US2019085354A1PendingUtilityA1

Isolated polynucleotides expressing or modulating micrornas or targets of same, transgenic plants comprising same and uses thereof

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Assignee: A B SEEDS LTDPriority: Feb 6, 2012Filed: Nov 30, 2018Published: Mar 21, 2019
Est. expiryFeb 6, 2032(~5.6 yrs left)· nominal 20-yr term from priority
Inventors:Rudy MaorIris NesherOrly Noivirt-BrikOsnat Yanai-Azulay
C12N 15/8261C12N 15/8273C12N 15/8243Y02A40/146C12N 15/8271C12N 15/8218
53
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Claims

Abstract

Disclosed are isolated polynucleotides expressing or modulating microRNAs or targets. Further disclosed are transgenic plants comprising an isolated polynucleotide expressing or modulating microRNAs or target for improving nitrogen use efficiency, abiotic stress tolerance, biomass, vigor or yield of a plant. The sequences of microRNAs and targets to be modulated are also disclosed.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A method of improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of a plant, the method comprising expressing within the plant an exogenous polynucleotide which downregulates an activity or expression of a gene encoding an RNAi molecule having a nucleic acid sequence at least 90% identical to SEQ ID NOs: 139, 57-79, 202-219, 126-138, 140-161, 236-255, 169-173, 260-261, 3953-5114, 5117-6277, 6278, 11905-11909, 11940-11955, 11959-11961, wherein said RNAi molecule regulates abiotic stress tolerance of the plant, thereby improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of the plant. 
     
     
         2 . A method of improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of a plant, the method comprising expressing within the plant an exogenous polynucleotide having a nucleic acid sequence at least 90% identical to SEQ ID NOs: 1-56, 174-201, 80-125, 220-235, 162-168, 256-259, 262-2086, 2087-3910, 3911, 11616, 11615, 11874, 11875-11904, 11910-11939, 11956, 11957 or 11958, wherein said nucleic acid sequence is capable of regulating abiotic stress tolerance of the plant, thereby improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of the plant. 
     
     
         3 . A transgenic plant exogenously expressing a polynucleotide having a nucleic acid sequence at least 90% identical to SEQ ID NOs: 1-56, 174-201, 80-125, 220-235, 162-168, 256-259, 262-2086, 2087-3910, 3911, 11616, 11615, 11874, 11875-11904, 11910-11939, 11956, 11957 or 11958, wherein said nucleic acid sequence is capable of regulating abiotic stress tolerance of the plant. 
     
     
         4 . The method of  claim 2  or transgenic plant of  claim 3 , wherein said exogenous polynucleotide encodes a precursor of said nucleic acid sequence. 
     
     
         5 . The method or transgenic plant of  claim 4 , wherein said precursor of said nucleic acid sequence is at least 60% identical to SEQ ID NO: 174-201, 220-235, 256-259, 2087-3910, 3911, 11875-11904, 11910-11939, 11615, 11956, 11957 or 11958. 
     
     
         6 . The method of  claim 2  or the transgenic plant of  claim 3 , wherein said exogenous polynucleotide encodes a mature miRNA. 
     
     
         7 . The method of  claim 2  or the transgenic plant of  claim 3 , wherein said exogenous polynucleotide is selected from the group consisting of SEQ ID NO: 1-56, 174-201, 80-125, 220-235, 162-168, 256-259, 262-2086, 2087-3910, 3911, 11616, 11615, 11874, 11875-11904, 11910-11939, 11956, 11957 or 11958. 
     
     
         8 . A nucleic acid construct comprising a nucleic acid sequence being at least 90% identical to SEQ ID NO: 139, 1-201, 202-235, 236-3910, 3911, 11616, 11615, 11874, 11875-11904, 11910-11939, 11956, 11957, 11958, 11940-11955, 11905-11909, 11959-11961, wherein said nucleic acid sequence is capable of regulating abiotic stress tolerance of a plant and wherein said nucleic acid sequence is under the regulation of a cis-acting regulatory element. 
     
     
         9 . The nucleic acid construct of  claim 8 , wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 1-56, 174-201, 80-125, 220-235, 162-168, 256-259, 262-2086, 2087-3910, 3911, 11616, 11615, 11874, 11875-11904, 11910-11939, 11956, 11957 or 11958. 
     
