利用RT-PCR技术检测吊瓜病毒种类的研究

Detection of Main Causal Viruses in Trichosanthes kirilowii by RT-PCR

小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)、西瓜花叶病毒(Watermelon mosaicvirus,WMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、南瓜花叶病毒(Squash mosaic virus,SqMV)和番木瓜环斑病毒(Papaya ringspot virus,PRSV)是危害葫芦科作物最严重、最广泛的5种病毒。根据已知的5种病毒序列设计特异性引物,对感染病毒的吊瓜总RNA进行RT-PCR扩增。结果表明长兴吊瓜的病毒种类主要为PRSV,感染率为100%,无复合病毒感染。吊瓜植株不同取样部位(茎尖、卷须、嫩叶、茎、花、子房、老叶等)的RT-PCR表明老叶是较好的病毒检测取样部位,而嫩叶是吊瓜脱毒最佳的外植体取样部位。对茎尖的PRSV表达量高过幼叶的原因也进行了分析,认为可能是病毒的表达量与植株的代谢强度有关。

Zucchini yellow mosaic virus（ZYMV）,Watermelon mosaic virus（WMV）,Cucumber mosaic virus（CMV）,Squash mosaic virus（SqMV） and Papaya ringspot virus（PRSV） are the most serious pathogens infecting cucurbitaceous crops.Five pairs of specific primers were designed based on the conserved sequences of the 5 viruses,and RT-PCR was applied to detect total RNA which was extracted from Trichosanthes kirilowii Maxim.leaves infected virus.The results showed that the causal virus of T.kirilowii in Changxing was PRSV with 100% incidence rate as no multiplex virus infection was detected.The RT-PCR results of different sampling parts of T.kirilowii suggested that the most suitable part for virus detection was old leave and the most virus-free part was young leave.The cause of more PRSV expressed in shoot tip than in young leaves was discussed,and it was possibly that the expression of virus was related to plant＇s metabolism intensity.