Ji Y H, Cai Z D, Zhou X W, Liu Y M, Xiong R Y, Zhao T M, Yu W G, Tao X R, Zhou Y J
Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P. R. China.
Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P. R. China.
Plant Dis. 2012 Aug;96(8):1229. doi: 10.1094/PDIS-03-12-0258-PDN.
Common bean (Phaseolus vulgaris) is one of the most economically important vegetable crops in China. In November 2011, symptoms with thickening and crumpling of leaves and stunting were observed on common bean with incidence rate of 50 to 70% in the fields of Huaibei, northern Anhui Province, China. Diseased common bean plants were found to be infested with large population of whiteflies (Bemisia tabaci), which induced leaf crumple symptoms in healthy common beans, suggesting begomovirus etiology. To identify possible begomoviruses, 43 symptomatic leaf samples from nine fields were collected and total DNA of each sample was extracted. PCR was performed using degenerate primers PA and PB to amplify a specific region covering AV2 gene of DNA-A and part of the adjacent intergenic region (2). DNA fragments were successfully amplified from 37 out of 43 samples and PCR amplicons of 31 samples were used for sequencing. Sequence alignments among them showed that the nucleotide sequence identity ranged from 99 to 100%, which implied that only one type of begomovirus might be present. Based on the consensus sequences, a primer pair MB1AbF (ATGTGGGATCCACTTCTAAATGAATTTCC) and MB1AsR (GCGTCGACAGTGCAAGACAAACTACTTGGGGACC) was designed and used to amplify the circular viral DNA genome. The complete genome (Accession No. JQ326957) was 2,781 nucleotides long and had the highest sequence identity (over 99%) with Tomato yellow leaf curl virus (TYLCV; Accession Nos. GQ352537 and GU199587). These samples were also examined by dot immunobinding assay using monoclonal antibody against TYLCV and results confirmed that TYLCV was present in the samples. These results demonstrated that the virus from common bean is an isolate of TYLCV, a different virus from Tomato yellow leaf curl China virus (TYLCCNV). TYLCV is a devastating pathogen causing significant yield losses on tomato in China since 2006 (4). The virus has also been reported from cowpea in China (1) and in common bean in Spain (3). To our knowledge, this is the first report of TYLCV infecting common bean in China. References: (1) F. M. Dai et al. Plant Dis. 95:362, 2011. (2) D. Deng et al. Ann. Appl. Biol. 125:327, 1994. (3) J. Navas-Castillo et al. Plant Dis. 83:29, 1999. (4) J. B. Wu et al. Plant Dis. 90:1359, 2006.
普通菜豆(Phaseolus vulgaris)是中国经济价值最重要的蔬菜作物之一。2011年11月,在中国安徽省北部淮北地区的菜豆田中,观察到叶片增厚、皱缩以及植株矮化的症状,发病率为50%至70%。发现患病的普通菜豆植株上有大量烟粉虱(Bemisia tabaci),这些烟粉虱能使健康的普通菜豆产生叶片皱缩症状,表明病因是双生病毒。为鉴定可能的双生病毒,从9个田块采集了43份有症状的叶片样本,并提取了每个样本的总DNA。使用简并引物PA和PB进行PCR,以扩增覆盖DNA-A的AV2基因和部分相邻基因间隔区的特定区域(2)。43个样本中有37个成功扩增出DNA片段,对其中31个PCR扩增产物进行了测序。它们之间的序列比对显示,核苷酸序列同一性在99%至100%之间,这意味着可能只存在一种双生病毒。基于一致序列,设计了引物对MB1AbF(ATGTGGGATCCACTTCTAAATGAATTTCC)和MB1AsR(GCGTCGACAGTGCAAGACAAACTACTTGGGGACC),用于扩增环状病毒DNA基因组。完整基因组(登录号JQ326957)长2781个核苷酸,与番茄黄化曲叶病毒(TYLCV;登录号GQ352537和GU199587)具有最高的序列同一性(超过99%)。还使用抗TYLCV的单克隆抗体通过斑点免疫结合试验对这些样本进行了检测,结果证实样本中存在TYLCV。这些结果表明,来自普通菜豆的病毒是TYLCV的一个分离株,与番茄黄化曲叶中国病毒(TYLCCNV)是不同的病毒。自2006年以来,TYLCV是一种毁灭性病原体,在中国导致番茄产量大幅损失(4)。在中国,该病毒也在豇豆中被报道过(1),在西班牙的普通菜豆中也有报道(3)。据我们所知,这是TYLCV侵染中国普通菜豆的首次报道。参考文献:(1)F. M. Dai等人,《植物病害》95:362,2011年。(2)D. Deng等人