侵染我国芋的杆状DNA病毒分子鉴定及特异性检测  

Molecular identification and specific detection of Badnavirus from taro grown in China

作　　者：施世明[1] 王彦芬[1] 王国平[1,2] 王利平[1] 徐文兴[1] 洪霓[1,2] 

机构地区：[1]华中农业大学植物科技学院,湖北省作物病害监测与安全控制重点实验室,武汉430070 [2]华中农业大学,农业微生物学国家重点实验室,武汉430070

出　　处：《植物病理学报》2013年第6期590-595,共6页Acta Phytopathologica Sinica

基　　金：国家农业行业专项计划(nyhyzx200903017-08)

摘　　要：采用已报道杆状DNA病毒属（Badnavirus）的通用检测引物BadnaFP／BadnaRP，从7份芋样品中扩增到Badnavirus病毒的RT／RNaseH基因保守区域，获得12个克隆序列，长度为576bp。序列分析结果显示，来自不同样品的克隆间核苷酸和氨基酸序列相似性分别为78．8％～99．5％和81．3％～99．5％；来自同一样品扩增产物的克隆间在序列上也存在较大差异，核苷酸和氨基酸序列相似性分别为89．6％～100％和92．7％～100％。在系统进化树中，本研究所获序列与Badnavirus病毒的亲缘关系相对较近，聚在同一簇，表明此病毒为Badnavirus属病毒，但与芋杆状病毒（Tarobacillifo硎virus，TaBV）序列相似性较低，核苷酸和氨基酸相似性分别为58．0％～62．2％和58．5％～64．1％，推测为杆状DNA病毒属的一个新种。根据所测定序列设计了用于该病毒检测的引物P197／P433，对51份芋样品检测结果表明，该引物可有效检测来源于我国芋的Badna—virus病毒。The universal degenerate primers BadnaFP/BadnaRP were used for the detection of badnaviruses in taro grown in China. The target fragment of 576 bp, covering the conserved region of genes encoding reverse transcriptase （RT） and ribonuclease H （RNase H）, was amplified from seven samples. Sequence analysis of 12 clones of PCR products from those samples showed that the clones from different samples had similarities of 78. 8%-99.5% and 81.3%-99.5% at nucleotide （nt） and amino acid （aa） levels, and the clones from the same sample had similarities of 89.6%-100% and 92.7%-100% at nt and aa levels, respectively. Phylogenetic analysis revealed that all the isolates belonged to a member of Badnavirus. However, the obtained sequences shared only 58.0%-62.2% nt and 58.5%-64.1% aa similarities with sequences of Taro bacilliform virus, suggesting that it might be a new tentative species in the genus Badnavirus. Primers P197/P433 based on the obtained sequences were used for detecting this virus from 51 taro samples. The results showed that the designed primers could be used for effective and specific detection of the badnaviruses in taro grown in China.

关 键 词：芋 杆状DNA病毒 PCR 序列分析 

分 类 号：S432.41[农业科学—植物病理学]