紫花苜蓿花粉管通道法转基因技术初探

Preliminary Exploration of Alfalfa Transgene Technique via Pollen-tube Pathway

以紫花苜蓿品种阿尔冈金为受体材料，以含gus基因的植物双元表达载体pPZP221为外源基因供体进行了花粉管通道法转基因研究。于授粉后6h、9h、12h、20h、24h、28h、32h、48h滴加质粒DNA溶液，荚果成熟后收获转化种子，分别使用2套引物对T0代植株进行PCR检测，结果表明：20～48h所得到的T0代植株均具有较高的转化频率，其中授粉后32h导入质粒DNA所获得的转化率最高，为16．7％；PCR检测初步证明外源基因已整合到受体植物基因组中。

In order to establish a simple, practical technique with high transformation frequency in alfalfa （Medicago sativa L. ）, the pollen-tube pathway method was used for alfalfa transformation. The alfalfa cultivar ‘Algonquin＇ was used as a recipient and the plant binary vector pPZP221 containing gus gene was used as donor DNA. The plasmid DNA solution was applied to the stigma at 6h, 9h, 12h, 20h, 24h, 28h, 32h and 48h after hand-pollination, respectively. PCR results showed that the transformation rate of To seedlings from 20h to 48h was relatively high and the highest rate was up to 16.7% at 32h after pollination. This results suggested that foreign DNA was inserted into the genome of transgenic To seedlings.