CAJ | 학술논문

采用基因枪法将小麦反义Mlo基因导入扬麦158和济麦20的幼胚愈伤组织中,在含除草剂的分化培养基上经两轮筛选,获得抗性再生植株.PCR检测、PCR-Southern杂交、基因组DNA斑点杂交和除草剂BASTA抗性分析结果证实已获得转基因扬麦158和济麦20阳性植株,荧光定量表达分析亦证明Mlo基因发生沉默.对T0和T1代转基因植株的白粉病抗性鉴定表明,有6个转基因株系高抗白粉病.对T1代转基因小麦接种白粉菌后孢子发育的显微观察结果显示, Mlo反义基因的导入明显加快了乳突的形成和维持时间,有效抑制了吸器的发育,因而使转Mlo反义基因材料表现抗病性.
채용기인창법장소맥반의Mlo기인도입양맥158화제맥20적유배유상조직중,재함제초제적분화배양기상경량륜사선,획득항성재생식주.PCR검측、PCR-Southern잡교、기인조DNA반점잡교화제초제BASTA항성분석결과증실이획득전기인양맥158화제맥20양성식주,형광정량표체분석역증명Mlo기인발생침묵.대T0화T1대전기인식주적백분병항성감정표명,유6개전기인주계고항백분병.대T1대전기인소맥접충백분균후포자발육적현미관찰결과현시, Mlo반의기인적도입명현가쾌료유돌적형성화유지시간,유효억제료흡기적발육,인이사전Mlo반의기인재료표현항병성.
The antisense wheat Mlo gene, Ta-Mlo, was transformed into wheat (Tritivum aestivum L.) varieties Yangmai 158 and Jimai 20 via biolistic transformation using immature embryo calli as explants. After two rounds of bialaphos selection and regen-eration, herbicide-resistant plants were obtained, which were subsequently confirmed by PCR, PCR-Southern hybridization, ge-nomic dot hybridization, and BASTA resistance analysis. The results showed that the Ta-Mlo antisense transgenic Yangmai 158 and Jimai 20 plants were obtained. The real time fluorescence quantitative PCR analysis proved that the transcript of Ta-Mlo was knocked down in these transgenic plants. The disease resistance test showed that the six transgenic lines appeared highly resis-tance to powdery mildew pathogen Blumeria graminis f. sp. tritici (Bgt). The transgenic lines showed distinct acceleration of the production and stabilization of papillae, and effective suppression to further development of haustoria of Bgt. Therefore, the transgenic lines showed high resistance to Bgt.