DNA methylome analysis reveals novel insights into active hypomethylated regulatory mechanisms of temperature-dependent flower opening in Osmanthus fragrans

IF 8.7 1区 农林科学 Q1 Agricultural and Biological Sciences
Shiwei Zhong, Huijun Zhu, Wenle Li, Dan Wu, Yunfeng Miao, Bin Dong, Yiguang Wang, Zhen Xiao, Qiu Fang, Jinping Deng, Hongbo Zhao
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Abstract

s Short-term ambient low temperature (ALT) stimulation is necessary for Osmanthus fragrans to facilitate continued flower opening after floral bud development reaches maturity. DNA methylation, a vital epigenetic modification, regulates various biological processes in response to temperature fluctuations. However, its role in temperature-driven flower opening remains elusive. In this study, we identified the pivotal timeframe during which O. fragrans promptly detected temperature cues. Using whole-genome bisulfite sequencing (WGBS), we explored global DNA hypomethylation during this phase, with the most significant changes occurring in CHH sequence contexts. Auxin transport inhibitor (TIBA) application revealed that ALT-induced endogenous auxin accumulation promoted peduncle elongation. In our mRNA-seq analysis, we discovered the differentially expressed genes (DEGs) with hypo-differentially methylated regions (hypo-DMRs) were mainly enriched in auxin and temperature response, RNA processing, carbohydrate and lipid metabolism. The transcripts of three DNA demethylase genes (OfROS1a, OfDML3, OfDME) showed upregulation. Furthermore, all DNA methylase genes, except for OfCMT2b, also displayed increased expression, specifically with two of them, OfCMT3a and OfCMT1, being associated with hypo-DMRs. Promoter assays showed that OfROS1a, with promoters containing low temperature and auxin responsive elements, were activated by ALT and exogenous IAA at low concentrations but inhibited at high concentrations. Overexpression OfROS1 reduced endogenous auxin levels but enhanced the expression of genes related to auxin response and spliceosome in petunia. Furthermore, OfROS1 promoted sucrose synthesis in petunia corollas. Our data characterized the rapid response of active DNA hypomethylation to ALT and suggested a possible epiregulation of temperature-dependent flower opening in O. fragrans.
DNA 甲基组分析揭示了桂花随温度开放的活性低甲基化调控机制的新见解
s 短期环境低温(ALT)刺激是桂花在花芽发育成熟后继续开花所必需的。DNA 甲基化是一种重要的表观遗传修饰,可调节各种生物过程以应对温度波动。然而,它在温度驱动的花朵开放过程中的作用仍然难以捉摸。在这项研究中,我们确定了芳香木迅速检测温度线索的关键时间段。利用全基因组亚硫酸氢盐测序(WGBS),我们探索了这一阶段的全球DNA低甲基化情况,其中最显著的变化发生在CHH序列上下文中。应用叶绿素转运抑制剂(TIBA)发现,ALT 诱导的内源叶绿素积累促进了花序梗的伸长。在mRNA-seq分析中,我们发现具有低差异甲基化区域(hypo-DMRs)的差异表达基因(DEGs)主要富集在辅助素和温度响应、RNA加工、碳水化合物和脂质代谢等领域。三个 DNA 去甲基化酶基因(OfROS1a、OfDML3 和 OfDME)的转录本出现上调。此外,除 OfCMT2b 外,所有 DNA 甲基化酶基因的表达量都有所增加,其中两个基因(OfCMT3a 和 OfCMT1)与低 DMRs 有关。启动子分析表明,OfROS1a 的启动子含有低温和叶绿素反应元件,在低浓度时会被 ALT 和外源 IAA 激活,但在高浓度时会被抑制。过量表达 OfROS1 会降低矮牵牛的内源辅素水平,但会增强与辅素响应和剪接体相关的基因的表达。此外,OfROS1 还能促进矮牵牛花冠中蔗糖的合成。我们的数据描述了活性 DNA 低甲基化对 ALT 的快速反应,并提出了对 O. fragrans 花朵开放的温度依赖性的可能外显调控。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Horticulture Research
Horticulture Research Biochemistry, Genetics and Molecular Biology-Biochemistry
CiteScore
11.20
自引率
6.90%
发文量
367
审稿时长
20 weeks
期刊介绍: Horticulture Research, an open access journal affiliated with Nanjing Agricultural University, has achieved the prestigious ranking of number one in the Horticulture category of the Journal Citation Reports ™ from Clarivate, 2022. As a leading publication in the field, the journal is dedicated to disseminating original research articles, comprehensive reviews, insightful perspectives, thought-provoking comments, and valuable correspondence articles and letters to the editor. Its scope encompasses all vital aspects of horticultural plants and disciplines, such as biotechnology, breeding, cellular and molecular biology, evolution, genetics, inter-species interactions, physiology, and the origination and domestication of crops.
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