根癌农杆菌介导的香蕉高效遗传转化系统的建立  

Establishment of a High Efficient Agrobacterium tumefaciens-Mediated Transformation System for Banana

作　　者：胡春华[1] 魏岳荣[1] 易干军[1] 黄秉智[1] 黄永红[1] 

机构地区：[1]广东省农业科学院果树研究所,广州510640

出　　处：《分子植物育种》2010年第1期172-178,共7页Molecular Plant Breeding

基　　金：国家科技支撑项目(2008BAD92B06);广东省科技攻关项目(2007A020200003-3);广东省农科院院长基金(20090104)共同资助

摘　　要：为建立根癌农杆菌介导的香蕉高效遗传转化系统,本研究以雄性花诱导产生的贡蕉胚性悬浮系为转化受体,从潮霉素筛选浓度和筛选方法两方面进行了优化。研究结果表明,贡蕉悬浮系在M2液体培养下潮霉素的适合筛选浓度为5mg/L。将液体共培养7d后的贡蕉胚性悬浮系转接到M2液体筛选培养基进行培养,每10d继代一次。GUS组织染色表明,经过3代抗性筛选的ECS几乎全为转化细胞。经过3个月的胚诱导培养获得成熟抗性体细胞胚,平均抗性体细胞胚得率为4580个/mLPCV。同时,转化苗的PCR检测结果表明,gus基因已整合到贡蕉基因组中。本研究表明液体筛选系统有利于转化胚性悬浮细胞的增殖,大大地提高了香蕉转基因的转化效率。In this research, we employed male-flower-derived embryogenic cell suspensions （ECS） ofMusa acuminata var. Pisang Mas （AA） as transformation receptor, and optimized the genetic transformation system of banana from aspects of hygromycin selection concentration and selection method in order to establish a high efficient A - grobacterium tumefaciens-mediated genetic transformation system for banana. The results showed that the suitable selection concentration of hygromycin on M2 liquid medium cultivation of ECS was 5 mg/L. The infected ECS were co-cultivated for 7 days and then cultivated in M2 selective liquid medium and subcultured every 10 days. Histochemical GUS assay demonstrated that almost all the ECS were transformed cells after 3 generations subculture resistance screening. We successfully obtained the maturation resistance somatic embryos by embryonal induction on semi-solid selection medium for 3-month selection. And the rate of average resistance somatic embryos was 4 580 mature somatic embryos per 1 mL packed cell volume （PCV） of ECS. Simultaneously, PCR amplification of regenerated plants indicated that gus gene had been integrated into banana genome. The results suggested that the liquid screening system be facilitate to ECS proliferation, which would greatly improve transformation efficiency of banana.

关 键 词：香蕉 根癌农杆菌介导转化法 液体共培养 

分 类 号：S668.1[农业科学—果树学]