     
         10 . The nucleic acid construct of  claim 8 , wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 139, 57-79, 202-219, 126-138, 140-161, 236-255, 169-173, 260-261, 3953-5114, 5117-6277, 6278, 11905-11909, 11959-11961, 11940-11955. 
     
     
         11 . The nucleic acid construct of  claim 8 , wherein said cis-acting regulatory element comprises a promoter. 
     
     
         12 . The nucleic acid construct of  claim 11 , wherein said promoter comprises a tissue-specific promoter. 
     
     
         13 . The nucleic acid construct of  claim 12 , wherein said tissue-specific promoter comprises a root specific promoter. 
     
     
         14 . A transgenic plant exogenously expressing a polynucleotide which downregulates an activity or expression of a gene encoding an RNAi molecule having a nucleic acid sequence at least 90% identical to SEQ ID NOs: 139, 57-79, 202-219, 126-138, 140-161, 236-255, 169-173, 260-261, 3953-5114, 5117-6277, 6278, 11905-11909, 11940-11955, 11959-11961. 
     
     
         15 . A transgenic plant exogenously expressing a polynucleotide which downregulates an activity or expression of a gene encoding an RNAi molecule having a nucleic acid sequence at least 90% identical to SEQ ID NOs: 139, 57-79, 202-219, 126-138, 140-161, 236-255, 169-173, 260-261, 3953-5114, 5117-6277, 6278, 11905-11909, 11940-11955, 11959-11961. 
     
     
         16 . An isolated polynucleotide which downregulates an activity or expression of a gene encoding an RNAi molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 139, 57-79, 202-219, 126-138, 140-161, 236-255, 169-173, 260-261, 3953-5114, 5117-6277, 6278, 11905-11909, 11940-11955, 11959-11961. 
     
     
         17 . An isolated polynucleotide which downregulates an activity or expression of a gene encoding an RNAi molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 139, 1-201, 202-235, 236-3910, 3911, 3953-5114, 5117-6277, 6278, 11616, 11615, 11874, 11875-11904, 11910-11939, 11956, 11957, 11958, 11905-11909, 11940-11955, 11959-11961. 
     
     
         18 . The method of  claim 1 , the transgenic plant of  claim 14  or the isolated polynucleotide of  claim 16 , wherein said polynucleotide encodes a miRNA-Resistant Target as set forth in SEQ ID NO: 11258-11359. 
     
     
         19 . The transgenic plant of  claim 15 , wherein said polynucleotide encodes a miRNA-Resistant Target as set forth in SEQ ID NO: 11091-11257. 
     
     
         20 . The method of  claim 1 , the transgenic plant of  claim 14  or the isolated polynucleotide of  claim 16 , wherein said isolated polynucleotide encodes a target mimic as set forth in SEQ ID NO: 11564-11613. 
     
     
         21 . The transgenic plant of  claim 15 , wherein said polynucleotide encodes a target mimic as set forth in SEQ ID NO: 11437-11513. 
     
     
         22 . A nucleic acid construct comprising the isolated polynucleotide of  claim 16  or  17  under the regulation of a cis-acting regulatory element. 
     
     
         23 . The nucleic acid construct of  claim 22 , wherein said cis-acting regulatory element comprises a promoter. 
     
     
         24 . The nucleic acid construct of  claim 23 , wherein said promoter comprises a tissue-specific promoter. 
     
     
         25 . The nucleic acid construct of  claim 24 , wherein said tissue-specific promoter comprises a root specific promoter. 
     
     
         26 . The method of  claim 1  or  2 , further comprising growing the plant under limiting nitrogen conditions. 
     
     
         27 . The method of  claim 1  or  2 , further comprising growing the plant under abiotic stress. 
     
     
         28 . The method of  claim 27 , wherein said abiotic stress is selected from the group consisting of salinity, drought, water deprivation, flood, etiolation, low temperature, high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, nutrient excess, atmospheric pollution and UV irradiation. 
     
     
         29 . The method of  claim 1  or  2 , or the plant of  claim 3  or  14 , being a monocotyledon. 
     
