摘要
在河北石家庄市的番茄上分离得到番茄黄化曲叶病毒(tomato yellow leaf curl virus,TYLCV)及其伴随卫星DNA(Betasatellite DNA,又称DNAβ),对其进行分子鉴定及序列分析,明确TYLCV石家庄分离物及其伴随DNAβ分子结构特征及其进化起源。根据已报道的TYLCV及其伴随DNAβ序列设计引物,分别扩增TYLCV石家庄分离物及其伴随DNAβ的全长序列并测序。在NCBI上选取国内外有代表性的TYLCV序列和DNAβ序列,使用MEGA 6.0软件和DNAMAN软件对石家庄TYLCV分离物及其伴随DNAβ进行遗传变异分析。结果表明:TYLCV石家庄分离物全长为2781 bp,共有6个ORFs,将该分离物定名为TYLCV-SJZ2021。基因组序列比对发现,TYLCV-SJZ2021与山东省的分离物同源性最高(99%);与2015年本实验室报道的TYLCV石家庄分离物相似度为98.67%,存在37处碱基差异,均属于TYLCV以色列株系。TYLCV-SJZ2021的伴随DNAβ全长1343 bp,其上有病毒在细胞间移动相关蛋白的BC1编码区,序列比对发现该DNAβ与河南TYLCV伴随DNAβ分子最为相似(98%)。从分子水平上鉴定了TYLCV-SJZ2021及其伴随DNAβ的分类地位,为防治该病毒病害及其抗病育种奠定了基础。
Tomato yellow leaf curl virus(TYLCV)was isolated from tomatoes in Shijiazhuang City,Hebei Province,and it has an associated DNA betasatellite(also called DNAβ).Molecular identification and sequence analysis were carried out to determine genomic characterization and evolutionary origin of TYLCV Shijiazhuang isolate and its DNAβ.According to the genome sequence of the reported TYLCV and its DNAβ,the primer was designed to amplify full-length genome of TYLCV Shijiazhuang isolate and its DNAβand sequenced,re spectively.Select representative sequences of TYLCV isolates and its DNAβat home and abroad,the sequences were analyzed for genetic variation by MEGA 6.0 software and DNAMAN software.The results of sequencing showed that the total length of the TYLCV Shijiazhuang isolate was 2781 bp,with 6 ORFs,and it named TYLCV-SJZ2021.Genome sequence comparison showed that TYLCV-SJZ2021 had the highest homology(99%)with Shandong isolate,and its similarity with the reported by our lab in 2015 was 98.67%,with 37 base differences.The DNAβof TYLCV-SJZ2021 has a total length of 1343 bp,on which there is the BC1 coding region of the protein related to the movement of the virus between cells.Sequence comparison shows that TYLCV-SJZ2021 DNAβis the most similar to the reported Henan DNAβ(98%).The TYLCV-SJZ2021 and its DNAβin the virus classification status was identified at the molecular level,aad laying the foundation for the prevention and control of the virus disease and resistance breeding in this study.
作者
韩旭良
康忱
田哲娟
王鹏
李亚栋
赵雪芳
闫洪波
刘美霞
吴志明
HAN Xu-liang;KANG Chen;TIAN Zhe-juan;WANG Peng;LI Ya-dong;ZHAO Xue-fang;YAN Hong-bo;LIU Mei-xia;WU Zhi-ming(Hebei University of Economics and Business,Shijiazhuang 050051,China;Institute of Cash Crops,Hebei Academy of Agriculture and Forestry Sciences,Shijiazhuang 050051,China;Shengcheng Street Office of Shouguang,Shouguang 262700,China)
出处
《河北农业科学》
2023年第2期74-82,共9页
Journal of Hebei Agricultural Sciences
基金
河北省重点研发计划项目(19226342D)
河北省农林科学院创新工程项目(2022KJCXZX-JZS-1)。
关键词
番茄黄化曲叶病毒
DNAΒ
分子鉴定
序列分析
Tomato yellow leaf curl virus(TYLCV)
Associated DNA betasatellite
Sequence analysis
Molecular identification
作者简介
韩旭良(1996-),男,河北石家庄人,硕士研究生在读,研究方向为蔬菜抗病分子遗传育种。E-mail:1732070525@qq.com;通讯作者:刘美霞(1982-),女,山东寿光人,中级农艺师,主要从事农业技术推广工作。E-mail:lmxzwt@163.com;通讯作者:吴志明(1971-),男,河北临漳人,研究员,博士,主要从事蔬菜抗病分子遗传育种研究。Tel:0311-87652260,E-mail:Zhiming71@126.com。
