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RNAi-mediated resistance to rice black-streaked dwarf virus in transgenic rice

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Abstract

Rice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus in the family Reoviridae, causes significant economic losses in rice production in China and many other Asian countries. Development of resistant varieties by using conventional breeding methods is limited, as germplasm with high level of resistance to RBSDV have not yet been found. One of the most promising methods to confer resistance against RBSDV is the use of RNA interference (RNAi) technology. RBSDV non-structural protein P7-2, encoded by S7-2 gene, is a potential F-box protein and involved in the plant-virus interaction through the ubiquitination pathway. P8, encoded by S8 gene, is the minor core protein that possesses potent active transcriptional repression activity. In this study, we transformed rice calli using a mini-twin T-DNA vector harboring RNAi constructs of the RBSDV genes S7-2 or S8, and obtained plants harboring the target gene constructs and the selectable marker gene, hygromycin phosphotransferase (HPT). From the offspring of these transgenic plants, we obtained selectable marker (HPT gene)-free plants. Homozygous T5 transgenic lines which harbored either S7-2-RNAi or S8-RNAi exhibited high level resistance against RBSDV under field infection pressure from indigenous viruliferous small brown planthoppers. Thus, our results showed that RNA interference with the expression of S7-2 or S8 genes seemed an effective way to induce high level resistance in rice against RBSD disease.

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References

  • Azuhata F, Uyeda I, Kimura I, Shikata E (1993) Close similarity between genome structures of rice black-streaked dwarf and maize rough dwarf viruses. J Gen Virol 74:1227–1232

    Article  CAS  PubMed  Google Scholar 

  • Bai FW, Yan J, Qu ZC, Zhang HW, Xu J, Ye MM, Shen DL (2002) Phylogenetic analysis reveals that a dwarfing disease on different cereal crops in China is due to rice black streaked dwarf virus (RBSDV). Virus Genes 25:201–206

    Article  CAS  PubMed  Google Scholar 

  • Baulcombe D (2005) RNA silencing. Trends Biochem Sci 30:290–293

    Article  CAS  PubMed  Google Scholar 

  • Bubner B, Baldwin IT (2004) Use of real-time PCR for determining copy number and zygosity in transgenic plants. Plant Cell Rep 23:263–271

    Article  CAS  PubMed  Google Scholar 

  • Chen YK, Lohuis D, Goldbach R, Prins M (2004) High frequency induction of RNA-mediated resistance against Cucumber mosaic virus using inverted repeat constructs. Mol Breed 14:215–226

    Article  Google Scholar 

  • Dale PJ (1992) Spread of engineered genes to wild relatives. Plant Physiol 100:13–15

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Darbani B, Eimanifar A, Stewart CN, Camargo WN (2007) Methods to produce marker-free transgenic plants. Biotechnol J 2:83–90

    Article  CAS  PubMed  Google Scholar 

  • Ding J, Jia J, Yang L, Wen H, Zhang C, Liu W, Zhang D (2004) Validation of a rice specific gene, sucrose phosphate synthase, used as the endogenous reference gene for qualitative and real-time quantitative PCR detection of transgenes. J Agric Food Chem 52:3372–3377

    Article  CAS  PubMed  Google Scholar 

  • Doyle JJ (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Firth AE, Atkins JF (2009) Analysis of the coding potential of the partially overlapping 3’ORF in segment 5 of the plant fijiviruses. Virol J 6:32–36

    Article  PubMed  PubMed Central  Google Scholar 

  • German MA, Kandel-Kfir M, Swarzberg D, Matsevitz T, Granot D (2003) A rapid method for the analysis of zygosity in transgenic plants. Plant Sci 164:183–187

    Article  CAS  Google Scholar 

  • Hibino H (1996) Biology and epidemiology of rice viruses. Annu Rev Phytopathol 34:249–274

    Article  CAS  PubMed  Google Scholar 

  • Ishii M, Yoshimura S (1973) Epidemiological studies on the rice black-streaked dwarf virus in Kanto-Tosan district, Japan. J Cent Agric Exp Stat 17:61–121

    Google Scholar 

  • Isogai M, Uyeda I, Lee B (1998) Detection and assignment of proteins encoded by rice black streaked dwarf fijivirus S7, S8, S9 and S10. J Gen Virol 79:1487–1494

    Article  CAS  PubMed  Google Scholar 

  • Kalantidis K, Psaradakis S, Tabler M, Tsagris M (2002) The occurrence of CMV-specific short RNAs in transgenic tobacco expressing virus-derived double-stranded RNA is indicative of resistance to the virus. Mol Plant-Microbe Interact 15:826–833

    Article  CAS  PubMed  Google Scholar 

  • Kong H, Leebens-Mack J, Ni W, Ma H (2004) Highly heterogeneous rates of evolution in the SKP1 gene family in plants and animals: functional and evolutionary implications. Mol Biol Evol 21:117–128

