番茄JMJ524基因调控SlGLD1的DNA甲基化分析

生物技术通报 ›› 2016, Vol. 32 ›› Issue (12): 79-85.

番茄JMJ524基因调控SlGLD1的DNA甲基化分析

杨伟,潘宇,苏承刚,张兴国,李金华

西南大学园艺园林学院南方山地园艺学重点实验室，重庆 400715

收稿日期:2016-05-12 出版日期:2016-12-25 发布日期:2016-12-07
作者简介:杨伟，男，硕士研究生，研究方向：蔬菜分子生物学与基因工程；E-mail：shucaiyuanyi@163.com
基金资助:
国家自然科学基金项目（31301779），重庆市基础与前沿研究计划一般项目（cstc2015jcyjA80030），中央高校基本业务费专项基金项目（XDJK2014B045，XDJK2016E148）

Analysis of the DNA Methylation of SlGLD1 Regulated by JMJ524 in Tomato

YANG Wei,PAN Yu,SU Cheng-gang,ZHANG Xing-guo,LI Jin-hua

College of Horticulture and Landscape Architecture，Southwestern University，Key Laboratory of Horticultural Science in Southern Mountainous Region，Chongqing 400715

Received:2016-05-12 Published:2016-12-25 Online:2016-12-07

摘要/Abstract

摘要： 旨在分析番茄JMJ524基因如何通过影响SlGLD1 的DNA甲基化程度来调控其表达，从而导致番茄的赤霉素不敏感的矮化。利用生物信息学和BSP（Bisulfite sequencing PCR）分析JMJ524抑制系和野生型的SlGLD1的DNA甲基化程度。结果显示，分离出SlGLD1的启动子序列，发现其启动子区域含有3个GA响应的顺式作用元件；SlGLD1的启动子区域和编码区的DNA是高度甲基化的，而且在不同组织部位和突变体中的甲基化模式是不一样的；在基因的编码区发现了两个GC岛，推测SlGLD1的甲基化可能发生在基因的编码区；针对这两个GC岛进行的BSP分析结果表明：SlGLD1的这两个区段的DNA CpG是高度甲基化的，而且在JMJ524抑制系中，DNA的整体甲基化，特别是CpG甲基化程度显著低于野生型对照M82。JMJ524抑制表达导致了SlGLD1的基因甲基化程度降低，从而激活这个基因的表达，可能是导致番茄植株矮化的一个因素。

关键词: JmjC, GLD1, DNA甲基化, 番茄

Abstract: This work is to reveal the mechanism of JMJ524 modulatinges the SlGLD1 expression by affecting the DNA methylation of SlGLD1，and leading to the dwarf of tomato insensitive to gibberellin. Bioinformatics and BSP（Bisulfite sequencing PCR）were employed to analyze the DNA methylation of SlGLD1 in inhibitory line and wild line of JMJ524. Results showed that the SlGLD1 promoter sequence was isolated，which contained 3 GA response cis-elements. The DNAs of SlGLD1 promoter and coding sequence were highly methylated，moreover，the methylation pattern varied at the different tissues and mutants. Two GC islands were found in this coding region of the gene，thus it was inferred that the methylation of SlGLD1 might happen in the region. Further，the analysis by BSP of the 2 GC islands indicated that the DNA CpG in the 2 regions of SlGLD1 was highly methylated. The whole methylation of DNA in the inhibitory line of JMJ524，especially CpG methylation was significantly lower than wild-type M82. The inhibited expression by JMJ524 resulted in the decrease of gene methylation of SlGLD1，therefore the expression of the gene was activated，which might be the one of factors causing the dwarf of tomato plant.

Key words: JmjC, GLD1, DNA methylation, tomato

中图分类号:

10.13560/j.cnki.biotech.bull.1985.2016.12.014

杨伟,潘宇,苏承刚,张兴国,李金华. 番茄JMJ524基因调控SlGLD1的DNA甲基化分析[J]. 生物技术通报, 2016, 32(12): 79-85.

YANG Wei,PAN Yu,SU Cheng-gang,ZHANG Xing-guo,LI Jin-hua. Analysis of the DNA Methylation of SlGLD1 Regulated by JMJ524 in Tomato[J]. Biotechnology Bulletin, 2016, 32(12): 79-85.

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