc cagactccta cgggaggcag 360cagtagggaa tcttccgcaa tggacgaaag tctgacggag caacgccgcg tgagtgatga 420aggttttcgg atcgtaaaac tctgttgtta gggaagaaca agtaccgttc gaatagggcg 480gcaccttgac ggtacctaac cagaaagcca cggctaacta cgtgccagca gccgcggtaa 540tacgtaggtg gcaagcgttg tccggaatta ttgggcgtaa agcgcgcgca ggcggtttct 600taagtctgat gtgaaagccc ccggctcaac cggggagggt cattggaaac tggggaactt 660gagtgcagaa gaggagagtg gaattccacg tgtagcggtg aaatgcgtag agatgtggag 720gaacaccagt ggcgaaggcg actctctggt ctgtaactga cgctgaggcg cgaaagcgtg 780gggagcgaac aggattagat accctggtag tccacgccgt aaacgatgag tgctaagtgt 840tagagggttt ccgcccttta gtgctgcagc aaacgcatta agcactccgc ctggggagta 900cggtcgcaag actgaaactc aaaggaattg acgggggccc gcacaagcgg tggagcatgt 960ggtttaattc gaagcaacgc gaagaacctt accaggtctt gacatcctct gacaacccta 1020gagatagggc ttccccttcg ggggcagagt gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc 1140attcagttgg gcactctaag gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg 1200tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatggg cagaacaaag 1260ggcagcgaag ccgcgaggct aagccaatcc cacaaatctg ttctcagttc ggatcgcagt 1320ctgcaactcg actgcgtgaa gctggaatcg ctagtaatcg cggatcagca tgccgcggtg 1380aatacgttcc cgggccttgt acacaccgcc cgtcacacca cgagagtttg taacacccga 1440agtcggtgag gtaacctttt ggagccagcc gccgaaggtg ggacagatga ttggggtgaa 1500gtcgtaacaa ggtagccgta tcggaaggtg cggctggatc acctccttt 154981549DNABacillus megaterium 8tcggagagtt tgatcctggc tcaggatgaa cgctggcggc gtgcctaata catgcaagtc 60gagcgaactg attagaagct tgcttctatg acgttagcgg cggacgggtg agtaacacgt 120gggcaacctg cctgtaagac tgggataact tcgggaaacc gaagctaata ccggatagga 180tcttctcctt catgggagat gattgaaaga tggtttcggc tatcacttac agatgggccc 240gcggtgcatt agctagttgg tgaggtaacg gctcaccaag gcaacgatgc atagccgacc 300tgagagggtg atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc 360agtagggaat cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa 420ggctttcggg tcgtaaaact ctgttgttag ggaagaacaa gtacgagagt aactgctcgt 480accttgacgg tacctaacca gaaagccacg gctaactacg tgccagcagc cgcggtaata 540cgtaggtggc aagcgttatc cggaattatt gggcgtaaag cgcgcgcagg cggtttctta 600agtctgatgt gaaagcccac ggctcaaccg tggagggtca ttggaaactg gggaacttga 660gtgcagaaga gaaaagcgga attccacgtg tagcggtgaa atgcgtagag atgtggagga 720acaccagtgg cgaaggcggc tttttggtct gtaactgacg ctgaggcgcg aaagcgtggg 780gagcaaacag gattagatac cctggtagtc cacgccgtaa acgatgagtg ctaagtgtta 840gagggtttcc gccctttagt gctgcagcta acgcattaag cactccgcct ggggagtacg 900gtcgcaagac tgaaactcaa aggaattgac gggggcccgc acaagcggtg gagcatgtgg 960tttaattcga agcaacgcga agaaccttac caggtcttga catcctctga caactctaga 1020gatagagcgt tccccttcgg gggacagagt gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc 1140atttagttgg gcactctaag gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg 1200tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatgga tggtacaaag 1260ggctgcaaga ccgcgaggtc aagccaatcc cataaaacca ttctcagttc ggattgtagg 1320ctgcaactcg cctacatgaa gctggaatcg ctagtaatcg cggatcagca tgccgcggtg 1380aatacgttcc cgggccttgt acacaccgcc cgtcacacca cgagagtttg taacacccga 1440agtcggtgga gtaaccgtaa ggagctagcc gcctaaggtg ggacagatga ttggggtgaa 1500gtcgtaacaa ggtagccgta tcggaaggtg cggctggatc acctccttt 154991544DNABacillus pumilus 9ggagagtttg atcctggctc aggacgaacg ctggcggcgt gcctaataca tgcaagtcga 60gcggacagaa gggagcttgc tcccggatgt tagcggcgga cgggtgagta acacgtgggt 120aacctgcctg taagactggg ataactccgg gaaaccggag ctaataccgg atagttcctt 180gaaccgcatg gttcaaggat gaaagacggt ttcggctgtc acttacagat ggacccgcgg 240cgcattagct agttggtggg gtaatggctc accaaggcga cgatgcgtag ccgacctgag 300agggtgatcg gccacactgg gactgagaca cggcccagac tcctacggga ggcagcagta 360gggaatcttc cgcaatggac gaaagtctga cggagcaacg ccgcgtgagt gatgaaggtt 420ttcggatcgt aaagctctgt tgttagggaa gaacaagtgc gagagtaact gctcgcacct 480tgacggtacc taaccagaaa gccacggcta actacgtgcc agcagccgcg gtaatacgta 540ggtggcaagc gttgtccgga attattgggc gtaaagggct cgcaggcggt ttcttaagtc 600tgatgtgaaa gcccccggct caaccgggga gggtcattgg aaactgggaa acttgagtgc 660agaagaggag agtggaattc cacgtgtagc ggtgaaatgc gtagagatgt ggaggaacac 720cagtggcgaa ggcgactctc tggtctgtaa ctgacgctga ggagcgaaag cgtggggagc 780gaacaggatt agataccctg gtagtccacg ccgtaaacga tgagtgctaa gtgttagggg 840gtttccgccc cttagtgctg cagctaacgc attaagcact ccgcctgggg agtacggtcg 900caagactgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc atgtggttta 960attcgaagca acgcgaagaa ccttaccagg tcttgacatc ctctgacaac cctagagata 1020gggctttccc ttcggggaca gagtgacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt 1080gagatgttgg gttaagtccc gcaacgagcg caacccttga tcttagttgc cagcatttag 1140ttgggcactc taaggtgact gccggtgaca aaccggagga aggtggggat gacgtcaaat 1200catcatgccc cttatgacct gggctacaca cgtgctacaa tggacagaac aaagggctgc 1260gagaccgcaa ggtttagcca atcccataaa tctgttctca gttcggatcg cagtctgcaa 1320ctcgactgcg tgaagctgga atcgctagta atcgcggatc agcatgccgc ggtgaatacg 1380ttcccgggcc ttgtacacac cgcccgtcac accacgagag tttgcaacac ccgaagtcgg 1440tgaggtaacc tttatggagc cagccgccga aggtggggca gatgattggg gtgaagtcgt 1500aacaaggtag ccgtatcgga aggtgcggct ggatcacctc cttt 1544101549DNABacillus koreensis 10tcggagagtt tgatcctggc tcaggacgaa cgctggcggc gtgcctaata catgcaagtc 60gagcggactt gttagaagct tgcttctaac aagttagcgg cggacgggtg agtaacacgt 120gggtaacctg cctgtaagat ggggataact ccgggaaacc ggagctaata ccgaataaca 180ctttcgctcg catgagcgga tgttaaaaga cggtttcggc tgtcacttac agatggaccc 240gcggcgcatt agctagttgg tgaggtaacg gctcaccaag gcgacgatgc gtagccgacc 300tgagagggtg atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc 360agtagggaat cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa 420ggttttcgga tcgtaaaact ctgttgttag ggaagaacaa gtacgagagt aactgctcgt 480accttgacgg tacctaacca gaaagccacg gctaactacg tgccagcagc cgcggtaata 540cgtaggtggc aagcgttgtc cggaattatt gggcgtaaag cgcgcgcagg cggttcctta 600agtctgatgt gaaagcccac ggctcaaccg tggagggtca ttggaaactg gggaacttga 660gtgcagaaga ggaaagcgga attccacgtg tagcggtgaa atgcgtagag atgtggagga 720acaccagtgg