    Article  CAS  PubMed  Google Scholar 

  • Li C, Song J, Jiang L (1999) Research progress on the maize rough dwarf virus disease. Plant Prot 25:34–37

    Google Scholar 

  • Liu H, Wei C, Zhong Y, Li Y (2007a) Rice black-streaked dwarf virus minor core protein P8 is a nuclear dimeric protein and represses transcription in tobacco protoplasts. FEBS Lett 581:2534–2540

    Article  CAS  PubMed  Google Scholar 

  • Liu H, Wei C, Zhong Y, Li Y (2007b) Rice black-streaked dwarf virus outer capsid protein P10 has self-interactions and forms oligomeric complexes in solution. Virus Res 127:34–42

    Article  CAS  PubMed  Google Scholar 

  • Liu XY, Yang J, Xie L, Li J, Song XJ, Chen JP, Zhang HM (2015) P5-2 of rice black-streaked dwarf virus is a non-structural protein targeted to chloroplasts. Arch Virol 160:1211–1217

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • McElroy D, Rothenberg M, Reece KS, Wu R (1990) Characterization of the rice (Oryza sativa) actin gene family. Plant Mol Biol 15:257–268

    Article  CAS  PubMed  Google Scholar 

  • Miki D, Itoh R, Shimamoto K (2005) RNA silencing of single and multiple members in a gene family of rice. Plant Physiol 138:1903–1913

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Missiou A, Kalantidis K, Boutla A, Tzortzakaki S, Tabler M, Tsagris M (2004) Generation of transgenic potato plants highly resistant to potato virus Y (PVY) through RNA silencing. Mol Breed 14:185–197

    Article  CAS  Google Scholar 

  • Park J, Lee YK, Kang BK, Chung WI (2004) Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants. Theor Appl Genet 109:1562–1567

    Article  CAS  PubMed  Google Scholar 

  • Prior FA, Tackaberry ES, Aubin RA, Casley WL (2006) Accurate determination of zygosity in transgenic rice by real-time PCR does not require standard curves or efficiency correction. Transgenic Res 15:261–265

    Article  CAS  PubMed  Google Scholar 

  • Ruan YL, Chen SX, Lin RF, Jiang WL, Jin DD (1984) Studies on rice black-streaked dwarf virus. J Zhejiang Agric Sci 4:185–187

    Google Scholar 

  • Shikata E, Kitagawa Y (1977) Rice black-streaked dwarf virus: its properties, morphology and intracellular localization. Virology 77:826–842

    Article  CAS  PubMed  Google Scholar 

  • Shimizu T, Nakazono-Nagaoka E, Akita F, Uehara-Ichiki T, Omura T, Sasaya T (2011) Immunity to Rice black streaked dwarf virus, a plant reovirus, can be achieved in rice plants by RNA silencing against the gene for the viroplasm component protein. Virus Res 160:400–403

    Article  CAS  PubMed  Google Scholar 

  • Shimizu T, Ogamino T, Hiraguri A et al (2013) Strong resistance against rice grassy stunt virus is induced in transgenic rice plants expressing double-stranded RNA of the viral genes for nucleocapsid or movement proteins as targets for RNA interference. Phytopathology 103:513–519

    Article  PubMed  Google Scholar 

  • Smalla K, Borin S, Heuer H, Gebhard F, van Elsas JD, Nielsen K (2000) Horizontal transfer of antibiotic resistance genes from transgenic plants to bacteria. In: Fairbairn C, Scoles G, McHughen A (eds) Proceedings of the 6th international symposium on the biosafety of GMOs. University Extension Press, University of Saskatchewan, pp 146–154

  • Sun F, Yuan X, Xu Q, Zhou T, Fan Y, Zhou Y (2013a) Overexpression of rice black-streaked dwarf virus p7-1 in Arabidopsis results in male sterility due to non-dehiscent anthers. PLoS ONE 8:e79514

    Article  PubMed  PubMed Central  Google Scholar 

  • Sun L, Xie L, Andika IB, Tan Z, Chen J (2013b) Non-structural protein P6 encoded by rice black-streaked dwarf virus is recruited to viral inclusion bodies by binding to the viroplasm matrix protein P9-1. J Gen Virol 94:1908–1916

    Article  CAS  PubMed  Google Scholar 

  • Sun Z, Zhang S, Xie L et al (2013c) The secretory pathway and the actomyosin motility system are required for plasmodesmatal localization of the P7-1 of rice black-streaked dwarf virus. Arch Virol 158:1055–1064

    Article  CAS  PubMed  Google Scholar 

  • Supyani S, Hillman BI, Suzuki N (2007) Baculovirus expression of the 11 mycoreovirus-1 genome segments and identification of the guanylyltransferase-encoding segment. J Gen Virol 88:342–350

    Article  CAS  PubMed  Google Scholar 

  • Syvanen M (1999) In search of horizontal gene transfer. Nat Biotechnol 17:833

    Article  CAS  PubMed  Google Scholar 

  • Voinnet O (2005) Induction and suppression of RNA silencing: insights from viral infections. Nat Rev Genet 6:206–220

