农杆菌介导CSPs基因转化甘蓝型油菜转化体系的研究

On System of Transforming CSPs Gene into Brassica napus Mediated by Agrobacterium

以甘蓝型油菜下胚轴为外植体,通过农杆菌介导法,对其遗传转化条件进行了研究,建立了转冷激蛋白(CSPs)的遗传转化体系。结果表明,甘蓝型油菜下胚轴经预培养2d后,用浓度为OD600=0.4的农杆菌菌液侵染90s,再经共培养和愈伤诱导培养,外源基因转化到油菜下胚轴外植体的转化效率高。然后通过PCR检测转化后在筛选培养基上诱导成苗的油菜,初步确定CSPs基因已整合到油菜再生植株中。这为以后其他基因转化到甘蓝型油菜的研究奠定一定的基础。

he system of genetic transformation into hypocotyl （as explant） of Brassica napus mediated by Agrobacterium was studied. The results demonstrate that there had high conversion efficiency for exogenous genes into the hypocotyl （explants） Brassica napus L after the hypocotyl was pre - cultured for 2d and then infected for 90s by Agrobacterium tumefaciens （OD600 = 0. g） with CSPs gene, and the infected hypocoty was co -cultured and cultured for induced - callus. The rape seedlings induced on the screening culture - medium were detected by PCR analysis, which proved that the CSPs gene had been integrated into the genome of regenerated rape. From above, it laid foundation for the research on other gene transformed into B. napus in future.