摘要
为优化苎麻遗传转化体系,以中苎1号无菌苗的叶盘为外植体,研究了卡那霉素、头孢霉素、农杆菌的浓度以及共培养和侵染的时间对苎麻遗传转化的影响。结果表明:在愈伤诱导阶段,卡那霉素最佳浓度为50 mg/L;在生根阶段,卡那霉素最佳浓度为30 mg/L,头孢霉素最佳浓度为500 mg/L;农杆菌最佳浓度为OD_(600)=0.6,最佳侵染时间为10 min,最佳共培养时间为36 h。最终得到3株转化植株,根据PCR检测,证实Bt抗虫基因已经成功整合到1株苎麻中。综上所述,本研究优化了苎麻遗传转化体系,可以为今后利用基因工程技术对苎麻进行遗传改良提供参考。
In order to optimize the genetic transformation system of ramie,this study used leaf discs of sterile seedlings of Zhongzhu No.1 as explants to investigate the effects of concentrations of kanamycin,cefotaxime and Agrobacterium,on the genetic transformation of ramie,as well as co-cultivation time,and infection time.The results showed that the optimal concentration of kanamycin was 50 mg/L during the callus induction stage and 30 mg/L during the rooting stage,while the optimal concentration of cefotaxime was 500 mg/L.The optimal Agrobacterium concentration was OD_(600)=0.6,the optimal infection time was 10 min,and the optimal co-cultivation time was 36 h.Finally,three transformed plants were obtained,and PCR detection confirmed that the Bt insect-resistant gene was successfully integrated into 1 ramie plant.In conclusion,this study optimized the genetic transformation system of ramie,providing a reference for future genetic improvement of ramie using genetic engineering technology.
作者
何鹏亮
伍旭东
朱宁静
蒋美艳
邢虎成
揭雨成
HE Pengliang;WU Xudong;ZHU Ningjing;JIANG Meiyan;XING Hucheng;JIE Yucheng(Ramie Institute of Hunan Agricultural University,Changsha Hunan 410128;Hunan Engineering Research Center for Grass Crop Germplasm Innovation and Utilization,Changsha Hunan 410128)
出处
《现代农业科技》
2025年第7期150-155,共6页
Modern Agricultural Science and Technology
基金
国家科技平台项目“国家饲草与饲纤兼用作物种质资源共享服务平台”(NICGR2018-072)
国家自然科学基金项目“矿区野生苎麻镉富集特征和耐性及分子标识与相关基因的克隆”(31371704)
国家自然科学基金项目“镉胁迫下苎麻萝卜根细胞壁的解毒策略”(31872877)。
关键词
苎麻
外植体
叶盘
农杆菌侵染
Bt抗虫基因
遗传转化
ramie
explant
leaf disc
Agrobacterium infection
Bt insect-resistant gene
genetic transformation
