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期 刊 ：植物保护学报 2006年

关键词：菜豆；种子；菜豆普通花叶病毒；分子检测；

Keywords:Common bean, seed, Bean common mosaic virus, molecular detection,

摘 要 ：为研究国家作物种质库中保存菜豆种质携带种传菜豆普通花叶病毒(BCMV)的状况,根据BCMV的cp基因序列设计一对特异性引物,从BCMV侵染的菜豆叶片上扩增出与理论大小一致的714bp的片段。在此基础上,应用改进CTAB法从菜豆干种子中提取总RNA并特异性扩增出相同大小的片段。对目的片段进行克隆、测序和序列同源性比较,结果表明,扩增片段的序列与其它BCMV的cp基因序列同源性达88%~98%,确证该方法可以从菜豆干种子中直接检测BCMV。利用建立的方法,分别从单粒菜豆种子样本和30粒菜豆种子样本中检测到BCMV,其检测灵敏度较ELISA方法更高。来自河北的紫芸豆单粒种子带毒率高达100%,而在不同地理来源的6份菜豆种质中,4份被检出携带BCMV,表明入库保存的菜豆种质和种子已普遍为BCMV所侵染。

Abstract:For understanding the status of common bean germplasm, conserved in the National Crop GeneBank (NCGB), infected by Bean common mosaic virus (BCMV), a pair of specific primers were designed based the sequence of cp gene of BCMV and were used to amplify a 714bp band in leaves infected by BCMV. Using this technique and improved CTAB procedure for extracting total RNA, a same band was amplified in seeds of common bean. The band was cloned and sequenced, and then the sequence was compared with other strains of BCMV. The result showed that the nucleotide identity was 88%-98% between sequence of cloned band and other BCMV strains, and also means that BCMV in dry seed of common bean was identified successfully using RT-PCR technique. By the procedure BCMV was detected in individual seed and 30 seeds samples. In bean variety, Ziyundou, propagated in Hebei Province all seeds were infected by BCMV and in six accessions from different provinces BCMV was detected in four samples. It was confirmed that the common bean germplasm and seeds conserved in NCGB were infected by BCMV generally.

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