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以高抗黑星病的早酥梨为试材,利用从梨抗黑星病抑制消减文库中筛选出病原菌诱导特异表达基因片段,通过RACE技术克隆获得其全序列,命名为PbzsREMORIN(GenBank登录号为HQ901373)。该基因全长897bp,开放阅读框(ORF)597bp,编码198个氨基酸;生物信息学分析表明该基因具有remorin家族保守结构域;实时荧光定量RT-PCR结果表明,该基因受梨黑星病病原菌的诱导;用基因枪法将基因与GFP的融合蛋白转化到洋葱鳞茎表皮细胞中,瞬时表达显示remorin蛋白定位于细胞膜、细胞质和细胞核中;将目的基因转入烟草品种NC89,共获得27个转基因烟草株系;离体叶片接种烟草青枯病病原菌结果表明,过量表达该基因增强了NC89对烟草青枯病病原菌的抗性。该基因可能在植物与病原菌互作过程中起重要作用。
Highly resistant to aspergillus oryzae pear as the test material, the use of anti-Scirpus suppression subtracted from the library screening pathogenic bacteria-specific expression of gene fragments obtained by RACE cloned full-length sequence, named PbzsREMORIN (GenBank accession number For HQ901373). The gene was 897bp in length and 597bp in open reading frame (ORF), encoding 198 amino acids. The bioinformatics analysis showed that the gene has the remorin family conserved domain. Real-time fluorescent quantitative RT-PCR showed that the gene was affected by the pathogen Pear The fusion protein of GFP and GFP was transformed into onion bulb epidermal cells by gene gun method. The transient expression of remorin protein was located in the cell membrane, cytoplasm and nucleus. The target gene was transferred into tobacco variety NC89, and 27 transgenic tobacco The results of pathogen inoculation with tobacco leaf wilt showed that overexpression of this gene enhanced the resistance of NC89 to tobacco bacterial wilt pathogens. The gene may play an important role in plant-pathogen interactions.