转基因甘蓝型油菜的小孢子培养及转基因的遗传分析

目的]研究甘蓝型油菜转查尔酮异构酶反义基因T1代植株及其小孢子成苗的遗传表达,为甘蓝型油菜的转基因和种质资源创新研究提供参考。方法]以甘蓝油菜转查尔酮异构酶反义基因T1代5个独立转基因植株为材料进行小孢子培养,通过GUS报告基因的组织化学染色鉴定对外源转基因在花粉植株中的遗传进行了分析。结果]5个独立转基因植株均获得了胚状体和花粉植株;转基因在花粉培养后代和有性自交后代中遗传行为一致;在每个独立转基因植株的花粉植株中GUS阳性和阴性的分离比均为3∶1,符合孟德尔遗传规律,表明有2个转基因拷贝整合到受体2条非同源染色体上;3个独立转基因植株自交后代中GUS阳性和阴性的分离比符合15∶1,同样表明有2个转基因拷贝整合到受体2条非同源染色体上。结论]通过转基因油菜的小孢子培养快速获得转基因纯系是可行的。

Research on the Transgenic Brassica napus Microspore Culturing and Genetic Analysis of Transgenic Character

Objective] The reference for the innovative research on the transformation and germplasm resource of Brassica napus was provided through the study on the Brassica napus plant of T1 generation transferred with the antisense gene of chalcone isomerase and its expression in the plantlet from microspore culturing. Method] The microspores from five independent transgenic plants were cultured and the genetic analysis of exogenous gene in pollen plant was carried out through the staining identification of GUS reporter gene. Results] The embryoids and pollen plant could be obtained from five independent transgenic plants. The genetic behavior of the generation from pollen culturing was consistent with the sexual self-offspring. The segregation ratio of GUS-positive reaction and GUS-negative reaction in pollen plant of each independent transgenic plant was 3∶1,which was matched with Mendelian law and the result indicated that there were two transgenic copies that had been integrated into the receptor,two non-homologous chromosomes. The segregation ratio of GUS-positive reaction and GUS-negative reaction in self-progeny of three independent transgenic plants was 15∶1,also showing that there were two transgenic copies that had been integrated into the receptor,two non-homologous chromosomes. Conclusion] It was feasible to quickly produce the double haploid line by means of the microspore culture of transgenic Brassica napus plant.