农杆菌介导转化蝴蝶兰遗传体系的研究

为建立蝴蝶兰植株的高效遗传转化体系,以蝴蝶兰花梗腋芽诱导的类原球茎体为受体材料,利用农杆菌侵染并在侵染前后辅以超声波及负压处理的方法将gus基因导入到蝴蝶兰中,经过近90 d的筛选,获得了69个抗卡那霉素抗性PLB,经X-gluc组织化学染色法观察检测,表达率最高达39%,并获得了50株再生蝴蝶兰,其中17株PCR检测呈阳性。初步建立了蝴蝶兰遗传转化体系。另外,我们在农杆菌侵染前后辅以超声波及负压处理,可以提高转化效率。

Study on the Agrobacterium-mediated Transformation in Phalaenopsis

Using agrobacterium-mediaed transformation,gus gene was transferred into protocorm-like body(PLB) induced by the peduncles of Phalaenopsis.After 90 days of selective culture,69 Kanamycin-resistant PLBs were gained.The results of X-gluc histochemical assay showed that the gus expression efficiencies of resistant PLBs were up to 39 %.Of 50 regeneration plantlets produced from 69 Kanamycin-resistant PLBs,however,only 17 plantlets displayed gus gene PCR amplification detection positive results.In addition,we also discovered that the transformatoin efficiency of Phalaenopsis was improved by the treatments of ultrasonic and vacuum before the explants were infected by agrobacterium.At last,a effective genetic transformation system of Phalaenopsis was established.