     
         30 . The method of  claim 1  or  2 , or the plant of  claim 3  or  14 , being a dicotyledon. 
     
     
         31 . A method of improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of a plant, the method comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide having an amino acid sequence at least 80% homologous or identical to SEQ ID NOs: 9591-10364, wherein said polypeptide is capable of regulating abiotic stress tolerance of the plant, thereby improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of the plant. 
     
     
         32 . A transgenic plant exogenously expressing a polynucleotide encoding a polypeptide having an amino acid sequence at least 80% homologous or identical to SEQ ID NOs: 9591-10364, wherein said polypeptide is capable of regulating abiotic stress tolerance of the plant. 
     
     
         33 . A nucleic acid construct comprising a polynucleotide encoding a polypeptide having an amino acid sequence at least 80% homologous to SEQ ID NOs: 9591-10364, wherein said polypeptide is capable of regulating abiotic stress tolerance of the plant, and wherein said polynucleotide is under a transcriptional control of a cis-acting regulatory element. 
     
     
         34 . The method of  claim 31 , the transgenic plant of  claim 32  or the nucleic acid construct of  claim 33 , wherein said polynucleotide is selected from the group consisting of SEQ ID NO: 10365-10963. 
     
     
         35 . The method of  claim 31 , the transgenic plant of  claim 32  or the nucleic acid construct of  claim 33 , wherein said polypeptide is selected from the group consisting of SEQ ID NO: 9591-10364. 
     
     
         36 . The nucleic acid construct of  claim 33 , wherein said cis-acting regulatory element comprises a promoter. 
     
     
         37 . The nucleic acid construct of  claim 36 , wherein said promoter comprises a tissue-specific promoter. 
     
     
         38 . The nucleic acid construct of  claim 37 , wherein said tissue-specific promoter comprises a root specific promoter. 
     
     
         39 . The method of  claim 31 , further comprising growing the plant under limiting nitrogen conditions. 
     
     
         40 . The method of  claim 31 , further comprising growing the plant under abiotic stress. 
     
     
         41 . The method of  claim 40 , wherein said abiotic stress is selected from the group consisting of salinity, drought, water deprivation, flood, etiolation, low temperature, high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, nutrient excess, atmospheric pollution and UV irradiation. 
     
     
         42 . The method of  claim 31 , or the plant of  claim 32 , being a monocotyledon. 
     
     
         43 . The method of  claim 31 , or the plant of  claim 32 , being a dicotyledon. 
     
     
         44 . A method of improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of a plant, the method comprising expressing within the plant an exogenous polynucleotide which downregulates an activity or expression of a polypeptide having an amino acid sequence at least 80% homologous or identical to SEQ ID NOs: 6315-8129, wherein said polypeptide is capable of regulating abiotic stress tolerance of the plant, thereby improving abiotic stress tolerance, nitrogen use efficiency, biomass, vigor or yield of the plant. 
     
     
         45 . A transgenic plant exogenously expressing a polynucleotide which downregulates an activity or expression of a polypeptide having an amino acid sequence at least 80% homologous or identical to SEQ ID NOs: 6315-8129, wherein said polypeptide is capable of regulating abiotic stress tolerance of the plant. 
     
     
         46 . A nucleic acid construct comprising a polynucleotide which downregulates an activity or expression of a polypeptide having an amino acid sequence at least 80% homologous to SEQ ID NOs: 6315-8129, wherein said polypeptide is capable of regulating abiotic stress tolerance of a plant, said nucleic acid sequence being under the regulation of a cis-acting regulatory element. 
     
     
         47 . The method of  claim 44 , the transgenic plant of  claim 45  or the nucleic acid construct of  claim 46 , wherein said polynucleotide acts by a mechanism selected from the group consisting of sense suppression, antisense suppression, ribozyme inhibition, gene disruption. 
     
     
         48 . The nucleic acid construct of  claim 46 , wherein said cis-acting regulatory element comprises a promoter. 
     
     
         49 . The nucleic acid construct of  claim 48 , wherein said promoter comprises a tissue-specific promoter. 
     
     
         50 . The nucleic acid construct of  claim 49 , wherein said tissue-specific promoter comprises a root specific promoter.

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