“Gros Michel”香蕉胚性细胞悬浮系及遗传转化体系的建立  

Establishment of Embryogenic Cell Suspension and Genetic Transformation System of "Gros Michel" Banana

作　　者：邵秀红 吴少平[1,2] 窦同心 邓贵明[2] 盛鸥[2] 毕方铖[2] 胡春华[2] 易干军[2] 杨乔松[2] Shao Xiuhong;Wu Shaoping;Dou Tongxin;Deng Guiming;Sheng Ou;Bi Fangcheng;Hu Chunhua;Yi Ganjun;Yang Qiaosong(Horticulture and Landscape College,Hunan Agricultural University,Changsha,410128;Guangdong Key Laboratory of Tropical and Subtropical Fruit Tree Research,Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization,Ministry of Agriculture,Institute of Fruit Tree Research,Guangdong Academy of Agricultural Sciences,Guangzhou,510640)

机构地区：[1]湖南农业大学园艺园林学院,长沙410128 [2]广东省农业科学院果树研究所农业部南亚热带果树生物学与遗传资源利用重点实验室广东省热带亚热带果树研究重点实验室,广州510640

出　　处：《分子植物育种》2019年第3期846-854,共9页Molecular Plant Breeding

基　　金：广东省省级科技计划项目(2015B070701011;2016B020233002;2014B050502007;2012A020100007);广东省高层次人才特殊支持计划(2014TQ01N175);农业部"948"项目(2011-G16;2016-X22);国家香蕉产业技术体系项目(CARS-32-01);公益性行业(农业)科研专项经费项目(201403036);广东省现代农业产业共性技术创新团队建设项目(2017LM2148);广东省农业厅现代种业提升工程项目(粤农计(2017)64号);优势特色农作物品种及配套技术集成示范与推广项目;广东省自然科学基金博士启动纵向协同管理试点项目(2017A030310351)共同资助

摘　　要：本研究以"Gros Michel"香蕉未成熟雄花序为外植体,建立了其胚性细胞悬浮系及遗传转化体系。结果表明,90 d后可诱导产生浅黄色松散的胚性愈伤组织,经悬浮培养120 d后获得含有均质细胞团的胚性细胞悬浮系(ECS);将3个月龄的ECS置于胚诱导培养基30 d后有大量白色半透明状球形的体细胞胚发生,继代培养60 d后发育为成熟不透明的体细胞胚;将成熟的体胚在萌发培养基上培养10 d后有幼芽产生,转移至生根培养基30 d后得到幼苗。同时开展了含绿色荧光蛋白(GFP)的pCAMBIA0380载体的遗传转化研究,经过3次液体培养基筛选转移至半固体筛选培养基90 d后,在胚萌发培养基和生根培养基中不再添加抗生素。经对转化植株的根尖镜检及PCR鉴定,均为阳性植株。研究结果将为进一步开展"Gros Michel"优质抗病基因功能研究提供材料和技术支撑。In the research,embryogenic cell suspension and genetic transformation of "Gros Michel"banana were established with the immature male inflorescence as explants.The results showed that yellowish and loose embryogenic.callus could be developed after 90 days.Embryogenic cell suspensions (ECS)with homogeneous cell mass were obtained after 120 days of suspension culture.3-month-old ECS were cultured in embryo-inducing medium for 30 days,followed by a large number of white translucent spherical somatic embryos,which would be developed into mature opaque somatic embryos after subculture for 60 days.Mature somatic embryos were cultured on the germination medium for 10 days,and then buds were produced and transferred to the rooting medium for 30 days to obtain seedlings.At the same time,the genetic transformation of pCAMBIA0380 vector containing green fluorescent protein (GFP)was carried out.The embryogenic cells were selected after three times of liquid screening medium and transferred to semi-solid screening culture medium for 90 days,followed by culturing in embryo germination medium and rooting medium without any antibiotic.After root tip microscopic examination and PCR identification of the transformed plants,they were found to be positive.Those results would provide materials and technical support for the further research on the function of high-quality disease resistance gene in "Gros Michel".

关 键 词：香蕉 Gros Michel 胚性细胞悬浮系 遗传转化体系 

分 类 号：S668.1[农业科学—果树学]