裸燕麦成熟胚组织培养体系的优化及创伤胚转化体系的初探  

Optimization of Mature Embryo Culture System of Naked Oat and Preliminary Study on Transformation System of Traumatized Embryo

作　　者：马佳星 李庆华 刘玉海 南金生 融晓萍 路素香 韩冰[1] MA Jiaxing;LI Qinghua;LIU Yuhai;NAN Jinsheng;RONG Xiaoping;LU Suxiang;HAN Bing(Inner Mongolia Agricultural University/Key Laboratory of Wheat Germplasm lnnovation and Utilization Autonomous Region Higher School/Key Laboratory of Grassland Resources of Education Ministry/Key Laboratory of Biological Manufacturing of Inner Mongolia Autonomous Region,Hohhot 010018,China;Ulanqab Agricultural Technology Extension Center,Ulanqab 012000,China;Inner Mongolia Technology Promotion Center of Agricultural&Animal Husbandry,Hohhot 010020,China;Agricultural and Rural Bureau of Wanquan District,Zhangjiakou 076250,China)

机构地区：[1]内蒙古农业大学/麦类种质创新利用自治区高等学校重点实验室/草地资源教育部重点实验室/内蒙古自治区生物制造重点实验室,内蒙古呼和浩特010018 [2]内蒙古自治区乌兰察布市农业技术推广中心,内蒙古乌兰察布012000 [3]内蒙古自治区农牧业技术推广中心,内蒙古呼和浩特010020 [4]河北省张家口市万全区农业农村局,河北张家口076250

出　　处：《大麦与谷类科学》2024年第5期21-28,36,共9页Barley and Cereal Sciences

基　　金：国家重点研发计划(2022YFE0119800);内蒙古农业大学学院创新团队建设项目(TD202103)。

摘　　要：对8个裸燕麦品种的成熟胚进行诱导愈伤和分化培养,选用出愈率最高的品种裸燕麦1号,进一步对培养基进行优化。结果表明:在ML2诱导培养基中裸燕麦1号出愈率为87.33%,在DM1分化培养基中分化率较高。最佳培养体系为:使用ML2诱导培养基诱导愈伤,约15 d后进行继代培养,继代2次后转移至DM1分化培养基,待分化出叶片后转移至生根培养基。通过农杆菌介导法,使用带有绿色荧光蛋白GFP基因的pBI121质粒对裸燕麦1号创伤胚进行侵染,探究菌液浓度、选择剂浓度及移栽时间对转化率的影响。结果表明,裸燕麦创伤胚转化高效组合培养体系包括:农杆菌菌液OD600为0.8时进行侵染,卡那霉素选择剂的最适浓度为40 mg/L,创伤胚转化培养10 d后进行移栽。本试验以农杆菌侵染裸燕麦创伤胚建立的遗传转化体系具有可行性,为以后裸燕麦再生体系建立提供借鉴。Callus induction and differentiation culture in the mature embryos of 8 naked oat varieties were performed.Naked Oat No.1 was selected as the variety with the highest healing rate,and the medium was further optimized.The results showed that No.1 oat had the healing rate of 87.33% in ML2 induction medium and higher differentiation rate in DM1 differentiation medium.The optimal culture system was as follows:ML2 induction medium was used to induce callus,subculture was carried out after about 15 days,followed by two subcultures and then transferred to DM1 differentiation medium,and then transferred to rooting medium after differentiation of leaves.The pBI121 plasmid with GFP gene was used to infect the callus embryos of naked Oat No.1 by Agrobacterium-mediated method,and the effects of concentration of bacterial solution,concentration of selective agent and transplanting time on the transformation rate were investigated.The results showed that the efficient combined culture system for callus embryo transformation of naked oats included Agrobacterium solution(OD600)of 0.8 for infection,the optimal concentration of kanamycin selective agent was 40 mg/L,and the callus embryo was transplanted after 10 days of transformation culture.The Agrobacterium-mediated genetic transformation system of callus embryos of naked oats preliminarily established in this experiment is feasible,which provides a reference for the establishment of the regeneration system of naked oats in the future.

关 键 词：裸燕麦 组织培养 遗传转化 农杆菌介导 

分 类 号：S511[农业科学—作物学]