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首页> 外文期刊>Plant Cell, Tissue and Organ Culture: An International Journal on in Vitro Culture of Higher Plants >Transgenic Brassica napus L. lines carrying a two gene construct demonstrate enhanced resistance against Plutella xylostella and Sclerotinia sclerotiorum
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Transgenic Brassica napus L. lines carrying a two gene construct demonstrate enhanced resistance against Plutella xylostella and Sclerotinia sclerotiorum

机译：携带两个基因构建体的转基因甘蓝型油菜品系显示出对小菜蛾和小核盘菌的增强抗性

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Plant diseases and insect pests are serious threat to the growth and yield of oilseed rape. In this study, a binary vector carrying sporamin and chitinase PjChi-1 genes in tandem was introduced into Brassica napus cv. ZS 758 via Agrobacterium tumefaciens for dual resistance against disease and insect attack. Thirty-two regenerated plantlets exhibiting hygromycin resistance were selected following Agrobacterium-mediated transformation of 600 leaf petiole explants. Of these, 27 transformants were confirmed to carry the two transgenes as detected by polymerase chain reaction (PCR) with 4.5% transformation efficiency. Eight plantlets were randomly selected for further confirmation by Southern and northern blot hybridization analyses. Four plants carried single copy of the transgenes, while the remaining four plants carried either two or three copies of the transgenes. Moreover, expression of the sporamin transgene was detected by northern blot hybridization in transgenic lines, but not in wild-type plants. These eight T-0 plants were grown in vitro, and inoculated with the Lepidoptera larvae of Plutella xylostella and with spores of the fungal pathogen of Sclerotinia sclerotiorum. Transgenic plants exhibited high levels of resistance to P. xylostella and S. sclerotiorum when compared to untransformed wild-type plants. Genetic analysis of T-1 progeny confirmed Mendelian segregation of the introduced genes. Therefore, these transgenic lines demonstrate a promising potential for variety development of oilseed rape lines with enhanced resistance against both P. xylostella and S. sclerotiorum.
机译：植物病害和虫害严重威胁油菜的生长和产量。在这项研究中，将携带sporamin和几丁质酶PjChi-1基因串联的二元载体引入了甘蓝型油菜。 ZS 758通过根癌农杆菌对疾病和昆虫的双重抵抗。在农杆菌介导的600个叶柄外植体转化之后，选择了32株表现出对潮霉素抗性的再生小植株。其中，通过聚合酶链反应（PCR）检测到27个转化体带有两个转基因，转化效率为4.5％。随机选择八株小植株，以通过Southern和Northern印迹杂交分析进一步确认。四株植物带有单拷贝的转基因，而其余四株植物带有两个或三个拷贝的转基因。此外，通过Northern印迹杂交在转基因品系中检测了孢菌素转基因的表达，但在野生型植物中未检测到。这八株T-0植物在体外生长，并接种小菜蛾的鳞翅目幼虫和菌核盘菌的真菌病原体的孢子。与未转化的野生型植物相比，转基因植物对小菜蛾和核盘菌有很高的抗性。 T-1后代的遗传分析证实了导入基因的孟德尔分离。因此，这些转基因品系显示出具有发展潜力的油菜油菜品系，其具有增强的抗小菜蛾和核盘菌的抗性。

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