花粉管通道法转柱头外露棉DNA及转化体RAPD分析  

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

作　　者：周平兰[1] 梁满中[1] 高健[2] 陈良碧[1] 

机构地区：[1]湖南师范大学生命科学学院植物学系,中国长沙410081 [2]湖南科技大学生命科学学院生物系,中国湘潭411201

出　　处：《湖南师范大学自然科学学报》2004年第4期81-86,共6页Journal of Natural Science of Hunan Normal University

基　　金：湖南省自然科学基金重点资助项目(02ZY2058)

摘　　要：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(TCGGCACGCA)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.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 S462 (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 S462 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.

关 键 词：柱头外露 花粉管通道法 RAPD分析 基因位点 引物 供体 转化 受体 DNA CGC 

分 类 号：S511[农业科学—作物学] S562.032