转AcAMP-sn基因抗全蚀病小麦新种质的创制与鉴定

抗菌肽是一类具有广谱抗菌性的小分子多肽,在植物防卫反应中起着重要作用。本研究人工合成了洋葱抗菌肽基因AcAMP-sn,利用基因重组技术构建了该基因的单子叶植物表达载体pAHC25::AcAMP-sn,使AcAMP-sn基因的表达受玉米Ubiqutin启动子控制。采用基因枪介导法,将AcAMP-sn基因导入小麦品种扬麦18,利用PCR、半定量RT-PCR及荧光实时定量RT-PCR检测、分析转基因小麦T0~T4代目的基因及其表达量,并对转基因植株进行全蚀病的抗性鉴定。PCR检测结果表明,导入的AcAMP-sn基因能够在转基因小麦中遗传。与受体扬麦18相比,在5个转基因小麦T4代株系中,AcAMP-sn基因的表达量及全蚀病抗性均显著提高,且全蚀病菌的相对含量明显下降,说明AcAMP-sn基因的过表达可以增强转基因小麦对全蚀病的抗性。

Development and Characterization of AcAMP-sn Transgenic Wheat with Enhanced Resistance to Wheat Take-all

Antimicrobial peptides, small molecular weight proteins with broad spectrum antimicrobial activity, play important roles in plant defense responses. The open-reading-frame sequence of AcAMP-sn was synthesized and used to construct the gene transformation vector pAHC25::AcAMP-sn, in which AcAMP-sn gene was driven by maize ubiquitin promoter and should be highly expressed in monocot plants. The vector pAHC25::AcAMP-sn DNA was introduced into Yangmai 18 via particle bombardment. AcAMP-sn transgenic wheat plants were subjected to PCR, semi-RT-PCR, Q-RT-PCR analyses, and disease response assessments. PCR analyses revealed that theintroducedgene AcAMP-sn could be stably inherited in five transgenic wheat lines from T₀ to T₄ generations. Semi-RT-PCR and Q-RT-PCR analyses showed that the AcAMP-sn gene was highly expressed in transgenic wheat lines compared with untransformed Yangmai 18. Based on disease response assessments for T₄ generations, the significantly enhanced-resistance to take-all accompanied with decreased fungal abundance, in the five independent AcAMP-sn transgenic lines. Over-expression of AcAMP-sn gene in transgenic wheat plants confers increased resistance to wheat take-all.