Plant tolerance to low water, low nitrogen and cold ii

Abstract

Polynucleotides incorporated into nucleic acid constructs have been introduced into plants and were ectopically expressed. The encoded polypeptides of the invention have been shown to confer at least one regulatory activity and confer earlier flowering, longer floral organ retention, increased cold tolerance, greater tolerance to water deprivation, altered carbon-nitrogen balance sensing, increased low nitrogen tolerance, and/or increased tolerance to hyperosmotic stress as compared to a control plant.

1 . A nucleic acid construct comprising a recombinant nucleic acid sequence encoding a polypeptide, wherein:
 the polypeptide shares an amino acid identity with any of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48, wherein the percent amino acid identity is selected from the group consisting of at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 78.6%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 82.1%, at least about 83%, at least about 83.9%, at least about 84%, at least about 85%, at least about 85.7%, at least about 86%, at least about 87%, at least about 87.5%, at least about 88%, at least about 89%, at least about 89.3%, at least about 90%, at least about 91%, at least about 92%, at least about 92.9%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, and about 100% ; or   the recombinant nucleic acid sequence specifically hybridizes to the complement of the sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31 under stringent conditions comprising two wash steps at least as stringent as 6×SSC at 65° C. of 10-30 minutes for each wash step; or   the recombinant nucleic acid sequence specifically hybridizes to the complement of the sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31 under stringent conditions comprising two wash steps of 0.2 to 2.0×SSC at 50° C. to 65° C. for 10-30 minutes for each wash step;   wherein when the polypeptide is expressed in a plant, the polypeptide regulates transcription and confers at least one regulatory activity resulting in an altered trait in the plant as compared to a control plant;, and the altered trait is selected from the group consisting of: earlier flowering, longer floral organ retention, increased tolerance to low nitrogen conditions, altered carbon-nitrogen balance sensing, increased tolerance to cold, increased tolerance to water deficit conditions, increased tolerance to sucrose, and increased tolerance to hyperosmotic stress.   
     
     
         2 . The nucleic acid construct of  claim 1 , wherein the polypeptide has at least 78% identity to SEQ ID NO: 33. 
     
     
         3 . The nucleic acid construct of  claim 1 , wherein the stringent conditions comprising two wash steps of 0.5×SSC, 0.1% SDS at 65° C. of 10-30 minutes for each wash step. 
     
     
         4 . The nucleic acid construct of  claim 1 , wherein the polypeptide comprises SEQ ID NO: 33. 
     
     
         5 . The nucleic acid construct of  claim 1 , wherein the nucleic acid construct further comprises a constitutive, inducible, or tissue-specific promoter operably lined to the recombinant nucleic acid sequence. 
     
     
         6 . The nucleic acid construct of  claim 1 , wherein the nucleic acid construct is comprised within a recombinant host cell. 
     
     
         7 . A transgenic plant having earlier flowering, longer floral organ retention, greater tolerance to low nitrogen conditions, altered carbon-nitrogen balance sensing, greater tolerance to cold, greater tolerance to water deficit conditions, greater tolerance to sucrose, or greater tolerance to hyperosmotic stress, as compared to a control plant, wherein the transgenic plant comprises:
 a nucleic acid construct comprising a recombinant nucleic acid sequence encoding a polypeptide, wherein:   the polypeptide shares an amino acid identity with any of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48, wherein the percent amino acid identity is selected from the group consisting of at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 78.6%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 82.1%, at least about 83%, at least about 83.9%, at least about 84%, at least about 85%, at least about 85.7%, at least about 86%, at least about 87%, at least about 87.5%, at least about 88%, at least about 89%, at least about 89.3%, at least about 90%, at least about 91%, at least about 92%, at least about 92.9%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, and about 100%; or   the recombinant nucleic acid sequence specifically hybridizes to the complement of the sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31 under stringent conditions comprising two wash steps at least as stringent as 6×SSC at 65° C. of 10-30 minutes for each wash step; or   the recombinant nucleic acid sequence specifically hybridizes to the complement of the sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31 under stringent conditions comprising two wash steps of 0.2 to 2.0×SSC at 50° C. to 65° C. for 10-30 minutes for each wash step;   wherein when the polypeptide is expressed in a plant, the polypeptide regulates transcription and confers at least one regulatory activity resulting in an altered trait in the plant as compared to a control plant; and   the altered trait is selected from the group consisting of: earlier flowering, longer floral organ retention, increased tolerance to low nitrogen conditions, altered carbon-nitrogen balance sensing, increased tolerance to cold, increased tolerance to water deficit conditions, increased tolerance to sucrose, and increased tolerance to hyperosmotic stress.   
     
