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番茄斑萎病毒(Tomato spotted wilt virus,TSWV)寄主广泛,危害多种作物,近年发现侵染烟草引起严重经济损失。研究该病毒的分子检测方法,对健康烟草种苗培育和指导病害防控具有重要意义。本文根据TSWV核壳体蛋白基因(N基因)序列设计引物,用RT-PCR方法对采于云南疑似番茄斑萎病毒症状的5个烟草病害标样(102,514,自10,自16,自58)进行检测,PCR产物连接pMD载体并测序。测序结果表明:这5种烟草病害分离物与已报道TSWV N基因核酸序列相似性大于97.04%,所编码氨基酸序列的相似性达97.67%以上。这说明获得的5种病毒分离物均属于TSWV。本研究建立了侵染云南烟草的番茄斑萎病毒(TSWV)的RT-PCR检测方法。
Tomato spotted wilt virus (TSWV) hosts a wide range of crops and is harmful to many crops. In recent years, it has been found that the infection of tobacco causes serious economic losses. Studying the molecular detection method of this virus is of great significance for the cultivation of tobacco seedlings and the guidance of disease prevention and control. Based on the sequence of TSWV nucleocapsid protein gene (N gene), five tobacco disease standard samples (102,514, from 10, from 16, 58) collected from the suspected tomato spotted wilt virus in Yunnan were designed by RT-PCR. The PCR product was ligated into the pMD vector and sequenced. Sequencing results showed that the similarity of the five tobacco isolates to the reported TSWV N gene was greater than 97.04% and the similarity of the encoded amino acid sequences was over 97.67%. This shows that the obtained five virus isolates belong to TSWV. In this study, a RT-PCR method for the detection of tomato spotted wilt virus (TSWV) in Yunnan tobacco was established.