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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RTPCR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。
In order to identify the pathogen that caused the symptoms of Atractylodes macrocephala such as Atractylodes macrocephala in Jiangxian County, Shanxi Province, the methods of biological inoculation, non-sequence-dependent PCR amplification (SIA) and RTPCR were used to study the pathogens of Atractylodes macrocephala. The results showed that: The results indicated that the disease of Atractylodes macrocephala could be caused by virus, which indicated that the symptoms of Atractylodes macrocephala were reverse green spot, Atractylodes macrocephala was caused by the infection of Broad bean wilt virus 2 (BBWV2). In order to clarify the taxonomic status of BBWV2 Amyloid isolate (BBWV2-Am), BBWV2-Am RNA2 coat protein large subunit gene (LCP) and coat protein small subunit (SCP) sequences. Sequence homology analysis showed that the homologies of LCP gene and SCP gene with those of other published BBWV2 genes were 79.3% 87.2% and 80.1% -89.2%, respectively. The amino acid sequence homologies were 91.2% -95.7% and 89.4% -95.5%, respectively. Phylogenetic tree analysis showed that BBWV2-Am and BBWV2 rehmanniae BBWV2-Rg formed an independent Branch, the two kinship recently. This is the first report that BBWV2 infects Atractylodes.