Efficient silence system for verifying cucumber gene function in short term by down-regulated expression of endogenous target gene induced by ALSV virus

By constructing the ALSV2 virus silencing fragment plasmid and inoculating cucumber seeds using vacuum method, the operation process of cucumber gene function research was simplified, efficient gene silencing and rapid verification were achieved, and the problem of cucumber gene function research was solved.

CN120442708APending Publication Date: 2025-08-08NANJING AGRICULTURAL UNIVERSITY
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Patent Information

Application Number
CN202510525159.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-04-24
Publication Date
2025-08-08

AI Technical Summary

Technical Problem

In the prior art, the genetic transformation system for cucumber gene function research is cumbersome to operate, long cycles, and difficult, resulting in the ALSV-mediated VIGS system steps and high equipment costs, making it difficult to achieve efficient gene silencing.

Method used

By constructing an ALSV2 virus silencing fragment plasmid containing endogenous gene of interest, using Agrobacterium tumefaciens GV3101 to transform and prepare VIGS infectious solution, the cucumber germination seeds were inoculated by vacuum method, simplifying the operation process and achieving rapid gene function verification.

Benefits of technology

It realizes efficient silencing of cucumber gene function, simplifies RNA virus inoculation operation, and the gene silencing efficiency reaches 100%, and gene function verification is completed in a short period of time.

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Abstract

The invention discloses an RNA (Ribonucleic Acid) virus-mediated cucumber simple, rapid and efficient gene silencing technical system, the operation process of RNA virus inoculation is simplified, only an agrobacterium VIGS mixed infection solution containing ALSV1 and ALSV2 target gene silencing vectors needs to be prepared, seeds germinated by the inoculated cucumber are directly infected by utilizing a vacuumizing method, and the seeds are directly infected by utilizing an agrobacterium tumefaciens VIGS mixed infection solution containing ALSV1 and ALSV2 target gene silencing vectors. The cucumber seedling characterization verification gene function can be observed in 16-18 days, the cucumber gene silencing efficiency can reach 100% through the method, and a new path is developed for verification and evaluation of the cucumber gene function.
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