cgaaggcggc tttctggtct gtaactgacg ctgaggcgcg aaagcgtggg 780gagcaaacag gattagatac cctggtagtc cacgccgtaa acgatgagtg ctaagtgtta 840gagggtttcc gccctttagt gctgcagcta acgcattaag cactccgcct ggggagtacg 900gccgcaaggc tgaaactcaa aggaattgac gggggcccgc acaagcggtg gagcatgtgg 960tttaattcga agcaacgcga agaaccttac caggtcttga catcctttga ccactctaga 1020gatagagctt tccccttcgg gggacaaagt gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc 1140attaagttgg gcactctaag gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg 1200tcaaatcatc atgcccctta tgacctgggc tacacacgtg ctacaatgga tgatacaaag 1260ggttgcgaag ccgcgaggtg aagctaatct cataaaatca ttctcagttc ggattgtagg 1320ctgcaactcg cctacatgaa gctggaatcg ctagtaatcg cggatcagca tgccgcggtg 1380aatacgttcc cgggccttgt acacaccgcc cgtcacacca cgagagtttg taacacccga 1440agtcggtggg gtaaccgtaa ggagccagcc gcctaaggtg ggacagatga ttggggtgaa 1500gtcgtaacaa ggtagccgta tcggaaggtg cggctggatc acctccttt 1549111454DNABacillus drentensis 11cttggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcgaatc ttcaggagct tgctcctgtt ggttagcggc ggacgggtga gtaacacgtg 120ggcaacctgc ctgtaagact gggataacac cgggaaaccg gtgctaatac cggataatcc 180ttttcctctc atgaggaaaa gctgaaagtc ggtttcggct gacacttaca gatgggcccg 240cggcgcatta gctagttggt gaggtaacgg ctcaccaagg cgacgatgcg tagccgacct 300gagagggtga tcggccacac tgggactgag acacggccca gactcctacg ggaggcagca 360gtagggaatc ttccacaatg gacgaaagtc tgatggagca acgccgcgtg agcgatgaag 420gccttcgggt cgtaaagctc tgttgttagg gaagaacaag taccggagta actgccggta 480ccttgacggt acctaaccag aaagccacgg ctaactacgt gccagcagcc gcggtaatac 540gtaggtggca agcgttgtcc ggaattattg ggcgtaaagc gcgcgcaggc ggtcctttaa 600gtctgatgtg aaagcccacg gctcaaccgt ggagggtcat tggaaactgg gggacttgag 660tgcagaagag gaaagcggaa ttccacgtgt agcggtgaaa tgcgtagaga tgtggaggaa 720caccagtggc gaaggcggct ttctggtctg taactgacgc tgaggcgcga aagcgtgggg 780agcaaacagg attagatacc ctggtagtcc acgccgtaaa cgatgagtgc taagtgttag 840ggggtttccg ccccttagtg ctgcagctaa cgcattaagc actccgcctg gggagtacgg 900ccgcaaggct gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt 960ttaattcgaa gcaacgcgaa gaaccttacc aggtcttgac atcctctgac actcctagag 1020ataggacgtt ccccttcggg ggacagagtg acaggtggtg catggttgtc gtcagctcgt 1080gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta gttgccagca 1140ttcagttggg cactctaagg tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt 1200caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggat ggtacaaagg 1260gctgcaagac cgcgaggttt agccaatccc ataaaaccat tctcagttcg gattgcaggc 1320tgcaactcgc ctgcatgaag ccggaatcgc tagtaatcgc ggatcagcat gccgcggtga 1380atacgttccc gggccttgta cacaccgccc gtcacaccac gagagtttgt aacacccgaa 1440gtcggtgggg taac 1454121454DNABacillus subtilis 12cttggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcgaatc ttcaggagct tgctcctgtt ggttagcggc ggacgggtga gtaacacgtg 120ggcaacctgc ctgtaagact gggataacac cgggaaaccg gtgctaatac cggataatcc 180ttttcctctc atgaggaaaa gctgaaagtc ggtttcggct gacacttaca gatgggcccg 240cggcgcatta gctagttggt gaggtaacgg ctcaccaagg cgacgatgcg tagccgacct 300gagagggtga tcggccacac tgggactgag acacggccca gactcctacg ggaggcagca 360gtagggaatc ttccacaatg gacgaaagtc tgatggagca acgccgcgtg agcgatgaag 420gccttcgggt cgtaaagctc tgttgttagg gaagaacaag taccggagta actgccggta 480ccttgacggt acctaaccag aaagccacgg ctaactacgt gccagcagcc gcggtaatac 540gtaggtggca agcgttgtcc ggaattattg ggcgtaaagc gcgcgcaggc ggtcctttaa 600gtctgatgtg aaagcccacg gctcaaccgt ggagggtcat tggaaactgg gggacttgag 660tgcagaagag gaaagcggaa ttccacgtgt agcggtgaaa tgcgtagaga tgtggaggaa 720caccagtggc gaaggcggct ttctggtctg taactgacgc tgaggcgcga aagcgtgggg 780agcaaacagg attagatacc ctggtagtcc acgccgtaaa cgatgagtgc taagtgttag 840ggggtttccg ccccttagtg ctgcagctaa cgcattaagc actccgcctg gggagtacgg 900ccgcaaggct gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt 960ttaattcgaa gcaacgcgaa gaaccttacc aggtcttgac atcctctgac actcctagag 1020ataggacgtt ccccttcggg ggacagagtg acaggtggtg catggttgtc gtcagctcgt 1080gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta gttgccagca 1140ttcagttggg cactctaagg tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt 1200caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggat ggtacaaagg 1260gctgcaagac cgcgaggttt agccaatccc ataaaaccat tctcagttcg gattgcaggc 1320tgcaactcgc ctgcatgaag ccggaatcgc tagtaatcgc ggatcagcat gccgcggtga 1380atacgttccc gggccttgta cacaccgccc gtcacaccac gagagtttgt aacacccgaa 1440gtcggtgggg taac 1454131320DNAClostridium bifermentans 13gcttttgtat caaagctccg gcggtacagg atggacccgc gtctgattag ctagttggta 60aggtaatggc ttaccaaggc aacgatcagt agccgacctg agagggtgat cggccacact 120ggaactgaga cacggtccag actcctacgg gaggcagcag tggggaatat tgcacaatgg 180gcgaaagcct gatgcagcaa cgccgcgtga gcgatgaagg ccttcgggtc gtaaagctct 240gtcctcaagg aagataatga cggtacttga ggaggaagcc ccggctaact acgtgccagc 300agccgcggta atacgtaggg ggctagcgtt atccggaatt actgggcgta aagggtgcgt 360aggtggtttt ttaagtcaga agtgaaaggc tacggctcaa ccgtagtaag cttttgaaac 420tagagaactt gagtgcagga gaggagagta gaattcctag tgtagcggtg aaatgcgtag 480atattaggag gaataccagt agcgaaggcg gctctctgga ctgtaactga cactgaggca 540cgaaagcgtg gggagcaaac aggattagat accctggtag tccacgccgt aaacgatgag 600tactaggtgt cgggggttac ccccctcggt gccgcagcta acgcattaag tactccgcct 660gggaagtacg ctcgcaagag tgaaactcaa aggaattgac ggggacccgc acaagtagcg 720gagcatgtgg tttaattcga agcaacgcga agaaccttac ctaagcttga catcccactg 780acctctccct aatcggagat ttcccttcgg ggacagtggt gacaggtggt gcatggttgt 840cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgccttt 900agttgccagc attaagttgg gcactctaga gggactgccg aggataactc ggaggaaggt 960ggggatgacg tcaaatcatc atgcccctta tgcttagggc tacacacgtg ctacaatggg 1020tggtacagag ggttgccaag ccgcgaggtg gagctaatcc cttaaagcca ttctcagttc 1080ggattgtagg ctgaaactcg cctacatgaa gctggagtta ctagtaatcg cagatcagaa 1140tgctgcggtg aatgcgttcc cgggtcttgt acacaccgcc cgtcacacca tggaagttgg 1200gggcgcccga agccggttag ctaacctttt aggaagcggc cgtcgaaggt gaaaccaatg 1260actggggtga agtcgtaaca aggtagccgt atcggaaggt gcggctggat cacctccttt 1320141510DNAClostridium beijerinckii 14tattgagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcttaac acatgcaagt 60cgagcgatga agttccttcg ggaatggatt agcggcggac gggtgagtaa cacgtgggta 120acctgcctca tagaggggaa tagcctttcg aaaggaagat taataccgca taagattgta 180gtgccgcatg gcatagcaat taaaggagta atccgctatg agatggaccc gcgtcgcatt 240agctagttgg tgaggtaacg gctcaccaag gcgacgatgc gtagccgacc tgagagggtg 300atcggccaca ttgggactga gacacggccc agactcctac gggaggcagc agtggggaat 360attgcacaat gggggaaacc ctgatgcagc aacgccgcgt gagtgatgac ggtcttcgga 420ttgtaaagct ctgtcttcag ggacgataat gacggtacct gaggaggaag ccacggctaa 480ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggat ttactgggcg 540taaagggagc gtaggtggat atttaagtgg gatgtgaaat actcgggctt aacctgggtg 600ctgcattcca aactggatat ctagagtgca ggagaggaaa gtagaattcc tagtgtagcg 660gtgaaatgcg tagagattag gaagaatacc agtggcgaag gcgactttct ggactgtaac 720tgacactgag gctcgaaagc gtggggagca aacaggatta gataccctgg tagtccacgc 780cgtaaacgat gaatactagg tgtaggggtt gtcatgacct ctgtgccgcc gctaacgcat 840taagtattcc gcctggggag tacggtcgca agattaaaac tcaaaggaat tgacgggggc 900ccgcacaagc agcggagcat gtggtttaat tcgaagcaac gcgaagaacc ttacctagac 960ttgacatctc ctgaattacc cttaatcggg gaagcccttc ggggcaggaa gacaggtggt 1020gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac 1080ccttattgtt agttgctacc atttagttga gcactctagc gagactgccc gggttaaccg 1140ggaggaaggt ggggatgacg tcaaatcatc atgcccctta tgtctagggc tacacacgtg 1200ctacaatggc tggtacagag agatgctaaa ccgtgaggtg gagccaaact ttaaaaccag 1260tctcagttcg gattgtaggc tgaaactcgc ctacatgaag ctggagttgc tagtaatcgc 1320gaatcagaat gtcgcggtga atacgttccc gggccttgta cacaccgccc gtcacaccat 1380gagagttggc aatacccaaa gttcgtgagc taacgcgcaa gcggggcagc gacctaaggt 1440agggtcagcg attggggtga agtcgtaaca aggtagccgt aggagaacct gcggctggat 1500cacctccttt 1510151505DNAClostridium pasteurianum 15aattgagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcttaac acatgcaagt 60cgagcgagaa accttcgggt ttctagcggc ggacgggtga gtaacacgtg ggtaacctgc 120ctcaaagagg ggaatagcct cccgaaaggg agattaatac cgcataatat tacagcttcg 180catgaagcag taattaaagg agtaatccgc tttgagatgg acccgcggcg cattagctag 240ttggagaggt aacggctcac caaggcgacg atgcgtagcc gacctgagag ggtgatcggc 300cacattggaa ctgagacacg gtccagactc ctacgggagg cagcagtggg gaatattgca 360caatgggcga aagcctgatg cagcaacgcc gcgtgagtga tgacggtctt cggattgtaa 420agctctgtct tttgggacga taatgacggt accaaaggag gaagccacgg ctaactacgt 480gccagcagcc gcggtaatac gtaggtggca agcgttgtcc ggatttactg ggcgtaaagg 540atgtgtaggc ggatacttaa gtgagatgtg aaagccccgg gcttaacttg gggactgcat 600ttcaaactgg gtgtctagag tgcaggagag gaaagcggaa ttcctagtgt agcggtgaaa 660tgcgtagaga ttaggaagaa catcagtggc gaaggcggct ttctggactg taactgacgc 720tgaggcatga aagcgtgggg agcaaacagg attagatacc ctggtagtcc acgccgtaaa 780cgatgagtac taggtgtagg aggtatcgac tccttctgtg ccgcagtaaa cacaataagt 840actccgcctg ggaagtacgg tcgcaagatt aaaactcaaa ggaattgacg ggggcccgca 900caagcagcgg agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc tagacttgac 960atctcctgaa tagcgtagag atacgtgaag cccttcgggg caggaagaca ggtggtgcat 1020ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt 1080atcattagtt gctaccatta agttgagcac tctagtgaga ctgcccgggt taaccgggag 1140gaaggcgggg atgacgtcaa atcatcatgc cccttatgtc tagggctaca cacgtgctac 1200aatggtgaga acaacgagat gcaataccgc gaggtggagc caaacttgaa aactcatccc 1260agttcggatt gtaggctgaa attcgcctac atgaagttgg agttgctagt aatcgcgaat 1320cagaatgtcg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatgaga 1380gctggtaaca cccgaagtcc gtgaggtaac ctttatggag ccagcggccg aaggtgggat 1440tagtgattgg ggtgaagtcg taacaaggta gccgtaggag aacctgcggc tggatcacct 1500ccttt 1505161567DNALactobacillus paracasei 16tatgagagtt tgatcctggc tcaggatgaa cgctggcggc gtgcctaata catgcaagtc 60gaacgagttc tcgttgatga tcggtgcttg caccgagatt caacatggaa cgagtggcgg 120acgggtgagt aacacgtggg taacctgccc ttaagtgggg gataacattt ggaaacagat 180gctaataccg catagatcca agaaccgcat ggttcttggc tgaaagatgg cgtaagctat 240cgcttttgga tggacccgcg gcgtattagc tagttggtga ggtaacggct caccaaggcg 300atgatacgta gccgaactga gaggttgatc ggccacattg ggactgagac acggcccaaa 360ctcctacggg aggcagcagt agggaatctt ccacaatgga cgcaagtctg atggagcaac 420gccgcgtgag tgaagaaggc tttcgggtcg taaaactctg ttgttggaga agaatggtcg 480gcagagtaac tgttgtcggc gtgacggtat ccaaccagaa agccacggct aactacgtgc 540cagcagccgc ggtaatacgt aggtggcaag cgttatccgg atttattggg cgtaaagcga 600gcgcaggcgg ttttttaagt ctgatgtgaa agccctcggc ttaaccgagg aagcgcatcg 660gaaactggga aacttgagtg cagaagagga cagtggaact ccatgtgtag cggtgaaatg 720cgtagatata tggaagaaca ccagtggcga aggcggctgt ctggtctgta actgacgctg 780aggctcgaaa gcatgggtag cgaacaggat tagataccct ggtagtccat gccgtaaacg 840atgaatgcta ggtgttggag ggtttccgcc cttcagtgcc gcagctaacg cattaagcat 900tccgcctggg gagtacgacc gcaaggttga aactcaaagg aattgacggg ggcccgcaca 960agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag gtcttgacat 1020cttttgatca cctgagagat caggtttccc cttcgggggc aaaatgacag gtggtgcatg 1080gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaaccctta 1140tgactagttg ccagcattta gttgggcact ctagtaagac tgccggtgac aaaccggagg 1200aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac acgtgctaca 1260atggatggta caacgagttg cgagaccgcg aggtcaagct aatctcttaa agccattctc 1320agttcggact gtaggctgca actcgcctac acgaagtcgg aatcgctagt aatcgcggat 1380cagcacgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatgaga 1440gtttgtaaca cccgaagccg gtggcgtaac ccttttaggg agcgagccgt ctaaggtggg 1500acaaatgatt agggtgaagt cgtaacaagg tagccgtagg agaacctgcg gctggatcac 1560ctccttt 156717512DNAFontibacillus sp. (panacisegetis) 17ttagcggcgg acgggtgagt aacacgtagg taacctgcct gtaagactgg gataactagc 60ggaaacgtta gctaataccg gataatttat tttctcgcat ggggaagtaa tgaaagacgg 120agcaatctgt cacttgcaga tggacctgcg gcgcattagc tagttggtgg ggtaacggct 180caccaaggcg acgatgcgta gccgacctga gagggtgaac ggccacactg ggactgagac 240acggcccaga ctcctacggg aggcagcagt agggaatctt ccgcaatgga cgaaagtctg 300acggagcaac gccgcgtgag tgatgaaggt tttcggatcg taaagctctg ttgccaggga 360agaacgttcg gtagagtaac tgctaccgga gtgacggtac ctgagaagaa agccccggct 420aactacgtgc cagcagccgc ggtaatacgt agggggcaag cgttgtccgg aattattggg 480cgtaaagcgc gcgcaggcgg ctatttaagt ct 512181561DNAOceanobacillus oncorhynchi 18ttatggagag tttgatcttg gctcaggacg aacgctggcg gcgtgcctaa tacatgcaag 60tcgagcgcgg gaagcgaacg gaactcttcg gagggaagtt cgtggaacga gcggcggacg 120ggtgagtaac acgtaggcaa cctgcctgta agactgggat aactcgcgga aacgcgagct 180aataccggat aacactttct atcacctgat ggaaagttga aaggcggctt ttgctgtcac 240ttacagatgg gcctgcggcg cattagctag ttggtgaggt aacggctcac caaggcgacg 300atgcgtagcc gacctgagag ggtgatcggc cacactggga ctgagacacg gcccagactc 360ctacgggagg cagcagtagg gaatcttccg caatggacga aagtctgacg gagcaacgcc 420gcgtgagtga tgaaggtttt cggatcgtaa aactctgttg tcagggaaga acaagtacga 480tagtaactga tcgtaccttg acggtacctg accagaaagc cacggctaac tacgtgccag 540cagccgcggt aatacgtagg tggcaagcgt tgtccggaat tattgggcgt aaagcgctcg 600caggcggttc tttaagtctg atgtgaaatc ttgcggctca accgcaaacg tgcattggaa 660actggaggac ttgagtgcag aagaggagag tggaattcca cgtgtagcgg tgaaatgcgt 720agagatgtgg aggaacacca gtggcgaagg cgactctctg gtctgtaact gacgctgagg 780agcgaaagcg tggggagcga acaggattag ataccctggt agtccacgcc gtaaacgatg 840agtgctaggt gttagggggt ttccgcccct tagtgctgaa gttaacgcat taagcactcc 900gcctggggag tacggccgca aggctgaaac tcaaaagaat tgacggggac ccgcacaagc 960ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc ttgacatcct 1020ttgaccgctc tagagataga gttttccctt cggggacaaa gtgacaggtg gtgcatggtt 1080gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccttaatc 1140ttagttgcca gcatttagtt gggcactcta aggtgactgc cggtgacaaa ccggaggaag 1200gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg tgctacaatg 1260gacggaacaa agggaagcga acccgcgagg tccagcaaat cccataaaac cgttctcagt 1320tcggattgca ggctgcaact cgcctgcatg aagccggaat cgctagtaat cgcggatcag 1380catgccgcgg tgaatacgtt cccgggtctt gtacacaccg cccgtcacac cacgagagtt 1440cgtaacaccc gaagtcggtg aggtaacctt ttggagccag ccgccgaagg tgggacgaat 1500gattggggtg aagtcgtaac aaggtagccg tatcggaagg tgcggctgga tcacctcctt 1560t 1561191553DNAPaenibacillus lautus 19attggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcggact tgatggagtg cttgcactcc tgaaggttag cggcggacgg gtgagtaaca 120cgtaggcaac ctgccctcaa gactgggata actaccggaa acggtagcta ataccggata 180atttattttg cagcattgtg aaataatgaa aggcggagca atctgtcact tgaggatggg 240cctgcggcgc attagctagt tggtggggta acggcccacc aaggcgacga tgcgtagccg 300acctgagagg gtgaacggcc acactgggac tgagacacgg cccagactcc tacgggaggc 360agcagtaggg aatcttccgc aatgggcgaa agcctgacgg agcaacgccg cgtgagtgat 420gaaggttttc ggatcgtaaa gctctgttgc caaggaagaa cgtcttctag agtaactgct 480aggagagtga cggtacttga gaagaaagcc ccggctaact acgtgccagc agccgcggta 540atacgtaggg ggcaagcgtt gtccggaatt attgggcgta aagcgcgcgc aggcggttct 600ttaagtctgg tgtttaaacc cgaggctcaa cttcgggtcg cactggaaac tggggaactt 660gagtgcagaa gaggagagtg gaattccacg tgtagcggtg aaatgcgtag atatgtggag 720gaacaccagt ggcgaaggcg actctctggg ctgtaactga cgctgaggcg cgaaagcgtg 780gggagcaaac aggattagat accctggtag tccacgccgt aaacgatgaa tgctaggtgt 840taggggtttc gatacccttg gtgccgaagt taacacatta agcattccgc ctggggagta 900cggtcgcaag actgaaactc aaaggaattg acggggaccc gcacaagcag tggagtatgt 960ggtttaattc gaagcaacgc gaagaacctt accaagtctt gacatccctc tgaatcctct 1020agagatagag gcggccttcg ggacagaggt gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatttt agttgccagc 1140acttcgggtg ggcactctag aatgactgcc ggtgacaaac cggaggaagg cggggatgac 1200gtcaaatcat catgcccctt atgacttggg ctacacacgt actacaatgg ctggtacaac 1260gggaagcgaa gccgcgaggt ggagccaatc ctataaaagc cagtctcagt tcggattgca 1320ggctgcaact cgcctgcatg aagtcggaat tgctagtaat cgcggatcag catgccgcgg 1380tgaatacgtt cccgggtctt gtacacaccg cccgtcacac cacgagagtt tacaacaccc 1440gaagtcggtg gggtaaccct taggggagcc agccgccgaa ggtggggtag atgattgggg 1500tgaagtcgta acaaggtagc cgtatcggaa ggtgcggctg gatcacctcc ttt 1553201553DNAPaenibacillus chibensis 20cttggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcggagt tgatgaggtg cttgcacctc tgatgcttag cggcggacgg gtgagtaaca 120cgtaggtaac ctgcctgtaa gactgggata actaccggaa acggtagcta ataccggata 180atttattttc tctcctgggg agataatgaa agacggagca atctgtcact tacagatggg 240cctgcggcgc attagctagt tggtgaggta acggctcacc aaggcgacga tgcgtagccg 300acctgagagg gtgaacggcc acactgggac tgagacacgg cccagactcc tacgggaggc 360agcagtaggg aatcttccgc aatggacgaa agtctgacgg agcaacgccg cgtgagtgat 420gaaggttttc ggatcgtaaa gctctgttgc cagggaagaa cgtccggtag agtaactgct 480accggagtga cggtacctga gaagaaagcc ccggctaact acgtgccagc agccgcggta 540atacgtaggg ggcaagcgtt gtccggaatt attgggcgta aagcgcgcgc aggcggtcac 600ttaagtctgg tgtttaaggc caaggctcaa ccttggttcg cactggaaac tgggtgactt 660gagtgcagaa gaggagagtg gaattccacg tgtagcggtg aaatgcgtag atatgtggag 720gaacaccagt ggcgaaggcg actctctggg ctgtaactga cgctgaggcg cgaaagcgtg 780gggagcaaac aggattagat accctggtag tccacgccgt aaacgatgaa tgctaggtgt 840taggggtttc gatacccttg gtgccgaagt taacacatta agcattccgc ctggggagta 900cggtcgcaag actgaaactc aaaggaattg acggggaccc gcacaagcag tggagtatgt 960ggtttaattc gaagcaacgc gaagaacctt accaagtctt gacatccctc tgaatcctct 1020agagatagag gcggccttcg ggacagaggt gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatttt

