摘要
本研究对信前胡烟草花叶病毒进行鉴定、原核蛋白表达和序列分析,为信前胡脱毒苗的培育和鉴定提供理论依据.通过lncRNA测序和RT-PCR鉴定信前胡烟草花叶病毒蛋白基因,利用大肠杆菌重 组技术表达信前胡烟草花叶病毒蛋白,并采用生物信息学方法进行序列分析.信前胡烟草花叶病毒蛋白基因包括复制酶(ORF1)、运动蛋白(ORF2)和外壳蛋白(ORF3);信前胡烟草花叶病毒复制酶、运动蛋白和外壳蛋白均可通过RT-PCR检测到,并可实现大肠杆菌重组表达,表明信前胡烟草花叶病毒的lncRNA测序结果准确;信前胡烟草花叶病毒复制酶、运动蛋白和外壳蛋白基因cDNA总长度分别为3351 bp、807 bp和480 bp;信前胡烟草花叶病毒复制酶、运动蛋白和外壳蛋白与烟草花叶病毒的亲缘关系较近,同源性分别为99.64％、98.88％、98.74％.信前胡烟草花叶病毒(Peucedanum praeruptorm tobacco mosaic virus,PpTMV)在江西省上饶市信前胡主产区信州、广信、玉山、广丰、婺源、德兴、弋阳的7个区或县均有发生.这是首次报道在信前胡上检测到PpTMV.
Abstract
To provide theoretical basis for the cultivation and identification of virus-free seedlings of P.praeruptorum Dunn.cv.Xinzhou,its tobacco mosaic virus was identified,its protein was expressed and its sequence was analyzed.The protein gene of tobacco mosaic virus of P.praeruptorum Dunn.cv.Xinzhou was identified by lncRNA sequencing and RT-PCR,and its protein of tobacco mosaic virus was expressed by E.coli recombinant technology,and its gene sequence was analyzed by bioinformatics method.The protein genes of tobacco mosaic virus of P.praeruptorum Dunn.cv.Xinzhou included replicase(ORF1),movement protein(ORF2)and coat protein(ORF3).The replicating enzyme,movement protein and coat protein of tobacco mosaic virus could be detected by RT-PCR,and could be recombined and expressed in E.coli,indicating that the sequencing result of lncRNA of tobacco mosaic virus was accurate.The total length of the cDNA of the replicase,movement protein and coat protein genes of the tobacco mosaic virus was 3351 bp,807 bp and 480 bp,respectively.The replicase,movement protein and coat protein of tobacco mosaic virus from P.praeruptorum Dunn.cv.Xinzhou were closely related to tobacco mosaic virus,with the homology of 99.64％,98.88％and 98.74％respectively.Peucedanum praeruptom tobacco mosaic virus(PpTMV)has occurred in seven districts or counties in Xinzhou,Guangxin,Yushan,Guangfeng,Wuyuan,Dexing and Yiyang,the main producing areas of P.praeruptorum Dunn.cv.Xinzhou in Shangrao,Jiangxi Province.This article is the first report of PpTMV detected in P.praeruptorum Dunn.cv.Xinzhou.
基金项目
国家自然科学基金资助目(31960079)
江西省科技厅重点研发计划一般项目(20202BBG73010)
江西省教育厅科学技术研究项目(GJJ201704)
2022年上饶市科技专项项目(社发类)(饶科发[2023]5号)
2022年上饶市科技专项项目(社发类)(2022A008)
江西省现代农业产业技术体系建设专项(JXARS-13-赣东站)
上饶市科技局平台载体建设项目(2020J001)
上饶师范学院2023年大学生创新创业训练计划项目(2023-PX-18)
国家科技期刊平台
NSTL
万方数据