Mehta P, Wyman J A, Nakhla M K, Maxwell D P
Department of Entomology, University of Wisconsin, Madison, WI 53706.
J Econ Entomol. 1994 Oct;87(5):1285-90. doi: 10.1093/jee/87.5.1285.
The sweetpotato whitefly, Bemisia tabaci (Gennadius), is an important pest worldwide. A new biotype of sweetpotato whitefly, biotype B, causes damage by direct feeding and by the transmission of plant viruses, such as geminiviruses. In the Mediterranean area, tomato yellow leaf curl geminivirus (TYLCV) is the most serious disease of tomatoes. Another whitefly-transmitted geminivirus, tomato mottle geminivirus (ToMoV), is presently a serious problem in tomato production in west-central and southwestern Florida. Because of the increasing incidence of whitefly-transmitted geminiviruses, it is necessary to develop rapid and simple diagnostic methods for the detection of viruliferous whiteflies. The polymerase chain reaction is a sensitive and specific technique for the detection and identification of plant pathogens. Polymerase chain reaction methods were used successfully to amplify 1.1-kb DNA fragments from individual viruliferous B. tabaci carrying either TYLCV or ToMoV, and no amplified DNA fragments were obtained when nonviruliferous B. tabaci adults were processed similarly. Southern hybridization analysis proved that fragments amplified from viruliferous B. tabaci adults were viral DNA. This polymerase chain reaction-based detection method is sensitive enough to detect TYLCV and ToMoV in individual viruliferous B. tabaci in mixed samples of up to 25 (1 viruliferous: 24 nonviruliferous) and 10 (1 viruliferous: 9 nonviruliferous) individuals, respectively. The potential uses of this polymerase chain reaction-based detection method in epidermiological studies of geminiviruses are discussed.
烟粉虱,Bemisia tabaci(根纳季),是一种世界性的重要害虫。烟粉虱的一个新生物型,即B型烟粉虱,通过直接取食和传播植物病毒(如双生病毒)造成危害。在地中海地区,番茄黄化曲叶双生病毒(TYLCV)是番茄最严重的病害。另一种由粉虱传播的双生病毒,番茄斑驳双生病毒(ToMoV),目前是佛罗里达州中西部和西南部番茄生产中的一个严重问题。由于粉虱传播的双生病毒发病率不断上升,有必要开发快速简便的诊断方法来检测带毒粉虱。聚合酶链反应是一种用于检测和鉴定植物病原体的灵敏且特异的技术。聚合酶链反应方法成功地从携带TYLCV或ToMoV的单个带毒烟粉虱中扩增出1.1-kb的DNA片段,而对无毒烟粉虱成虫进行类似处理时未获得扩增的DNA片段。Southern杂交分析证明,从带毒烟粉虱成虫中扩增出的片段是病毒DNA。这种基于聚合酶链反应的检测方法灵敏度足以分别在多达25只(1只带毒:24只无毒)和10只(1只带毒:9只无毒)个体的混合样本中的单个带毒烟粉虱中检测到TYLCV和ToMoV。讨论了这种基于聚合酶链反应的检测方法在双生病毒流行病学研究中的潜在用途。