    Article  CAS  PubMed  Google Scholar 

  • Wang ZH, Fang SG, Xu JL, Sun LY, Li DW, Yu JL (2003) Sequence analysis of the complete genome of rice black-streaked dwarf virus isolated from maize with rough dwarf disease. Virus Genes 27:163–168

    Article  PubMed  Google Scholar 

  • Wang BX, Jiang L, Chen LM et al (2010) Screening of rice resources against rice black-streaked dwarf virus and mapping of resistant QTL. Acta Agron Sin 36:1258–1264

    Article  CAS  Google Scholar 

  • Wang Q, Tao T, Zhang Y, Wu W, Li D, Yu J, Han C (2011) Rice black-streaked dwarf virus P6 self-interacts to form punctate, viroplasm-like structures in the cytoplasm and recruits viroplasm-associated protein P9-1. Virol J 8:8–24

    Article  Google Scholar 

  • Wang Q, Tao T, Han Y et al (2013) Nonstructural protein P7-2 encoded by rice black-streaked dwarf virus interacts with SKP1, a core subunit of SCF ubiquitin ligase. Virol J 10:1–12

    Article  Google Scholar 

  • Waterhouse PM, Wang MB, Lough T (2001) Gene silencing as an adaptive defence against viruses. Nature 411:834–842

    Article  CAS  PubMed  Google Scholar 

  • Wu J, Ni Y, Liu H, Rao L, Zhou Y, Zhou X (2013) Development and use of three monoclonal antibodies for the detection of rice black-streaked dwarf virus in field plants and planthopper vectors. Virol J 10:114

    Article  PubMed  PubMed Central  Google Scholar 

  • Yang J, Zhang HM, Ying L et al (2014) Rice black-streaked dwarf virus genome segment S5 is a bicistronic mRNA in infected plants. Arch Virol 159:307–314

    Article  CAS  PubMed  Google Scholar 

  • Yu H, Liu Q, Wang L et al (2005) Agrobacterium-mediated transformation of rice mature embryos and regeneration of transgenic plants with Metr gene. Yi Chuan 27:772–778

    CAS  PubMed  Google Scholar 

  • Zechendorf B (1994) What the public thinks about biotechnology. Nat Biotechnol 12:870–875

    Article  CAS  Google Scholar 

  • Zhang H, Chen J, Lei J, Adams MJ (2001a) Sequence analysis shows that a dwarfing disease on rice, wheat and maize in China is caused by rice black-streaked dwarf virus. Eur J Plant Pathol 107:563–567

    Article  CAS  Google Scholar 

  • Zhang HM, Chen J, Adams M (2001b) Molecular characterisation of segments 1 to 6 of rice black-streaked dwarf virus from China provides the complete genome. Arch Virol 146:2331–2339

    Article  CAS  PubMed  Google Scholar 

  • Zhang L, Wang Z, Wang X, Li D, Han C, Zhai Y, Yu J (2005) Two virus-encoded RNA silencing suppressors, P14 of Beet necrotic yellow vein virus and S6 of Rice black streak dwarf virus. Chin Sci Bull 50:305–310

    CAS  Google Scholar 

  • Zhang C, Liu Y, Liu L et al (2008) Rice black streaked dwarf virus P9-1, an α-helical protein, self-interacts and forms viroplasms in vivo. J Gen Virol 89:1770–1776

    Article  CAS  PubMed  Google Scholar 

  • Zheng T, Yang J, Zhong W et al (2012) Novel loci for field resistance to black-streaked dwarf and stripe viruses identified in a set of reciprocal introgression lines of rice (Oryza sativa L.). Mol Breed 29:925–938

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the National Key Research and Development Program (2016YFD0102001-006), the National Natural Science Foundation of China (31670313), Six Big Talent Peak of Jiangsu Province (NY018), Key Project of Jiangsu Education Department of China (15KJA180010), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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    1. Key Laboratory of Plant Functional Genomics of the Ministry of Education, Jiangsu Key Laboratory of Crop Genetics and Physiology/Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou, 225009, China

      Mohamed M. S. Ahmed, Shiquan Bian, Muyue Wang, Jing Zhao, Bingwei Zhang, Qiaoquan Liu, Changquan Zhang, Shuzhu Tang, Minghong Gu & Hengxiu Yu

    2. Department of Crop Protection, Faculty of Agriculture, University of Khartoum, 13314, Khartoum North, Sudan

      Mohamed M. S. Ahmed

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    1. Mohamed M. S. Ahmed
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    Mohamed M. S. Ahmed, Shiquan Bian, Muyue Wang, Jing Zhao and Bingwei Zhang have contributed equally to this work.

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    Ahmed, M.M.S., Bian, S., Wang, M. et al. RNAi-mediated resistance to rice black-streaked dwarf virus in transgenic rice. Transgenic Res 26, 197–207 (2017). https://doi.org/10.1007/s11248-016-9999-4

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