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.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Cho EK, Goodman RM (1979) Strains of soybean mosaic virus: classification based on virulence in resistant soybean cultivars. Phytopathology 69:467–470
Cho EK, Goodman RM (1982) Evaluation of resistance in soybeans to soybean mosaic virus strains. Crop Sci 22:1133–1136
Choi BK, Koo JM, Ahn HJ, Yum HJ, Choi CW, Ryu KH, Chen P, Tolin SA (2005) Emergence of Rsv-resistance breaking soybean mosaic virus isolates from Korean soybean cultivars. Virus Res 112:42–51
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
Desbiez C, Lecoq H (2004) The nucleotide sequence of watermelon mosaic virus (WMV, Potyvirus) reveals interspecific recombination between two related potyviruses in the 5′ part of the genome. Arch Virol 149:1619–1632
Furutani N, Hidaka S, Kosaka Y, Shizukawa Y, Kanematsu S (2006) Coat protein gene-mediated resistance to soybean mosaic virus in transgenic soybean. Breed Sci 56:119–124
Gagarinova AG, Babu M, Poysa V, Hill JH, Wang A (2008) Identification and molecular characterization of two naturally occurring soybean mosaic virus isolates that are closely related but differ in their ability to overcome Rsv4 resistance. Virus Res 138:50–56
Gore MA, Hayes AJ, Jeong SC, Yue YG, Buss GR, Maroof MAS (2002) Mapping tightly linked genes controlling potyvirus infection at the Rsv1 and Rpv1 region in soybean. Genome 45:592–599
Gao L, Ding X, Li K, Liao W, Zhong Y, Ren R, Liu Z, Adhimoolam K, Zhi H (2015) Characterization of soybean mosaic virus resistance derived from inverted repeat-SMV-HC-Pro genes in multiple soybean cultivars. Theor Appl Genet 128:1489–1505
Hill JH, Whitham SA (2014) Control of virus diseases in soybeans. Adv Virus Res 90:355–390
Halbert SE, Irwin ME, Goodman RM (1981) Alate aphid (Homoptera: Aphididae) species and their relative importance as field vectors of soybean mosaic virus. Ann Appl Biol 97:1–9
Hayes AJ, Ma G, Buss GR, Maroof MAS (2000) Molecular marker mapping of RSV4, a gene conferring resistance to all known strains of soybean mosaic virus. Crop Sci 40:1434–1437
Hill JH, Lucas BS, Benner HI, Tachibana H, Hammond RB, Pedigo LP (1980) Factors associated with the epidemiology of soybean mosaic virus in Iowa. Phytopathology 70:536–540
Iino Y, Sugimoto A, Yamamoto M (1991) Whose overexpression inhibits sexual development, encodes a ribonuclease. Embo J 10:221–226
Ishida I, Ogawa T, Yoshioka M, Hori T, Ohtani T (1995) Production of plants tolerant to multiple viral infections by genetic manipulations. Acta Hortic 10:927–928
Jeong SC, Maroof MAS (2004) Detection and genotyping of SNPs tightly linked to two disease resistance loci, Rsv1 and Rsv3 of soybean. Plant Breed 123:305–310
Koo JM, Choi BK, Ahn HJ, Yum HJ, Choi CW (2005) First report of an Rsv resistance-breaking isolate of soybean mosaic virus in Korea. Plant Pathol 54:573–573
Lamontagne B, Larose S, Boulanger J, Elela SA (2001) The RNase III family: a conserved structure and expanding functions in eukaryotic dsRNA metabolism. Yeast 45:154–158
Liu JZ, Braun E, Qiu WL, Shi YF, Marcelino-Guimarães FC, Navarre D, Hill JH, Whitham SA (2014) Positive and negative roles for soybean MPK6 in regulating defense responses. Mol Plant-Microbe Interact 27:824–834
Luan H, Shine MB, Cui X, Chen X, Ma N, Kachroo P, Zhi H, Kachroo A (2016) The Potyviral P3 protein targets eukaryotic elongation factor 1A to promote the unfolded protein response and viral pathogenesis. Plant Physiol 172:221–234
Lim HS, Ko TS, Hobbs HA, Lambert KN, Yu JM, McCoppin NK, Korban SS, Hartman GL, Domier LL (2007) Soybean mosaic virus helper component-protease alters leaf morphology and reduces seed production in transgenic soybean. Plants Phytopathol 97:366–372
Maroof MAS, Tucker DM, Skoneczka JA, Bowman BC, Tripathy S, Tolin SA (2010) Fine mapping and candidate gene discovery of the soybean mosaic virus resistance gene Rsv4. Plant Genome 3:14–22
Nicholson AW (1999) Function, mechanism and regulation of bacterial ribonucleases. FEMS Microb Rev 23:371–390
Ogawa T, Toguri T, Kudoh H, Okamura M, Momma T, Yoshioka M, Hagiwara Y, Sano T (2005) Double-stranded RNA-specific ribonuclease confers tolerance against chrysanthemum stunt viroid and tomato spotted wilt virus in transgenic chrysanthemum plants. Breed Sci 55:49–55
