香蕉遗传转化体系优化  

Optimization of the Banana Genetic Transformation System

作　　者：甘珊珊 王静毅[1] 程运江[2] 刘菊华[1] GAN Shanshan;WANG Jingyi;CHENG Yunjiang;LIU Juhua(Institute of Tropical Bioscience and Biotechnology,CATAS/Key Laboratory of Tropical Crops Biology and Genetic Resources(Ministry of Agriculture and Rural Affairs)/National Key Laboratory for Tropical Crop Breeding,Sanya Research Institute/Hainan Key Laboratory for Protection and Utilization of Tropical Bioresources,Hainan Institute for Tropical Agricultural Resources,Haikou,Hainan 571101,China;College of Horticulture and Forestry Science,Huazhong Agricultural University/National R&D Center for Citrus Preservation/Key Laboratory of Horticultural Crop Biology and Germplasm Genetic Improvement(Fruit Crops),Ministry of Agriculture and Rural Affairs/National Key Laboratory for Germplasm Innovation&Utilization of Horticultural Crops,Wuhan,Hubei 430070,China)

机构地区：[1]中国热带农业科学院热带生物技术研究所/农业农村部热带作物生物学与遗传资源利用重点实验室/三亚研究院热带作物生物育种全国重点实验室/海南热带农业资源研究院海南省热带农业生物资源保护与利用重点实验室,海南海口571101 [2]华中农业大学园艺林学学院/国家柑橘保鲜技术研发专业中心/农业农村部园艺作物生物学与种质创制(果树)重点实验室/果蔬园艺作物种质创新与利用全国重点实验室,湖北武汉430070

出　　处：《热带农业科学》2024年第10期17-24,共8页Chinese Journal of Tropical Agriculture

基　　金：海南省自然科学基金(No.321RC638);中国热带农业科学院国家热带农业科学中心科技创新团队(No.CATAS CXTD202310);现代农业产业技术体系(No.CARS-31)。

摘　　要：香蕉(Musa spp.)高度不育和多倍体特性严重阻碍常规育种方法的应用,而香蕉遗传转化技术存在基因型依赖性强、转化效率偏低等问题。以贡蕉(Musa acuminata cv.Mas)多芽体为起始转化材料,利用根癌农杆菌介导β-葡萄糖苷酸酶基因(GUS)介导转化,通过研究外植体转化状态、不同转化载体及乙酰丁香酮(acetosyringone,AS)孵育时间、农杆菌菌液浓度、预培养时间、不同农杆菌菌株和共培养方式对香蕉遗传转化过程的影响,优化遗传转化条件,以期获得高效的香蕉遗传转化体系。结果表明,以贡蕉多芽体薄切片为外植体,黑暗条件下预培养1 d,使用以p CAMBIA1304构建的植物表达载体EHA105农杆菌制备侵染液时,AS孵育0~4 h,侵染液农杆菌OD600为0.4~0.6,侵染后将多芽体切片放置在固体MS培养基(添加200μmol/L AS)上进行共培养,再生植株转化效率较好。通过优化遗传转化体系的条件,提高香蕉遗传转化率,为解决产业发展关键技术难题及精准定向分子育种提供技术支撑,为香蕉产业的健康可持续发展提供重要保障。The highly sterile and polyploid characteristics of banana(Musa spp.)severely hinder the application of conven-tional breeding methods.However,banana genetic transformation technology suffers from strong genotype dependence and low transformation efficiency.In this study,Agrobacterium tumefaciens was utilized to mediate the transformation of the GUS gene,and multiple bud clumps of the Musa acuminata cv.Mas(AA)were used as the starting transformation material.To optimize the genetic transformation conditions by studying the effects of the transformation status of the explants,different transformation vectors and incubation times of acetosyringone(AS),concentrations of Agrobacterium tumefaciens,pre-cultivation times,different Agrobacterium strains and co-cultivation methods on the process of banana genetic transfor-mation were studied,to obtain a highly efficient genetic transformation system for banana.The results revealed that the multi-ple bud slices of M.acuminata cv.Mas(AA)were used as explants and were pre-cultured for 1 d.When the infiltration solu-tion was prepared by using Agrobacterium EHA105 containing the plant expression vector constructed with pCAMBIA1304,the AS was incubated for 0-4 h,and the OD600 of A.tumefaciens infestation solution was 0.4-0.6.The infiltrated multiple bud slices were placed in a solid MS media(with the addition of 200μmol·L-1 AS)for co-culture,and the transformation effi-ciency of regenerated plants was better.Optimizing the conditions of the genetic transformation system to improve the genetic transformation rate of bananas will provide vital technical support for solving the crucial technical problems in the develop-ment of the banana industry and the precisely directed molecular breeding of bananas.It will provide a critical guarantee for the healthy and sustainable development of the banana industry.

关 键 词：香蕉 遗传转化体系 多芽体 优化 转化率 

分 类 号：S668.1[农业科学—果树学]