光暗条件下大蒜DNA甲基化差异的初步的研究何艳霞.pdfVIP

植物生理学通讯 第43卷 第1期，2007年2月 8 5 光暗条件下大蒜DNA甲基化差异的初步研究 * 何艳霞，王子成 ，曹红平，张成婉 河南大学农业生物技术研究所，河南开封475001 提要：用甲基敏感扩增多态性(MSAP)技术分析光暗条件下生长的大蒜DNA甲基化水平差异。结果表明，用 8对引物组 合扩增出343条带，完全一致的带型240条，不一致的103条。在不一致的条带中，光照条件下相同的47条带与黑暗条件 下相同的带型有差异，与黑暗相比光照下出现的去甲基化带28条，有56条差异带在2个处理内个体间也有差异。总的趋 势是光照引起大蒜DNA去甲基化。 关键词：DNA甲基化；甲基敏感扩增多态性；大蒜 Initial Study of DNA Methylation Difference of Garlic (A llium sativum L.) Growing in Light and Darkness * HE Yan-Xia, WANG Zi-Cheng , CAO Hong-Ping, ZHANG Cheng-Wan Institute of Agricultural Biotechnology, H enan University, Kaifeing, H enan 475001, China Abstract: The variation patterns of DNA methylation in Garlic (A llium sativum L.) genome under the condition of light and darkness was studied using MSAP technique to reveal the effect of light to the level of plant DNA methylation. Total of 343 fragments were identified by eight primer combinations and of which 240 were shared by all the samples in the two conditions. It was shown that 47 consistent fragments in light condition appeared different comparing with those in darkness. Comparing with the darkness condition, the 28 fragments appeared to be demethylated in light condition, and 56 fragments appeared to be varied among individuals in the same condition. The overall results showed that light could induce DNA demethylation in Garlic. Key words: DNA methylation; methylation sensitive amplified polymorphism; garlic DNA甲基化在高等生物的生命活动中具有维 甲基敏感扩增多态性 持基因组稳定及调节生长发育等功能(Chan等 amplified polymorphism，MSAP)技术是基于扩增 2005)。DNA甲基化程度与一系列的生物过程相 片段长度多态性(AFLP)技术，以对甲基化敏感的 关，如基因组稳定、X染色体失活、基因的转 酶HapII和MspI代替MseI作高频内切酶，以EcoRI 录调节和转座子元件的活性、基因沉默、基因组 为低频内切酶进行AFLP反应。这种方法适用于 印迹等(Bird和Wolffe 1999；Besyor 2000；Bender 检测材料DNA序列未发生变化情况下的DNA甲基 2004；Gehring等2004；Wassenegger 2002)。作 化水平变化。 为一种表观遗传修饰，DNA甲基化在不改变碱基 本文以遗传基础一致、光和暗条件下