# US 9,516,824 B2 · Method for modulating the number of archesporial cells in a developing anther > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,516,824 B2 - Document ID: US9516824B2 - Status: Granted - Kind: B2 - Application number: 14/373,319 - Filed: 2013-02-13 - Published: 2014-12-04 - Granted: 2016-12-13 - Earliest claimed: 2012-02-14 - Assignees: The Board of Trustees of the Leland Stanford Junior University - Applicants: The Board of Trustees of the Leland Stanford Junior University - Inventors: Timothy Kelliher; Virginia Walbot - Web page: https://patentgrep.com/patent/US9516824B2 ## CPC - A01H 3/04: by treatment with chemicals - A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES ## Abstract Certain embodiments provide a method of altering the number of archesporial cells in a developing anther of a plant in certain embodiments, the method comprises exposing the anther to redox-modulatory conditions prior to differentiation of germline cells in the anther, thereby changing the redox potential of cells in the anther and altering the number of archesporial cells in the anther. This method may be employed to increase or decrease the number of archesporial cells in a developing anther, and may be employed to produce male sterile plants. ## Claims ### Claim 1 A method of altering the amount of pollen in a developing anther of a plant, comprising: (a) identifying a plant comprising an immature anther at a stage of development in which archesporial cells are being specified or are not yet specified; (b) locally exposing the immature anther of the plant of (a) to redox-modulatory conditions prior to or during specification of archesporial cells in the anther, thereby changing the redox potential of cells in said anther and altering the number of progenitor cells that differentiate into archesporial cells in said anther, and (c) growing the plant such that the immature anther develops into a mature anther that has at least epidermal cells, subepidermal cells, and either an increase in pollen or a decrease in pollen, relative to a control anther of the same stage as the immature anther of (a) that has not been exposed to the redox-modulatory conditions and (i) wherein a plant having increased pollen production and/or enlarged anther size is the result of locally exposing said anther to hypoxic conditions or to a reducing agent at a concentration that lowers the amount of reactive oxygen species in cells of said anther, as compared to a control plant not expose to said hypoxic conditions or reducing agent; or (ii) wherein a plant having reduced pollen production is the result of locally exposing said anther to an oxidizing agent at a concentration that increases the amount of reactive oxygen species in cells of said anther, as compared to a control plant not exposed to said oxidizing agent. ### Claim 2 (depends on 1) The method of claim 1, wherein the exposing step (b) comprises locally exposing the anther to hypoxic conditions or to a reducing agent at a concentration that lowers the amount of reactive oxygen species in said cells of the anther, thereby lowering the amount of reactive oxygen species in said cells and increasing the number of archesporial cells. ### Claim 3 (depends on 2) The method of claim 2, wherein increasing the number of archesporial cells in said anther results in a plant having larger anther size and/or higher pollen production, relative to a control plant that has not been exposed to hypoxic conditions or to said reducing agent. ### Claim 4 (depends on 2) The method of claim 2, wherein said exposing comprises placing said anther in an environment that contains less than 10% oxygen. ### Claim 5 (depends on 1) The method of claim 1, wherein the exposing step (b) comprises contacting said anther with an oxidizing agent at a concentration that increases the amount of reactive oxygen species in said cells, thereby increasing the amount of reactive oxygen species in said cells and decreasing the number of archesporial cells. ### Claim 6 (depends on 5) The method of claim 5, wherein decreasing the number of archesporial cells in said anther results in a plant having smaller anther size and/or lower pollen production than a control plant that has not been subjected to said applying. ### Claim 7 (depends on 5) The method of claim 5, wherein decreasing the number of archesporial cells results in a male sterile plant. ### Claim 8 (depends on 5) The method of claim 5, wherein said oxidizing agent is a peroxide. ### Claim 9 (depends on 1) The method of claim 1, wherein the exposing step (b) comprises exposing said developing anther to a gas. ### Claim 10 (depends on 1) The method of claim 1, wherein the exposing step (b) comprises spraying said developing anther with a liquid that comprises a redox-modulatory compound. ### Claim 11 (depends on 10) The method of claim 10, wherein said redox-modulatory compound is dissolved in said liquid. ### Claim 12 (depends on 10) The method of claim 10, wherein said redox-modulatory compound is in or on a particle that is present in said liquid. ### Claim 13 (depends on 1) The method of claim 1, wherein said exposing comprises placing a solid form of a redox-modulatory compound on said developing anther. ### Claim 14 (depends on 1) The method of claim 1, wherein said plant is a monocot. ### Claim 15 (depends on 1) The method of claim 1, wherein said plant is a dicot. ### Claim 16 A method of making a male sterile plant comprising: (a) identifying a plant comprising one or more immature anthers at a stage of development in which archesporial cells are being specified or are not yet specified; (b) locally exposing the anthers of the plant of (a) to an oxidizing agent at a concentration that increases the amount of reactive oxygen species in cells in said anthers prior to or during specification of archesporial cells in the anthers, wherein the oxidizing agent increases the amount of reactive oxygen species in said cells and decreases the number of progenitor cells that differentiate into archesporial cells; and (c) cultivating a male sterile plant, wherein the male sterile plant contains a mature anther that has at least epidermal cells, subepidermal cells, and no pollen, wherein male sterility of said plant results from the treatment of step (b). ### Claim 17 (depends on 16) The method of claim 16, further comprising crossing said male sterile plant with another plant to produce a hybrid plant. ### Claim 18 (depends on 17) The method of claim 17, wherein said hybrid plant has hybrid vigor relative to its parents. ### Claim 19 (depends on 18) The method of claim 18, wherein said plant is rice or corn.