TA29-barnase基因转化菜心

The pTA29-barnase Chimeric Gene Transformation of Brassica campestris L. subsp. chinensis Makino var. parachinensis Mediated by Agrobacterium

摘要:

利用根癌农杆菌导入法, 以菜心带柄子叶为外植体, 对TA29-barnase基因转化菜心进行研究。获得转化植株,进行PCR、Southern blotting杂交和半定量RT-PCR检测, 表明目的基因已经整合到转化植株中, 并且目的基因在转基因植株花蕾中得到表达, 但是表达水平在不同转基因植株间存在差别; 转基因植株开花后, 均表现雄性不育, 不能产生花粉或产生没有活力的少量花粉, 自交不能结实; 用未转化植株正常花粉对雄性不育植株进行授粉, 能够正常结实; 保持系(未转化植株)与不育株杂交后代中雄性不育株与可育株的比例为1:1, 在杂交后代植株子叶期, 喷洒10 mg/L的PPT可以完全杀死可育株; 利用其他菜心品种为父本与不育株进行杂交, 获得的F1植株在生长势和产量方面表现优势, 表明开展菜心优势育种具有一定的潜力。

Abstract:

In order to induce male sterility of Brassica campestris L. subsp. chinensis Makino var. parachinensis, we introduced the chimeric pTA29-barnase gene into it by Agrobacteriumtume faciens transformation. We obtained the transgenic plants, and determined them by PCR, Southern blotting and RT-PCR analysis. Results indicated that the RNase (barnase) gene had been transferred into genome of plant, and its expression level was different among transformation plants. All transgenic plants were male sterile; there was no vigor or a little pollen without fertility in the anther of transgenic plants. The transgenic plants failed to produce seeds under the condition self-control pollination, but hybrid seeds set were obtained when these transgenic plants were cross-pollinated artificially with normal pollen from untransformed plants. Progeny from cross-pollinated maintainer line with transgenic plants segregated in the 1:1 for male sterility and male fertility, and these phenotypes corresponded directly to the presence or absence of the chimeri TA29-barnase gene. The male fertile plants of co-separated progenies could die by spraying 10 mg/L PPT in cotyledon seedling stage. The hybrid F1 between male sterility and other varieties showed heterosis in yield and growth. All these show that it is an efficient method to induce male sterility in Brassica campestris L. subsp. chinensis Makino var. parachinensis by TA29-barnase gene, there is potential on heterosis breeding of Brassica campestris L. subsp. chinensis Makino var. parachinensis.