| 乔宁1，魏家鹏1，李美芹*1，刘永光1，姜会霞1，竺晓平2，田素波3.两种甜瓜病毒寿光分离物的分子检测与鉴定[J].植物保护,2015,41(5):130-133. |
| 两种甜瓜病毒寿光分离物的分子检测与鉴定 |
| Molecular detection and identification of two melon viruses from Shouguang |
| 投稿时间：2014-08-16 修订日期：2014-10-26 |
| DOI： |
| 中文关键词: 甜瓜坏死斑点病毒 瓜类褪绿黄化病毒 RT-PCR扩增 序列分析 |
| 英文关键词:MNSV CCYV RT-PCR sequence analysis |
| 基金项目:公益性行业(农业)科研专项(201303028); 山东省自然科学基金(ZR2012CM032); 山东省科技发展计划(2010GNC10915); 潍坊市科技发展计划(201301159，2013YD177); 山东省农业良种工程现代种业企业科技培育计划 |
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| 中文摘要: |
| 甜瓜坏死斑点病毒(Melon necrotic spot virus, MNSV)及瓜类褪绿黄化病毒(Cucurbit chlorotic yellows virus, CCYV)近年来在瓜类种植区均大面积发生, 危害较为严重, 已成为制约甜瓜生产的重要因素。本研究广泛收集疑似感染MNSV及 CCYV的甜瓜病叶, 从中提取植物总RNA进行RT-CR扩增, 将产物分别连接到pEASY T1 Simple克隆载体上, 对含有目的片段的重组子进行测序及比对分析。结果显示得到了与预期结果一致的DNA序列, 其扩增产物大小分别为673 bp (MNSV)和877 bp (CCYV)。同源性分析结果表明，MNSV和CCYV寿光分离物的核苷酸序列与中国其他地区或一些国家已报道的分离物同源性达99%～100%。 |
| 英文摘要: |
| Melon necrotic spot virus (MNSV) and Cucurbit chlorotic yellows virus (CCYV) have seriously affected the yield and quality of melons in large melon growing areas in recent years. In this study, diseased leaves infected by MNSV and CCYV were widely collected from Shouguang, Shandong Province. Total RNA were extracted from these leaf tissues and RT-PCR was performed. The PCR products were inserted into pEASY T1 Simple cloning vector, and the expected DNA fragments with the size of 673 bp (MNSV) and 877 bp (CCYV) were sequenced and analyzed by BLAST search in GenBank, respectively. Homology analysis showed that nucleotide sequence of MNSV and CCYV isolates from Shouguang had 99%-100% identity with those isolates from other parts of China or some countries previously reported. |
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