GENOME WIDE IDENTIFICATION AND CHARACTERIZATION OF GENE EXPRESSION REGULATORY ELEMENTS IN ZEA MAYS FOR USE IN PLANTS

Abstract

The present invention provides non-coding regulatory element polynucleotide molecules isolated from the lipid transfer protein (LTP) gene of Oryza sativa and useful for expressing transgenes in plants. The invention further discloses compositions, polynucleotide constructs, transformed host cells, transgenic plants and seeds containing the Oryza sativa regulatory polynucleotide sequences, and methods for preparing and using the same.

Claims

1-30. (canceled)

31. A polynucleotide molecule comprising: i) a regulatory polynucleotide molecule comprising a nucleic acid sequence selected from the group consisting of: (a) a nucleic acid sequence that exhibits 98% or greater identity to SEQ ID NO:9617 and has promoter activity; (b) a fragment comprising at least 95 contiguous nucleotides of SEQ ID NO: 9617 and has promoter activity; and (c) the nucleic acid sequence of SEQ ID NO:9617; and ii) a heterologous transcribable polynucleotide molecule, wherein the regulatory polynucleotide molecule is operably linked to the heterologous transcribable polynucleotide molecule.

32. A polynucleotide molecule comprising: i) a regulatory polynucleotide molecule comprising a nucleic acid sequence selected from the group consisting of: (a) a nucleic acid sequence that exhibits 98% or greater identity to SEQ ID NO:9617 and has promoter activity; (b) a fragment comprising at least 95 contiguous nucleotides of SEQ ID NO: 9617 and has promoter activity; and (c) the nucleic acid sequence of SEQ ID NO:9617; and ii) a heterologous transcribable polynucleotide molecule, wherein the regulatory polynucleotide molecule is operably linked to the heterologous transcribable polynucleotide molecule, wherein said regulatory polynucleotide molecule is SEQ ID NO:9617.

33. The polynucleotide molecule of claim 31, wherein said regulatory polynucleotide molecule is a promoter.

34. A polynucleotide molecule comprising: i) a regulatory polynucleotide molecule comprising a nucleic acid sequence selected from the group consisting of: (a) a nucleic acid sequence that exhibits 98% or greater identity to SEQ ID NO:9617 and has promoter activity; (b) a fragment comprising at least 95 contiguous nucleotides of SEQ ID NO: 9617 and has promoter activity; and (c) the nucleic acid sequence of SEQ ID NO:9617; and ii) a heterologous transcribable polynucleotide molecule, wherein the regulatory polynucleotide molecule is operably linked to the heterologous transcribable polynucleotide molecule, wherein said regulatory polynucleotide molecule is a promoter and wherein said promoter is further described as a promoter that exhibits a constitutive expression pattern.

35. The polynucleotide molecule of claim 33, wherein said promoter is further described as a promoter that preferentially expresses in a tissue selected from the group consisting of: embryo, endosperm, internode, kernel, leaf, pollen, root, silk, inflorescent meristem, germinating seedling, shoot apical meristem, spikelet floral meristem, spikelet pair meristem, tassel, ovule, stem, callus, cob, shoot, radicle, ear.

36. The polynucleotide molecule of claim 33, wherein said promoter is further described as a promoter that preferentially expresses at a stage selected from the group consisting of: day, night, V1 through V15 leaf stage, 1 through 40 days after pollination, tasseling stage, harvest, 1 through 45 days after planting, emergence.

37. The polynucleotide molecule of claim 33, wherein said promoter is further described as a promoter that preferentially expresses under a stress condition selected from the group consisting of: low nitrogen, high nitrogen, cold, drought, excess water, nutrient deprivation, light deprivation.

38. A chimeric molecule comprising: i) a regulatory polynucleotide molecule comprising a nucleic acid sequence selected from the group consisting of: (a) a nucleic acid sequence that exhibits 98% or greater identity to SEQ ID NO:9617 and has promoter activity; (b) a fragment comprising at least 95 contiguous nucleotides of SEQ ID NO: 9617 and has promoter activity; and (c) the nucleic acid sequence of SEQ ID NO:9617; and ii) a heterologous transcribable polynucleotide molecule, wherein the regulatory polynucleotide molecule is operably linked to the heterologous transcribable polynucleotide molecule.

39. The chimeric molecule of claim 38, wherein said transcribable polynucleotide molecule is a gene of agronomic interest.

40. The chimeric molecule of claim 38, wherein said transcribable polynucleotide molecule is a gene controlling the phenotype of a trait selected from the group consisting of: herbicide tolerance, insect control, modified yield, fungal disease resistance, virus resistance, nematode resistance, bacterial disease resistance, plant growth and development, starch production, modified oils production, high oil production, modified fatty acid content, high protein production, fruit ripening, enhanced animal and human nutrition, biopolymers, environmental stress resistance, pharmaceutical peptides and secretable peptides, improved processing traits, improved digestibility, enzyme production, flavor, nitrogen fixation, hybrid seed production, fiber production, and biofuel production.

