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

doi:10.3969/j.issn.1004-1524.2018.03.08

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (3): 406-412.DOI: 10.3969/j.issn.1004-1524.2018.03.08

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

张趁华^1,2, 郑红英², 严丹侃³, 韩科雷^2,4, 彭杰军², 鲁宇文², 林林², 章东方³, 陈剑平^1,2,*, 燕飞^2,*

1.福建农林大学生命科学学院,福建福州350002;
2.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,浙江杭州310021;
3.安徽省农业科学院植物保护与农产品质量安全所,安徽合肥230031;
4.南京农业大学植物保护学院,江苏南京 210095

收稿日期:2017-11-30 出版日期:2018-03-20 发布日期:2018-03-21
通讯作者: *陈剑平,E-mail: jpchen2001@126.com;燕飞,E-mail: fei.yan@mail.zaas.ac.cn
作者简介:张趁华(1984—),女,河北石家庄人,博士,从事细胞生物学研究。E-mail: zchcxj@126.com
基金资助:
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)

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

ZHANG Chenhua^1,2, ZHENG Hongying², YAN Dankan³, HAN Kelei^2,4, PENG Jiejun², LU Yuwen², LIN Lin², ZHANG Dongfang³, CHEN Jianping^1,2,*, YAN Fei^2,*

1. College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
2. 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, Hangzhou 310021, China;
3. Institute of Plant Protection and Agricultural Product Quality and Safety, Anhui Academy of Agricultural Sciences, Hefei 230031, China;
4. College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China

Received:2017-11-30 Online:2018-03-20 Published:2018-03-21

摘要/Abstract

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, ClYVV),并获得了ClYVV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ClYVV-HF。获得的ClYVV-HF基因组由9 585个核苷酸组成,不包括poly(A)尾巴。ClYVV-HF的全部核苷酸序列与之前报道的来自日本的分离株(AB011819、NC_003536)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ClYVV-HF衍生的小干扰RNA的表征。vsiRNAs的大小为21~22 nt,vsiRNAs的5' 末端碱基偏向于A / U。ClYVV-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,ClYVV可通过汁液摩擦接种多种豆科作物。这是有关中国ClYVV分离株全部核苷酸序列的首次报道。

关键词: 蚕豆, ClYVV, 小干扰RNA

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(ClYVV) existed in the broad bean samples and the complete genome ClYVV-HF was obtained by direct sequencing of the amplified genomic region, subsequently. The ClYVV genome of China consists of 9 585 nucleotides excluding the poly(A) tail, the full nucleotide sequence of ClYVV-HF shared a nucleotide identity of 95.45% and 95.10% with that of isolates from Japan(AB011819, NC_003536) and an isolate from Korea (KF975894) reported previously. Characterization of ClYVV-derived small interfering RNAs was further analyzed. The prevalent vsiRNAs size was 21-22 nt, and the 5'-terminal base of vsiRNAs was biased towards A/U. The ClYVV-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 ClYVV could be used to frictionally inoculate many leguminous crops through juice. This is the first report about the full nucleotide sequence of ClYVV isolate from China.

Key words: broad bean, ClYVV, siRNA

中图分类号:

Q945.8

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. 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[J]. 浙江农业学报, 2018, 30(3): 406-412.

ZHANG Chenhua, ZHENG Hongying, YAN Dankan, HAN Kelei, PENG Jiejun, LU Yuwen, LIN Lin, ZHANG Dongfang, CHEN Jianping, YAN Fei. Identification of Clover yellow vein virus infecting broad bean in China by deep sequencing and characterization of virus-derived small interfering RNAs[J]. , 2018, 30(3): 406-412.

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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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