Methods and compositions for producing male sterile plants

摘要

Methods of making a targeted modification in a male fertility gene in the genome of a plant are disclosed. The methods involve contacting a plant cell with an engineered double-strand-break-inducing agent capable of inducing a double-strand break in a target sequence in the male fertility gene and identifying a cell comprising an alteration in the target sequence. Also disclosed are plants, plant cells, plant parts, and seeds comprising a male fertility gene with an alteration in a male fertility gene. Nucleic acid molecules comprising male fertility genes with at least one targeted modification therein, optimized nucleic acid molecules encoding endonucleases that are engineered double-strand-break-inducing agents and expression cassettes, host cells, and plants comprising one or more of the nucleic acid molecules are further disclosed.

主权项

1. A method for making a targeted modification in a male fertility gene in the genome of a maize plant, said method comprising:
(a) contacting at least one plant cell comprising, in a MS45 male fertility gene, a target sequence comprising SEQ ID NO: 20 with an engineered meganuclease that is capable of inducing a double-strand break at the target sequence; and (b) identifying at least one cell from step (a) comprising an alteration in its genome at the target sequence wherein the alteration is selected from the group consisting of (i) replacement of at least one nucleotide, (ii) a deletion of at least one nucleotide, (iii) an insertion of at least one nucleotide, and (iv) any combination of (i) through (iii); and (c) regenerating a plant from the at least one plant cell comprising the alteration in its genome, wherein the alteration of the male fertility gene is a null mutation.