摘要
为了提高百合花粉管通道法遗传转化效率,采用pBI121-GFP-FT载体,对外源基因携带形式、导入方式和导入时间进行了研究。对质粒和农杆菌两种形式携带的外源基因的研究结果表明:菌液形式携带外源基因有利于百合花粉管通道法遗传转化。采用切割柱头授粉后涂抹外源基因、常规授粉后涂抹外源基因以及子房注射外源基因3种导入方式进行百合遗传转化,结果表明常规授粉后涂抹外源基因更适宜百合花粉管通道法遗传转化。对授粉后0 h涂抹外源基因和授粉后延迟3 h涂抹外源基因2种处理进行比较研究,结果表明授粉后0 h涂抹外源基因更适宜于百合花粉管通道法遗传转化。将试验所得种子进行荧光观察,转基因植株有绿色荧光。PCR检测结果表明GFP-FT基因已经整合到百合基因组中。
In order to improve transgenic efficiency,the expression vector pBI121-GFP-FT was transformed into lily using pollen-tube pathway method and the effects of exogenous vector type,introduction way and introduction time（after pollination）were discussed.The effects of Agrobacterium and plasmid were compared.The results showed that exogenous vector type of Agrobacterium was better for lily transformation.Three introduction methods were carried out,including cutting stigma pollination and conventional pollination followed by exogenous vector and injecting exogenous vector into ovary.The results indicated that smearing exogenous vector after conventional pollination was suitable for lily transformation.Different introduction times after pollination for 0 h and 3 h were considered,results indicated that 0 h was the optimization introduction time.The green fluorescence was observed in the seeds under fluorescent microscope.Results of PCR also proved that the GFP-FT gene had integrated into lily genome.
出处
《黑龙江农业科学》
2011年第8期15-18,共4页
Heilongjiang Agricultural Sciences
基金
国家"863"计划资助项目(2008AA10Z156)
"十一五"国家科技支撑计划资助项目(2009BADB8B04)
北京市教育委员会产学研合作资助项目