agttgccagc 1140atttcggatg ggcactctag aatgactgcc ggtgacaaac cggaggaagg cggggatgac 1200gtcaaatcat catgcccctt atgacttggg ctacacacgt actacaatgg ccagtacaac 1260gggaagcgaa atcgcgagat ggagccaatc ctatcaaagc tggtctcagt tcggattgca 1320ggctgcaacc cgcctgcatg aagtcggaat tgctagtaat cgcggatcag catgccgcgg 1380tgaatacgtt cccgggtctt gtacacaccg cccgtcacac cacgagagtt tacaacaccc 1440gaagtcggtg gggtaacccg caagggagcc agccgccgaa ggtggggtag atgattgggg 1500tgaagtcgta acaaggtagc cgtatcggaa ggtgcggctg gatcacctcc ttt 1553211552DNAPaenibacillus cookii 21cttggagagt ttgatcctgg ctcaggacga acgctggcgg cgtgcctaat acatgcaagt 60cgagcggagt tgatggggag cttgctctcc tgagacttag cggcggacgg gtgagtaaca 120cgtaggcaac ctgcccgtaa gaccgggata actaccggaa acggtagcta ataccggata 180atttatcgct tcgcatggag cggtaatgaa agacggagca atctgtcact tacggatggg 240cctgcggcgc attagctagt tggtgaggta acggctcacc aaggcgacga tgcgtagccg 300acctgagagg gtgaacggcc acactgggac tgagacacgg cccagactcc tacgggaggc 360agcagtaggg aatcttccgc aatgggcgaa agcctgacgg agcaacgccg cgtgagtgat 420gaaggttttc ggatcgtaaa gctctgttgc cagggaagaa cgtcgggtag agtaactgct 480atccgagtga cggtacctga gaagaaagcc ccggctaact acgtgccagc agccgcggta 540atacgtaggg ggcaagcgtt gtccggaatt attgggcgta aagcgcgcgc aggcggtcac 600ttaagtctgg tgtttaaggc tagggctcaa ctctagttcg cactggaaac tgggtgactt 660gagtgcagaa gaggaaagtg gaattccacg tgtagcggtg aaatgcgtag agatgtggag 720gaacaccagt ggcgaaggcg actttctggg ctgtaactga cgctgaggcg cgaaagcgtg 780gggagcaaac aggattagat accctggtag tccacgccgt aaacgatgaa tgctaggtgt 840taggggtttc gatacccttg gtgccgaagt taacacatta agcattccgc ctggggagta 900cggtcgcaag actgaaactc aaaggaattg acggggaccc gcacaagcag tggagtatgt 960ggtttaattc gaagcaacgc gaagaacctt accaggtctt gacatccctc tgaatcctct 1020agagatagag gcggccttcg ggacagagga gacaggtggt gcatggttgt cgtcagctcg 1080tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac ccttgatttt agttgccagc 1140acattaaggt gggcactcta gaatgactgc cggtgacaaa ccggaggaag gcggggatga 1200cgtcaaatca tcatgcccct tatgacctgg gctacacacg tactacaatg gccagtacaa 1260cgggaagcga agtcgcgaga cggagccaat cctatcaaag ctggtctcag ttcggattgc 1320aggctgcaac ccgcctgcat gaagtcggaa ttgctagtaa tcgcggatca gcatgccgcg 1380gtgaatacgt tcccgggtct tgtacacacc gcccgtcaca ccacgagagt ttacaacacc 1440cgaagtcggt ggggtaaccg caaggagcca gccgccgaag gtggggtaga tgattggggt 1500gaagtcgtaa caaggtagcc gtatcggaag gtgcggctgg atcacctcct tt 1552221330DNAPaenibacillus sp. (chitinolyticus) 22ctgtggctta cggatgggcc tgcggcgcat tagctagttg gtgaggtaac ggctcaccaa 60ggcgacgatg cgtagccgac ctgagagggt gaacggccac actgggactg agacacggcc 120cagactccta cgggaggcag cagtagggaa tcttccgcaa tggacgcaag tctgacggag 180caacgccgcg tgagtgatga aggttttcgg atcgtaaagc tctgttgcca gggaagaacg 240ccaaggagag taactgctct ttgggtgacg gtacctgaga agaaagcccc ggctaactac 300gtgccagcag ccgcggtaat acgtaggggg caagcgttgt ccggaattat tgggcgtaaa 360gcgcgcgcag gcggtttttt aagtctggtg tttaatcccg aggctcaacc tcggttcgca 420ccggaaactg ggagactgga gtgcaggaga ggaaagtgga attccacgtg tagcggtgaa 480atgcgtagag atgtggagga acaccagtgg cgaaggcgac tttctggcct gtaactgacg 540ctgaggcgcg aaagcgtggg gagcaaacag gattagatac cctggtagtc cacgccgtaa 600acgatgaatg ctaggtgtta ggggtttcga tacccttggt gccgaagtta acacagtaag 660cattccgcct ggggagtacg ctcgcaagag tgaaactcaa aggaattgac ggggacccgc 720acaagcagtg gagtatgtgg tttaattcga agcaacgcga agaaccttac caggtcttga 780catccctctg accggcttag agataagcct ttccttcggg acagaggtga caggtggtgc 840atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc 900ttgaacttag ttgccagcag gtaaagctgg gcactctaag ttgactgccg gtgacaaacc 960ggaggaaggc ggggatgacg tcaaatcatc atgcccctta tgacctgggc tacacacgta 1020ctacaatggc cggtacaacg ggaagcgaag gagcgatccg gagccaatcc tagaaaagcc 1080ggtctcagtt cggattgcag gctgcaactc gcctgcatga agtcggaatt gctagtaatc 1140gcggatcagc atgccgcggt gaatacgttc ccgggtcttg tacacaccgc ccgtcacacc 1200acgagagttt acaacacccg aagtcggtga ggtaaccgca aggagccagc cgccgaaggt 1260ggggtagatg attggggtga agtcgtaaca aggtagccgt atcggaaggt gcggctggat 1320cacctccttt 133023866DNAPaenibacillus sp. (P1PXP2) 23cagagggatg tcaagacctg gtaaggttct tcgcgttgct tcgaattaaa ccacatactc 60cactgcttgt gcgggtcccc gtcaattcct ttgagtttca gtcttgcgac cgtactcccc 120aggcggaatg cttaatgtgt taacttcggc accaagggta tcgaaacccc taacacctag 180cattcatcgt ttacggcgtg gactaccagg gtatctaatc ctgtttgctc cccacgcttt 240cgcgcctcag cgtcagttac agcccagaga gtcgccttcg ccactggtgt tcctccacat 300atctacgcat ttcaccgcta cacgtggaat tccactctcc tcttctgcac tcaagtcacc 360cagtttccag tgcgaaccaa ggttgagcct tggccttaaa caccagactt aaatgaccgc 420ctgcgcgcgc tttacgccca ataattccgg acaacgcttg ccccctacgt attaccgcgg 480ctgctggcac gtagttagcc ggggctttct tctcaggtac cgtcactccg atagcagtta 540ctctaccgga cgttcttccc tggcaacaga gctttacgat ccgaaaacct tcatcactca 600cgcggcgttg ctccgtcagg ctttcgccca ttgcggaaga ttccctactg ctgcctcccg 660taggagtctg ggccgtgtct cagtcccagt gtggccgttc accctctcag gtcggctacg 720catcgtcgcc ttggtgagcc gttacctcac caactagcta atgcgccgca ggcccatccg 780caagtgacag attgctccgt ctttcatcat cccctcagga gaggaaatga gatatccggt 840attagctcac gtttccgtgg gttatc 866241533DNAPseudomonas sp. 24ctgaagagtt tgatcatggc tcagattgaa cgctggcggc aggcctaaca catgcaagtc 60gagcggatga cgggagcttg ctccttgatt cagcggcgga cgggtgagta atgcctagga 120atctgcctgg tagtggggga caacgtttcg aaaggaacgc taataccgca tacgtcctac 180gggagaaagc aggggacctt cgggccttgc gctatcagat gagcctaggt cggattagct 240agtaggtgag gtaatggctc acctaggcga cgatccgtaa ctggtctgag aggatgatca 300gtcacactgg aactgagaca cggtccagac tcctacggga ggcagcagtg gggaatattg 360gacaatgggc gaaagcctga tccagccatg ccgcgtgtgt gaagaaggtc ttcggattgt 420aaagcacttt aagttgggag gaagggcagt aagctaatac cttgctgttt tgacgttacc 480gacagaataa gcaccggcta actctgtgcc agcagccgcg gtaatacaga gggtgcaagc 540gttaatcgga attactgggc gtaaagcgcg cgtaggtggt tcgttaagtt ggatgtgaaa 600gccccgggct caacctggga actgcatcca aaactggcga gctagagtat ggtagagggt 660ggtggaattt cctgtgtagc ggtgaaatgc gtagatatag gaaggaacac cagtggcgaa 720ggcgaccacc tggactgata ctgacactga ggtgcgaaag cgtggggagc aaacaggatt 780agataccctg gtagtccacg ccgtaaacga tgtcaactag ccgttggaat ccttgagatt 840ttagtggcgc agctaacgca ttaagttgac cgcctgggga gtacggccgc aaggttaaaa 900ctcaaatgaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa 960cgcgaagaac cttaccaggc cttgacatgc agagaacttt ccagagatgg attggtgcct 1020tcgggaactc tgacacaggt gctgcatggc tgtcgtcagc tcgtgtcgtg agatgttggg 1080ttaagtcccg taacgagcgc aacccttgtc cttagttacc agcacgttat ggtgggcact 1140ctaaggagac tgccggtgac aaaccggagg aaggtgggga tgacgtcaag tcatcatggc 1200ccttacggcc tgggctacac acgtgctaca atggtcggta cagagggttg ccaagccgcg 1260aggtggagct aatctcacaa aaccgatcgt agtccggatc gcagtctgca actcgactgc 1320gtgaagtcgg aatcgctagt aatcgcaaat cagaatgttg cggtgaatac gttcccgggc 1380cttgtacaca ccgcccgtca caccatggga gtgggttgca ccagaagtag ctagtctaac 1440cttcgggggg acggttacca cggtgtgatt catgactggg gtgaagtcgt aacaaggtag 1500ccgtagggga acctgcggct ggatcacctc ctt 1533251523DNAStreptomyces griseus 25acggagagtt tgatcctggc tcaggacgaa cgctggcggc gtgcttaaca catgcaagtc 60gaacgatgaa gcctttcggg gtggattagt ggcgaacggg tgagtaacac gtgggcaatc 120tgcccttcac tctgggacaa gccctggaaa cggggtctaa taccggataa cactctgtcc 180cgcatgggac ggggttaaaa gctccggcgg tgaaggatga gcccgcggcc tatcagcttg 240ttggtggggt aatggcctac caaggcgacg acgggtagcc ggcctgagag ggcgaccggc 300cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg gaatattgca 360caatgggcga aagcctgatg cagcgacgcc gcgtgaggga tgacggcctt cgggttgtaa 420acctctttca gcagggaaga agcgagagtg acggtacctg cagaagaagc gccggctaac 480tacgtgccag cagccgcggt aatacgtagg gcgcaagcgt tgtccggaat tattgggcgt 540aaagagctcg taggcggctt gtcacgtcgg atgtgaaagc ccggggctta accccgggtc 600tgcattcgat acgggctagc tagagtgtgg taggggagat cggaattcct ggtgtagcgg 660tgaaatgcgc agatatcagg aggaacaccg gtggcgaagg cggatctctg ggccattact 720gacgctgagg agcgaaagcg tggggagcga acaggattag ataccctggt agtccacgcc 780gtaaacgttg ggaactaggt gttggcgaca ttccacgtcg tcggtgccgc agctaacgca 840ttaagttccc cgcctgggga gtacggccgc aaggctaaaa ctcaaaggaa ttgacggggg 900cccgcacaag cagcggagca tgtggcttaa ttcgacgcaa cgcgaagaac cttaccaagg 960cttgacatat accggaaagc atcagagatg gtgcccccct tgtggtcggt atacaggtgg 1020tgcatggctg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1080cccttgttct gtgttgccag catgcccttc ggggtgatgg ggactcacag gagactgccg 1140gggtcaactc ggaggaaggt ggggacgacg tcaagtcatc atgcccctta tgtcttgggc 1200tgcacacgtg ctacaatggc cggtacaatg agctgcgatg ccgcgaggcg gagcgaatct 1260caaaaagccg gtctcagttc ggattggggt ctgcaactcg accccatgaa gtcggagttg 1320ctagtaatcg cagatcagca ttgctgcggt gaatacgttc ccgggccttg tacacaccgc 1380ccgtcacgtc acgaaagtcg gtaacacccg aagccggtgg cccaacccct tgtgggaggg 1440agctgtcgaa ggtgggactg gcgattggga cgaagtcgta acaaggtagc cgtaccggaa 1500ggtgcggctg gatcacctcc ttt 152326299PRTBacillus megaterium 26Met Leu Thr Asp Leu His Ile Ala Val Ile Gly Gly Asp Ala Arg Gln1 5 10 15Leu Glu Val Ile Arg Lys Leu Ile Gln Leu Asp Ala Lys Thr Ser Leu 20 25 30Ile Gly Phe Asp Gln Leu Asp His Gly Phe Thr Gly Ala Thr Lys Tyr 35 40 45Gln Ile Glu Glu Leu Asp Phe Ser Asp Val Asp Ala Ile Ile Leu Pro 50 55 60Val Pro Gly Thr Asn His Glu Gly Gln Val Asp Thr Ile Phe Ser Asn65 70 75 80Glu Lys Val Val Leu Thr Glu Glu Ile Leu Lys Lys Thr Pro Glu His 85 90 95Cys Ile Ile Tyr Ser Gly Ile Ser Asn Gly Tyr Leu Asn Glu Leu Val 100 105 110Lys Thr Thr Asn Arg Lys Leu Val Gln Leu Phe Glu Arg Asp Asp Val 115 120 125Ala Ile Tyr Asn Ser Ile Pro Thr Val Glu Gly Thr Ile Met Leu Val 130 135 140Ile Gln His Thr Asp Phe Thr Ile His Gly Ser Asn Ile Ser Val Leu145 150 155 160Gly Leu Gly Arg Val Gly Met Ser Val Ala Arg Ser Phe Ala Ala Leu 165 170 175Gly Ala Asn Val Lys Val Gly Ala Arg Lys Ser Glu His Leu Ala Arg 180 185 190Ile Ala Glu Met Gly Leu Gln Pro Phe Tyr Leu Ser Glu Leu Asp Lys 195 200 205Glu Ile Ala Asp Ser Asp Ile Cys Ile Asn Thr Ile Pro Tyr Pro Ile 210 215 220Leu Thr Ala Lys Thr Leu Ser Asn Val Pro Thr His Ala Leu Ile Ile225 230 235 240Asp Leu Ala Ser Lys Pro Gly Gly Thr Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Ile Lys Ala Ile Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Gln Ile Val Ala Asn Val Leu Val Asn Leu Leu 275 280 285Lys Asp Ala Ala Asp Ala Arg Glu Glu Lys Lys 290 29527297PRTBacillus subtilis 27Met Leu Thr Gly Leu Lys Ile Ala Val Ile Gly Gly Asp Ala Arg Gln1 5 10 15Leu Glu Ile Ile Arg Lys Leu Thr Glu Gln Gln Ala Asp Ile Tyr Leu 20 25 30Val Gly Phe Asp Gln Leu Asp His Gly Phe Thr Gly Ala Val Lys Cys 35 40 45Asn Ile Asp Glu Ile Pro Phe Gln Gln Ile Asp Ser Ile Ile Leu Pro 50 55 60Val Ser Ala Thr Thr Gly Glu Gly Val Val Ser Thr Val Phe Ser Asn65 70 75 80Glu Glu Val Val Leu Lys Gln Asp His Leu Asp Arg Thr Pro Ala His 85 90 95Cys Val Ile Phe Ser Gly Ile Ser Asn Ala Tyr Leu Glu Asn Ile Ala 100 105 110Ala Gln Ala Lys Arg Lys Leu Val Lys Leu Phe Glu Arg Asp Asp Ile 115 120 125Ala Ile Tyr Asn Ser Ile Pro Thr Val Glu Gly Thr Ile Met Leu Ala 130 135 140Ile Gln His Thr Asp Tyr Thr Ile His Gly Ser Gln Val Ala Val Leu145 150 155 160Gly Leu Gly Arg Thr Gly Met Thr Ile Ala Arg Thr Phe Ala Ala Leu 165 170 175Gly Ala Asn Val Lys Val Gly Ala Arg Ser Ser Ala His Leu Ala Arg 180 185 190Ile Thr Glu Met Gly Leu Val Pro Phe His Thr Asp Glu Leu Lys Glu 195 200 205His Val Lys Asp Ile Asp Ile Cys Ile Asn Thr Ile Pro Ser Met Ile 210 215 220Leu Asn Gln Thr Val Leu Ser Ser Met Thr Pro Lys Thr Leu Ile Leu225 230 235 240Asp Leu Ala Ser Arg Pro Gly Gly Thr Asp Phe Lys Tyr Ala Glu Lys 245 250 255Gln Gly Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Gln Ile Leu Ala Asn Val Leu Ser Lys Leu Leu 275 280 285Ala Glu Ile Gln Ala Glu Glu Gly Lys 290 29528296PRTPaenibacillus cookii 28Met Leu Thr Gly Leu Thr Ile Ala Ile Ile Gly Gly Asp Ala Arg Gln1 5 10 15Leu Glu Ile Ile Arg Lys Leu Thr Glu Gln His Ala Asp Ile Tyr Leu 20 25 30Ala Gly Phe Asp Gln Leu Asp Asp Gly Phe Thr Gly Thr Val Lys Cys 35 40 45Lys Ile Asp Glu Ile Pro Phe Gln Lys Ile Asp Ser Ile Ile Leu Pro 50 55 60Val Ser Ala Thr Thr Gly Glu Gly Val Val Ser Thr Val Phe Ser Asn65 70 75 80Glu Glu Val Val Leu Lys Gln Ser Tyr Leu Glu Arg Thr Pro Glu His 85 90 95Cys Val Ile Tyr Ser Gly Ile Ser Asn Ala Tyr Leu Glu Gly Ile Ala 100 105 110Ser Glu Ala Gly Arg Lys Leu Val Lys Leu Phe Glu Arg Asp Asp Ile 115 120 125Ala Ile Phe Asn Ser Ile Pro Thr Val Glu Gly Thr Ile Met Met Ala 130 135 140Ile Gln His Thr Asp Tyr Thr Ile His Gly Ser Asn Val Ala Val Leu145 150 155 160Gly Met Gly Arg Thr Gly Met Thr Ile Ala Arg Thr Phe Ala Ala Leu 165 170 175Gly Ala Lys Val Lys Val Gly Ala Arg Ser Ser Ala His Leu Ala Arg 180 185 190Ile Thr Glu Met Gly Leu Ser Pro Phe Gln Leu Glu Glu Leu Thr Glu 195 200 205His Val Asn Asp Ile Asp Ile Cys Ile Asn Thr Val Pro Ser Leu Ile 210 215 220Leu Asn Gln Ser Val Leu Ser Arg Met Thr Pro Lys Thr Leu Ile Leu225 230 235 240Asp Leu Ala Ser Arg Pro Gly Gly Thr Asp Phe Lys Tyr Ala Glu Lys 245 250 255Gln Gly Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Gln Ile Ile Ala Asn Val Leu Ser Lys Leu Leu 275 280 285Ala Asp Leu Lys Lys Glu Gly Lys 290 29529299PRTBacillus licheniformis 29Met Leu Thr Gly Leu Thr Ile Ala Ile Ile Gly Gly Asp Ala Arg Gln1 5 10 15Leu Glu Ile Ile Arg Lys Leu Thr Glu Gln Asp Ala Lys Val Phe Leu 20 25 30Ile Gly Phe Asp Gln Leu Asp His Gly Phe Thr Gly Ala Thr Lys Leu 35 40 45Lys Leu Asn Glu Leu Asp Phe Gly Thr Ile Asp Ser Ile Ile Leu Pro 50 55 60Val Ser Gly Thr Ser Met Glu Gly Thr Val Ala Thr Val Phe Ser Asn65 70 75 80Glu Lys Val Val Leu Lys Gln Glu His Leu Glu Lys Thr Lys Pro His 85 90 95Cys Ala Ile Tyr Ser Gly Ile Ser Asn Gln Tyr Leu Asp Gly Met Ala 100 105 110Lys Gly Ala Asn Arg Arg Leu Ile Lys Leu Phe Glu Arg Asp Asp Ile 115 120 125Ala Ile Tyr Asn Ser Ile Pro Thr Val Glu Gly Ala Ile Met Met Ala 130 135 140Ile Gln His Thr Asp Phe Thr Ile His Gly Ser Asn Val Met Val Leu145 150 155 160Gly Leu Gly Arg Thr Gly Met Ser Ile Ser Arg Thr Phe Ser Ala Leu 165 170 175Gly Ala Arg Val Lys Val Gly Ala Arg Asp Ser Ala His Leu Ala Arg 180 185 190Ile Met Glu Met Gly Leu Thr Pro Phe His Thr Asn Glu Leu Ala Glu 195 200 205His Val Glu Asn Ile Asp Ile Cys Ile Asn Thr Ile Pro Ser Leu Ile 210 215 220Leu Asp Lys His Val Leu Ser Arg Met Thr Pro Arg Thr Leu Ile Leu225 230 235 240Asp Leu Ala Thr Arg Pro Gly Gly Thr Asp Phe Asp Phe Ala Glu Lys 245 250 255Gln Gly Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Gln Ile Ile Ala Asn Val Leu Cys Asn Leu Leu 275 280 285Ser Glu Leu Thr Thr Asp Arg Lys Gly Leu Ser 290 29530298PRTPaenibacillus lautus 30Met Leu Thr Gly Val Arg Thr Val Phe Val Gly Gly Asp Ala Arg Gln1 5 10 15Ile Glu Val Ile Arg Lys Cys Ala Glu Met Asp Ala Ser Val Met Ile

