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作物杂志,2024, 第1期: 233–238 doi: 10.16035/j.issn.1001-7283.2024.01.031

所属专题： 玉米专题； 杂粮作物

• 植物保护 • 上一篇    

江苏淮安市高粱上玉米黄花叶病毒的分子检测与鉴定

王海涛(), 任春梅, 董岩, 李硕, 程兆榜, 季英华()   

  1. 江苏省农业科学院植物保护研究所/江苏省食品质量安全重点实验室/国家重点实验室培育基地，210014，江苏南京
  • 收稿日期:2022-12-16 修回日期:2023-09-22 出版日期:2024-02-15 发布日期:2024-02-20
  • 通讯作者: 季英华，主要从事植物分子病毒学研究，E-mail：jiyinghua@jaas.ac.cn
  • 作者简介:王海涛，主要从事植物分子病毒学研究，E-mail：wanghaitao9105@163.com
  • 基金资助:
    国家自然科学基金(32001871);国家自然科学基金(32072506);江苏省自然科学基金(BK20200283)

Molecular Detection and Identification of Maize Yellow Mosaic Virus on Sorghum in Huai’an, Jiangsu

Wang Haitao(), Ren Chunmei, Dong Yan, Li Shuo, Cheng Zhaobang, Ji Yinghua()   

  1. Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences / Key Laboratory of Food Quality and Safety of Jiangsu Province/State Key Laboratory Breeding Base, Nanjing 210014, Jiangsu, China
  • Received:2022-12-16 Revised:2023-09-22 Online:2024-02-15 Published:2024-02-20
  • Contact: Ji Yinghua

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摘要：

为掌握江苏省高粱病毒病的发生情况，从江苏淮安采集高粱疑似病毒病样品20份，进行了分子检测与鉴定。结果发现，使用玉米黄花叶病毒（maize yellow mosaic virus，MaYMV）引物可扩增出与目的片段一致的条带，检出率为80%。利用MaYMV外壳蛋白（CP）基因引物，以阳性样本RNA为模板进行扩增、克隆、酶切验证和测序分析，并构建进化树发现，淮安分离物CP基因与江苏分离物CP基因的进化关系较近。这些结果表明江苏淮安高粱受到了MaYMV侵染，这是该病毒在我国高粱上的首次报道。

关键词: 玉米黄花叶病毒, 高粱病毒病, 分子检测, 进化树分析

Abstract:

In order to understand the incidence of sorghum virus disease in Jiangsu province, the 20 suspected sorghum virus disease samples were collected from Huai’an city, and molecular detection and identification were carried out. The results showed that the bands consistent with the target fragment could be amplified by using the maize yellow mosaic virus (MaYMV) primer, and the detection rate was 80%. We used MaYMV coat protein (CP) gene primers to perform amplification, cloning, enzyme digestion and sequencing analysis using a positive RNA as template. The phylogenetic tree analysis showed that CP genes of Huai’an isolate and Jiangsu isolate had a close evolutionary relationship. These results indicated that this is the first report of MaYMV infection in China sorghum.

Key words: Maize yellow mosaic virus, Sorghum virus disease, Molecular detection, Phylogenetic tree analysis

表1

检测中所使用的病毒引物

病毒名称Virus name 引物名称Primer name 引物序列Primer sequence (5’-3’)
玉米黄花叶病毒Maize yellow mosaic virus MaYMV（280bp F） CAAAGGACTCTCTCACGGGC
MaYMV（280bp R） CCTCCGATTTGATTGGACGC
玉米相关单份体病毒1 Maize-associated totivirus 1 MATV（567bp F） AGTGTTAAATATGAGTGGGC
MATV（567bp R） AGCCATCCGATGATGTATCTT
玉米条纹病毒Maize stripe tenuivirus MSpV（303bp F） CGAATGATGCGAGCAAGGC
MSpV（303bp R） CACTTGACAGCTCATCTGCG
玉米黄矮病毒Maize yellow dwarf virus RMV MYDV（RMVCP-F） ATGAATTTGGGAGGCCG
MYDV（RMVCP-R） CTATTTCGGATTTTGGGTATGAA
玉米矮花叶病毒Maize dwarf mosaic virus MDMV（454bp F） TATCGAACACCCGAGCAACC
MDMV（454bp R） CGTGTGTTTGAACCACGGAC
高粱花叶病毒Sorghum mosaic virus SrMV（430bp F） AAGCTCAACGAGACGCAGAA
SrMV（430bp R） ATGAGCCCACTTGCGACTAC
高粱绿斑病毒Sorghum chlorotic spot virus SrCMV（276bp F） GCAGTTCGCCATGCGTTTAT
SrCMV（276bp R） CGCTTTCAACCCTTCTCCCT
甘蔗条纹花叶病毒Sugarcane streak mosaic virus SCSMV（575bp F） CGGTCGATGGACAAGGAACA
SCSMV（575bp R） ACGAACCGAGAACTGAACCC
甘蔗花叶病毒Sugarcane mosaic virus SCMV（433bp F） CAGGGGGCCAAAGAGACAAA
SCMV（433bp R） TACGCTTCAGCTGCATCACT
约翰草花叶病毒Johnsongrass mosaic virus JGMV（703bp F） CACCCGCTGCAAATCAAACA
JGMV（703bp R） GGTGTGCGCGAATTGATCTC
狐尾草花叶病毒Foxtail mosaic virus FoMV（338bp F） GACGAGGAGTTGGTACGCAT
FoMV（338bp R） ACGTATGGGTCGAGAAGTGC
甘蔗黄叶病毒Sugarcane yellow leaf virus SCYLV（317bp F） ACGTAGAGCACGTGTTGGAG
SCYLV（317bp R） TCCTCTTGACGGGGAAGCTA
大麦黄矮病毒Barley yellow dwarf virus BYDV（426bp F） CAGGCGGGACTGAGGTATTC
BYDV（426bp R） TGGCCGTCATCAAACTGACA
水稻黑条矮缩病毒Rice black streaked dwarf virus RBSDV（733bp F） CATTGTCAAATCGCCCCACG
RBSDV（733bp R） TACCGCGCTCCAAGTTTGTT
南方水稻黑条矮缩病毒Southern rice black streaked dwarf virus SRBSDV（562bp F） CCCACATCGCGTCATCTCAA
SRBSDV（562bp R） TGGTCGTAACCGCCATAGTG

图1

田间采集的高粱病样症状

图2

采集的高粱疑似病毒样中MaYMV的RT-PCR检测 M：DNA分子量标准；(a) 不同病毒引物检测高粱样品，(b) 高粱疑似病样检测。

图3

淮安MaYMV CP的扩增、克隆和酶切验证 M：DNA marker；(a) 1表示MaYMV CP基因的扩增，2表示阴性；(b) CP基因克隆载体的双酶切验证。

图4

淮安MaYMV CP核苷酸序列进化树分析

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