根癌农杆菌介导甘蔗遗传转化Bt(cry1Ab)基因  

Genetic Transformation of Bt (cry1Ab) Gene into Sugarcane (Saccharum officinarum L.) Mediated by Agrobacterium tumefaciens

作　　者：李晓梅[1] 王闵霞 秦廷豪[1] 阳翠[1] 安琪[1] 张军[1] 

机构地区：[1]四川省植物工程研究院生物技术研究所,资中612400 [2]四川省农业科学研究院生物与核技术研究所,成都610000

出　　处：《生物技术通报》2013年第2期100-105,共6页Biotechnology Bulletin

基　　金：四川省科技计划项目(2010CJY0047)

摘　　要：以自育甘蔗品种"川蔗23号"诱导的胚性愈伤为受体材料,首次用较低的根癌农杆菌侵染浓度(OD600=0.1左右)与较短的侵染时间(1.5 min)成功实现甘蔗遗传转化Bt(cry1Ab)基因。结果表明,胚性愈伤分化与小芽生根最适潮霉素筛选浓度分别为20 mg/L和30 mg/L;愈伤组织胚性的一致性是影响筛选阶段愈伤再生的重要因素,培养40 d的愈伤组织为转化最适受体;转化材料经连续的潮霉素抗性筛选后,获得65株抗性植株,通过特异性引物的PCR扩增检测,有2株获得阳性条带,初步表明目的基因已整合到甘蔗染色体基因组中。In this research, embryonic callus induced from sugarcane （ Saccharum offwinarum L. ） cultivar ＂Chuanzhe 23＂ was selected as recipient material. We successfully transformed anti-insect gene Bt （ crylAb ） into sugarcane mediated by Agrobactenum tumefaciens EHA105 when the explants infected with lower concentration ofAgrobactenum tumefaciens （ OD ~ 0.1 ） and shorter infusion time （ 1.5 min ） . The optimal concentrations of hygromycin for the selection of transformed shoots and roots induction respectively were 20 mg/L and 30 mg/L. The calli cultured for 40 d were optimal for transformation ; 65 transformants were obtained after successive hygromycin resistance selection. The hyg-resistant plants were detected by PCR analysis, and 2 plants showed positive, which initially proved that the target gene had been integrated into sugarcane genome.

关 键 词：甘蔗 遗传转化 Bt(cry1Ab)基因 

分 类 号：S566.1[农业科学—作物学]