# US 11,078,492 B2 · Isolated polynucleotides and polypeptides, and methods of using same for increasing nitrogen use efficiency, yield, growth rate, vigor, biomass, oil content, and/or abiotic stress tolerance > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,078,492 B2 - Document ID: US11078492B2 - Status: Granted - Kind: B2 - Application number: 16/124,286 - Filed: 2018-09-07 - Published: 2019-02-28 - Granted: 2021-08-03 - Earliest claimed: 2011-11-28 - Assignees: Evogene Ltd. - Applicants: Evogene Ltd. - Inventors: David Panik; Basia Judith Vinocur; Hagai Karchi - Web page: https://patentgrep.com/patent/US11078492B2 ## CPC - C12N 15/8261: {with agronomic (input) traits, e.g. crop yield} - C12N 15/8243: {involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine} - C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition} - C12N 15/8271: {for stress resistance, e.g. heavy metal resistance} - C12N 15/8273: {for drought, cold, salt resistance} - Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract Provided are methods of increasing nitrogen use efficiency, yield, biomass, growth rate, vigor, oil content, fiber yield, fiber quality and/or abiotic stress tolerance of a plant by expressing within the plant an exogenous polynucleotide comprising a nucleic acid sequence at least 80% identical to SEQ ID NO: 387, 1-386, 388-469, 763-3704 and 3705; or an exogenous polynucleotide encoding a polypeptide at least 80% identical to SEQ ID NO: 602, 470-601, 603-762, 3706-5858, 5860-5910, 5912, 5914-5923, 5925-6046 or 6047. Also provided is an isolated polynucleotide comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 387, 1-386, 388-469, 763-3704 and 3705, which can be used to increase the nitrogen use efficiency, yield, biomass, growth rate, vigor, oil content, fiber yield, fiber quality and/or abiotic stress tolerance of a plant. ## Claims ### Claim 1 A method of increasing nitrogen use efficiency, yield, biomass, growth rate, and/or tolerance to nitrogen deficiency of a plant, wherein the nitrogen deficiency is a growth condition including an applied level of nitrogen which is below a level needed for a metabolism, growth, reproduction and/or viability of a control plant of the same species, the method comprising: (a) expressing within the plant a heterologous polynucleotide comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence as set forth by SEQ ID NO: 475, or a homologous polypeptide having at least 83% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475, and (b) selecting said plant expressing said heterologous polynucleotide for an increased nitrogen use efficiency, yield, biomass, growth rate, and/or tolerance to nitrogen deficiency as compared to a control plant of the same species which is grown under the same growth conditions, thereby increasing the nitrogen use efficiency, yield, biomass, growth rate, and/or the tolerance to nitrogen deficiency of the plant. ### Claim 2 (depends on 1) The method of claim 1, wherein said amino acid sequence is selected from the group consisting of SEQ ID NO: 475, 3726, 3727, 3728, 3731, 3732, 3733, 3734, 3735, 3736, 3737, 3738, 3739 and 5643. ### Claim 3 (depends on 1) The method of claim 1, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 264, 216, 6, 788-793, 796-804 and 3242. ### Claim 4 (depends on 1) The method of claim 1, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 264, 216, and 6. ### Claim 5 (depends on 1) The method of claim 1, wherein said homologous polypeptide comprises an amino acid sequence having at least 95% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475. ### Claim 6 (depends on 1) The method of claim 1, wherein said heterologous polynucleotide encodes a polypeptide comprising an amino acid sequence as set forth by SEQ ID NO: 475. ### Claim 7 (depends on 1) The method of claim 1, further comprising growing the plant expressing said heterologous polynucleotide under nitrogen-limiting conditions which comprise 40-50% of the Nitrogen content present under optimal growth conditions of a control plant of the same species. ### Claim 8 (depends on 1) The method of claim 1, wherein said homologous polypeptide comprises an amino acid sequence having at least 98% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475. ### Claim 9 (depends on 1) The method of claim 1, wherein the nitrogen deficiency comprises 0-10% of the Nitrogen content present under optimal growth conditions of a control plant of the same species. ### Claim 10 A method of increasing tolerance of a plant to nitrogen deficiency, wherein the nitrogen deficiency is a growth condition including an applied level of nitrogen which is below a level needed for a metabolism, growth, reproduction and/or viability of a control plant of the same species, the method comprising: (a) expressing within the plant a heterologous polynucleotide comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence as set forth by SEQ ID NO: 475, or a homologous polypeptide having at least 83% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475, and (b) selecting said plant expressing said heterologous polynucleotide for an increased tolerance to nitrogen deficiency as compared to a control plant of the same species which is grown under the same growth conditions, thereby increasing the tolerance of the plant to nitrogen deficiency. ### Claim 11 (depends on 10) The method of claim 10, wherein said amino acid sequence is selected from the group consisting of SEQ ID NO: 475, 3726, 3727, 3728, 3731, 3732, 3733, 3734, 3735, 3736, 3737, 3738, 3739 and 5643. ### Claim 12 (depends on 10) The method of claim 10, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 264, 216, 6, 788-793, 796-804 and 3242. ### Claim 13 (depends on 10) The method of claim 10, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 264, 216, and 6. ### Claim 14 (depends on 10) The method of claim 10, wherein said homologous polypeptide comprises an amino acid sequence having at least 95% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475. ### Claim 15 (depends on 10) The method of claim 10, wherein said homologous polypeptide comprises an amino acid sequence having at least 98% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475. ### Claim 16 (depends on 10) The method of claim 10, wherein said heterologous polynucleotide encodes a polypeptide comprising an amino acid sequence as set forth by SEQ ID NO: 475. ### Claim 17 A method of increasing leaf number, rosette area, rosette diameter, plot coverage, growth rate of leaf number, growth rate of plot coverage and/or growth rate of rosette diameter of a plant, comprising: (a) expressing within the plant a heterologous polynucleotide comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence as set forth by SEQ ID NO: 475, or a homologous polypeptide having at least 83% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475, and (b) selecting said plant expressing said heterologous polynucleotide for an increased leaf number, rosette area, rosette diameter, plot coverage, growth rate of leaf number, growth rate of plot coverage and/or growth rate of rosette diameter as compared to a control plant of the same species which is grown under the same growth conditions, thereby increasing the leaf number, rosette area, rosette diameter, plot coverage, growth rate of leaf number, growth rate of plot coverage and/or growth rate of rosette diameter of the plant. ### Claim 18 (depends on 17) The method of claim 17, wherein said amino acid sequence is selected from the group consisting of SEQ ID NO: 475, 3726, 3727, 3728, 3731, 3732, 3733, 3734, 3735, 3736, 3737, 3738, 3739 and 5643. ### Claim 19 (depends on 17) The method of claim 17, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 264, 216, 6, 788-793, 796-804 and 3242. ### Claim 20 (depends on 17) The method of claim 17, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 264, 216, and 6. ### Claim 21 (depends on 17) The method of claim 17, wherein said homologous polypeptide comprises an amino acid sequence having at least 95% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475. ### Claim 22 (depends on 17) The method of claim 17, wherein said homologous polypeptide comprises an amino acid sequence having at least 98% sequence identity and conservative amino acid substitution(s) with respect to the amino acid sequence set forth by SEQ ID NO: 475. ### Claim 23 (depends on 17) The method of claim 17, wherein said heterologous polynucleotide encodes a polypeptide comprising an amino acid sequence as set forth by SEQ ID NO: 475.