花粉管通道法转基因技术在甜瓜品种河套蜜瓜上的应用

为建立甜瓜(Cucumis melo L.)简便易行转化频率高的转基因技术,以甜瓜品种河套蜜瓜为受体材料,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.自交授粉后分别于1、2、3、4、5、6、7、8、9、10 h,切去柱头上端1/3部分,立即滴加质粒DNA溶液,果实成熟后收获转化种子.对T0代植株进行PCR检测,结果表明不同时间处理所得到的T0代植株均具有较高的转化频率,其中授粉后7 h滴加DNA溶液所获得的转化率为28.3%.对一部分PCR阳性的T0代植株基因组DNA进行Southern杂交分析,结果表明所有样品均出现特异性杂交条带,证明外源基因已整合到受体植物基因组中.

Establishment of a Melon Transformation Technique via the Pollen-tube Pathway Method

In order to establish a simple,practical technique with high transformation frequency in melon(Cucumis melo L.),the pollen-tube pathway method was used for melon transformation.The melon cultivar Hetao 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 cut surfaces of stigmas at 1,2,3,4,5,6,7,8,9,and 10 h after hand-pollination,respectively.The PCR results show that the transformation rate of all T0 seedlings is very high and the highest rate is up to 28.3% at 7 h after pollination.Southern blot analysis confirmed that foreign DNA was indeed inserted into the genome of transgenic T0 seedlings.