Abstract
Soybean mosaic virus (SMV) is one of the most destructive viral diseases in soybean and causes severe reduction of soybean yield and destroys the seed quality. However, the production of SMV resistant plants by transgenic is the most effective and economical means. Based on our previous yeast two-hybrid assay, the GmVma12 was selected as a strong candidate gene for further function characterization. Here we transformed soybean plants with a construct containing inverted repeat of-GmVma12 sequence to analyze the role of GmVma12 during SMV invasion. Totals of 33 T0 and 160 T1 plants were confirmed as positive transgenic plants through herbicide application, PCR detection and LibertyLink® strip screening. Based on the segregation ratio and Southern Blot data, T1 lines No. 3 and No. 7 were selected to generate T2 plants. After SMV-SC15 inoculation, 41 T1 and 38 T2 plants were identified as highly resistant, and their quantification disease levels were much lower than non-transformed plants. The transcript level of GmVma12 in T2 plants decreased to 70% of non-transformed plants. The expression level of SMV-CP transcript in T2 transgenic plants was lower than that in non-transformed plants and SMV CP protein in T2 plants could not be detected by Enzyme-linked Immunosorbent assay, which indicated that SMV production would be inhibited in transgenic plants. Moreover, coat mottles of T2 seeds were obliterated significantly. In conclusion, inverted repeat of the hairpin structure of GmVma12 interfered with the transcription of GmVma12, which can induce resistance to SMV in soybean. This research lays the foundation for the mechanism of SMV pathogenesis, and provides new ideas for SMV prevention and control.
Highlights
Soybean (Glycine max (L.) Merr.) originated from China has been cultivated for more than five thousand years, supplying plant fat and protein in human daily diet
This study demonstrated that interfering the transcription of GmVma12 could enhance the resistance to Soybean mosaic virus (SMV) in soybean plant
The cotyledons were laid on co-cultivation medium (CCM) with filter paper (Fig. 1c) and incubated for 5 d in dark
Summary
Soybean (Glycine max (L.) Merr.) originated from China has been cultivated for more than five thousand years, supplying plant fat and protein in human daily diet. Soybean mosaic virus (SMV) infection is prevalent from north spring soybean production area to the Yangtze River valley area in China (Li et al 2010). It was reported that the first transgenic soybean was produced in 1988 using Agrobacterium-mediated transformation and since this technology has been widely used in soybean transgenic research (Hinchee et al 1988). Using agrobacterium transformation to induce RNAi-mediated resistance to SMV is a popular strategy in China. The RNAi strategy was used to trigger robust resistance against three viruses in soybean plants by expressing several short inverted repeat of a portion of the virus sequences (Zhang et al 2011). The construct containing inverted repeat of SMV-HC-Pro was transformed into five soybean genotypes. The transgenic soybeans demonstrated strong resistant to SMV in transgenic plants (Gao et al 2015). The transgenic lines with silenced P3 cistron by RNAi showed significantly increased resistance to multiple potyvirus strains and isolates (Yang et al 2018)
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