花粉管通道法转柱头外露棉DNA及转化体RAPD分析

提取“异9-3”柱头外露棉总基因组DNA，通过花粉管通道法经微量注射导入抗虫棉707．1的胚囊内，D1代田间筛选到柱头外露的转化植株，遗传分析结果表明转化株柱头外露性状由一对隐性基因控制，50个随机引物进行PCR扩增，33个引物扩增出的DNA在供体、受体和转化体之间呈现出多态性，其中引物S462(TCG GCA CGC A)在供体和转化体之间出现相同产物而受体没有出现扩增产物，该引物有可能作为柱头外露系的分子标记．RAPD分析结果表明转化体与受体有较多相同基因位点，而与供体相同基因位点较少，转化体还有供体和受体都不具有的基因位点，这可能是外源DNA导入时引起碱基突变所致。

Transgenic Cotton Obtained Through Introducing Open Budvia the Pollen Tube Pathway Method and Their RAPD Analysis

Via pollen-tube pathway the genomic DNA of the open-bud cotton Y9-3 was introduced into the insect-resistant cotton 707-1. The variant plants with the characteristic of open-bud were selected from D2 generation. Inheritance analysis showed that the characteristic of open-bud was controlled by a pair of recessive gene. RAPD(Random Amplified Polymorphism DNA) was used to detect the genetic diversity among the donor, the receptor and the variant plants. On the optimized condition of PCR, total 33 out of 50 pairs of primers showed the polymorphism. The primer S_(462) (TCG GCA CGC A) amplified the same PCR products with model DNA of the donor and the variant, nothing was amplified in the receptor. So it was supposed that the primer S_(462) was the molecular marker of the open-bud. There were more similar gene positions in the variant and the receptor than that in the variant and the donor. And the variant had some especial gene positions because of gene mutation.