20 25 30Ala Gly Phe Glu Lys Leu Gln Asp Ser Phe Gln Gly Val Thr Arg Glu 35 40 45Pro Leu Thr Pro Glu Leu Leu Ser Asp Ala Asp Ala Leu Ile Leu Pro 50 55 60Val Val Gly Cys Asp Asp Glu Gly Arg Val Ser Ala Leu Phe Ser Glu65 70 75 80Gly Pro Leu Arg Leu Gln Glu Glu His Ile Ala Ala Met Pro Gly His 85 90 95Gly Val Ile Tyr Thr Gly Met Ala Lys Pro Tyr Leu Arg Ser Leu Cys 100 105 110Asp Lys Tyr Lys Ile Lys Leu Val Glu Ile Leu Glu Arg Asp Asp Val 115 120 125Ala Ile Tyr Asn Ser Ile Pro Thr Ala Glu Gly Ala Leu Met Met Ala 130 135 140Ile Gln Asn Thr Asp Phe Thr Ile His Gly Ser Thr Ser Met Val Leu145 150 155 160Gly Met Gly Arg Thr Gly Phe Thr Met Ala Arg Ser Leu Gln Gly Leu 165 170 175Gly Ala Lys Ile Arg Met Gly Val Arg Lys Ser Glu His Tyr Ala Arg 180 185 190Ala Glu Glu Met Gly Trp Lys Pro Phe Leu Val Arg Asp Leu Gly Ser 195 200 205Tyr Val Ser Asp Ile Asp Leu Leu Phe Asn Thr Ile Pro Thr Met Ile 210 215 220Val Thr Ala Gln Ile Ile Ser Lys Met Pro Arg Glu Ala Val Ile Ile225 230 235 240Asp Leu Ala Ser Ala Pro Gly Gly Cys Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Ile Lys Ala Met Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Ile Ile Met Ala Asn Thr Leu Val Glu Leu Ile 275 280 285Ser Glu Glu Ile Lys Ile Arg Glu Asp Ala 290 29531294PRTOceanobacillus oncorhynchi 31Met Ser Arg Arg Ile Ala Val Ile Gly Gly Asp Ala Arg Tyr Leu Glu1 5 10 15Leu Ile Lys Ile Leu Lys Ser Asn His Asp Asn Glu Val Ile Leu Cys 20 25 30Gly Phe Asp Lys Leu Glu Gln Gly Phe Thr Gly Leu Asn Glu Ser Ala 35 40 45Leu Asp Glu Leu Asp Gln Ser Lys Leu Asp Val Val Val Leu Pro Ile 50 55 60Thr Gly Thr Asp Ser Lys Gly Asn Val Glu Thr Val Phe Thr Asp Lys65 70 75 80Lys Ile His Leu Asp Glu Ala Trp Phe Gln Lys Leu His Ala Glu Cys 85 90 95Met Ile Phe Thr Gly Met Thr Asn Ala Tyr Leu Thr Ser Met Ala Glu 100 105 110Lys Ala Gly Val Thr Leu Val Pro Leu Leu Asp Arg Asp Asp Val Ala 115 120 125Ile Tyr Asn Ser Ile Pro Thr Ala Glu Gly Ala Ile Met Met Ala Phe 130 135 140Glu His Thr Asp Gln Thr Val His Ser Ser Arg Val Met Val Val Gly145 150 155 160Phe Gly Arg Val Gly Asn Thr Val Ala Asn Lys Phe Ser Ala Leu Gly 165 170 175Ala Lys Val Ser Val Cys Ala Arg Ser Ile Arg Asp Leu Ala Arg Ile 180 185 190Thr Glu Met Gly Leu Gln Ala Val Pro Leu His Glu Leu Ser Asn His 195 200 205Thr Glu Asn Cys Asp Ile Leu Ile Asn Thr Ile Pro Ser Leu Val Val 210 215 220Thr Lys Glu Ala Ile Gln Asn Leu Pro Thr Asn Ala Val Ile Ile Asp225 230 235 240Leu Ala Ser Lys Pro Gly Gly Thr Asp Phe Asp Phe Ala Lys Lys Arg 245 250 255Gly Ile Gln Ala Ile Leu Ala Arg Ser Leu Pro Gly Ile Val Ala Pro 260 265 270Arg Thr Ala Gly Lys Ile Leu Ala Asn Val Met Glu Gln Ile Leu Glu 275 280 285Glu Glu Arg Ala Ser Glu 29032299PRTBacillus amyloliquefaciens 32Met Leu Thr Gly Val Gln Ile Val Phe Leu Gly Gly Asp Ala Arg Gln1 5 10 15Ile Glu Val Ile Arg Lys Cys Ser Glu Met Asp Ala Thr Val Ser Val 20 25 30Val Gly Phe Asp Asn Leu Lys Glu Lys Leu Gln Gly Val Thr Arg Asp 35 40 45His Leu Thr Ala Glu Leu Leu Ala Ala Ala Asp Val Leu Val Leu Pro 50 55 60Val Val Gly Cys Asp Asp Asn Gly Ile Ile His Thr Gln Phe Ser Asn65 70 75 80Glu Ser Leu Lys Leu Gln Asp Glu His Met Ala Ala Leu Arg Arg Gly 85 90 95Cys Lys Val Tyr Thr Gly Met Ala Lys Pro Tyr Leu Arg Ser Leu Cys 100 105 110Ala His His Glu Ile Lys Leu Ile Glu Leu Leu Asp Arg Asp Glu Val 115 120 125Ala Ile Ser Asn Ser Ile Pro Thr Ser Glu Gly Ala Leu Val Met Ala 130 135 140Ile Gln Asn Thr Asp Phe Thr Ile His Gly Ser Asn Cys Met Val Leu145 150 155 160Gly Leu Gly Arg Thr Gly Phe Thr Met Ala Lys Ser Leu Gln Gly Leu 165 170 175Gly Ala Lys Val Lys Val Gly Val Arg Ser Glu Lys Asp Val Ala Arg 180 185 190Ala Glu Val Met Gly Trp Glu Pro Phe Leu Thr Arg Asp Leu Ala Asp 195 200 205His Val Arg Asn Ile Asp Leu Ile Phe Asn Thr Ile Pro Thr Met Ile 210 215 220Val Thr Ala Gln Ile Leu Ser Arg Met Pro Gln Ser Ala Val Ile Ile225 230 235 240Asp Leu Ala Ser Ala Pro Gly Gly Cys Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Ser Ile Ile Ala Asn Thr Leu Val Gln Leu Ile 275 280 285Ser Asp Glu Phe Lys Thr Arg Gly Asp Gly Gln 290 29533300PRTBacillus sp. 33Met Leu Thr Gly Leu Gln Ile Ala Val Ile Gly Gly Asp Ala Arg Gln1 5 10 15Leu Glu Val Ile Arg Lys Leu Thr Glu Leu Asp Ala Lys Leu Tyr Leu 20 25 30Val Gly Phe Glu Gln Leu Asp His Ala Phe Ser Gly Ala Val Lys Glu 35 40 45Lys Leu Asp Glu Val Asp Phe Thr Cys Ile Asp Ala Ile Ile Leu Pro 50 55 60Val Pro Gly Ala Gly Val Asp Gly Gln Ile Asp Thr Ile Phe Ser Asn65 70 75 80Glu Lys Ile Thr Ile Asn Glu Glu Ile Leu Lys Lys Thr Pro Gln His 85 90 95Cys Lys Ile Tyr Ser Gly Ile Asn Pro Pro Tyr Leu Gln Glu Ile Ser 100 105 110Thr Lys Ala Asp Arg Glu Val Val Gln Leu Phe Asn Arg Asp Asp Val 115 120 125Ala Ile Tyr Asn Ser Ile Pro Thr Val Glu Gly Ala Leu Met Met Ala 130 135 140Ile Gln His Thr Asp Phe Thr Ile His Gly Ser Asn Val Thr Val Leu145 150 155 160Gly Leu Gly Arg Thr Gly Met Ser Ile Ala Arg Ala Phe His Ala Leu 165 170 175Gly Ala Lys Val Lys Val Gly Ala Arg Lys Ser Glu His Ile Ala Arg 180 185 190Ile Thr Glu Met Gly Leu Thr Pro Phe His Leu Ser Asp Ile Glu Glu 195 200 205Ala Val Val Asp Thr Asp Ile Cys Ile Asn Thr Ile Pro Val Gln Val 210 215 220Val Val Ala Ser Val Ile Ala Lys Met Pro Val His Thr Leu Ile Ile225 230 235 240Asp Leu Ala Ser Lys Pro Gly Gly Thr Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Val Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Arg Ile Leu Ala Asn Val Leu Ser Gln Leu Ile 275 280 285Leu Ala Asn Phe Asp Glu Arg Glu Asp Lys Gln Ser 290 295 30034299PRTPaenibacillus chibensis 34Met Leu Thr Gly Val Gln Ile Val Phe Leu Gly Gly Asp Ala Arg Gln1 5 10 15Val Glu Val Ile Arg Lys Cys Ser Glu Met Asp Ala Thr Val Ser Val 20 25 30Val Gly Phe Asp Asn Leu Lys Gln Lys Leu Gln Gly Val Thr Arg Asp 35 40 45His Leu Thr Ala Glu Leu Leu Ala Ala Ala Asp Val Leu Val Leu Pro 50 55 60Val Val Gly Cys Asp Asp Asn Gly Asn Ile His Thr Gln Phe Ser Asn65 70 75 80Glu Pro Leu Lys Leu Gln Asp Glu His Met Ala Ser Leu Arg Lys Gly 85 90 95Cys Lys Val Tyr Thr Gly Met Ala Lys Pro Tyr Leu Arg Ser Leu Cys 100 105 110Ala Gln His Glu Ile Lys Leu Val Glu Leu Leu Asp Arg Asp Glu Val 115 120 125Ala Ile Ser Asn Ser Ile Pro Thr Ala Glu Gly Ala Leu Val Met Ala 130 135 140Ile Gln Asn Thr Asp Phe Thr Ile His Gly Ser Arg Cys Met Val Leu145 150 155 160Gly Leu Gly Arg Thr Gly Phe Thr Met Ala Lys Ser Leu Gln Gly Leu 165 170 175Gly Ala Lys Val Lys Val Gly Val Arg Ser Glu Lys Asp Val Ala Arg 180 185 190Ala Glu Val Met Gly Trp Glu Pro Phe Leu Thr Arg Asp Leu Gly Asp 195 200 205His Val Ser Asn Ile Asp Leu Ile Phe Asn Thr Ile Pro Thr Met Ile 210 215 220Val Thr Ala Gln Ile Leu Ser Lys Met Pro Leu Ser Ser Val Ile Ile225 230 235 240Asp Leu Ala Ser Ala Pro Gly Gly Cys Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Leu Ile Ile Ala Gly Ser Leu Val Gln Leu Ile 275 280 285Ser Asp Glu Phe Lys Thr Arg Gly Asp Gly Glu 290 29535299PRTBacillus flexus 35Met Leu Thr Asp Leu His Ile Ala Val Ile Gly Gly Asp Ala Arg Gln1 5 10 15Leu Glu Val Ile Arg Lys Leu Ile Gln Leu Asp Ala Lys Thr Ser Leu 20 25 30Ile Gly Phe Asp Gln Leu Asp His Gly Phe Thr Gly Ala Thr Lys Tyr 35 40 45Gln Ile Asp Glu Leu Asn Phe Ser Asp Val Asp Ala Ile Ile Leu Pro 50 55 60Val Pro Gly Thr Asn His Glu Gly Gln Val Asp Thr Ile Phe Ser Asn65 70 75 80Glu Lys Val Ile Leu Thr Glu Glu Ile Leu Ala Ser Thr Pro Ala His 85 90 95Cys Thr Ile Tyr Ser Gly Ile Ser Asn Asp Tyr Leu Asn Ser Leu Val 100 105 110Gln Lys Thr Asn Arg Thr Leu Ile Gln Leu Phe Glu Arg Asp Asp Val 115 120 125Ala Ile Tyr Asn Ser Ile Pro Thr Val Glu Gly Thr Ile Met Leu Val 130 135 140Ile Gln His Thr Asp Phe Thr Ile His Gly Ala Asn Ile Ser Val Leu145 150 155 160Gly Leu Gly Arg Val Gly Met Ser Val Ala Arg Ser Phe Ala Ala Leu 165 170 175Gly Ala Asn Val Lys Val Gly Ala Arg Lys Ser Glu His Leu Ala Arg 180 185 190Ile Ser Glu Met Gly Leu Thr Pro Phe His Leu Asn Asp Leu Ala Gln 195 200 205Glu Ile Thr Asp Ser Asp Ile Cys Ile Asn Thr Ile Pro Tyr Pro Val 210 215 220Leu Thr Ser Ser Val Leu Ala Asn Ile Pro Thr His Ala Leu Val Val225 230 235 240Asp Leu Ala Ser Lys Pro Gly Gly Thr Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Ile Lys Ala Ile Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Gln Ile Val Ala Asn Val Ile Val Thr Leu Leu 275 280 285Lys Glu Ala Ala Asp Gln Arg Glu Glu Lys Gln 290 29536300PRTBacillus firmus 36Met Leu Thr Gly Thr Gln Ile Ala Val Ile Gly Gly Asp Ala Arg Gln1 5 10 15Leu Glu Ile Ile Arg Lys Leu Thr Glu Leu Asp Ala Lys Leu Ser Leu 20 25 30Ile Gly Phe Glu Gln Leu Asp His Ala Phe Ser Gly Ala Val Lys Glu 35 40 45Lys Ile Asp Glu Val Asp Phe Ser His Ile Asp Ala Ile Ile Leu Pro 50 55 60Val Pro Gly Thr Gly Leu Glu Gly Gln Ile Glu Thr Ile Phe Ser Asn65 70 75 80Glu Lys Val Thr Leu Glu Glu Glu Ile Leu Ser Gln Thr Pro Ala His 85 90 95Cys Thr Val Tyr Ser Gly Ile Thr Asn Ser Tyr Leu Thr Gly Val Thr 100 105 110Lys Ser Ala Asp Arg Arg Leu Val Gln Leu Phe Glu Arg Asp Asp Val 115 120 125Ala Ile Tyr Asn Ser Ile Pro Thr Val Glu Gly Thr Ile Met Met Ala 130 135 140Ile Gln His Thr Asp Phe Thr Ile His Gly Ser Asn Ile Ala Val Ile145 150 155 160Gly Leu Gly Arg Val Gly Met Ser Val Ala Arg Thr Phe Arg Ala Leu 165 170 175Gly Ala Lys Val Lys Val Gly Ala Arg Lys Ser Glu His Ile Ala Arg 180 185 190Ile Thr Glu Met Gly Leu Thr Pro Phe Asn Leu Lys Glu Ile Glu Asp 195 200 205Ala Val Lys Asp Val Asp Ile Cys Ile Asn Thr Ala Pro His Leu Val 210 215 220Val Thr Ala Ser Val Ile Ser Lys Met Pro Thr His Thr Leu Ile Ile225 230 235 240Asp Leu Ala Ser Lys Pro Gly Gly Thr Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Val Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Gln Ile Leu Ala Asn Val Leu Ser Gln Leu Ile 275 280 285Met Glu Asp Leu Gln Lys Arg Lys Gly Asn Thr Lys 290 295 30037291PRTVirgilia halophilus 37Met Asn Leu Lys Asp Lys Lys Ile Leu Leu Ile Gly Gly Asp Glu Arg1 5 10 15Tyr Leu Glu Val Val Lys Lys Leu Asp Asp Leu Gly Ala Ser Val Val 20 25 30Leu Ala Gly Tyr Asp Lys Ala Gly Leu Ser Ser Gly Arg Val Gln Ile 35 40 45Ser Lys Leu Glu Asp Val Asn Phe Ser Asn Leu Tyr Ala Ile Leu Leu 50 55 60Pro Val Ser Gly Thr Asp Gly Glu Gly Asn Ile Thr Met Ser Ser Phe65 70 75 80Thr Asp Gln Gln Leu Cys Leu Thr Glu Gln Met Ile Ser Gln Leu Pro 85 90 95Pro Ser Cys Lys Ile Tyr Thr Gly Val Ser Gly Ser Phe Leu Lys Arg 100 105 110Met Gly Ser Lys Phe Gln Lys Glu Ile Ile Ser Ile Leu Ala Arg Glu 115 120 125Asp Ile Ala Ile Tyr Asn Ser Ile Pro Thr Ala Glu Gly Ala Leu Gln 130 135 140Leu Ala Met Glu Gln Thr Asp Tyr Thr Met His Ser Ala Ser Val Met145 150 155 160Val Leu Gly Phe Gly Lys Val Gly Met Thr Thr Ala Arg Leu Phe Ser 165 170 175Ala Val Gly Cys Asn Val Ser Val Ala Ile Arg Lys Asp Ser Ala Ala 180 185 190Ala Arg Val Arg Glu Met Gly Leu Lys Pro Leu Tyr Thr His His Leu 195 200 205Ser Glu Glu Ile Gly Gln Tyr Gln Ile Ile Ile Asn Thr Val Pro Asp 210 215 220Leu Val Leu Asp Glu Ser Leu Leu Asn Ile Val Ser Ser Lys Ala Leu225 230 235 240Ile Ile Asp Leu Ala Ser Ser Pro Gly Gly Val Asp Phe Ser Val Ala 245 250 255Asp Glu Leu Gly Ile Arg Thr Ile His Ala Leu Gly Leu Pro Gly Lys 260 265 270Val Ala Pro Lys Thr Ala Gly Ser Ile Ile Ala Asp Thr Phe Val Ser 275 280 285Leu Leu Ser 29038269PRTVirgibacillus halophilus 38Met Lys Thr Ile Val Thr Gly Phe Asn Lys Leu Asp Gln Gly Phe Thr1 5 10 15Gly Val Gln His Val Glu Phe Ala Glu Met Glu His Glu Asp Ile Asp 20 25 30Val Val Val Leu Pro Ile Thr Gly Thr Gln Lys Gly Gly Lys Val Glu 35 40 45Thr Val Phe Ser Asp Glu Glu Ile Val Leu Thr Lys Asp Trp Phe Glu 50 55 60Lys Phe Gln Arg Pro Thr Pro Val Phe Thr Gly Ile Ser Asn Gln Asp65 70 75 80Leu Asp Gly Met Val Lys Asn Ser Lys Ala Gln Ile Ile Pro Leu Leu 85 90

