新疆哈密瓜病毒的DAS-ELISA检测和分子鉴定

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

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(DAS–ELISA)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 virus,CGMMV)进行检测和分子鉴定。结果表明:在所检测的121个样品中,黄瓜花叶病毒(Cucumber mosaic virus,CMV)的检出率最高,为70.2%;西瓜花叶病毒(Watermelon mosaic virus,WMV)和小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)次之,分别为62.0%和37.2%,甜瓜坏死斑点病毒(Melon necrotic spot virus,MNSV)的检出率仅为2.5%,CGMMV、南瓜花叶病毒(Squash mosaic virus,Sq MV)、番木瓜环斑病毒(Papaya ring spot virus W,PRSV-W)和烟草坏死病毒(Tobacco necrosis virus,TNV)未检测到。CMV、WMV、ZYMV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ⅠB。ZYMV、WMV和MNSV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,ZYMV和WMV具有明显的地域分化现象。

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 Turpan,Shanshan 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 virus（CGMMV）the important quarantine virus infecting Hami melon. The results showed that the detection rates of Cucumber mosaic virus（CMV）,Watermelon mosaic virus（WMV）,Zucchini yellow mosaic virus（ZYMV）,and Melon necrotic spot virus（MNSV）amongst the 121 detected samples were 70.2%,62.0%,37.2%,and 2.5% respectively,whereas Squash mosaic virus（Sq MV）,Papaya ring spot virus（PRSV）,CGMMV and Tobacco necrosis virus（TNV）had not detected as positive. CMV,WMV 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 amplification,sequencing and phylogenetic analysis,CMV isolates from Hami melon in Xinjiang were categorized into subgroupⅠB. Although WMV,ZYMV and MNSV had high homology with other reported strains in CP gene nucleotide sequence,there were still some variations. ZYMV and WMV had a significant geographical differentiation.