马铃薯病毒诱导应答基因抑制消减杂交文库构建及分析

发布时间：2018-07-08 07:56

本文选题：马铃薯 + 病毒　；参考：《草业学报》2017年12期

【摘要】：马铃薯病毒积累引起的种薯退化是马铃薯生产中造成产量和品质下降的重要原因之一。本研究以马铃薯病毒病携带植株叶片c DNA为试验组(Tester)、脱毒种苗叶片c DNA为驱动组(Driver),采用抑制消减杂交(suppression subtractive hybridization,SSH)技术构建了马铃薯病毒诱导应答基因的c DNA文库;为验证文库构建效果,从文库中随机挑取了98个阳性克隆经PCR验证后测序,获得了45条高质量的有效非重复序列;经与Gen Bank进行同源比对后发现,其中14条非重复序列属于马铃薯病毒基因序列,22条与已知基因序列同源性较高,9条无同源参考基因;选取文库中出现频率较高的2个ESTs(expressed sequence tag,表达序列标签)用qRT-PCR技术分析发现,其表达量受马铃薯病毒侵染的诱导。结果表明,该SSH文库构建较为成功,为进一步筛选与马铃薯病毒致病、防御相关的应答基因,解析马铃薯与病毒互作的分子机理,利用生物技术手段培育抗病毒马铃薯奠定了基础。
[Abstract]:Seed potato degradation caused by the accumulation of potato virus is one of the important reasons for the decline of yield and quality in potato production. In this study, the cDNA library of potato virus-induced response gene was constructed by suppression subtractive hybridization (suppression subtractive) hybridization technique, using the leaves of potato virus carrying plants as test group (Tester) and virus-free seedling leaves as driving group (driver). In order to verify the effect of library construction, 98 positive clones were randomly selected from the library and sequenced by PCR. Among them, 14 non-repeat sequences belong to 22 sequences of potato virus genes with high homology with known genes and 9 non-homologous reference genes. Two ests (expressed sequence tag, expressed sequence tags with high frequency in the library were selected. The results were analyzed by qRT-PCR. Its expression was induced by potato virus infection. The results showed that the SSH library was successfully constructed. In order to further screen the responsive genes related to the pathogenesis and defense of potato virus, the molecular mechanism of the interaction between potato and virus was analyzed. Using biotechnology to cultivate anti-virus potato laid the foundation.
【作者单位】：甘肃省农业科学院生物技术研究所;
【基金】：甘肃省农科院青年创新基金(2013GAAS29) 甘肃省农业科学院创新团队(2015GAAS02);甘肃省农业科学院农业科技创新重大项目(2016GAAS59-02) 国家自然科学基金项目(31460388)资助
【分类号】：S435.32

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