A method for improving the green seedling differentiation rate of barley microspore callus after Agrobacterium infection and its application
By optimizing the induction, subculture, infection and differentiation steps in the barley microspore culture process and combining it with the specific culture medium composition, the problem of low differentiation rate after Agrobacterium infection was solved, efficient green seedling differentiation was achieved, and haploid molecular breeding was supported.
CN117502233BActive Publication Date: 2025-09-05SHANGHAI ACAD OF AGRI SCI
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Patent Information
- Application Number
- CN202311606873.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-11-28
- Publication Date
- 2025-09-05
- Estimated Expiration
- 2043-11-28
AI Technical Summary
Technical Problem
In the prior art, the green seedling differentiation rate of barley microspore callus after Agrobacterium infection is low, which affects the application of microspore regeneration technology in haploid molecular breeding.
Method used
By optimizing specific steps and culture medium composition, including barley microspore induction culture, subculture, Agrobacterium infection and negative pressure treatment, combined with the use of N6 and 2/3MS culture media, the differentiation rate of barley microspore callus after Agrobacterium infection was improved.
Benefits of technology
The green seedling differentiation rate of barley microspore callus tissue after Agrobacterium infection was significantly improved to 26.67-43.33%, meeting the needs of haploid molecular breeding.
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Abstract
The present invention relates to the field of barley planting technology, and in particular to a method for improving the green shoot differentiation rate of barley microspore callus after Agrobacterium infection and its application. The method for improving the green shoot differentiation rate of barley microspore callus after Agrobacterium infection of the present invention comprises the following steps: (1) inoculating barley microspores into an induction medium to obtain microspore callus; (2) inoculating the microspore callus into a solid induction medium for subculture to obtain subculture microspore callus; (3) mixing the subculture microspore callus with an Agrobacterium suspension, transferring the subculture to a co-culture medium for co-culture, and obtaining infected callus; (4) treating the infected callus with negative pressure, transferring the infected callus to a differentiation medium for differentiation culture, and obtaining barley microspore callus green shoots. The method of the present invention can significantly improve the green shoot differentiation rate of barley microspore callus after Agrobacterium infection.
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