SNAC1基因作为筛选标记基因用于棉花的遗传转化

Application of SNAC1 Gene as a Selection Marker Gene for Cotton (Gossypium hirsutum L.)Transformation

以SNAC1基因作为筛选标记基因,NaCl作为筛选剂,通过农杆菌介导法将SNAC1和GUS基因导入棉花细胞并得到胚性愈伤组织。经过PCR检测证实,外源基因已经整合到棉花基因组中,GUS染色证明GUS基因得到表达。研究了NaCl作为棉花转化细胞的筛选剂在农杆菌介导转化中的应用浓度及方法,即NaCl的筛选浓度在1.1%～1.5%(W/V)之间,在愈伤组织诱导初期适当低一点,随着愈伤组织的生长而加大筛选浓度。由于NaCl不利于胚的分化,经过2～3次继代筛选后要及时去除NaCl以促进胚的分化。

Employing SNAC1 gene as a selection marker gene and NaCl as a selection agent, SNAC1 and GUS genes were introduced into cotton genome via Agrobacterium. Transgenic calli were confirmed by PCR analysis, and expression of the GUS gene was showed with GUS staining. The concentration and method for employing NaCl as a selection agent had been studied. The reasonable selection concentration of NaCl should be 1.1%-1.5%（W/V）. The start concentration of NaCl in callus induction medium should be lower and the concentration increased as calli proliferated. Because NaCl is not beneficial for embryo differentiation and development, NaCl should be removed from the culture medium after the calli were subcultured for 2-3 times.