两种甜瓜病毒寿光分离物的分子检测与鉴定

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 vi-rus ,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-PCR 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%。%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 clo-ning vector,and the expected DNA fragments with the size of 673 bp (MNSV)and 877 bp (CCYV)were se-quenced and analyzed by BLAST search in GenBank,respectively.Homology analysis showed that nucleotide se-quence of MNSV and CCYV isolates from Shouguang had 99% - 100% identity with those isolates from other parts of China or some countries previously reported.