粉葛不同外植体的诱导与松散型愈伤组织增殖研究  

Study on Induction of Different Explants of Pueraria montana var.thomsonii(Bentham)M.R.Almeida and Loose Callus Proliferation

作　　者：袁名远 罗凯 刘玉 吴丽雯 孙慧茹 赵苓云 李宇 何美军 Yuan Mingyuan;Luo Kai;Liu Yu;Wu Liwen;Sun Huiru;Zhao Lingyun;Li Yu;He Meijun(College of Biological Science and Technology,Hubei Minzu University,Enshi 445000,China;Institute of Chinese Herbal Medicines,Hubei Academy of Agricultural Sciences/Key Laboratory of Chinese Herbal Medicine Biology and Cultivation,Ministry of Agriculture and Rural Affairs,Enshi 445000,China)

机构地区：[1]湖北民族大学生物科学与技术学院,湖北恩施445000 [2]湖北省农业科学院中药材研究所/农业农村部中药材生物学与栽培重点实验室,湖北恩施445000

出　　处：《山东农业科学》2023年第7期57-65,共9页Shandong Agricultural Sciences

基　　金：湖北省重点研发计划项目(2020BCA059);湖北省现代农业产业技术体系项目(HBHZD-ZB-2020-005);湖北省农业科学院青年基金项目(2022NKYJJ17);恩施州科技计划项目(D20210006,D202100018)。

摘　　要：为建立系统的粉葛松散型愈伤组织培养体系,比较分析了不同浓度的外源激素6-BA和NAA组合对粉葛叶片、叶柄和茎段三种外植体的诱导效果,并对具有松散形态的愈伤组织进行增殖培养,运用响应面法结合2,4-D对增殖条件进行系统优化,采用分光光度法检测此条件下诱导的愈伤组织中总黄酮和总酚的含量。结果表明,茎段和叶柄的最适诱导条件均为2.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA,诱导率最高分别达到96.30%和95.83%;叶片的最适诱导条件为2.0 mg·L^(-1)6-BA+0.5 mg·L^(-1)NAA,诱导率最高为42.86%。筛选出茎段作为松散型愈伤组织诱导和增殖的材料,增殖的初步条件为1.0 mg·L^(-1)6-BA+0.25 mg·L^(-1)NAA;响应面优化后的松散型愈伤组织最适增殖条件为1.10 mg·L^(-1)6-BA+0.25 mg·L^(-1)NAA+0.95 mg·L^(-1)2,4-D,此条件下愈伤组织主要为黄绿色,呈现较松散的状态,增殖系数最高达到9.34,总黄酮和总酚含量分别为25.07、27.44 mg·g^(-1)。In order to establish a systematic culture system of loose callus of Pueraria montana var.thomsonii(Bentham)M.R.Almeida,the induction effects of different concentrations of plant exogenous hor-mones 6-BA and NAA on leaf,petiole and stem explants were compared.Then,callus with loose morphology were cultured for proliferation,and response surface method combined with 2,4-D was used to optimize the proliferation conditions.The contents of total flavonoids and total phenols in induced callus under this condition were also determined by spectrophotometry.The results showed that the optimal induction conditions for stems and petioles were 2.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA with the highest induction rates as 96.30%and 95.83%,respectively.The optimal induction condition for leaves was 2.0 mg·L^(-1)6-BA+0.5 mg·L^(-1)NAA with the highest induction rate as 42.86%.The stems were selected as the material for the induction and prolifera-tion of loose callus,and the primary conditions for proliferation were 1.0 mg·L^(-1)6-BA+0.25 mg·L^(-1)NAA.After response surface optimization,the optimal proliferation conditions of loose callus were 1.10 mg·L^(-1)6-BA+0.25 mg·L^(-1)NAA+0.95 mg·L^(-1)2,4-D.Under these conditions,the callus was mainly yellowish green,and showed a relatively loose state,the highest proliferation coefficient was 9.34,and the contents of total fla-vonoids and total phenols were 25.07 mg·g^(-1)and 27.44 mg·g^(-1),respectively.

关 键 词：粉葛 松散型愈伤组织 外源激素 增殖 次生代谢产物 响应面法 

分 类 号：S567.239[农业科学—中草药栽培]