Increased multiple virus resistance in transgenic soybean overexpressing the double-strand RNA-specific ribonuclease gene PAC1
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Viruses constitute a major constraint to soybean production worldwide and are responsible for significant yield losses every year. Although varying degrees of resistance to specific viral strains has been identified in some soybean genetic sources, the high rate of mutation in viral genomes and mixed infections of different viruses or strains under field conditions usually hinder the effective control of viral diseases. In the present study, we generated transgenic soybean lines constitutively expressing the double-strand RNA specific ribonuclease gene PAC1 from Schizosaccharomyces pombe to evaluate their resistance responses to multiple soybean-infecting virus strains and isolates. Resistance evaluation over three consecutive years showed that the transgenic lines displayed significantly lower levels of disease severity in field conditions when challenged with soybean mosaic virus (SMV) SC3, a prevalent SMV strain in soybean-growing regions of China, compared to the non-transformed (NT) plants. After inoculation with four additional SMV strains (SC7, SC15, SC18, and SMV-R), and three isolates of bean common mosaic virus (BCMV), watermelon mosaic virus (WMV), and bean pod mottle virus (BPMV), the transgenic plants exhibited less severe symptoms and enhanced resistance to virus infections relative to NT plants. Consistent with these results, the accumulation of each virus isolate was significantly inhibited in transgenic plants as confirmed by quantitative real-time PCR and double antibody sandwich enzyme-linked immunosorbent assays. Collectively, our results showed that overexpression of PAC1 can increase multiple virus resistance in transgenic soybean, and thus provide an efficient control strategy against RNA viruses such as SMV, BCMV, WMV, and BPMV.
KeywordsSoybean Multiple virus resistance Soybean mosaic virus Bean common mosaic virus Watermelon mosaic virus Bean pod mottle virus
This work was supported by Grants from China National Novel Transgenic Organisms Breeding Project (2016ZX08004-004), Jilin Provincial Agricultural Science & Technology Innovation Project (CXGC2017JQ013) and National Natural Science foundation of China (31671764). We thank Prof. Haijian Zhi (Nanjing Agricultural University in China) for providing virus strains and isolates and Prof. Shifang Li (Chinese Academy of Agricultural Sciences, China) for providing the plasmid pBI121-PAC1. We would also like to thank Editage (www.editage.cn) for English language editing.
Yingshan Dong and Qiyun Li designed the experiments. Xiangdong Yang and Lu Niu conducted the experiments and drafted the manuscript. Jing Yang, Guojie Xing, Dongquan Guo, Qianqian Zhao, and Haiyun Li performed A. tumefaciens-mediated transformation experiments. Wei Zhang conducted the virus inoculation assays. Hongli He and Xiaofang Zhong participated in the qRT-PCR analyses. All authors participated in the manuscript revision.
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Conflict of interest
All authors declare that they have no conflict of interest in the publication of this paper.
- Cao X, Lu Y, Di D, Zhang Z, Liu H, Tian L, Zhang A, Zhang Y, Shi L, Guo B, Xu J, Duan X, Wang X, Han C, Miao H, Yu J, Da Li (2013) Enhanced virus resistance in transgenic maize expressing a dsRBA-specific endoribonuclease gene from E. coli. PLoS ONE 8:60829. https://doi.org/10.1371/journal.pone.0060829 CrossRefGoogle Scholar
- Ishida I, Ogawa T, Yoshioka M, Hori T, Ohtani T (1995) Production of plants tolerant to multiple viral infections by genetic manipulations. Acta Hortic 10:927–928Google Scholar
- Lamontagne B, Larose S, Boulanger J, Elela SA (2001) The RNase III family: a conserved structure and expanding functions in eukaryotic dsRNA metabolism. Yeast 45:154–158Google Scholar
- Rotondo G, Huang JY, Frendewey D (1997) Substrate structure requirements of the Pac1 ribonuclease from Schizosaccharmyces pombe. Publ RNA Soc 3:1182–1193Google Scholar
- Sinclair JB (1989) Compendium of soybean diseases. 3rd Ed. APS Press, Saint PaulGoogle Scholar
- Shukla DD, Ward CW, Brunt AA (1994) The Potyviridae. CAB International, WallingfordGoogle Scholar
- Zhou GC, Wu XY, Zhang YM, Wu P, Wu XZ, Liu LW, Wang Q, Hang YY, Yang JY, Shao ZQ, Wang B, Chen JQ (2014) A genomic survey of thirty soybean-infecting bean common mosaic virus (BCMV) isolates from China pointed BCMV as a potential threat to soybean production. Virus Res 191:125–133CrossRefGoogle Scholar