《植物生理学报》 2009, 45(6): 544-548

红豆杉脱分化过程中的遗传和表观遗传变异

李丽琴, 付春华, 赵春芳, 吴文娟, 余龙江*

华中科技大学生命科学与技术学院, 武汉430074

通信作者：余龙江；E-mail: yulj@hust.edu.cn；Tel: 027-87792264

摘　要：

采用扩增片段长度多态性(AFLP)和甲基化敏感扩增多态性(MSAP)技术分析红豆杉脱分化前后基因组 DNA 和 DNA 甲基化状态的变化。选用 32 个 AFLP 引物组合从红豆杉植株及其愈伤组织分别扩增出 1 834 个片段, 无多态性片段产生。 这说明红豆杉植株在诱导形成愈伤组织的过程中基因组 DNA 保持高度的遗传稳定性。另用 32 个 MSAP 引物组合从红豆 杉植株及其愈伤组织分别扩增出1 197个片段, 总扩增位点的甲基化水平由脱分化前的12.4%上升为16.2%, 表明红豆杉在 脱分化过程中的某些位点发生了甲基化。红豆杉脱分化前后的DNA甲基化模式也存在较大差异, 说明DNA甲基化对愈伤 组织形成有调控作用。

关键词：脱分化; 红豆杉; AFLP; MSAP

收稿：2009-02-11   修定：2009-05-21

资助：国家自然科学基金(20776058)和2006 年教育部新世纪优秀人才支持计划(NCET-06-0646)。

Genetic and Epigenetic Variations of Taxus media L. cv. Hicksii during Process of Dedifferentiation

LI Li-Qin, FU Chun-Hua, ZHAO Chun-Fang, WU Wen-Juan, YU Long-Jiang*

School of Biological Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China

Corresponding author: YU Long-Jiang; E-mail: yulj@hust.edu.cn; Tel: 027-87792264

Abstract:

The amplified fragment-length polymorphism (AFLP) and methylation-sensitive amplified polymorphism (MSAP) techniques were used to assess genetic and epigenetic variations of Taxus media during process of dedifferentiation. 1 834 bands were amplified from Taxus media plant and its callus with 32 pairs of AFLP primers, and no polymorphism fragments were detected. These indicated DNA genome of Taxus media kept highly genetic stability during the process of dedifferentiation. At the same time, 1 197 bands were amplified from Taxus media plant and its callus with 32 pairs of MSAP primers, methylation level rose from 12.4% to 16.2%, and epigenetic changes due to methylation were detected. What’s more, methylation patterns experienced markedly variation, which meant DNA methylation would play role during the process of dedifferentiation.

Key words: dedifferentiation; Taxus media; AFLP; MSAP