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侵染蚕豆ClYVV的鉴定及其衍生的小干扰RNA的深度测序鉴定研究

Identification of Clover yellow vein virus infecting broad bean in China by deep sequencing and characterization of virus-derived small interfering RNAs

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【作者】 张趁华郑红英严丹侃韩科雷彭杰军鲁宇文林林章东方陈剑平燕飞

【Author】 ZHANG Chenhua;ZHENG Hongying;YAN Dankan;HAN Kelei;PENG Jiejun;LU Yuwen;LIN Lin;ZHANG Dongfang;CHEN Jianping;YAN Fei;College of Life Sciences,Fujian Agriculture and Forestry University;Stake Key Laboratory Breeding Base for Sustainable Control of Plant Pest and Disease,Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province,Institute of Virology and Biotechnology,Zhejiang Academy of Agricultural Sciences;Institute of Plant Protection and Agricultural Product Quality and Safety,Anhui Academy of Agricultural Sciences;College of Plant Protection,Nanjing Agricultural University;

【机构】 福建农林大学生命科学学院浙江省农业科学院病毒学与生物技术研究所,浙江省和农业部植保生物技术重点实验室,省部共建浙江省植物有害生物防控国家重点实验室培育基地安徽省农业科学院植物保护与农产品质量安全所南京农业大学植物保护学院

【摘要】 对安徽省合肥市采集到的具有明显典型花叶症状的蚕豆叶片进行深度测序,鉴定到样品中存在三叶草黄脉病毒(Clover yellow vein virus,Cl YVV),并获得了Cl YVV的完整基因组序列,随后通过直接测序扩增基因组区域获得完整的基因组Cl YVV-HF。获得的Cl YVV-HF基因组由9 585个核苷酸组成,不包括poly(A)尾巴。Cl YVV-HF的全部核苷酸序列与之前报道的来自日本的分离株(AB011819、NC003536)和来自韩国的分离株(KF975894)分别具有95.45%和95.10%的核苷酸同源性。进一步分析了Cl YVV-HF衍生的小干扰RNA的表征。vsiRNAs的大小为2122 nt,vsiRNAs的5’末端碱基偏向于A/U。Cl YVV-HF来源的小干扰RNA大多来源于其基因组链中的7个热点,其中5个热点以21 nt vsiRNAs为主。生物学实验表明,Cl YVV可通过汁液摩擦接种多种豆科作物。这是有关中国Cl YVV分离株全部核苷酸序列的首次报道。

【Abstract】 The broad bean samples causing prominent mosaic on leaves were collected in Hefei City,Anhui Province in China,and viruses in samples were detected by deep sequencing. The results showed that Clover yellow vein virus( Cl YVV) existed in the broad bean samples and the complete genome Cl YVV-HF was obtained by direct sequencing of the amplified genomic region,subsequently. The Cl YVV genome of China consists of 9 585 nucleotides excluding the poly( A) tail,the full nucleotide sequence of Cl YVV-HF shared a nucleotide identity of 95. 45% and 95. 10%with that of isolates from Japan( AB011819,NC003536) and an isolate from Korea( KF975894) reported previously. Characterization of Cl YVV-derived small interfering RNAs was further analyzed. The prevalent vsiRNAs size was21-22 nt,and the 5’-terminal base of vsiRNAs was biased towards A/U. The Cl YVV-derived small interfering RNAs were most derived from its genomic-strand RNAs,among the 7 hot spots across the genome,5 hot spots were predominated by 21 nt vsiRNAs. Biological experiments showed that Cl YVV could be used to frictionally inoculate many leguminous crops through juice. This is the first report about the full nucleotide sequence of Cl YVV isolate from China.

【关键词】 蚕豆ClYVV小干扰RNA
【Key words】 broad beanClYVVsiRNA
【基金】 公益性行业(农业)科研项目子课题(201303028);浙江省科技计划项目(2017C32040);莆田市科技资助项目(2016S3001)
  • 【文献出处】 浙江农业学报 ,Acta Agriculturae Zhejiangensis , 编辑部邮箱 ,2018年03期
  • 【分类号】S436.43
  • 【被引频次】3
  • 【下载频次】124
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