培养基类型和植物生长物质浓度对王枣子愈伤组织诱导和继代的影响  

Effects of Medium Type and Plant Growth Substance Concentration on Callus Induction and Subculture of Isodon Amethystoides

作　　者：张媛媛 朱智慧 张苗苗 苏勇 方政宇 孙伟[2] 薛建平[1] Zhang Yuanyuan;Zhu Zhihui;Zhang Miaomiao;Su Yong;Fang Zhengyu;Sun Wei;Xue Jianping(Anhui Provincial Engineering Laboratory for Efficient Utilization of Featured Resource Plants,College of Life Sciences,Huaibei Normal University,Huaibei 235000,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]淮北师范大学生命科学学院安徽省特色资源植物利用工程实验室 [2]中国中医科学院中药研究所

出　　处：《世界科学技术-中医药现代化》2020年第8期2851-2856,共6页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：安徽省教育厅高校科技平台创新团队项目(KJ2015TD001):采煤塌陷区生态修复于利用,负责人:薛建平

摘　　要：目的优化王枣子愈伤组织的诱导和探究疏松愈伤组织的继代条件,为王枣子悬浮细胞培养奠定初步基础。方法以王枣子无菌苗幼叶为外植体,采用单因素试验研究不同培养基类型对愈伤组织诱导的影响;采用L9(32)正交试验设计,研究2,4-D、KT不同浓度配比对王枣子疏松愈伤组织的增殖培养的影响。结果愈伤组织的诱导率在MS、1/2MS和B5培养基上呈上升趋势。MS的出愈时间最晚,1/2MS的出愈时间次之;在B5培养基上出愈时间最早,且愈伤组织的质地和生长状态较好。正交试验的结果显示,愈伤组织的在B5培养基上添加2,4-D 2.5 mg·L-1+KT 1.0 mg·L-1增殖倍数最小为19.78倍,添加2,4-D 1.5 mg·L-1+KT 0.2 mg·L-1达到最大增殖倍数27.34倍;王枣子疏松愈伤组织增殖试验中2,4-D(P<0.01)的影响效应比KT(P=0.091)大。结论B5培养基是最佳的王枣子愈伤组织诱导培养基,B5培养基中添加2,4-D 1.5 mg·L-1+KT 0.5 mg·L-1为疏松愈伤组织增殖的最佳配比。Objective To optimize the systems for callus induction and loose callus subculture of I.amethystoides and to lay the initial foundation for its suspension cell culture.Methods Using the young leaves of Isodon amethystoides aseptic explants as explants and the effects of different medium types(MS,1/2 MS and B5)on callus induction were studied by single factor experiment;the effect of different concentration of 2,4-D and KT on the proliferation culture of loose callus was studied by L9(32)orthogonal design.Results The induction rate of callus showed an increased trend in the three basic media of MS,1/2 MS and B5.The callus production time of MS was the last,and that of 1/2 MS was the second,while that of callus on B5 medium was the earliest,and the texture and growth state of callus were better than the other two media.The results of orthogonal experiment showed that the proliferation-fold of callus was 19.78 times on medium supplemented with 2,4 D 2.5 mg·L-1 and KT 1.0 mg·L-1 on B5 medium,while it reached the maximum multiplication rate of 27.34 times with the addition of 2,4 D 1.5 mg·L-1 and KT 0.2 mg·L-1.The effect of 2,4-D(P<0.01)in the proliferation of loose callus was greater than that of KT(P=0.091).Conclusion B5 medium is the best medium for callus induction of I.amethystoides and the B5 medium supplemented with 2,4-D 1.5 mg·L-1 and KT 0.5 mg·L-1 is the best proportion for the proliferation of loose callus.

关 键 词：王枣子 愈伤组织 诱导 继代 培养基类型 植物生长物质 

分 类 号：S567.239[农业科学—中草药栽培]