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园艺学报 ›› 2016, Vol. 43 ›› Issue (6): 1107-1116.doi: 10.16420/j.issn.0513-353x.2016-0144

• 蔬菜 • 上一篇    下一篇

新疆哈密瓜病毒的DAS-ELISA检测和分子鉴定

潘亚南1,2,*,韩 剑1,2,*,吴海波3,吉艳玲4,王纯利5,刘 芳5,罗 明1,2,**,张祥林6   

  1. (1新疆农业大学农学院,乌鲁木齐 830052;2新疆自治区高校农林有害生物监测与安全防控重点实验室,乌鲁木齐 830052;3新疆农业科学院哈密瓜研究中心,乌鲁木齐 830091;4吐鲁番市农业技术推广中心,新疆吐鲁番838000;5鄯善县农业技术推广中心,新疆吐鲁番 838200;6新疆出入境检验检疫局,乌鲁木齐 830063)
  • 出版日期:2016-06-25 发布日期:2016-06-25

Detection of Viruses Infecting Hami Melon and Their Molecular Identification in Xinjiang

PAN Ya-nan1,2,*,HAN Jian1,2,*,WU Hai-bo3,JI Yan-ling4,WANG Chun-li5,LIU Fang5,LUO Ming1,2,**,and ZHANG Xiang-lin6   

  1. (1Agronomy College,Xinjiang Agricultural University,Urumqi 830052,China;2Key Laboratory of Harmful Biological Monitoring and Safety Prevention of Xinjiang Autonomous Region Forestry Universities,Urumqi 830052,China;3Hami Melon Research Center,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China;4Turpan Agricultural Technology Extension Center,Turpan,Xinjiang 838000,China;5Shanshan Agricultural Technology Extension Center,Turpan,Xinjiang 838200,China;6Xinjiang Entry-Exit Inspection and Quarantine Bureau,Urumqi 830063,China)
  • Online:2016-06-25 Published:2016-06-25

摘要:

为了明确当前新疆哈密瓜病毒病的主要毒原种类及其遗传分化,为哈密瓜病毒病害的防治提供科学依据,对吐鲁番、鄯善、昌吉等哈密瓜主产区的病毒病调查、采样,采用双抗体夹心酶联免疫吸附法(DASELISA)和RT-PCR技术对侵染哈密瓜的7种主要病毒及瓜类重要检疫性病毒——黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virusCGMMV)进行检测和分子鉴定。结果表明:在所检测的121样品中,黄瓜花叶病毒(Cucumber mosaic virusCMV)的检出率最高,为70.2%;西瓜花叶病毒(Watermelon mosaic virusWMV)和小西葫芦黄花叶病毒(Zucchini yellow mosaic virusZYMV)次之,分别为62.0%37.2%,甜瓜坏死斑点病毒(Melon necrotic spot virusMNSV)的检出率仅为2.5%CGMMV、南瓜花叶病毒(Squash mosaic virusSqMV)、番木瓜环斑病毒(Papaya ring spot virus WPRSV-W)和烟草坏死病毒(Tobacco necrosis virusTNV)未检测到。CMVWMVZYMV在田间分布广泛,2种及3种病毒的复合侵染发生普遍(60.19%)。对各病毒分离物进行CP基因扩增、序列测定和系统发育分析,确定了CMV新疆哈密瓜分离物归属于CMV亚组ⅠBZYMVWMVMNSV与已报道的病毒株系CP基因核苷酸序列具有较高的同源性,但存在一定的变异,ZYMVWMV具有明显的地域分化现象。

关键词: 哈密瓜, 病毒, 反转录PCR, 复合侵染, 遗传变异

Abstract:

This research aims at identifying the currently prevailing viruses infecting Hami melon in Xinjiang and their genetic variations for providing scientific basis to prevent these virus diseases. Field survey was conducted in TurpanShanshan and Changji and samples with viral symptoms were collected for the lab tests. Both DAS-ELISA and RT-PCR were applied for the detection and molecular identification of 7 common viruses as well as Cucumber green mottle mosaic virusCGMMVthe important quarantine virus infecting Hami melon. The results showed that the detection rates of Cucumber mosaic virusCMV),Watermelon mosaic virusWMV),Zucchini yellow mosaic virusZYMV),and Melon necrotic spot virusMNSVamongst the 121 detected samples were 70.2%62.0%37.2%and 2.5% respectivelywhereas Squash mosaic virus SqMV),Papaya ring spot virusPRSV),CGMMV and Tobacco necrosis virusTNVhad not detected as positive. CMVWMV and ZYMV are widely distributed in the field. Mixed infection with two or three different viruses on the same individual host was common with an occurrence of 60.19%. Based on the CP gene amplificationsequencing and phylogenetic analysisCMV isolates from Hami melon in Xinjiang were categorized into subgroupB. Although WMVZYMV and MNSV had high homology with other reported strains in CP gene nucleotide sequencethere were still some variations. ZYMV and WMV had a significant geographical differentiation.

Key words: Hami melon, virus, RT-PCR, mixed infection, genetic variation

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