95Asp Arg Asp Asp Val Ala Ile Tyr Asn Ser Ile Pro Thr Ala Glu Gly 100 105 110Thr Ile Met Met Ala Met Glu His Thr Asp Tyr Thr Ile His Ser Ser 115 120 125Arg Val Ile Val Ala Gly Phe Gly Arg Val Gly His Thr Val Ala Asn 130 135 140Lys Phe Ser Ala Leu Gly Ala Lys Val Ser Val Ala Ala Ser Ser Ile145 150 155 160His Asp Ile Ala Arg Ile Asn Glu Met Gly Leu Phe Ala Ile Thr Met 165 170 175Lys Glu Leu Ala Lys Ala Ala Ala Asp Cys Asp Ile Leu Ile Asn Thr 180 185 190Ile Pro Ala Pro Val Ile Asn Lys Glu Ala Ile Ser Gln Leu Pro His 195 200 205His Ala Leu Ile Phe Asp Leu Ala Ser Lys Pro Gly Gly Thr Asp Phe 210 215 220Asp Tyr Ala Lys Arg Arg Gly Ile Lys Ala Ile Leu Ser Glu Ser Leu225 230 235 240Pro Gly Val Val Ala Pro Lys Thr Ala Gly Lys Ile Leu Ala Asp Val 245 250 255Ile Ile Gln Ile Leu Ser Gln Arg Lys Gly Phe Glu Gln 260 26539299PRTPaenibacillus azoreducens 39Met Leu Thr Gly Val Gln Ile Val Phe Leu Gly Gly Asp Ala Arg Gln1 5 10 15Ile Glu Val Ile Arg Lys Cys Ser Glu Met Asp Ala Thr Val Ser Val 20 25 30Val Gly Phe Asp Asn Leu Lys Glu Lys Leu Gln Gly Val Thr Arg Asp 35 40 45Gln Leu Thr Gly Glu Leu Leu Ala Gly Ala Asp Val Leu Val Leu Pro 50 55 60Val Val Gly Cys Asp Asp Asn Gly Ile Ile His Thr Gln Phe Ser Asn65 70 75 80Glu Ser Leu Lys Leu Gln Asp Glu His Met Ala Ser Leu Arg Arg Gly 85 90 95Cys Lys Val Tyr Thr Gly Met Ala Lys Pro Tyr Leu Arg Ser Leu Cys 100 105 110Ala His His Glu Ile Arg Leu Val Glu Leu Leu Asp Arg Asp Glu Val 115 120 125Ala Ile Ser Asn Ser Ile Pro Thr Ala Glu Gly Ala Leu Val Met Ala 130 135 140Ile Gln Asn Thr Asp Phe Thr Ile His Gly Ser Asp Cys Met Val Leu145 150 155 160Gly Leu Gly Arg Thr Gly Phe Thr Met Ala Lys Ser Leu Gln Gly Leu 165 170 175Gly Ala Arg Val Lys Val Gly Val Arg Ser Glu Arg Asp Phe Ala Arg 180 185 190Ala Glu Val Met Gly Trp Glu Pro Phe Leu Thr Arg Asp Leu Ala Asp 195 200 205Tyr Val Arg Ser Ile Asp Leu Ile Phe Asn Thr Ile Pro Thr Met Ile 210 215 220Val Thr Ala Gln Ile Leu Ser Arg Met Pro Gln Asn Thr Val Ile Ile225 230 235 240Asp Leu Ala Ser Ala Pro Gly Gly Cys Asp Phe Arg Tyr Ala Glu Lys 245 250 255Arg Gly Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Ile Val Ala 260 265 270Pro Lys Thr Ala Gly Ser Ile Ile Ala Asn Ser Leu Val Gln Met Ile 275 280 285Ser Asp Glu Phe Lys Thr Arg Gly Asp Gly Glu 290 29540296PRTArtificial SequenceConsensus DpaA sequence 40Thr Gly Lys His Ile Ala Val Ile Gly Gly Asp Ala Arg Gln Leu Glu1 5 10 15Leu Ile Arg Lys Leu Val Glu Leu Gly Ala Lys Val Ser Leu Val Gly 20 25 30Phe Asp Gln Leu Asp His Gly Phe Thr Gly Ala Thr Lys Ser Ser Ser 35 40 45Leu Glu Glu Ala Leu Ser Asp Val Asp Val Ile Ile Leu Pro Val Pro 50 55 60Gly Thr Asn Asp Glu Gly Asn Val Asp Thr Val Phe Ser Asn Glu Lys65 70 75 80Leu Val Leu Thr Glu Glu Leu Leu Glu Leu Thr Pro Glu His Cys Thr 85 90 95Ile Phe Ser Gly Ile Ala Asn Pro Tyr Leu Lys Glu Leu Ala Lys Glu 100 105 110Thr Asn Arg Lys Leu Val Glu Leu Phe Glu Arg Asp Asp Val Ala Ile 115 120 125Leu Asn Ser Ile Pro Thr Ala Glu Gly Ala Ile Met Met Ala Ile Glu 130 135 140His Thr Pro Ile Thr Ile His Gly Ser Asn Val Leu Val Leu Gly Phe145 150 155 160Gly Arg Thr Gly Met Thr Leu Ala Arg Thr Leu Lys Ala Leu Gly Ala 165 170 175Asn Val Thr Val Gly Ala Arg Lys Ser Ala His Leu Ala Arg Ile Thr 180 185 190Glu Met Gly Leu Ser Pro Phe His Leu Ser Glu Leu Ala Glu Glu Val 195 200 205Gly Lys Ile Asp Ile Ile Phe Asn Thr Ile Pro Ala Leu Val Leu Thr 210 215 220Lys Glu Val Leu Ser Lys Met Pro Pro Glu Ala Leu Ile Ile Asp Leu225 230 235 240Ala Ser Lys Pro Gly Gly Thr Asp Phe Glu Tyr Ala Glu Lys Arg Gly 245 250 255Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Lys Val Ala Pro Lys 260 265 270Thr Ala Gly Gln Ile Leu Ala Asn Val Leu Ser Gln Leu Leu Ala Glu 275 280 285Asp Leu Ile Ala Arg Lys Glu Asn 290 29541296PRTArtificial SequenceDpaA consensus sequence 41Thr Gly Lys His Ile Ala Val Ile Gly Gly Asp Ala Arg Gln Leu Glu1 5 10 15Leu Ile Arg Lys Leu Val Glu Leu Gly Ala Lys Val Ser Leu Val Gly 20 25 30Phe Asp Gln Leu Asp His Gly Phe Thr Gly Ala Thr Lys Ser Ser Ser 35 40 45Leu Glu Glu Ala Leu Ser Asp Val Asp Val Ile Ile Leu Pro Val Pro 50 55 60Gly Thr Asn Asp Glu Gly Asn Val Asp Thr Val Phe Ser Asn Glu Lys65 70 75 80Leu Val Leu Thr Glu Glu Leu Leu Glu Leu Thr Pro Glu His Cys Thr 85 90 95Ile Phe Ser Gly Ile Ala Asn Pro Tyr Leu Lys Glu Leu Ala Lys Glu 100 105 110Thr Asn Arg Lys Leu Val Glu Leu Phe Glu Arg Asp Asp Val Ala Ile 115 120 125Leu Asn Ser Ile Pro Thr Ala Glu Gly Ala Ile Met Met Ala Ile Glu 130 135 140His Thr Pro Ile Thr Ile His Gly Ser Asn Val Leu Val Leu Gly Phe145 150 155 160Gly Arg Thr Gly Met Thr Leu Ala Arg Thr Leu Lys Ala Leu Gly Ala 165 170 175Asn Val Thr Val Gly Ala Arg Lys Ser Ala His Leu Ala Arg Ile Thr 180 185 190Glu Met Gly Leu Ser Pro Phe His Leu Ser Glu Leu Ala Glu Glu Val 195 200 205Gly Lys Ile Asp Ile Ile Phe Asn Thr Ile Pro Ala Leu Val Leu Thr 210 215 220Lys Glu Val Leu Ser Lys Met Pro Pro Glu Ala Leu Ile Ile Asp Leu225 230 235 240Ala Ser Lys Pro Gly Gly Thr Asp Phe Glu Tyr Ala Glu Lys Arg Gly 245 250 255Ile Lys Ala Leu Leu Ala Pro Gly Leu Pro Gly Lys Val Ala Pro Lys 260 265 270Thr Ala Gly Gln Ile Leu Ala Asn Val Leu Ser Gln Leu Leu Ala Glu 275 280 285Asp Leu Ile Ala Arg Lys Glu Asn 290 29542201PRTBacillus megaterium 42Met Ser Leu Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15Cys Thr Tyr Asp Ala Val Met Pro Glu Ile Glu Lys Leu Val Asn Leu 20 25 30Gly Ala Glu Val Leu Pro Val Val Ser Tyr Thr Val Gln Ser Thr Asn 35 40 45Thr Arg Phe Gly Asp Gly Glu Asp Trp Val Lys Lys Ile Glu Glu Leu 50 55 60Thr Gly His Ala Val Ile Asn Thr Ile Val Lys Ala Glu Pro Leu Gly65 70 75 80Pro Lys Ile Pro Leu Asp Cys Met Val Val Ala Pro Ile Thr Gly Asn 85 90 95Thr Met Ser Lys Phe Ala Asn Ala Met Thr Glu Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Thr Leu Arg Asn Asn Lys Pro Val Val Leu Gly Ile 115 120 125Ser Thr Asn Asp Ala Leu Gly Leu Asn Gly Val Asn Leu Met Arg Leu 130 135 140Met Ala Thr Lys Asn Ile Tyr Phe Ile Pro Phe Gly Gln Asp Asp Pro145 150 155 160Val Leu Lys Pro Asn Ser Met Val Ala Arg Met Thr Met Leu Ser Asp 165 170 175Thr Val Tyr Ala Ala Leu Glu Asp Lys Gln Ile Gln Pro Val Ile Val 180 185 190Glu Arg Phe Arg Asp Gly Gln Glu Ser 195 20043200PRTBacillus subtilis 43Met Ser Ser Leu Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser1 5 10 15His Cys Thr Tyr Glu Ala Val Phe Pro Gln Ile Glu Ala Leu Val Asn 20 25 30Glu Gly Ala Glu Val Arg Pro Val Val Thr Phe Asn Val Lys Ser Thr 35 40 45Asn Thr Arg Phe Gly Glu Gly Ala Glu Trp Val Lys Lys Ile Glu Glu 50 55 60Leu Thr Gly Tyr Glu Ala Ile Asp Ser Ile Val Lys Ala Glu Pro Leu65 70 75 80Gly Pro Lys Leu Pro Leu Asp Cys Met Val Ile Ala Pro Leu Thr Gly 85 90 95Asn Ser Met Ser Lys Leu Ala Asn Ala Met Thr Asp Ser Pro Val Leu 100 105 110Met Ala Ala Lys Ala Thr Ile Arg Asn Asn Arg Pro Val Val Leu Gly 115 120 125Ile Ser Thr Asn Asp Ala Leu Gly Leu Asn Gly Thr Asn Leu Met Arg 130 135 140Leu Met Ser Thr Lys Asn Ile Phe Phe Ile Pro Phe Gly Gln Asp Asp145 150 155 160Pro Phe Lys Lys Pro Asn Ser Met Val Ala Lys Met Asp Leu Leu Pro 165 170 175Gln Thr Ile Glu Lys Ala Leu Leu His Gln Gln Leu Gln Pro Ile Leu 180 185 190Val Glu Asn Tyr Gln Gly Asn Asp 195 20044196PRTPaenibacillus cookii 44Met Ser Leu Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15Cys Thr Tyr Glu Glu Val Phe Pro Gln Ile Glu Ala Leu Ile Ser Gln 20 25 30Gly Ala Glu Val Arg Pro Val Val Thr Ser Thr Val Gln Ser Thr Asp 35 40 45Thr Arg Phe Gly Glu Gly Gly Asp Trp Val Arg Lys Ile Glu Glu Ala 50 55 60Thr Gly Phe Glu Ala Ile Asp Ser Ile Val Lys Ala Glu Pro Leu Gly65 70 75 80Pro Lys Leu Pro Leu Asp Cys Met Val Ile Ala Pro Leu Thr Gly Asn 85 90 95Ser Met Ser Lys Leu Ala Asn Ala Met Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Thr Ile Arg Asn Gly Arg Pro Val Val Leu Gly Ile 115 120 125Ser Thr Asn Asp Gly Leu Gly Leu Asn Gly Thr Asn Leu Met Arg Leu 130 135 140Met Ser Ala Lys Asn Ile Tyr Phe Ile Pro Phe Gly Gln Asp Asp His145 150 155 160Val Lys Lys Pro Thr Ser Leu Val Ala Arg Met Asp Leu Leu Pro Ile 165 170 175Thr Val Glu Lys Ala Leu Leu His Gln Gln Val Gln Pro Val Leu Val 180 185 190His His His Glu 19545196PRTBacillus licheniformis 45Met Ser Ile Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15Cys Thr Tyr Asp Ala Val Phe Pro Gln Ile Glu Ala Leu Ile Asn Lys 20 25 30Gly Ala Glu Val Arg Pro Val Val Thr His Thr Val Lys Ser Thr Asp 35 40 45Thr Arg Phe Gly Glu Gly Glu Glu Trp Val Arg Arg Ile Glu Glu Leu 50 55 60Thr Gly Phe Glu Val Ile Asp Ser Ile Pro Lys Ala Glu Pro Leu Gly65 70 75 80Pro Lys Thr Pro Leu Asp Cys Met Val Val Ala Pro Leu Thr Gly Asn 85 90 95Ser Met Ser Lys Leu Ala Asn Ala Gln Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Thr Met Arg Asn Ser Arg Pro Val Val Leu Gly Ile 115 120 125Ser Thr Asn Asp Ala Leu Gly Leu Asn Gly Val Asn Leu Met Arg Leu 130 135 140Met Ala Ala Lys Asn Val Tyr Phe Ile Pro Phe Gly Gln Asp Asp Pro145 150 155 160Tyr Lys Lys Pro Asn Ser Leu Val Ala Lys Met Asp Leu Leu Val Pro 165 170 175Ala Val Glu Glu Ala Leu Ser His Lys Gln Ile Gln Pro Ile Leu Val 180 185 190His Asn Asp Gln 19546196PRTPaenibacillus lautus 46Met Asn Trp Asn Gly Ile Thr Val Gly Tyr Ala Leu Thr Gly Ser His1 5 10 15Cys Thr Leu Glu Glu Val Met Pro Gln Ile Gln Arg Phe Lys Asp Gly 20 25 30Gly Ala Asn Val Val Pro Ile Val Ser Ser Thr Ile Met Thr Thr Asp 35 40 45Thr Arg Phe Gly Thr Ser Glu Asn Trp Gln Lys Gln Leu Lys Asp Ile 50 55 60Thr Gly Asn Asp Ile Ile Ser Thr Ile Val Glu Ala Glu Pro Leu Gly65 70 75 80Pro Ser Lys Leu Leu Asp Val Leu Val Ile Ala Pro Cys Thr Gly Asn 85 90 95Thr Thr Ser Lys Leu Ala Asn Ala Met Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Gln Met Arg Asn Cys Arg Pro Leu Val Leu Ala Ile 115 120 125Ser Thr Asn Asp Gly Leu Gly Leu Asn Ala Ala Asn Ile Ala Lys Leu 130 135 140Leu Val Thr Lys Asn Ile Tyr Phe Val Pro Tyr Gly Gln Asp Asn Pro145 150 155 160Gln Gln Lys Pro Asn Ser Leu Val Ala Lys Met Asn Leu Ile Pro Glu 165 170 175Ala Cys Tyr Ala Ala Leu Glu Gly Lys Gln Leu Gln Pro Met Ile Val 180 185 190Glu Tyr Ser Arg 19547201PRTOceanobacillus oncorhynchi 47Met Thr Phe Lys Asn Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15His Thr Leu Pro His Ile Phe Pro Ile Ile Glu Glu Leu Ile Glu Gln 20 25 30Gly Ala Glu Val Ile Pro Phe Ile Thr Glu Met Val Gln Tyr Thr Asp 35 40 45Thr Lys His Gly Lys Ala Ala Asp Asn Val Lys Arg Leu Glu Lys Ala 50 55 60Ala Asn His Pro Ile Ile Thr Ser Ile Pro Asp Ala Glu Pro Tyr Gly65 70 75 80Pro Asp Lys Pro Leu Asp Val Met Val Ile Ala Pro Leu Thr Gly Asn 85 90 95Ser Met Ser Lys Leu Ala Asn Ala His Thr Asp Asn Pro Val Leu Met 100 105 110Ala Ala Lys Ser Thr Leu Arg Asn Glu His Pro Leu Leu Leu Ala Leu 115 120 125Thr Thr Asn Asp Ala Leu Gly Leu Asn Ala Lys Asn Leu Ala Val Leu 130 135 140Leu Asn Ala Lys His Ile Tyr Phe Val Pro Phe Gly Gln Asp Asn Pro145 150 155 160His Gln Lys Pro Ser Ser Leu Ser Ala Asn Leu Asp Gln Leu Ile Pro 165 170 175Ala Ala Glu Ala Ala Leu Lys Gly Lys Gln Ile Gln Pro Ile Ile Val 180 185 190Pro Tyr Ser Thr Lys Asn Val Leu Lys 195 20048198PRTBacillus amyloliquefaciens 48Met Asn Trp Gln Gly Lys Thr Val Gly Tyr Ala Val Thr Gly Ser His1 5 10 15Cys Thr Leu Glu Glu Ile Met Pro Gln Val Lys Arg Phe Val Glu Ala 20 25 30Gly Ala Asn Val Val Pro Ile Ala Ser Gly Ser Val Gln Val Thr Asp 35 40 45Thr Arg Phe Gly Thr Ala Gln Asn Trp Leu Gln Gln Leu Lys Asp Ile 50 55 60Thr Gly Asn Asp Ile Ile Thr Thr Ile Val Glu Ala Glu Pro Leu Gly65 70 75 80Pro Ser Lys Leu Leu Asp Val Leu Val Ile Ala Pro Cys Thr Gly Asn 85 90 95Thr Thr Ser Lys Leu Ala Asn Ala Met Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Gln Met Arg Asn Gln Arg Pro Leu Val Leu Ala Ile 115 120 125Ser Thr Asn Asp Gly Leu Gly Leu Asn Ala Ser Asn Ile Ala Lys Leu 130 135 140Leu Ile Thr Lys Asn Ile Tyr Phe Val Pro Phe Gly Gln Asp Asn Pro145 150 155 160Phe Gln Lys Pro Asn Ser Leu Val Ala Gln Met Asp Leu Ile Pro Glu 165