41. The chimeric molecule of claim 43, wherein said herbicide tolerance gene is selected from the group consisting of genes that encode for: phosphinothricin acetyltransferase, glyphosate resistant EPSPS, hydroxyphenyl pyruvate dehydrogenase, dalapon dehalogenase, bromoxynil resistant nitrilase, anthranilate synthase, glyphosate oxidoreductase and glyphosate-N-acetyl transferase.

42. A transgenic plant cell stably transformed with a polynucleotide molecule comprising: i) a regulatory polynucleotide molecule comprising a nucleic acid sequence selected from the group consisting of: (a) a nucleic acid sequence that exhibits 98% or greater identity to SEQ ID NO:9617 and has promoter activity; (b) a fragment comprising at least 95 contiguous nucleotides of SEQ ID NO: 9617 and has promoter activity; and (c) the nucleic acid sequence of SEQ ID NO:9617; and ii) a heterologous transcribable polynucleotide molecule, wherein the regulatory polynucleotide molecule is operably linked to the heterologous transcribable polynucleotide molecule.

43. A transgenic plant stably transformed with a polynucleotide molecule comprising: i) a regulatory polynucleotide molecule comprising a nucleic acid sequence selected from the group consisting of: (a) a nucleic acid sequence that exhibits 98% or greater identity to SEQ ID NO:9617 and has promoter activity; (b) a fragment comprising at least 95 contiguous nucleotides of SEQ ID NO: 9617 and has promoter activity; and (c) the nucleic acid sequence of SEQ ID NO:9617; and ii) a heterologous transcribable polynucleotide molecule, wherein the regulatory polynucleotide molecule is operably linked to the heterologous transcribable polynucleotide molecule.

44. A seed of said transgenic plant of claim 43, wherein the seed comprises the polynucleotide molecule.

45. A progeny of the plant of claim 43, wherein the progeny comprises the polynucleotide molecule.

46. The transgenic plant cell of claim 42, wherein said plant cell is from a monocotyledonous plant selected from the group consisting of wheat, maize, rye, rice, corn, oat, barley, turfgrass, sorghum, millet and sugarcane.

47. The transgenic plant of claim 43, wherein said plant is a monocotyledonous plant selected from the group consisting of wheat, maize, rye, rice, corn, oat, barley, turfgrass, sorghum, millet and sugarcane.

48. The seed of the transgenic plant of claim 47, wherein the seed comprises the polynucleotide molecule.

49. The transgenic plant cell of claim 42, wherein said plant cell is from a dicotyledonous plant selected from the group consisting of tobacco, tomato, potato, soybean, cotton, canola, sunflower and alfalfa.

50. The transgenic plant of claim 43, wherein said plant is a dicotyledonous plant selected from the group consisting of tobacco, tomato, potato, soybean, cotton, canola, sunflower and alfalfa.

51. The seed of the transgenic plant of claim 50, wherein the seed comprises the polynucleotide molecule.

52. A method of inhibiting weed growth in transgenic glyphosate-tolerant crop plants comprising planting the transgenic plants transformed with an expression cassette comprising: a) the chimeric molecule of claim 38, active in a plant cell and wherein the heterologous transcribable polynucleotide molecule is a glyphosate tolerance gene; and b) applying glyphosate to the field at an application rate that inhibits the growth of weeds, wherein the growth and yield of the transgenic crop plant is not substantially affected by the glyphosate application.

53. A method of conferring insect resistance in transgenic crop plants comprising planting transgenic plants transformed with an expression cassette comprising the chimeric molecule of claim 38 active in a plant cell, wherein the heterologous transcribable polynucleotide molecule comprises an insect resistance gene.

54. A method of conferring drought tolerance to transgenic crop plants comprising planting transgenic plants transformed with an expression cassette comprising the chimeric molecule of claim 38 active in a plant cell, wherein the heterologous transcribable polynucleotide molecule comprises a drought tolerance gene.

55. A method of conferring low nitrogen tolerance to transgenic crop plants comprising planting transgenic plants transformed with an expression cassette comprising the chimeric molecule of claim 38 active in a plant cell, wherein the heterologous transcribable polynucleotide molecule comprises a low nitrogen tolerance gene.

56. A method of conferring cold tolerance to transgenic crop plants comprising planting transgenic plants transformed with an expression cassette comprising the chimeric molecule of claim 38 active in a plant cell, wherein the heterologous transcribable polynucleotide molecule comprises a cold tolerance gene.

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