【摘要】 滇重楼(Paris polyphylla var. yunnanensis)是多年生草本植物,其药用部位为地下根茎,是一种重要的中药材。为明确侵染滇重楼的病毒病原,对采自云南省曲靖市的滇重楼进行透射电镜观察、DASELISA和RT-PCR技术检测。在透射电镜下观察到杆状病毒粒子,大小为(200～300)nm×(20～36)nm。对采集到的7份病样进行DAS-ELISA检测,结果表明,其中5份病样呈阳性。通过RT-PCR技术扩增、克隆获得1个全长为6 357 bp的曲靖分离物PMMoV-QJ的全基因组序列。将PMMoV-QJ的全基因组序列与国内外已经报道的10个PMMoV分离物进行同源性分析比对,其同源性为99.42%～99.81%。基于全基因序列的系统发育分析表明,PMMoV-QJ与韩国分离物亲缘关系密切。本研究明确了该分离物的亲缘关系,从分子水平鉴定该分离物,为后续滇重楼病毒病的预防提供理论依据。更多还原

【Abstract】 Paris polyphyllavar. yunnanensis is a perennial herb,and its medicinal part is underground rhizomes. It is an important chinese medicinal material. In order to clarify the viral pathogens infecting the Paris polyphylla,the transmission electron microscopy,DAS-ELISA and RT-PCR techniques were performed on the Paris polyphylla collected from Qujing,Yunnan province. Baculovirus particles were observed under transmission electron microscopy and were(200～300)nm ×(20～36)nm in size. The results of DAS-ELISA on 7 samples collected showed that 5 of them were positive. A whole genome sequence of a 6 357 bp Qujing isolate(PMMoV-QJ,accession number MK784568)was cloned by RT-PCR amplification. The homologous sequence of PMMoV-QJ were analyzed by homology with 10 PMMoV isolates reported at home and abroad,and the homology was between 99.42% and 99.81%. Phylogenetic analysis based on the whole gene sequence showed that PMMoV-QJ was closely related to Korean isolates. This study clarifies the genetic relationship of the isolate,and identifies the isolate from the molecular level,which provides a theoretical basis for the prevention of the subsequent viral disease.更多还原