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当前位置: 首页 > 文章 > 冬种马铃薯卷叶病田间调查及RT-PCR检测技术研究 湖北农业科学 2010,49 (9) 96-99
Position: Home > Articles > Inspection and Studies on RT-PCR Detection Technology of Potato Leafroll Virus in Winter Potato Hubei Agricultural Sciences 2010,49 (9) 96-99

冬种马铃薯卷叶病田间调查及RT-PCR检测技术研究

作  者：
陈兆贵;林于绵;黄成
单  位：
惠州学院生命科学系
关键词：
马铃薯;卷叶病毒病;田间调查;RT-PCR检测
摘  要：
对惠州市冬种马铃薯卷叶病发病情况进行调查研究,建立了马铃薯卷叶病毒(PLRV)RT-PCR检测技术。结果表明,马铃薯卷叶病发病率为3.0%～29.7%。比较LiCl、异硫氰酸胍和试剂盒3种方法提取马铃薯卷叶病毒总RNA,以异硫氰酸胍和试剂盒提取效果较好,LiCl次之;根据卷叶病毒外壳蛋白基因序列设计引物,从感染马铃薯卷叶病毒的马铃薯叶片中提取出病毒RNA,合成cDNA并运用RT-PCR技术进行体外扩增,得到一条长度约336bp的特异PCR扩增产物,与理论设计的外壳蛋白基因大小一致,而健康叶片RNA扩增未得到任何产物。应用RT-PCR技术对马铃薯试管苗和种薯进行了卷叶病毒检测。
译  名：
Inspection and Studies on RT-PCR Detection Technology of Potato Leafroll Virus in Winter Potato
作  者：
CHEN Zhao-gui,LIN Yu-mian,HUANG Cheng(Department of Life Science,Huizhou University,Huizhou 516007,Guangdong,China)
关键词：
potato;leafroll virus (PLRV);investigation in field;RT-PCR detection
摘  要：
The occurrence of potato leafroll virus(PLRV) of winter potato in Huizhou city was investigated,and the RT-PCR detection technology of PLRV was set up.The results showed that the PLRV disease rate was 3.0%-29.7%.Three extracting methods of total RNA:LiCl,Isothiocyanate-phenol-chloroform and RNAiso Plus extracting methods were compared in this report.All in all,the experiment results showed that the method of RNAiso Plus and Isothiocyanate-phenol-chloroform were better than LiCl extracting method.A pair of DNA primers(P1 and P2) were designed and synthesized based on the nucleotide sequence of the coat protein(CP) gene of PLRV.The PLRV RNA which was used for cDNA synthesis was directly extracted from virus-infected potato leaves.A specific PCR fragment about 336bp was obtained by reverse transcription and polymerase chain reaction(RT-PCR) amplification,which had the expected length of PLRV CP gene,at the same time,no fragment was obtained from the control.PLRV was detected in tube seedling and seed by RT-PCR technology.

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