title: Hairpin RNA Targeting Multiple Viral Genes Confers Strong Resistance to Rice Black-Streaked Dwarf Virus.
abstract: Rice black-streaked dwarf virus (RBSDV) belongs to the genus Fijivirus in the family of Reoviridae and causes severe yield loss in rice-producing areas in Asia. RNA silencing, as a natural defence mechanism against plant viruses, has been successfully exploited for engineering virus resistance in plants, including rice. In this study, we generated transgenic rice lines harbouring a hairpin RNA (hpRNA) construct targeting four RBSDV genes, S1, S2, S6 and S10, encoding the RNA-dependent RNA polymerase, the putative core protein, the RNA silencing suppressor and the outer capsid protein, respectively. Both field nursery and artificial inoculation assays of three generations of the transgenic lines showed that they had strong resistance to RBSDV infection. The RBSDV resistance in the segregating transgenic populations correlated perfectly with the presence of the hpRNA transgene. Furthermore, the hpRNA transgene was expressed in the highly resistant transgenic lines, giving rise to abundant levels of 21-24 nt small interfering RNA (siRNA). By small RNA deep sequencing, the RBSDV-resistant transgenic lines detected siRNAs from all four viral gene sequences in the hpRNA transgene, indicating that the whole chimeric fusion sequence can be efficiently processed by Dicer into siRNAs. Taken together, our results suggest that long hpRNA targeting multiple viral genes can be used to generate stable and durable virus resistance in rice, as well as other plant species.
authors.affiliation: Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R &amp; D Center, Nanjing 210014, China. qiezi324@163.com.|Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China. qiezi324@163.com.
authors.forename: Fangquan
authors.initials: F
authors.lastname: Wang
authors.affiliation: Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R &amp; D Center, Nanjing 210014, China. liwqtomorrow@163.com.|Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China. liwqtomorrow@163.com.
authors.forename: Wenqi
authors.initials: W
authors.lastname: Li
authors.affiliation: Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R &amp; D Center, Nanjing 210014, China. zhujinyanrain@163.com.|Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China. zhujinyanrain@163.com.
authors.forename: Jinyan
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R &amp; D Center, Nanjing 210014, China. weidadefan@sohu.com.|Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China. weidadefan@sohu.com.
authors.forename: Fangjun
authors.initials: F
authors.lastname: Fan
authors.affiliation: Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R &amp; D Center, Nanjing 210014, China. wangjunjaas@aliyun.com.|Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China. wangjunjaas@aliyun.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R &amp; D Center, Nanjing 210014, China. wgzhong0503@139.com.|Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China. wgzhong0503@139.com.
authors.forename: Weigong
authors.initials: W
authors.lastname: Zhong
authors.affiliation: CSIRO Agriculture, GPO Box 1600, Canberra, ACT 2601, Australia. ming-bo.wang@csiro.au.
authors.forename: Ming-Bo
authors.initials: MB
authors.lastname: Wang
authors.affiliation: CSIRO Agriculture, GPO Box 1600, Canberra, ACT 2601, Australia. qing.liu@csiro.au.
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.affiliation: CSIRO Agriculture, GPO Box 1600, Canberra, ACT 2601, Australia. qianhao.zhu@csiro.au.
authors.forename: Qian-Hao
authors.initials: QH
authors.lastname: Zhu
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China. zhoutong@jaas.ac.cn.
authors.forename: Tong
authors.initials: T
authors.lastname: Zhou
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China. 15005167120@163.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Lan
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China. yjzhou@jaas.ac.cn.
authors.forename: Yijun
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R &amp; D Center, Nanjing 210014, China. yangjie168@aliyun.com.|Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China. yangjie168@aliyun.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
mesh_terms: D036022:Capsid Proteins / Q000235:genetics*; D012275:Oryza / Q000235:genetics* / Q000276:immunology / Q000821:virology; D057865:Plant Immunity / Q000235:genetics*; D034622:RNA Interference; D012087:Reoviridae / Q000235:genetics* / Q000472:pathogenicity; D019076:Transgenes
chemical_list: D036022:Capsid Proteins; C071651:S10 protein, rice gall dwarf virus
keywords: RBSDV; RNAi; hpRNA; multiple viral genes; strong resistance