170 175Ala Cys Tyr Ala Ala Leu Glu Gly Lys Gln Leu Gln Pro Met Ile Leu 180 185 190Gln Arg Val Phe Ser Ala 19549199PRTBacillus sp. 49Met Asn Leu Lys Gly Lys Lys Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15Cys Thr Tyr Asp Ala Val Phe Pro Glu Ile Glu Lys Leu Val Gly Ala 20 25 30Gly Ala Glu Val Ile Pro Val Val Thr Phe Thr Val Gln Asn Thr Val 35 40 45Thr Arg Phe Gly Asp Gly Glu Asp Trp Ile Lys Arg Ile Glu Glu Val 50 55 60Thr Gly Asn Lys Val Ile Asp Ser Ile Val Lys Ala Glu Pro Leu Gly65 70 75 80Pro Lys Ile Pro Leu Asp Cys Met Val Val Ala Pro Leu Thr Gly Asn 85 90 95Ser Met Ser Lys Phe Ala Asn Ala Met Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Thr Leu Arg Asn Glu Lys Pro Val Val Leu Gly Ile 115 120 125Ser Thr Asn Asp Ala Leu Gly Leu Asn Gly Thr Asn Leu Met Arg Leu 130 135 140Met Ser Thr Lys Asn Ile Tyr Phe Ile Pro Phe Gly Gln Asp Asp Pro145 150 155 160Val Lys Lys Pro Asn Ser Met Val Ala Arg Met Thr Ala Leu Ser Asp 165 170 175Thr Ile Val Lys Ala Ile Asn Gly Glu Gln Ile Gln Pro Val Ile Val 180 185 190Glu Arg Tyr Lys Asp Gly Asn 19550198PRTPaenibacillus chibensis 50Met Asn Trp Gln Gly Lys Thr Val Gly Tyr Ala Ile Thr Gly Ser His1 5 10 15Cys Thr Leu Glu Glu Ile Met Pro Gln Val Lys Arg Phe Val Asp Glu 20 25 30Gly Ala Lys Val Val Pro Ile Val Ser Asn Ser Val Gln Val Thr Asp 35 40 45Thr Arg Phe Gly Thr Ala Gln Asn Trp Leu Gln Gln Leu Lys Asp Ile 50 55 60Thr Gly Asn Asp Ile Ile Ser Ser Ile Val Asp Ala Glu Pro Leu Gly65 70 75 80Pro Ser Lys Leu Leu Asp Val Leu Val Ile Ala Pro Cys Thr Gly Asn 85 90 95Thr Thr Ser Lys Leu Ala Asn Ala Met Thr Asp Thr Pro Val Leu Met 100 105 110Ala Ala Lys Ala Gln Met Arg Asn Leu Arg Pro Leu Val Leu Ala Ile 115 120 125Ser Thr Asn Asp Gly Leu Gly Leu Asn Ala Ala Asn Ile Ala Lys Leu 130 135 140Leu Val Thr Lys Asn Ile Tyr Phe Val Pro Phe Gly Gln Asp Asn Pro145 150 155 160Leu Gln Lys Pro Asn Ser Leu Val Ala Gln Met Asp Leu Ile Pro Glu 165 170 175Ala Cys Tyr Ala Ala Leu Glu Gly Arg Gln Leu Gln Pro Met Ile Leu 180 185 190Gln Arg Ile Phe Ser Ala 19551197PRTBacillus flexus 51Met Ser Leu Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15Cys Thr Tyr Asp Ala Val Met Pro Glu Ile Glu Lys Leu Val Asn Leu 20 25 30Gly Ala Glu Val Met Pro Val Val Ser Tyr Thr Val Gln Ser Thr Asn 35 40 45Thr Arg Phe Gly Asp Gly Glu Asp Trp Ile Arg Lys Ile Glu Glu Val 50 55 60Thr Gly Asn Ser Val Ile Asn Thr Ile Val Lys Ala Glu Pro Leu Gly65 70 75 80Pro Lys Ile Pro Leu Asp Cys Met Val Val Ala Pro Ile Thr Gly Asn 85 90 95Thr Met Ser Lys Phe Ala Asn Ala Met Thr Glu Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Thr Leu Arg Asn Asn Lys Pro Val Val Leu Gly Ile 115 120 125Ser Thr Asn Asp Ala Leu Gly Leu Asn Gly Val Asn Leu Met Arg Leu 130 135 140Met Ala Thr Lys Asn Ile Tyr Phe Ile Pro Phe Gly Gln Asp Asp Pro145 150 155 160Val Ser Lys Pro Asn Ser Met Val Ala Arg Met Pro Met Leu Ser Asp 165 170 175Thr Val Tyr Ala Ala Leu Glu Gly Lys Gln Ile Gln Pro Val Val Val 180 185 190Glu Arg Phe Arg Asp 19552199PRTBacillus firmus 52Met Ser Leu Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15Cys Thr Tyr Asp Ala Val Phe Pro Glu Ile Glu Arg Leu Val Leu Ala 20 25 30Gly Ala Glu Val Leu Pro Val Val Thr Phe Thr Val Lys Ser Thr Glu 35 40 45Thr Arg Phe Gly Lys Gly Glu Asp Trp Val Gln Arg Ile Glu Asp Leu 50 55 60Thr Gly Asn Lys Val Ile Asp Ser Ile Val Lys Ala Glu Pro Leu Gly65 70 75 80Pro Lys Ile Pro Leu Asp Cys Met Val Ile Ala Pro Leu Thr Gly Asn 85 90 95Thr Met Ser Lys Phe Ala Asn Ala Met Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Thr Leu Arg Asn Gly Lys Pro Val Val Leu Gly Ile 115 120 125Ser Thr Asn Asp Ala Leu Gly Leu Asn Gly Val Asn Leu Met Arg Leu 130 135 140Met Ala Thr Lys Asn Ile Tyr Phe Ile Pro Tyr Gly Gln Asp Asp Pro145 150 155 160Val Lys Lys Pro Asn Ser Met Val Ala Arg Met Thr Ala Leu Tyr Asp 165 170 175Thr Val Ile His Ala Met Glu Gly Lys Gln Leu Gln Pro Val Leu Val 180 185 190Glu Arg Tyr Lys Asp Glu Ser 19553196PRTVirgibacillus halophilus 53Met Ser Leu Asp Gly Lys Arg Ile Gly Phe Gly Leu Thr Ala Ser His1 5 10 15Cys Thr Tyr Glu Ala Val Phe Pro Glu Met Glu Arg Leu Ile Asn Met 20 25 30Gly Ala Glu Val Val Pro Val Val Thr Tyr Asn Val Lys Asn Val Asp 35 40 45Thr Lys Phe Gly Lys Ala Ser Asp His Ile Lys Arg Leu Glu Glu Ile 50 55 60Thr Asn Lys Glu Val Val Ala Thr Ile Pro Asp Ala Glu Pro Leu Gly65 70 75 80Pro Ile Thr Pro Leu Asp Cys Met Val Ile Ala Pro Leu Thr Gly Asn 85 90 95Ser Met Ser Arg Leu Ala Asn Ala Ile Thr Asp Ser Pro Pro Leu Met 100 105 110Ala Ala Lys Ala Thr Met Arg Asn Gln Asn Pro Val Val Leu Gly Ile 115 120 125Ser Thr Asn Asp Ala Leu Gly Leu Asn Gly Val Asn Leu Met Lys Leu 130 135 140Met Ala Ser Lys Met Ile Tyr Phe Ile Pro Phe Gly Gln Asp Asp Pro145 150 155 160Val Lys Lys Pro Asn Ser Leu Val Ser Asp Met Thr Leu Leu Pro Glu 165 170 175Thr Ile Glu Ser Ala Leu Asn Gly Asn Gln Leu Gln Pro Val Leu Ile 180 185 190Pro Phe Gln Ser 19554198PRTPaenibacillus azoreducens 54Met Asn Trp Gln Gly Lys Thr Val Gly Tyr Ala Ile Thr Gly Ser His1 5 10 15Cys Thr Leu Glu Glu Ile Met Pro Gln Val Lys Arg Phe Val Asp Glu 20 25 30Gly Ala Lys Val Val Pro Ile Val Ser Asn Thr Val Gln Val Thr Asp 35 40 45Thr Arg Phe Gly Thr Ala His Asn Trp Leu Gln Arg Leu Lys Asp Ile 50 55 60Thr Gly Ser Glu Leu Ile Ser Thr Ile Val Glu Ala Glu Pro Leu Gly65 70 75 80Pro Ser Lys Leu Leu Asp Val Leu Val Ile Ala Pro Cys Thr Gly Asn 85 90 95Thr Thr Ser Lys Leu Ala Asn Ala Ile Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Gln Met Arg Asn Leu Arg Pro Leu Val Leu Ala Ile 115 120 125Ser Thr Asn Asp Gly Leu Gly Leu Asn Ala Ala Asn Ile Ala Lys Leu 130 135 140Leu Val Ala Lys Asn Ile Tyr Phe Val Pro Phe Gly Gln Asp Asn Pro145 150 155 160His Gln Lys Pro Asn Ser Leu Val Ala Gln Met Asp Leu Ile Pro Glu 165 170 175Ala Cys Tyr Ala Ala Leu Glu Gly Arg Gln Leu Gln Pro Met Leu Leu 180 185 190Gln Arg Ile Phe Ser Ala 19555202PRTArtificial SequenceDpaB consensus sequenceMISC_FEATURE(3)..(4)X is any amino acidMISC_FEATURE(21)..(22)X is any amino acidMISC_FEATURE(24)..(24)X is any amino acidMISC_FEATURE(26)..(26)X is any amino acidMISC_FEATURE(29)..(29)X is any amino acidMISC_FEATURE(32)..(33)X is any amino acidMISC_FEATURE(38)..(38)X is any amino acidMISC_FEATURE(40)..(40)X is any amino acidMISC_FEATURE(42)..(43)X is any amino acidMISC_FEATURE(47)..(47)X is any amino acidMISC_FEATURE(54)..(57)X is any amino acidMISC_FEATURE(59)..(62)X is any amino acidMISC_FEATURE(64)..(65)X is any amino acidMISC_FEATURE(68)..(70)X is any amino acidMISC_FEATURE(72)..(73)X is any amino acidMISC_FEATURE(76)..(76)X is any amino acidMISC_FEATURE(83)..(84)X is any amino acidMISC_FEATURE(94)..(94)X is any amino acidMISC_FEATURE(98)..(98)X is any amino acidMISC_FEATURE(119)..(119)X is any amino acidMISC_FEATURE(122)..(123)X is any amino acidMISC_FEATURE(128)..(128)X is any amino acidMISC_FEATURE(139)..(140)X is any amino acidMISC_FEATURE(143)..(144)X is any amino acidmisc_feature(146)..(147)Xaa can be any naturally occurring amino acidMISC_FEATURE(154)..(154)X is any amino acidMISC_FEATURE(162)..(163)X is any amino acidMISC_FEATURE(171)..(171)X is any amino acidMISC_FEATURE(173)..(173)X is any amino acidMISC_FEATURE(176)..(181)X is any amino acidMISC_FEATURE(184)..(184)X is any amino acidMISC_FEATURE(186)..(186)X is any amino acidMISC_FEATURE(188)..(188)X is any amino acidMISC_FEATURE(191)..(192)X is any amino acidMISC_FEATURE(194)..(202)X is any amino acid 55Met Met Xaa Xaa Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser1 5 10 15His Cys Thr Tyr Xaa Xaa Val Xaa Pro Xaa Ile Glu Xaa Leu Val Xaa 20 25 30Xaa Gly Ala Glu Val Xaa Pro Xaa Val Xaa Xaa Thr Val Gln Xaa Thr 35 40 45Asp Thr Arg Phe Gly Xaa Xaa Xaa Xaa Trp Xaa Xaa Xaa Xaa Glu Xaa 50 55 60Xaa Thr Gly Xaa Xaa Xaa Ile Xaa Xaa Ile Val Xaa Ala Glu Pro Leu65 70 75 80Gly Pro Xaa Xaa Pro Leu Asp Cys Met Val Ile Ala Pro Xaa Thr Gly 85 90 95Asn Xaa Met Ser Lys Leu Ala Asn Ala Met Thr Asp Ser Pro Val Leu 100 105 110Met Ala Ala Lys Ala Thr Xaa Arg Asn Xaa Xaa Pro Val Val Leu Xaa 115 120 125Ile Ser Thr Asn Asp Ala Leu Gly Leu Asn Xaa Xaa Asn Leu Xaa Xaa 130 135 140Leu Xaa Xaa Thr Lys Asn Ile Tyr Phe Xaa Pro Phe Gly Gln Asp Asp145 150 155 160Pro Xaa Xaa Lys Pro Asn Ser Leu Val Ala Xaa Met Xaa Leu Leu Xaa 165 170 175Xaa Xaa Xaa Xaa Xaa Ala Leu Xaa Gly Xaa Gln Xaa Gln Pro Xaa Xaa 180 185 190Val Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 195 20056196PRTArtificial SequenceDpaB consensus sequence 56Met Ser Leu Lys Gly Lys Arg Ile Gly Phe Gly Leu Thr Gly Ser His1 5 10 15Cys Thr Tyr Asp Glu Val Met Pro Glu Ile Glu Lys Leu Val Asp Glu 20 25 30Gly Ala Glu Val Thr Pro Ile Val Ser Tyr Thr Val Gln Thr Thr Asp 35 40 45Thr Arg Phe Gly Lys Ala Glu Glu Trp Ile Lys Lys Ile Glu Glu Ile 50 55 60Thr Gly Asn Lys Val Ile Asn Thr Ile Val Glu Ala Glu Pro Leu Gly65 70 75 80Pro Lys Lys Leu Leu Asp Cys Met Val Ile Ala Pro Cys Thr Gly Asn 85 90 95Thr Met Ala Lys Leu Ala Asn Ala Ile Thr Asp Ser Pro Val Leu Met 100 105 110Ala Ala Lys Ala Thr Leu Arg Asn Gln Arg Pro Val Val Leu Ala Ile 115 120 125Ser Thr Asn Asp Ala Leu Gly Leu Asn Ala Lys Asn Leu Gly Arg Leu 130 135 140Leu Asn Thr Lys Asn Ile Tyr Phe Val Pro Phe Gly Gln Asp Asp Pro145 150 155 160Val Lys Lys Pro Asn Ser Leu Val Ala Arg Met Asp Leu Leu Ile Asp 165 170 175Thr Val Glu Glu Ala Leu Glu Gly Lys Gln Leu Gln Pro Val Leu Ile 180 185 190Glu Tyr Phe Arg 19557218PRTClostridium beijerinckii 57Met Lys Ile Tyr Gly Ile Asn Gly Ser Pro Arg Lys Asn Lys Asn Thr1 5 10 15Ala Thr Leu Leu Gln Lys Ala Leu Asp Gly Val Lys Glu Ala Ala Lys 20 25 30Asp Lys Glu Ile Glu Thr Glu Ile Ile Asn Leu Tyr Asp Leu Asn Tyr 35 40 45Thr Gly Cys Ile Ser Cys Phe Ala Cys Lys Arg Leu Gly Ser Asn Ser 50 55 60Tyr Gly Lys Cys Ala Val Lys Asp Asp Leu Gln Glu Val Leu Glu Lys65 70 75 80Val Ser Gln Ser Asp Gly Leu Ile Phe Ser Ser Pro Val Tyr Phe Ser 85 90 95Asn Val Thr Gly Lys Phe Leu Ser Phe Leu Glu Arg Leu Leu Phe Pro 100 105 110Tyr Leu Val Tyr Asp Asn Asn Gly Thr Ser Leu Ala Pro Lys Arg Met 115 120 125Pro Thr Ala Phe Ile Tyr Thr Met Asn Val Lys Glu Glu Val Met Lys 130 135 140Gln Ile Gly Tyr Leu Lys Thr Phe Glu Arg Met Glu Ser Asn Ile Gly145 150 155 160His Ile Phe Thr Lys Pro Leu Val Met Tyr Ser Asn Asn Thr Tyr Gln 165 170 175Phe Asp Asp Tyr Ser Lys Tyr Lys Val Glu Ser Phe Ser Glu Glu Glu 180 185 190Lys Ala Ala His Arg Lys Ile Gln Phe Pro Leu Asp Cys Gln Lys Ala 195 200 205Phe Glu Leu Gly Ala Asn Leu Ile Lys His 210 21558193PRTClostridium beijerinckii 58Met Ser Lys Val Val Ile Phe Asn Gly Ser Pro Arg Lys Asn Gly Tyr1 5 10 15Thr Thr Lys Leu Leu Glu Gln Val Ala Lys Gly Ala Lys Ser Lys Gly 20 25 30Ala Glu Ile Ile Glu Phe Asp Leu Asn Asp Ser Gly Ile Arg Gly Cys 35 40 45Gln Gly Cys Phe Tyr Cys Arg Thr His Asp Gly Cys Ala Val Lys Asp 50 55 60Tyr Leu Gln Pro Met Tyr Lys Ala Ile Thr Glu Ala Asp Ala Ile Val65 70 75 80Phe Gly Ser Pro Ile Tyr Tyr Tyr Gln Ile Thr Gly Gln Ala Lys Ile 85 90 95Trp Leu Asp Arg Thr Phe Pro Met Val Gly Asp Asn Phe Ala Pro Arg 100 105 110His Ser Gly Lys Lys Leu Ile Thr Ile Phe Thr Gln Gly Ser Gln Asn 115 120 125Pro Glu Met Gly Ala Glu Gly Ile Lys Phe Val Asn Asn Met Leu Ala 130 135 140Ala Tyr Gly Trp Lys Leu Glu Asp Ser Ile Leu Cys Cys Gly Thr Thr145 150 155 160Asn Phe His Ser Glu Lys Leu Gly Arg Tyr Asp Ser Asn Leu Glu Gln 165 170 175Phe Glu Glu Leu Ser Leu Arg Ala Phe Lys Asp Gly Glu Asn Leu Val 180 185 190Arg59183PRTClostridium beijerinckii 59Met Ser Lys Val Val Ile Phe Lys Gly Ser Pro Arg Lys Asn Gly Tyr1 5 10 15Thr Ala Arg Leu Leu Glu Gln Val Ala Lys Gly Ala Lys Ser Lys Gly 20 25 30Ala Glu Val Ile Glu Phe Asp Leu Asn Asp Ser Gly Ile Arg Gly Cys 35 40 45Gln Gly Cys Met Tyr Cys Arg Thr His Asp Gly Cys Ala Val Asn Asp 50 55 60Tyr Leu Gln Pro Met Tyr Ala Ala Ile Lys Glu Ala Asp Ala Ile Val65 70 75 80Phe Gly Ser Pro Ile Tyr Tyr Tyr Thr Ile Thr Gly Gln Ser Lys Val 85 90 95Trp Phe Asp Arg Thr Phe Pro Met Ile Gly Asn Asp Tyr Lys Ala Lys 100 105 110Tyr Pro Gly Lys Lys Leu Ile Thr Ile Phe Thr Gln Gly Asn Pro Asp 115 120 125Pro Lys Ile Gly Ala Glu Gly Val Lys Phe Ala Asn Asn Met Leu Glu 130 135 140Glu Leu Gly Trp Lys Leu Glu Asp Ser Ile His Tyr Cys Gly Thr Ser145 150 155 160His Asn Pro Asp Leu Ala Met Phe Asp Glu Leu Ser Leu Arg Ala Phe 165 170 175Lys Asp Gly Glu Asn Leu Ala

