甘薯羽状斑驳病毒海南分离物全基因组序列克隆及分析  

Complete Genome Sequencing and Analysis of Sweet potato feathery mottle virus(SPFMV)Isolates from Hainan

作　　者：李嘉莹 沈文涛[2] 庹德财[2] 朱国鹏[1] 黎小瑛[2] 言普[2] 周鹏[1,2] Li Jiaying;Shen Wentao;Tuo Decai;Zhu Guopeng;Li Xiaoying;Yan Pu;Zhou Peng(College of Horticulture,Hainan University,Haikou,570228;Hainan Academy of Tropical Agricultural Resource,Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou,571101)

机构地区：[1]海南大学园艺学院,海口570228 [2]中国热带农业科学院热带生物技术研究所,海南热带农业资源研究院,海口571101

出　　处：《分子植物育种》2020年第8期2445-2451,共7页Molecular Plant Breeding

基　　金：海南省重点研发计划(ZDYF2019030)资助。

摘　　要：为了明确海南本地甘薯羽状斑驳病毒(sweet potato feathery mottle virus,SPFMV)种群的遗传进化关系及致病机理,本研究利用Small RNA测序技术对采自海南的甘薯样品进行鉴定,获得与SPFMV序列具有相似性的85个contigs,与NC_001841相似性较高。试验设计了5对共8条引物,分别为SPFMV-1F/1R、SPFMV-11F/1R、SPFMV-2F/2R、SPFMV-3F/3R和SPFMV-3F/33R PCR,扩增获得3987 bp、3710 bp、1996 bp、3369 bp和4730 bp目的片段。在此基础上再分段设计引物,进行基因全序列PCR扩增,序列拼接获得SPFMV-HN基因组全序列,其包含完整编码阅读框。本研究结果首次报道SPFMV海南分离物基因组全长,并且在P1区预测到另一个外框元件(pretty interesting sweet potato potyviral ORF,PISPO),聚合酶滑移机制可以产生带有额外核苷酸的转录变体,这些核苷酸可以翻译为P1N-PISPO和P3N-PIPO。本研究结果为进一步探究SPFMV致病分子机理和培育抗病甘薯品种奠定研究基础。In order to understand the genetic evolution and pathogenic mechanism of native sweet potato feather mottle virus(SPFMV)population in Hainan,the sweet potato samples collected from Hainan were identified by small RNA sequencing,then 85 contigs were obtained with having high similarity to sequences of SPFMV and NC_001841.First,small fragment primers were designed,primer combinations used as SPFMV-1F/1R,SPFMV-11F/1R,SPFMV-2F/2R,SPFMV-3F/3R and SPFMV-3F/33R,target fragments of 3987 bp,1996 bp and 4730 bp were amplified by PCR,respectively.The primers were designed in sections to perform full-length PCR amplification of the SPFMV-HN genome.The complete sequence of the SPFMV-HN genome was obtained with sequence splicing,including the complete coding reading frame,which is the first full report of the genome of SPFMV isolates from Hainan in China.And another frame element(periodic input same period output,PISPO)was predicted in the P1 region,and the polymerase slip mechanism could generate transcript variants with additional nucleotides that could be translated into P1N-PISPO and P3N-PIPO.This results could provide a basis for further understanding the pathogenic molecular mechanism of SPFMV-HN and the cultivation of disease-resistant sweet potato variety.

关 键 词：甘薯羽状斑驳病毒 SMALL RNA测序技术 全基因测序 序列分析 

分 类 号：S435.31[农业科学—农业昆虫与害虫防治]