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中国农学通报

中国农学通报 ›› 2024, Vol. 40 ›› Issue (21): 106-113.doi: 10.11924/j.issn.1000-6850.casb2023-0643

• 生物科学 • 上一篇    下一篇

信前胡烟草花叶病毒lncRNA测序鉴定、原核蛋白表达及其序列分析

尹明华1,2,3,4(), 郭依瑶1, 李启诺1, 罗泽全1, 林文龙1, 熊姿雯1   

  1. 1 上饶师范学院生命科学学院，江西上饶 334001
    2 上饶农业技术创新研究院，江西上饶 334001
    3 上饶市药食同源植物资源保护与利用重点实验室，江西上饶 334001
    4 上饶市薯芋类作物种质保存与利用重点实验室，江西上饶 334001
  • 收稿日期:2023-09-12 修回日期:2024-01-15 出版日期:2024-07-25 发布日期:2024-07-11
  • 作者简介:

    尹明华，女，1973年出生，江西永新人，教授，硕士，从事植物生物技术研究工作。通信地址：334001 江西省上饶市信州区志敏大道401号 上饶师范学院生命科学学院，Tel：0793-8153721，E-mail：。

  • 基金资助:
    国家自然科学基金资助目“怀玉山三叶青“超低温疗法脱毒”促进离体根膨大的机理研究”(31960079); 江西省科技厅重点研发计划一般项目“三叶青种质资源离体保存及其核心种质筛选、脱毒和大棚基质栽培关键技术研究与示范”(20202BBG73010); 江西省教育厅科学技术研究项目“怀玉山三叶青烟草花叶病毒CP和MP基因以及烟草病毒增殖蛋白基因cDNA全长克隆、序列信息和定量表达分析”(GJJ201704); 2022年上饶市科技专项项目（饶科发[2023]5号社发类）“怀玉山三叶青超低温疗法脱毒苗种质资源离体保存及其高山大棚基质栽培关键技术研究”(2022A008); 江西省现代农业产业技术体系建设专项“牧草种质资源离体保存”(JXARS-13-赣东站); 上饶市科技局平台载体建设项目“上饶市药食同源植物资源保护与利用重点实验室”(2020J001); 上饶师范学院2023年大学生创新创业训练计划项目“信前胡烟草花叶病毒lncRNA测序鉴定、原核蛋白表达及其序列分析”(2023-PX-18)

lncRNA Sequencing Identification, Prokaryotic Protein Expression and Sequence Analysis of Peucedanum praeruptorum Dunn. cv. Xinzhou Tobacco Mosaic Virus

YIN Minghua1,2,3,4(), GUO Yiyao1, LI Qinuo1, LUO Zequan1, LIN Wenlong1, XIONG Ziwen1   

  1. 1 College of Life Sciences, Shangrao Normal University, Shangrao, Jiangxi 334001
    2 Shangrao Agricultural Technology Innovation Research Institute, Shangrao, Jiangxi 334001
    3 Key Laboratory of Protection and Utilization of Medicinal and Edible Plant Resources in Shangrao City, Shangrao, Jiangxi 334001
    4 Key Laboratory of Germplasm Conservation and Utilization of Potato and Taro Crops in Shangrao City, Shangrao, Jiangxi 334001
  • Received:2023-09-12 Revised:2024-01-15 Published:2024-07-25 Online:2024-07-11

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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。

关键词: 信前胡, 烟草花叶病毒, lncRNA测序鉴定, RT-PCR, 原核蛋白表达, 序列分析

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.

Key words: P. praeruptorum Dunn. cv. Xinzhou, tobacco mosaic virus, lncRNA sequencing identification, RT-PCR, prokaryotic protein expression, sequence analysis