# US 10,480,000 B2 · Methods of increasing crop yield under abiotic stress > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,480,000 B2 - Document ID: US10480000B2 - Status: Granted - Kind: B2 - Application number: 15/326,437 - Filed: 2015-07-15 - Published: 2018-10-04 - Granted: 2019-11-19 - Earliest claimed: 2014-07-15 - Assignees: Ceres, Inc. - Applicants: Ceres, Inc. - Inventors: Roger I. Pennell; Wuyi Wang; Chuan-Yin Wu; Dwarkesh Parihar; Paresh Verma; Vijay R. Kumar; Shridhar J. Rao - Web page: https://patentgrep.com/patent/US10480000B2 ## CPC - C12N 15/8222: {Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation} - C12N 15/8201: {Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation} - 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 Methods and materials for increasing abiotic stress tolerance in plants are disclosed. For example, nucleic acids encoding abiotic stress tolerance-increasing polypeptides are disclosed as well as methods for using such nucleic acids to transform plant cells. Also disclosed are plants having increased tolerance to abiotic stress and methods of increasing plant yield under abiotic stress conditions. ## Claims ### Claim 1 A method of increasing plant yield in a plant grown under drought stress, osmotic stress, or nitrogen deficiency, said method comprising: (a) growing plants transformed with an exogenous nucleic acid under drought stress, osmotic stress, or nitrogen deficiency, said exogenous nucleic acid comprising a heterologous promoter operably linked to a nucleotide sequence encoding a polypeptide having at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2; and (b) selecting a transformed plant from said transformed plants that expresses said polypeptide and has increased yield when grown under drought stress, osmotic stress, or nitrogen deficiency as compared to the corresponding yield of a control plant of the same species that does not comprise said exogenous nucleic acid and grown under identical stress conditions. ### Claim 2 (depends on 1) The method according to claim 1, wherein growing said transformed plants is under drought stress, and said drought stress is selected from the group consisting of pre-flowering drought stress and post-flowering drought stress. ### Claim 3 (depends on 2) The method according to claim 2, wherein growing said transformed plants is under osmotic stress, and said osmotic stress is selected from the group consisting of a soil electric conductivity between 4 dS/m and 5 dS/m, and a soil conductivity between 6 dS/m and 7 dS/m. ### Claim 4 (depends on 1) The method according to claim 1, wherein growing said transformed plants is under nitrogen deficiency, and said nitrogen deficiency is selected from the group consisting of a 50 Kg per hectare nitrogen application, and a 75 Kg per hectare nitrogen application. ### Claim 5 (depends on 1) The method according to claim 1, further comprising harvesting biomass from said selected transformed plant. ### Claim 6 (depends on 1) The method according to claim 1, wherein said polypeptide has at least 97% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2. ### Claim 7 A method of producing a plant with tolerance to drought stress, osmotic stress, or nitrogen deficiency, said method comprising: (a) expressing in a plurality of plants transformed with an exogenous nucleic acid comprising a nucleotide sequence encoding a polypeptide, said exogenous nucleic acid comprising a heterologous promoter operably linked to the nucleotide sequence, and the polypeptide having at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2; (b) selecting from said plurality of transformed plants a transformed plant that expresses said polypeptide and has increased tolerance to drought stress, osmotic stress, or nitrogen deficiency as compared to a control plant of the same species lacking said exogenous nucleic acid and grown under identical stress conditions; and (c) growing the selected transformed plant having increased tolerance to drought stress, osmotic stress, or nitrogen deficiency as compared to a control plant of the same species lacking said exogenous nucleic acid and grown under identical stress conditions. ### Claim 8 (depends on 7) The method according to claim 7, wherein said selected transformed plant has said increased tolerance to drought stress, and said drought stress is selected from the group consisting of pre-flowering drought stress and post-flowering drought stress. ### Claim 9 (depends on 7) The method according to claim 7, wherein said selected transformed plant has said increased tolerance to osmotic stress, and said osmotic stress is selected from the group consisting of a soil electric conductivity between 4 dS/m and 5 dS/m, and a soil conductivity between 6 dS/m and 7 dS/m. ### Claim 10 (depends on 7) The method according to claim 7, wherein said selected transformed plant has said increased tolerance to nitrogen deficiency stress, and said nitrogen deficiency stress is selected from the group consisting of a 50 Kg per hectare nitrogen application, and a 75 Kg per hectare nitrogen application. ### Claim 11 (depends on 7) The method according to claim 7, wherein the polypeptide has 97% or greater amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2. ### Claim 12 (depends on 1, 7) The method according to claim 1 or claim 7, wherein said exogenous nucleic acid has 95% or greater nucleotide sequence identity to the nucleotide sequence of SEQ ID NO: 1. ### Claim 13 (depends on 1, 7) The method according to claim 1 or claim 7, wherein said plant is selected from the group consisting of Panicum virgatum, Sorghum bicolor, Miscanthus giganteus, Saccharum sp., Populus balsamifera, Zea mays, Glycine max, Brassica napus, Triticum aestivum, Gossypium hirsutum, Oryza sativa, Helianthus annuus, Medicago sativa, Beta vulgaris, and Pennisetum glaucum. ### Claim 14 A method of producing a transgenic plant with increased yield grown under drought stress, osmotic stress, or nitrogen deficiency stress, said method comprising: (a) growing plant cells transformed with an exogenous nucleic acid, wherein said exogenous nucleic acid comprising a polynucleotide having a nucleotide sequence encoding a polypeptide having at least 95% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 2, and wherein said nucleotide sequence is operably linked to a heterologous promoter; (b) producing transgenic plants from said transformed plant cells; and (c) selecting for a transgenic plant from said transgenic plants that expresses said polypeptide in the selected transgenic plant and which exhibits increased yield when grown under drought stress, osmotic stress, or nitrogen deficiency stress as compared to a control plant of the same plant species that does not comprise said exogenous nucleic acid and grown under identical stress conditions. ### Claim 15 (depends on 14) The method of claim 14, wherein said polypeptide has at least 97% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 2.