Ross JP (1983) Effect of soybean mosaic on component yields from blends of mosaic resistant and susceptible soybeans. Crop Sci 23:343–346
Rotondo G, Huang JY, Frendewey D (1997) Substrate structure requirements of the Pac1 ribonuclease from Schizosaccharmyces pombe. Publ RNA Soc 3:1182–1193
Reddy MSS, Ghabrial SA, Redmond CT, Dinkins RD, Collins GB (2001) Resistance to bean pod mottle virus in transgenic soybean lines expressing the capsid polyprotein. Phytopathology 91:831–838
Sinclair JB (1989) Compendium of soybean diseases. 3rd Ed. APS Press, Saint Paul
Shukla DD, Ward CW, Brunt AA (1994) The Potyviridae. CAB International, Wallingford
Sano T, Nagayama A, Ogawa T, Ishida I, Okada Y (1997) Transgenic potato expressing a double-stranded RNA-specific ribonuclease is resistant to potato spindle tuber viroid. Nat Biotechnol 15:1290–1294
Saghai Maroof MA, Jeong SC, Gunduz I, Tucker DM, Buss GR, Tolin SA (2008) Pyramiding of soybean mosaic virus resistance genes by marker-assisted selection. Crop Sci 48:517–526
Shi A, Chen P, Li D, Zheng C, Zhang B, Hou A (2009) Pyramiding multiple genes for resistance to soybean mosaic virus in soybean using molecular markers. Mol Breed 23:113–124
Telzur N, Abbo S, Myslabodski D, Mizrahi Y (1999) Modified CTAB procedure for DNA isolation from Epiphytic cacti of the Genera Hylocereus and Selenicereus (Cautaceae). Plant Mol Biol Rep 17:249–254
Toguri T, Ogawa T, Kakitani M, Tukahara M, Yoshioka M (2003) Agrobacterium-mediated transformation of chrysanthemum (Dendranthema grandiflora) plants with a disease resistance gene (pac1). Plant Biotechnol 20:121–127
Tougou M, Furutani N, Yamagishi N, Shizukawa Y, Takahata Y, Hidaka S (2006) Development of resistant transgenic soybeans with inverted repeat-coat protein genes of soybean dwarf virus. Plant Cell Rep 25:1213–1218
Watanabe Y, Ogawa T, Takahashi H, Ishida I, Takeuchi Y, Yamamoto M, Okada Y (1995) Resistance against multiple plant viruses in plants mediated by a double stranded-RNA specific ribonuclease. Febs Lett 372:165–168
Yang Y, Lin J, Zheng G, Zhang M, Zhi H (2014) Recombinant soybean mosaic virus is prevalent in Chinese soybean fields. Arch Virol 159:1793–1796
Yan F, Zheng Y, Zhang W, Xiao H, Shifang LI, Cheng Z (2006) Obtained transgenic wheat expressing pac1 mediated by Agrobacterium is resistant against barley yellow dwarf virus. Chin Sci Bull 51:2362–2368
Yang Y, Zheng G, Han L, Dagang W, Yang X, Yuan Y, Huang S, Zhi H (2013) Genetic analysis and mapping of genes for resistance to multiple strains of soybean mosaic virus in a single resistant soybean accession PI 96983. Theor Appl Genet 126:1783–1791
Yang X, Niu L, Zhang W, He H, Yang J, Xing G, Guo D, Du Q, Qian X, Yao Y, Li Q, Dong Y (2017) Robust RNAi-mediated resistance to infection of seven potyvirids in soybean expressing an intron hairpin NIb RNA. Transgenic Res 26:665–676
Yang X, Niu L, Zhang W, Yang J, Xing G, He H, Guo D, Du Q, Qian X, Yao Y, Li Q, Dong Y (2018) RNAi-mediated SMV P3 cistron silencing confers significantly enhanced resistance to multiple Potyvirus strains and isolates in transgenic soybean. Plant Cell Rep 37:103–114
Zhang L, French R, Langenberg WG, Mitra A (2001) Accumulation of barley stripe mosaic virus is significantly reduced in transgenic wheat plants expressing a bacterial ribonuclease. Transgenic Res 10:13–19
Zhang X, Sato S, Ye X, Dorrance AE, Morris TJ, Clemente TE, Qu F (2011) Robust RNAi-based resistance to mixed infection of three viruses in soybean plants expressing separate short hairpins from a single transgene. Phytopathology 101:1264–1269
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 191:125–133
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.
Conflict of interest
All authors declare that they have no conflict of interest in the publication of this paper.
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Yang, X., Niu, L., Zhang, W. et al. Increased multiple virus resistance in transgenic soybean overexpressing the double-strand RNA-specific ribonuclease gene PAC1. Transgenic Res 28, 129–140 (2019). https://doi.org/10.1007/s11248-018-0108-8