A TRV-VIGS-based Approach for High Throughput Gene Function Verification in Soybean (Glycine max)
SUN Tian-Jie1*, MA Nan1*, SUN Li-Yong2, WANG Meng-Xuan1, SUN Xi-Zhe1, ZHANG Jie1, WANG Dong-Mei1**
1. State Key Laboratory of North China Crop Improvement and Regulation/ Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology/ College of Life Sciences,Hebei Agricultural University,Baoding 071001,China; 2. Collaborative and Innovation Center of Hebei,Shijiazhuang 050000,China
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.
孙天杰, 麻楠, 孙立永, 王梦璇, 孙希哲, 张洁, 王冬梅. 一种基于TRV-VIGS的高通量大豆基因功能验证方法[J]. 农业生物技术学报, 2020, 28(11): 2080-2090.
SUN Tian-Jie, MA Nan, SUN Li-Yong, WANG Meng-Xuan, SUN Xi-Zhe, ZHANG Jie. A TRV-VIGS-based Approach for High Throughput Gene Function Verification in Soybean (Glycine max). 农业生物技术学报, 2020, 28(11): 2080-2090.
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