一种基于TRV-VIGS的高通量大豆基因功能验证方法

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Abstract Virus-induced gene silencing (VIGS) is an efficient and convenient reverse genetics method for rapid gene function study. The application of the VIGS is often restricted due to the incompatibility of viruses and plants. Tobacco rattle virus (TRV) -mediated VIGS technology achieves gene silencing on a variety of plant species, but its application is limited by plant damage and low silencing efficiency caused by conventional inoculation operation that applies agroinfiltration in plant leaves. In this study, soybean (Glycine max) was used as a material to validate a modified TRV-mediated VIGS protocol that used Agrobacterium tumefaciens to infect from the root after soil irrigation. The results showed that the A. tumefaciens carrying the VIGS plasmids that infected soybean seedlings implemented effectively silence in all plants tested in the same treatment batch, silencing efficiency of targeted genes were above 67%, and reached 98% of the highest. Further examination on the soybean ternately compound leaves at different positions of the treated plants revealed that the first to the third-round of ternately compound leaves had a satisfactory silencing effects, of which the first-round leaves had the highest silencing efficiency. The genes that were effectively silenced in this experiment included non-specific lipid-transfer protein (LTP), pathogenesis-related protein 1 (PR1), thaumatin-like protein (TLP), dehydrin-like protein (DHN), xyloglucan endotransglucosylase (XTH), and cysteine-rich transmembrane protein (CYSTM). The method was proved to be simple and feasible, and a large number of gene silenced plants were obtained in a short period of time. This study established an efficient system for rapid gene function verification in soybean.

Key words： Virus-induced gene silencing (VIGS) Tobacco rattle virus Silencing efficiency Soybean Systematic silencin

Received: 29 March 2020

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S184

Corresponding Authors: **dongmeiwang63@126.com

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http://journal05.magtech.org.cn/Jwk_ny/EN/ OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2020/V28/I11/2080

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