茅苍术试管苗继代培养的ISSR及MSAP分析  

Test-tube Seedlings of Atractylodes lancea(Thunb.) DC: ISSR and MSAP Analysis

作　　者：张成才 向增旭[1] Zhang Chengcai;Xiang Zengxu(College of Horticulture,Nanjing Agricultural University,Nanjing 210095)

机构地区：[1]南京农业大学园艺学院,南京210095

出　　处：《中国农学通报》2021年第12期72-78,共7页Chinese Agricultural Science Bulletin

基　　金：现代农业产业技术体系建设专项资金“国家中药材产业技术体系”(CARS-21)。

摘　　要：为探究茅苍术试管苗在长期继代培养中的遗传稳定性与DNA甲基化变化,保证工厂化生产种苗品质。以茅苍术幼嫩顶芽为初始材料,建立高效离体快繁体系进行长期继代培养,并运用ISSR、MSAP分子标记技术对不同继代次数的试管苗进行分析试验。结果显示不同继代次数的10个样本遗传多样性较低,具有遗传稳定性较高。其中,第1代与第10代样本之间遗传相似系数最低,亲缘关系较远;从继代第4代开始,不同引物扩增图谱条带在个别位点发生变化,且随着继代次数增加变化愈明显。经过长期继代培养后,茅苍术试管苗的甲基化水平有所下降,去甲基化模式和甲基化模式并存,但以去甲基化模式为主。长期继代培养后出现变异现象可能是培养条件、培养时间的不同导致基因被重新活化和表达或抑制、去甲基化模式高于甲基化模式造成的。本研究结果为茅苍术种质资源保存和工厂化生产提供了一定的理论依据。The purpose of this study is to explore the genetic stability and DNA methylation changes of test tube plantlets of Atractylodes lancea(Thunb.) DC in long-term subculture, so as to ensure the quality of plantlets produced in factory. We established an efficient in vitro rapid propagation system for long-term subculture with young terminal buds of Atractylodes lancea and analyzed the test tube plantlets with different subculture times by ISSR and MSAP. The results showed that 10 samples with different generations had low genetic diversity and high genetic stability. Among them, the genetic similarity coefficient between the first generation and the tenth generation samples was the lowest, and the genetic relationship was far away. From the fourth generation of subculture, the bands of different primer amplification patterns changed in individual loci,and the change was more obvious with the increase of subculture times. After long-term subculture, the methylation level of test tube plantlet of A. lancea decreased, and the demethylation mode and methylation mode coexisted, but the demethylation mode was the main mode. The mutation phenomenon after long-term subculture might be caused by different culture conditions and culture time, resulting in gene reactivation and expression or inhibition, and demethylation mode was higher than methylation mode. The results of this study provide a theoretical basis for germplasm conservation and industrial production of A. lancea.

关 键 词：茅苍术 继代培养 简单序列间重复 甲基化敏感扩增多态性 DNA甲基化 

分 类 号：S567.23[农业科学—中草药栽培]