转TaDREB3a基因大豆材料抗旱性鉴定

Identification of drought resistance of transgenic TaDREB3a soybean materials

研究通过分子生物学方法证明TaDREB3a基因已整合至大豆基因组中,对T1~T3连续3代大豆植株作筛选鉴定并分析遗传稳定性,初步评价室内和田间抗旱表现。通过除草剂筛选获得T1代135株抗性植株,经PCR检测其中96株扩增出基因目的条带,获得15个阳性TaDREB3a过表达大豆T1代株系;T2代转基因大豆株系经PEG模拟干旱处理和PCR鉴定获得阳性TaDREB3a过表达大豆T2代株系共10个;T3代转基因大豆株系经PCR鉴定、半定量PCR鉴定、Bar基因试纸条检测、Southern blot分析、Western blot分析,获得6个转基因阳性株系,证实TaDREB3a基因已整合于大豆基因组,可转录完整目的mRNA,其中4个株系检测到目的蛋白表达。盆栽干旱试验显示,TaDREB3a过表达可改善转基因大豆干旱条件下生长状况,转基因植株株高和荚数明显优于对照组植株;大田干旱试验结果显示,TaDREB3a过表达株系提高大豆抗旱性、改善干旱条件下地上形态,减少产量损失。

In this study, it proved that TaDREB3a gene had been integrated into the soybean genome through molecular biological method. The detections for identifying the transgenic soybean plants from T1 to T3 progenies and their heredity stabilities were conducted. Additionally, the agronomic performance of transgenic soybeans under drought stress in the lab testing or in the field, were also evaluated. A total of 135 T1 resistant plants were obtained by PPT screening, and of them, 96 positive over-expressing soybeans were further identified by PCR. Furthermore, the 15 transgenic soybean lines were screened under the detections by PEG simulation and PCR, and finally, a number of six T3 transgenic plants were obtained by PCR, semi- RT- PCR, Bar gene test paper, Southern blot and Western blot. It proved that TaDREB3a gene had been integrated into the soybean genome, and could express the target mRNA, and four of the lines could stably detect the expressions of the target protein.Drought experimental analysis in the pot showed that over-expressed TaDREB3a in soybean plants could remarkably improve the growing status of the transgenic soybeans under the drought stress. The plant height and number of pods of transgenic plants were significantly higher than that of in the control plants. Also, the similar results among the field testing exhibited that the overexpression TaDREB3a transgenic soybeans could enhance the drought resistance, optimize the growing status and reduce yield losses than the control plants.