侵染白术的蚕豆萎蔫病毒2号的分子检测与序列分析  

Molecular Identification and Sequence Analysis of Broad Bean Wilt Virus 2 Isolates from Atractylodes Macrocephala Koidz

作　　者：牛颜冰[1] 时晓丽[1] 张西梅[1] 赵慧琪 赵保佳 

机构地区：[1]山西农业大学生命科学学院,太谷030801

出　　处：《病毒学报》2015年第1期58-64,共7页Chinese Journal of Virology

基　　金：山西省自然科学基金(2012011032-5);山西农业大学"中青年学术带头人及学术骨干"专项资金

摘　　要：为鉴定引起山西绛县白术花叶、黄化等症状的病原，利用生物学接种、非序列依赖性PCR扩增（SIA）和RT-PCR的研究方法，对白术病样进行了研究，结果表明：接种感病白术病汁液的指示植物昆诺藜表现退绿枯斑，经单斑分离后转接健康白术植株呈现出与病样类似的症状，初步证明白术病害可能为病毒引起的；SIA检测表明感病白术为蚕豆萎蔫病毒2号（Broad bean wilt virus 2，BBWV2）侵染所致；为明确BBWV2白术分离物（BBWV2-Am）的分类地位，进一步克隆了BBWV2-Am RNA2外壳蛋白大亚基基因（LCP）和外壳蛋白小亚基基因（SCP）序列；序列同源性分析表明，LCP基因和SCP基因与已发表的BBWV2其它株系相应基因核苷酸序列的同源性分别为79.3%～87.2%和80.1%～89.2%，氨基酸序列同源性分别为91.2%～95.7%和89.4%～95.5%；系统进化树分析显示，BBWV2-Am与BBWV2地黄分离物（BBWV2-Rg）形成一个独立分支，二者亲缘关系最近。这是BBWV2侵染白术的首次报道。To identity the pathogen that causes the mosaic and yellowing symptoms on Atractylodes macrocephala Koidz in Jiangxian, Shanxi Province, biological inoculation, sequence-independent amplification （SIA）, RT-PCR and other identification methods were used. The results showed that the chlorotic and necrosis symptoms occurred in the indicator plant Chenopodium quinoa after it was infected with the pathogen, and the same symptoms appeared after the reinoculation of healthy Atractylodes macrocephala Koidz; this reflected that the disease was likely to be caused by a virus. The results of SIA and sequencing showed that Broad bean wilt virus 2 （BBWV2） was present in severely mosaic Atractylodes macrocephala Koidz leaves. To further characterize the BBWV2 isolate from Atractylodes macrocephala （BBWV2-Am）, the polyprotein partial gene encoded by BBWV2-Am RNA2 was cloned and sequenced. Sequence alignments showed that the nucleotide sequence identity of BBWV2-Am SCP and LCP genes ranged from 79.3% to 87.2% and from 80.1% to 89.2% compared to other BBWV2 strains, respectively; the deduced amino acid sequence similarities of the two gene products ranged from 91.2% to 95.7% and from 89.4% to 95.5%, respectively, compared to those of other BBWV2 strains. Phylogenetic comparisons showed that BBWV2-Am was most likely to be related to BBWV2-Rg, but formed an independent branch. This is the first report of BBWV2 in Atractylodes macrocephala Koidz.

关 键 词：白术 生物学接种 非序列依赖性PCR扩增 蚕豆萎蔫病毒2号 

分 类 号：S432.41[农业科学—植物病理学]