     
         8 . The transgenic plant of  claim 7 , wherein the polypeptide has at least 78% identity to SEQ ID NO: 33. 
     
     
         9 . The transgenic plant of  claim 7 , wherein the stringent conditions comprising two wash steps of 0.5×SSC, 0.1% SDS at 65° C. of 10-30 minutes for each wash step 
     
     
         10 . The transgenic plant of  claim 7 , wherein the transformed plant is a eudicot. 
     
     
         11 . The transgenic plant of  claim 7 , wherein the transformed plant is a legume. 
     
     
         12 . The transformed plant of  claim 7 , wherein the transformed plant is a transformed seed comprising the recombinant nucleic acid sequence. 
     
     
         13 . A method for increasing the tolerance of a plant to low nitrogen conditions, hyperosmotic stress or cold as compared to a control plant, the method comprising:
 (a) providing a nucleic acid construct comprising a recombinant nucleic acid sequence encoding a polypeptide, wherein:   the polypeptide shares an amino acid identity with any of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48, wherein the percent amino acid identity is selected from the group consisting of at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 78.6%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 82.1%, at least about 83%, at least about 83.9%, at least about 84%, at least about 85%, at least about 85.7%, at least about 86%, at least about 87%, at least about 87.5%, at least about 88%, at least about 89%, at least about 89.3%, at least about 90%, at least about 91%, at least about 92%, at least about 92.9%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, and about 100%; or   the recombinant nucleic acid sequence specifically hybridizes to the complement of the sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31 under stringent conditions comprising two wash steps at least as stringent as 6×SSC at 65° C. of 10-30 minutes for each wash step; or   the recombinant nucleic acid sequence specifically hybridizes to the complement of the sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31 under stringent conditions comprising two wash steps of 0.2 to 2.0×SSC at 50° C. to 65° C. for 10-30 minutes for each wash step;   wherein when the polypeptide is expressed in a plant, the polypeptide regulates transcription and confers at least one regulatory activity resulting in an altered trait in the plant as compared to a control plant, and the altered trait is selected from the group consisting of: earlier flowering, longer floral organ retention, increased tolerance to low nitrogen conditions, altered carbon-nitrogen balance sensing, increased tolerance to cold, increased tolerance to water deficit conditions, increased tolerance to sucrose, and increased tolerance to hyperosmotic stress; and   (b) transforming a target plant with the nucleic acid construct to produce a transformed plant having greater tolerance to low nitrogen conditions, hyperosmotic stress or cold than the control plant.   
     
     
         14 . The method of  claim 13 , wherein the methods further comprises the step of:
 (c) selecting a transgenic plant that ectopically expresses the polypeptide, and/or has greater tolerance to low nitrogen conditions, hyperosmotic stress or cold than the control plant.   
     
     
         15 . The method of  claim 13 , wherein the plant is more tolerant than the control plant to 8° C. during germination or growth, to MS medium with 20 mg/l of NH 4 NO 3  as the nitrogen source, to 9.4% sucrose, or removal from growth medium and drying for two hours at the seedling stage. 
     
     
         16 . The method of  claim 13 , wherein the polypeptide has at least about 78% identity to SEQ ID NO: 33. 
     
     
         17 . The method of  claim 13 , wherein the stringent conditions comprising two wash steps of 0.5×SSC, 0.1% SDS at 65° C. of 10-30 minutes for each wash step. 
     
     
         18 . The method of  claim 13 , wherein the nucleic acid construct comprises any of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, or 31. 
     
     
         19 . The method of  claim 13 , wherein the method steps further comprise selfing or crossing the transformed plant with itself or another plant, respectively, to produce a transformed seed.