18060182PRTClostridium beijerinckii 60Met Ser Lys Val Val Ile Phe Lys Gly Ser Pro Arg Lys Asn Gly Tyr1 5 10 15Thr Thr Lys Leu Leu Asp Gln Val Ala Lys Gly Ala Lys Ser Lys Gly 20 25 30Ala Glu Val Ile Glu Phe Asp Leu Asn Asp Thr Gly Ile Arg Gly Cys 35 40 45Gln Gly Cys Phe Tyr Cys Arg Thr His Asp Gly Cys Ala Val Asn Asp 50 55 60Tyr Leu Gln Pro Met Tyr Lys Ala Ile Ala Glu Ala Asp Ala Ile Val65 70 75 80Phe Gly Ser Pro Ile Tyr Met Phe Gln Ile Thr Ser Gln Ala Lys Thr 85 90 95Cys Leu Asp Arg Thr Phe Pro Met Val Glu Glu Leu Pro Asn Lys Phe 100 105 110Ile Pro Arg His Pro Gly Lys Lys Leu Ile Thr Val Phe Ala Gln Gly 115 120 125Ser Leu Asp Pro Lys Lys Gly Ala Glu Ala Ile Lys Tyr Val Asn Asn 130 135 140Ile Phe Asp Val Phe Gly Trp Lys Leu Glu Asp Cys Ile His Tyr Cys145 150 155 160Gly Thr Asp Asp Glu Val Phe Asn Glu Leu Ser Leu Arg Ala Phe Lys 165 170 175Asp Gly Glu Asn Leu Ala 18061193PRTClostridium beijerinckii 61Met Asn Ile Ile Gly Ile Ser Ala Ser Ser Arg Lys Glu Gly Asn Thr1 5 10 15Ala Trp Ile Val Asn Lys Ile Leu Glu Gly Ala Lys Glu Gln Gly Ala 20 25 30Glu Thr Gln Tyr Phe Asp Phe Asn Asn Leu Asp Ile Lys Pro Cys Gln 35 40 45Gly Cys Trp Ala Cys His Lys Gly Asp Gln Gly Cys Val Ile Lys Asp 50 55 60Asp Met Gln Lys Leu Asn Asp Ala Ile Asp Arg Ala Asn Val Ile Val65 70 75 80Phe Gly Ser Pro Ile Tyr Met Met Gln Met Ser Ala Gln Gly Lys Ile 85 90 95Ile Ile Asp Arg Met Phe Ala Arg Phe Ser Pro Arg Tyr Ser Pro Tyr 100 105 110Phe Lys Glu Glu Ser Ala Ala Glu Lys Arg Leu Val Leu Thr Phe Asn 115 120 125Gln Gly Asn Pro Asp Pro Glu Leu Phe Lys Ser Tyr Ile Asp Tyr Thr 130 135 140Lys His Met Phe Glu Leu Leu Glu Phe Asp Val Thr Glu Val Pro Val145 150 155 160Val Thr Gly Leu Arg Asn Gly Pro Ala Asn Glu Arg Glu Asp Leu Asn 165 170 175Ile Met Leu Lys Asp Val Gly Lys Thr Ile Val Ser Glu Gly Ile Ser 180 185 190Lys62208PRTClostridium beijerinckii 62Met Lys Val Leu Leu Ile Asn Gly Ser Pro Lys Ala Lys Gly Cys Thr1 5 10 15Tyr Thr Thr Leu Cys Glu Val Ala Asp Glu Leu Glu Lys Glu Asn Ile 20 25 30Glu Thr Glu Ile Phe Gln Ile Gly Asn Lys Pro Ile Ser Gly Cys Ile 35 40 45Asp Cys Gly Gly Cys Tyr Lys Ser Gly Glu Gly Lys Cys Val Phe Ser 50 55 60Asp Asp Ile Val Asn Ile Ala Leu Glu Lys Ala Lys Glu Ala Asp Gly65 70 75 80Phe Ile Phe Gly Ser Pro Val His Tyr Ala Ala Pro Ser Gly Ser Ile 85 90 95Thr Ser Phe Leu Asp Arg Phe Phe Tyr Ala Gly Asn Cys Phe Ala His 100 105 110Lys Pro Gly Ala Ala Val Val Ser Cys Arg Arg Gly Gly Ala Ala Ser 115 120 125Ala Phe Asp Gln Leu Asn Lys Tyr Phe Thr Ile Ser Asn Met Pro Val 130 135 140Val Ser Ser Gln Tyr Trp Asn Met Val His Gly Asn Thr Pro Glu Glu145 150 155 160Val Lys Gln Asp Leu Glu Gly Met Gln Thr Met Arg Met Leu Gly Lys 165 170 175Asn Met Ala Trp Leu Leu Lys Ser Ile Asp Ala Gly Lys Lys Ala Gly 180 185 190Ile Ser Leu Pro Glu Ser Glu Pro Arg Val Ala Thr Asn Phe Ile Arg 195 200 20563201PRTArchaeoglobus fulgidus 63Met Lys Leu Leu Ala Ile Asn Gly Ser Pro Asn Lys Arg Asn Thr Leu1 5 10 15Phe Leu Leu Glu Val Ile Ala Glu Glu Val Lys Lys Leu Gly His Glu 20 25 30Ala Glu Ile Ile His Leu Lys Asp Tyr Glu Ile Lys Glu Cys Lys Gly 35 40 45Cys Asp Ala Cys Leu Lys Gly Asp Cys Ser Gln Lys Asp Asp Ile Tyr 50 55 60Lys Val Leu Glu Lys Met Gln Glu Ala Asp Ala Ile Val Ile Gly Thr65 70 75 80Pro Thr Tyr Phe Gly Asn Val Thr Gly Ile Val Lys Asn Leu Ile Asp 85 90 95Arg Ser Arg Met Ala Arg Met Gly Asn Tyr Arg Leu Arg Asn Arg Val 100 105 110Phe Ala Pro Val Val Thr Ser Gly Leu Arg Asn Gly Gly Ala Glu Tyr 115 120 125Ala Ala Met Ser Leu Ile Val Tyr Ala Leu Gly Gln Ala Met Leu Pro 130 135 140Val Ser Ile Val Glu Asn Pro Ile Thr Thr Gly Thr Phe Pro Val Gly145 150 155 160Val Ile Gln Gly Asp Ala Gly Trp Arg Ser Val Lys Lys Asp Glu Ile 165 170 175Ala Ile Asn Ser Ala Lys Ala Leu Ala Lys Arg Ile Val Glu Val Ala 180 185 190Glu Ala Thr Lys Asn Leu Arg Glu Ser 195 20064193PRTMethanocaldococcus jannaschii, 64Met Lys Val Ile Gly Ile Ser Gly Ser Pro Arg Pro Glu Gly Asn Thr1 5 10 15Thr Leu Leu Val Arg Glu Ala Leu Asn Ala Ile Ala Glu Glu Gly Ile 20 25 30Glu Thr Glu Phe Ile Ser Leu Ala Asp Lys Glu Leu Asn Pro Cys Ile 35 40 45Gly Cys Asn Met Cys Lys Glu Glu Gly Lys Cys Pro Ile Ile Asp Asp 50 55 60Val Asp Glu Ile Leu Lys Lys Met Lys Glu Ala Asp Gly Ile Ile Leu65 70 75 80Gly Ser Pro Val Tyr Phe Gly Gly Val Ser Ala Gln Leu Lys Met Leu 85 90 95Met Asp Arg Ser Arg Pro Leu Arg Ile Gly Phe Gln Leu Arg Asn Lys 100 105 110Val Gly Gly Ala Val Ala Val Gly Ala Ser Arg Asn Gly Gly Gln Glu 115 120 125Thr Thr Ile Gln Gln Ile His Asn Phe Phe Leu Ile His Ser Met Ile 130 135 140Val Val Gly Asp Asn Asp Pro Thr Ala His Tyr Gly Gly Thr Gly Val145 150 155 160Gly Lys Ala Pro Gly Asp Cys Lys Asn Asp Asp Ile Gly Leu Glu Thr 165 170 175Ala Arg Asn Leu Gly Lys Lys Val Ala Glu Val Val Lys Leu Ile Lys 180 185 190Lys65225PRTPeptoclostridium difficile 65Met Ile Ile Thr Val Ile Asn Gly Ser Pro Arg Lys Asn Gly Ala Thr1 5 10 15Ser Lys Val Leu Thr Tyr Leu Tyr Lys Asp Ile Glu Arg Leu Ile Pro 20 25 30Asp Val Lys Ile Asn Tyr Phe Asp Leu Ser Glu Val Asn Pro Ser Tyr 35 40 45Cys Ile Gly Cys Leu Asn Cys Tyr Lys Met Gly Lys Cys Ile Asn Gln 50 55 60Asn Asp Lys Val Glu Tyr Ile His Asp Ile Ile Thr Lys Ser Asp Gly65 70 75 80Val Ile Phe Gly Ser Pro Thr Tyr Gly Ser Ser Val Thr Gly Leu Phe 85 90 95Lys Val Phe Thr Asp Arg Ala His Met Met Leu Glu Arg Leu Leu Tyr 100 105 110Arg Lys Pro Cys Ile Ala Val Thr Thr Tyr Glu Asn Ala Arg Gly Ser 115 120 125Lys Ala Ile Ser Phe Ile Lys Ser Met Val Leu Asp Ser Gly Gly Tyr 130 135 140Val Cys Gly Ser Leu Ser Ile Lys Thr Gly Phe Asn Gln Asn Pro Ile145 150 155 160Thr Glu Lys Val Glu Ser Lys Ile Gln Lys Val Ser Lys Lys Phe Ile 165 170 175Tyr Cys Ile Glu Glu Lys Lys Asn Pro Pro Val Leu Ser Gln Ile Tyr 180 185 190Asn Phe Ile Ala Ile Asn Ala Val Leu Lys Pro Met Ala Phe Lys Asp 195 200 205Ile Glu Gln Tyr Lys Gly Ile Ile Asp Arg Trp Glu Glu Gln Gly Ile 210 215 220Ile22566191PRTMethanosarcina thermophila 66Met Lys Ile Thr Gly Ile Ser Gly Ser Pro Arg Lys Gly Gln Asn Cys1 5 10 15Glu Lys Ile Ile Gly Ala Ala Leu Glu Val Ala Lys Glu Arg Gly Phe 20 25 30Glu Thr Asp Thr Val Phe Ile Ser Asn Glu Glu Val Ala Pro Cys Lys 35 40 45Ala Cys Gly Ala Cys Arg Asp Gln Asp Phe Cys Val Ile Asp Asp Asp 50 55 60Met Asp Glu Ile Tyr Glu Lys Met Arg Ala Ala Asp Gly Ile Ile Val65 70 75 80Ala Ala Pro Val Tyr Met Gly Asn Tyr Pro Ala Gln Leu Lys Ala Leu 85 90 95Phe Asp Arg Ser Val Leu Leu Arg Arg Lys Asn Phe Ala Leu Lys Asn 100 105 110Lys Val Gly Ala Ala Leu Ser Val Gly Gly Ser Arg Asn Gly Gly Gln 115 120 125Glu Lys Thr Ile Gln Ser Ile His Asp Trp Met His Ile His Gly Met 130 135 140Ile Val Val Gly Asp Asn Ser His Phe Gly Gly Ile Thr Trp Asn Pro145 150 155 160Ala Glu Glu Asp Thr Val Gly Met Gln Thr Val Ser Glu Thr Ala Lys 165 170 175Lys Leu Cys Asp Val Leu Glu Leu Ile Gln Lys Asn Arg Asp Lys 180 185 19067289PRTArtificial SequenceConsensus Isf sequenceMISC_FEATURE(4)..(6)X is any amino acidMISC_FEATURE(8)..(8)X is any amino acidMISC_FEATURE(14)..(14)X is any amino acidMISC_FEATURE(16)..(16)X is any amino acidMISC_FEATURE(18)..(19)X is any amino acidMISC_FEATURE(22)..(28)X is any amino acidMISC_FEATURE(34)..(34)X is any amino acidMISC_FEATURE(37)..(37)X is any amino acidMISC_FEATURE(39)..(39)X is any amino acidMISC_FEATURE(41)..(43)X is any amino acidMISC_FEATURE(45)..(45)X is any amino acidMISC_FEATURE(47)..(47)X is any amino acidMISC_FEATURE(49)..(50)X is any amino acidMISC_FEATURE(52)..(53)X is any amino acidMISC_FEATURE(55)..(55)X is any amino acidMISC_FEATURE(58)..(59)X is any amino acidMISC_FEATURE(61)..(64)X is any amino acidMISC_FEATURE(68)..(70)X is any amino acidMISC_FEATURE(72)..(74)X is any amino acidMISC_FEATURE(77)..(79)X is any amino acidMISC_FEATURE(81)..(86)X is any amino acidMISC_FEATURE(90)..(90)X is any amino acidMISC_FEATURE(92)..(92)X is any amino acidMISC_FEATURE(97)..(97)X is any amino acidMISC_FEATURE(99)..(102)X is any amino acidMISC_FEATURE(106)..(106)X is any amino acidMISC_FEATURE(108)..(110)X is any amino acidMISC_FEATURE(113)..(136)X is any amino acidMISC_FEATURE(140)..(140)X is any amino acidMISC_FEATURE(152)..(154)X is any amino acidMISC_FEATURE(156)..(166)X is any amino acidMISC_FEATURE(168)..(175)X is any amino acidMISC_FEATURE(177)..(181)X is any amino acidMISC_FEATURE(183)..(185)X is any amino acidMISC_FEATURE(188)..(200)X is any amino acidMISC_FEATURE(202)..(205)X is any amino acidMISC_FEATURE(207)..(209)X is any amino acidMISC_FEATURE(211)..(211)X is any amino acidMISC_FEATURE(213)..(216)X is any amino acidMISC_FEATURE(236)..(241)X is any amino acidMISC_FEATURE(243)..(244)X is any amino acidMISC_FEATURE(246)..(251)X is any amino acidMISC_FEATURE(253)..(255)X is any amino acidMISC_FEATURE(257)..(261)X is any amino acidMISC_FEATURE(263)..(265)X is any amino acidMISC_FEATURE(267)..(272)X is any amino acidMISC_FEATURE(276)..(276)X is any amino acid 67Met Met Lys Xaa Xaa Xaa Ile Xaa Gly Ser Pro Arg Lys Xaa Gly Xaa1 5 10 15Thr Xaa Xaa Leu Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Lys Glu Ala Ala 20 25 30Lys Xaa Leu Ile Xaa Gly Xaa Glu Xaa Xaa Xaa Phe Xaa Leu Xaa Asp 35 40 45Xaa Xaa Ile Xaa Xaa Cys Xaa Gly Cys Xaa Xaa Cys Xaa Xaa Xaa Xaa 50 55 60Ser Asn Ser Xaa Xaa Xaa Cys Xaa Xaa Xaa Asp Asp Xaa Xaa Xaa Ile65 70 75 80Xaa Xaa Xaa Xaa Xaa Xaa Glu Ala Asp Xaa Ile Xaa Phe Gly Ser Pro 85 90 95Xaa Tyr Xaa Xaa Xaa Xaa Thr Gly Gln Xaa Lys Xaa Xaa Xaa Asp Arg 100 105 110Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 115 120 125Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu Ile Thr Xaa Phe Ile Tyr Thr 130 135 140Met Asn Val Lys Glu Glu Val Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa145 150 155 160Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Met 165 170 175Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile Thr Xaa Xaa Xaa Xaa Xaa 180 185 190Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Pro Xaa Xaa 195 200 205Xaa Lys Xaa Asp Xaa Xaa Xaa Xaa Lys Val Ser Lys Lys Phe Ile Tyr 210 215 220Cys Ile Glu Glu Lys Lys Asn Pro Pro Val Leu Xaa Xaa Xaa Xaa Xaa225 230 235 240Xaa Leu Xaa Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Ala Xaa Xaa Xaa Gly 245 250 255Xaa Xaa Xaa Xaa Xaa Val Xaa Xaa Xaa Ile Xaa Xaa Xaa Xaa Xaa Xaa 260 265 270Gly Ile Ser Xaa Pro Glu Ser Glu Pro Arg Val Ala Thr Asn Phe Ile 275 280 285Arg

