Gene mutant for controlling male sterility of tobacco and method for creating male sterility line of tobacco

The creation of tobacco male sterile lines through CRISPR/Cas9 gene editing and molecular marking technology has solved the problems of lack of materials and difficult to guarantee seed quality in tobacco breeding, and achieved efficient and low-cost breeding and seed purity control.

CN120424958APending Publication Date: 2025-08-05YUNNAN ACAD OF TOBACCO AGRI SCI
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Patent Information

Application Number
CN202510568393.4
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-04-30
Publication Date
2025-08-05

AI Technical Summary

Technical Problem

The lack of excellent nuclear male sterile line materials in tobacco breeding makes it difficult to guarantee seed quality. Inbred line transplantation has problems of low purity and poor consistency, and traditional breeding methods are costly and inefficient. The application of modern molecular marking technology in tobacco breeding has not yet been fully utilized.

Method used

The two paralogic genes NtCYP704B1-T and NtCYP704B1-S of the tobacco male sterility-related gene NtCYP704B1 were simultaneously mutated by CRISPR/Cas9 gene editing technology, and the NtCYP704B1-T/S double-entangled sterile line was created, and molecular marker assisted breeding was used to perform molecular marker assisted breeding, and target single plants were screened.

Benefits of technology

The control of male sterility of tobacco is achieved, breeding efficiency and seed purity are improved, labor and material investment is reduced, and seed quality is ensured.

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Abstract

The invention belongs to the field of plant biotechnology breeding, and relates to a gene mutant for controlling male sterility of tobacco and a method for creating a male sterility line of tobacco. According to the gene mutant, two para-line homologous genes NtCYP704B1-T and NtCYP704B1-S of a tobacco male sterility related gene NtCYP704B1 are subjected to simultaneous mutation; compared with a gene NtCYP704B1-T with a nucleotide sequence as shown in SEQ ID NO.1, the nucleotide sequence of the NtCYP704B1-T mutant gene is characterized in that a basic group T is inserted behind the 102th basic group; compared with a gene NtCYP704B1-S with a nucleotide sequence as shown in SEQ ID NO.2, the nucleotide sequence of the NtCYP704B1-S mutant gene is characterized in that a basic group A is inserted behind the 100th basic group; the nucleotide sequence of the NtCYP704B1-T mutant gene and the nucleotide sequence of the NtCYP704B1-S mutant gene are as shown in SEQ ID NO. 5 and SEQ ID NO. 6 respectively. Two para-line homologous genes of NtCYP704B1 are simultaneously subjected to site-directed mutagenesis in common tobacco through a CRISPR / Cas9 gene editing technology, and the NtCYP704B1-T / S double-process sterile line is created, so that complete male sterility can be caused. A primer combination is developed aiming at the NtCYP704B1-T / S double-process sterile line and is used for fertility gene identification of plants and screening of target single plants in molecular marker-assisted breeding.
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