抗全蚀病的转PvPGIP2-TaLTP5双价基因小麦的创制及鉴定

Development and Characterization of Both PvPGIP2 and TaLTP5 Transgenic Wheat Lines with Resistance to Take-all

【目的】全蚀病是小麦的毁灭性病害。创制转PvPGIP2和TaLTP5双价基因小麦,分析外源PvPGIP2和TaLTP5在转基因小麦中的遗传与表达情况,选育抗全蚀病的转基因小麦新种质。【方法】采用基因重组技术构建了PvPGIP2和TaLTP5双价表达转基因载体pA25-PvPGIP2-TaLTP5,利用基因枪介导法将该载体转入小麦品种扬麦18,采用PCR、RT-PCR与qRT-PCR方法分析转基因小麦T0—T4植株中目的基因及其表达量,并对T3和T4逐株进行全蚀病接种与抗性鉴定。【结果】创制并选育出稳定的抗全蚀病转PvPGIP2和TaLTP5小麦株系6个。PvPGIP2和TaLTP5能够在这6个转基因小麦株系中稳定遗传,并高水平表达。对转PvPGIP2和TaLTP5小麦的全蚀病抗性鉴定结果表明,与受体扬麦18相比,转PvPGIP2和TaLTP5小麦对全蚀病抗性明显提高。【结论】创制了转PvPGIP2和TaLTP5抗全蚀病的小麦新种质,其对全蚀病具有一定的抗性。

[ Objective ] Take-all is a destructive disease of wheat production worldwide. The aim of this study is to develop and select stable PvPGIP2 and TaLTP5 transgenic wheat lines with resistance to take-all pathogen Gaeumannomyces graminis. [Method] Gene recombination technology was used to prepare the transformation vector pA25-PvPGIP2-TaLTP5 expressing both PvPGIP2 and TaLTPS. Particle bombardment method was used to introduce both PvPGIP2 and TaLTP5 into wheat cultivar Yangmai 18. PCR, RT-PCR and qRT-PCR methods were used to detect the presence and transcript levels of PvPGIP2 and TaLTP5 in the transgenic wheat plants of T0-T4 generations. G graminis mycelia plug inoculation and take-all severity and the disease index were used to score the resistance degrees of these transgenic wheat lines and non-transgenic wheat Yangmai 18 at young seedling-stage. [Result] The results indicated that the introduced PvPGIP2 and TaLTP5 genes were stably inherited, and could be highly expressed in six transgenic wheat lines. The transgenic wheat plants expressing PvPGIP2 and TaLTP5 showed significantly enhanced resistance to G graminis compared with non-transgenic wheat Yangmai 18. [Conclusion] These results suggested that the introduced PvPGIP2 and TaLTP5 genes can be used for improving wheat resistance to take-all.