# US 10,246,723 B2 · Method for propagating sterile male plant line

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- Number: US 10,246,723 B2
- Document ID: US10246723B2
- Status: Granted
- Kind: B2
- Application number: 14/437,633
- Filed: 2013-10-18
- Published: 2016-05-26
- Granted: 2019-04-02
- Earliest claimed: 2012-10-23
- Assignees: China Agricultural University
- Applicants: China Agricultural University
- Inventors: Jinsheng Lai; Haiming Zhao; Weibin Song; Yang Cui
- Web page: https://patentgrep.com/patent/US10246723B2

## CPC

- C12N 15/8289: {Male sterility}
- A01H 1/00: Processes for modifying genotypes {; Plants characterised by associated natural traits} (A01H4/00 takes precedence)
- A01H 1/023: {Male sterility}
- A01H 1/045: {using molecular markers}
- A01H 6/4684: {Zea mays [maize]}
- C12N 15/00: Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor (mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00)
- C12N 15/821: {Non-antibiotic resistance markers, e.g. morphogenetic, metabolic markers}
- C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]}
- C12N 15/8287: {for fertility modification, e.g. apomixis}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES

## Abstract

A method for propagating a sterile male plant line includes: (a) providing a first plant that comprises a homozygous recessive male sterility allele; (b) providing a second plant comprising the homozygous recessive allele and a construct, where the construct exists in a heterozygous state and comprises i) a first nucleotide sequence that will restore the male fertility of the first plant after expression, and ii) a second nucleotide sequence that is able to affect the grain shape or the endosperm nutrient material composition in a heterozygous state, which allows for distinguishing the grains with or without said construct by observation through naked eyes or devices; the first nucleotide sequence and the second nucleotide sequence are tightly connected with each other and coexist in plants; and (c) fertilizing the female gametes of the first plant with the male gametes of the second plant.

## Claims

### Claim 1

A nucleotide construct comprising:
a first nucleotide sequence comprising a wild type allele of a male fertility regulatory gene Ms45; and
a second nucleotide sequence, wherein the second nucleotide sequence comprises a nucleotide sequence that inhibits the expression of the protein presented by SEQ ID No:5, wherein the nucleotide sequence that inhibits the expression of the protein presented by SEQ ID No:5 comprises SEQfor-X-SEQrev;
wherein the SEQfor comprises the nucleotide sequence of positions 14-276 of SEQ ID No:2; the SEQrev comprises the nucleotide sequence of positions 401-663 of SEQ ID NO:2; and the X is an intron forming a hairpin structure and comprises the nucleotide sequence of positions 277-400 of SEQ ID No:2.

### Claim 2 (depends on 1)

The nucleotide construct of claim 1, wherein the first nucleotide sequence comprises an Ms45 expression element which results in expression of protein Ms45 presented by SEQ ID No:4, wherein the Ms45 expression element comprises SEQ ID No:1.

### Claim 3 (depends on 1)

The nucleotide construct of claim 1, wherein the first nucleotide sequence comprises the Ms45 expression element of SEQ ID No:1 or the Ms45 encoding sequence of SEQ ID No:8.