辽西李属坏死环斑病毒检测及其多样性分析

采用RT-PCR对辽西地区采集的45份核果叶片样品进行了李属坏死环斑病毒(PNRSV)的鉴定,其中9份样品为阳性。以阳性样品总RNA为模板,克隆了PNRSV基因组RNA3近全长序列。序列长度在1 612~1 619 bp之间,一致性为93.8%~100%。以本研究获得的9个和Gen Bank登录的42个PNRSV近全长RNA3序列为基础,截取cp基因、mp基因和近全长RNA3序列分别构建系统发育树,三者结果一致:均可将51个PNRSV分离物分为4个组,即PV32、PV96、PE5和本文报道的一个新的PNRSV组,9个辽西分离物分别属于PV32组或PV96组。本研究明确了我国辽西地区PNRSV的遗传多样性,证明了PNRSV基因组RNA3在研究PNRSV遗传多样性中的适用性,同时发现了一个新的PNRSV组,对于PNRSV遗传多样性研究意义重大。

Detection and genetic diversity analysis of Prunus necrotic ringspot virus in West Liaoning

Forty-five leaf-samples of stone fruit crops collected in West Liaoning area were detected for Prunus necrotic ringspot virus (PNRSV) by RT-PCR, and nine out of the 45 were positive. Using total RNAs of the positive samples as templates, nearly full-length RNA genome segment 3 (RNA3) of PNRSV were cloned and sequenced. The size of the obtained RNA3 fragments ranged from 1 612 to 1 619 bp and shared 93.8%-100% sequence identity. Another 42 RNA3 sequences of PNRSV were retrieved from GenBank for phylogenetic analysis, and the sequences of cp gene, mp gene and RNA3 were selected, respectively, to construct phylogenic trees, and consistent results were obtained. Phylogenic analysis indicated that the 51 PNRSV isolates could be classified into four groups, including PV32, PV96, PE5 and a possible novel group, the nine isolates obtained in West Liaoning belong to group PV32 and PV96, respectively. Our study presented the genetic diversity of PNRSV in West Liaoning and the suitability of using RNA3 for PNRSV diversity analysis. Meanwhile, a novel PNRSV group was identified, which would be important for further understanding the genetic diversity of PNRSV.