农杆菌介导的天绿香遗传转化影响因素的研究  

The Transformation of Insect Antifreezing Protein for Sauropus androgynus Mediated by Agrobacterium

作　　者：肖艳华[1] 曾幼玲[1] 兰海燕[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《植物研究》2008年第5期564-567,573,共5页Bulletin of Botanical Research

基　　金：新疆生物资源基因工程重点实验室开放课题(XJDX0201-200503);新疆高校创新研究群体基金项目(XJEDU2004G02)

摘　　要：通过农杆菌介导的方法将新疆准噶尔小胸鳖甲抗冻蛋白基因Mpafp149导入天绿香中，对影响农杆菌转化频率的各种因素进行了研究。结果表明农杆菌介导天绿香遗传转化的潮霉素最佳筛选浓度为8mg·L^-1，预培养时间2d，共培养时间2d，菌液浓度OD600为0．2～0．25，浸染时间为8Min时适宜天绿香不定芽的诱导，有利于天绿香转基因植株的获得。通过PCR检测，初步证明外源基因，Mpafp149已整合到天绿香基因组中。In order to improve the S. androgynus cold tolerance, genetic transformation system of S. androgynus was established in this study. The cDNA Mpafp149 of Microdera punctipennis dzung used as target gene to obtain the cold tolerant transgenic S. androgynus plants. The stems of S. artca was androgynus were transformed with Agrobacterium tumefaciens EHA105 strain which harbors plasmid pCAMBIA- 1302-Mpafp149. The transformation parameters of Agrobacterium concentration, pre-cuhure period, coculture period, immersion time and concentrations of antibiotics （hygromycin） were optimized selectively. The results showed that the better physiological and transforming condition was pre-cuhured and cocultured for 2 days respectively, immersed in OD600 0.2 - 0.25 liquid bacterium for 8 min. It was primarily proved by PCR that the Mpafp149 was integrated into the Sauropus androgynus and the To transgenic S. androgynus plants were obtained successfully.

关 键 词：天绿香 昆虫抗冻蛋白基因 农杆菌介导 遗传转化 

分 类 号：Q943.2[生物学—植物学]