Claims

1. A composition comprising cells of microbial species including or consisting of each of Bacillus amyloliquefaciens, Bacillus firmus, Bacillus flexus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus koreensis, Bacillus drentensis, Bacillus subtilis, Clostridium bifermentans, Clostridium beijerinckii, Clostridium pasteurianum, Lactobacillus paracasei, Fontibacillus sp. (panacisegetis), Oceanobacillus oncorhynchi, Paenibacillus lautus, Paenibacillus azoreducens, Paenibacillus chibensis, Paenibacillus cookii, Paenibacillus sp. (chitinolyticus), Paenibacillus sp. (P1XP2), Pseudomonas sp., and Streptomyces griseus.

2. The composition of claim 1, comprising cells of microbial species including or consisting of microbes with 16S rDNA nucleic acid sequences having at least 99% sequence identity to each of SEQ ID NOs: 3-25.

3. The composition of claim 2, comprising cells of microbial species including or consisting of microbes with 16S rDNA nucleic acid sequences of each of SEQ ID NOs: 3-25.

4. A composition comprising American Type Culture Collection deposit number PTA-125924.

5. The composition of claim 1, further comprising one or more of chitin, chitosan, glucosamine, amino acids, and liquid fertilizer.

6. A method comprising contacting soil, plants, plant parts, or seeds with the composition of claim 1.

7. The method of claim 6, further comprising contacting the soil, plants, plant parts, or seeds with: one or more of chitin, chitosan, glucosamine, and amino acids; a liquid fertilizer; and/or one or more pesticides, one or more fungicides, one or more herbicides, one or more insecticides, one or more plant hormones, one or more plant elicitors, or combinations of two or more thereof.

8-9. (canceled)

10. The method of claim 6, further comprising activating the microbial species in the composition prior to contacting the soil, plants, plant parts, or seeds with the composition.

11. A composition comprising the composition of claim 1 and a carrier or a seed.

12. The composition of claim 11, wherein the carrier comprises urea, potash, ammonium phosphate, ammonium nitrate, clay, peat, coal, inorganic soil, charcoal, sawdust, wheat/soy/oat brain, compost, coco coir, perlite, vermiculite, bentonite, Azomite.RTM., kaolin, silicates, pumice, talc, a liquid fertilizer or a liquid dust control chemical.

13. The composition of claim 11, wherein the seed comprises corn seed, sunflower seed, canola seed, wheat seed, cucumber seed, tomato seed, rice seed, and/or cotton seed.

14. The composition of claim 11, further comprising one or more insecticide and/or fungicide.

15. A method for co-formulating a microbe with a carrier or seed, comprising: identifying a microbe that comprises one or more dipicolinic acid (DPA) synthase genes, a microbe that expresses one or more DPA synthase proteins, and/or a microbe that produces DPA; selecting the microbe for co-formulation with a carrier or seed; and co-formulating the selected microbe with the carrier or the seed.

16. (canceled)

17. The method of claim 15, wherein the one or more DPA synthase genes comprises a DPA synthase subunit A (DpaA) gene and/or or a DPA synthase subunit B (DpaB) gene.

18. The method of claim 17, wherein: the DpaA gene encodes a DpaA protein with at least 20% sequence identity to any one of the amino acid sequences of SEQ ID NOs: 26-41 or wherein the DpaB gene encodes a DpaB protein with at least 20% sequence identity to any one of the amino acid sequences of SEQ ID NOs: 42-56; the DpaA gene encodes a DpaA protein with at least 60% sequence identity to any one of the amino acid sequences of SEQ ID NOs: 26-41 or wherein the DpaB gene encodes a DpaB protein with at least 60% sequence identity to any one of the amino acid sequences of SEQ ID NOs: 42-56; or the DpaA gene encodes a DpaA protein comprising or consisting of any one of the amino acid sequences of SEQ ID NOs: 26-41 or wherein the DpaB gene encodes a DpaB protein comprising or consisting of any one of the amino acid sequences of SEQ ID NOs: 42-56.

19-24. (canceled)

25. The method of claim 15, wherein identifying a microbe that comprises one or more dipicolinic acid (DPA) synthase genes comprises detecting nucleic acids or nucleic acid sequences that encode the one or more DPA synthase genes.

26-27. (canceled)

28. The method of claim 15, wherein identifying a microbe that expresses the one or more DPA synthase proteins comprises an immunoassay or mass spectrometry.

29. (canceled)

30. The method of claim 15, wherein identifying a microbe that produces DPA comprises detecting DPA in the microbe or in medium containing the microbe.

31-32. (canceled)

33. The method of claim 15, wherein identifying a microbe that produces DPA comprises identifying a microbe that comprises one or more EtfA or Isf gene and/or expresses one or more EtfA or Isf proteins.

34. The method of claim 33, wherein the Isf protein: comprises at least 20% sequence identity to any one of the amino acid sequences of SEQ ID NOs: 57-66; comprises at least 60% sequence identity to any one of the amino acid sequences of SEQ ID NOs: 57-66; or comprises or consists of any one of the amino acid sequences of SEQ ID NOs: 57-66.

35-36. (canceled)

37. The method of claim 15, wherein the selected microbe has increased viability when co-formulated with a carrier or seed compared to a microbe that does not comprise one or more dipicolinic acid (DPA) synthase genes, does not express one or more DPA synthase proteins, and/or does not produce DPA.

38. The method of claim 15, wherein co-formulating the selected microbe with the carrier or seed comprises contacting one or more selected microbes with the carrier or seed.

39. The method of claim 38, wherein the one or more selected microbes are in a liquid medium or are in a solid or dry form.

40. (canceled)

41. The method of claim 38, further comprising contacting the carrier or seed with one or more microbes that do not comprise one or more DPA synthase genes, do not express one or more DPA proteins, and/or do not produce DPA.

42. The method of claim 15, wherein the carrier comprises a dry or solid carrier or a liquid carrier.

43. The method of claim 42, wherein the dry or solid carrier comprises a dry fertilizer, a soil-derived substance, an organic substance, an inert material, or a mixture of two or more thereof or wherein the liquid carrier comprises a liquid fertilizer or a liquid dust control chemical.

44-47. (canceled)