XYLEM NAC DOMAIN 1(EjXND1)通过负向调节枇杷果实中的 EjHB1-EjPRX12 模块来缓解冷诱导的木质化。

Zihao Liang, Yanna Shi, Yiqing Huang, Jiao Lu, Mengxue Zhang, Xizhi Cao, Ruoqian Hu, Dongdong Li, Wenbo Chen, Changqing Zhu, Di Wu, Kunsong Chen
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引用次数: 0

摘要

简介木质素是次生细胞壁的主要成分,在植物生长和抵御压力(如低温和虫害)方面发挥作用。此外,木质素还会在肉质果实中积累,对果实质量产生负面影响。红肉枇杷对温度敏感,会出现低温诱导的木质化现象。目前已开发出多种技术,如低温调节(LTC)处理,以缓解冷害症状:本研究旨在阐明冷诱导枇杷果实木质化的调控机制:方法:通过分析转录组分离出目标基因。方法:通过转录组分析分离出目标基因,分别在烟草叶片和枇杷果实中采用瞬时转基因方法分析基因功能,并在肝草中进行稳定过表达。通过体外蛋白-蛋白相互作用实验、双荧光素酶实验、EMSA等方法对调控机制进行了研究:结果:本研究发现木质部NAC结构域转录因子EjXND1是枇杷果实木质化的抑制因子。结果:本研究发现木质部 NAC 结构域转录因子 EjXND1 是枇杷果实木质化的抑制因子,它能与木质素激活因子 EjHB1 相互作用,导致 EjHB1 对 EjPRX12 启动子的激活作用减弱。此外,在 EjXDN1 的启动子中还发现了两个高度甲基化的区域。其中一个区域的甲基化水平与 EjXND1 的表达呈负相关。此外,研究还表明,该区域的高甲基化会削弱 EjXND1 激活因子与其启动子的结合亲和力:结论:EjXND1通过靶向EjHB1-EjPRX12模块,在改良低温调节(mLTC)处理中发挥作用,从而缓解红肉枇杷果实中由低温引起的木质化。
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XYLEM NAC DOMAIN 1 (EjXND1) relieves cold-induced lignification by negatively regulating the EjHB1-EjPRX12 module in loquat fruit.

Introduction: Lignin is a principal constituent of the secondary cell wall, which plays a role in both plant growth and defensing against stress, such as low temperature and pest infestation. Additionally, it also accumulates in fleshy fruits and negatively affects fruit quality. Red-fleshed loquat is temperature sensitive and exhibits cold-induced lignification. A number of technologies have been developed, for example, Low Temperature Conditioning (LTC) treatment, which has been applied in order to relieve the symptom of cold injury.

Objectives: The present study seeks to elucidate the regulatory mechanism underlying cold-induced lignification in loquat fruit.

Methods: The target genes were isolated through the analysis of transcriptome. The gene function was analyzed by transient transgenic method in tobacco leaves and loquat fruit, respectively, as well as stable overexpression in liverwort. The regulatory mechanism study was achieved by in vitro protein-protein interaction assays, dual-luciferase assay, and EMSA.

Results: In the present study, the Xylem NAC Domain transcription factor EjXND1 was identified as a repressor of loquat fruit lignification. It was demonstrated that EjXND1 could interact with the characterized lignin activator EjHB1, resulting in a diminution of the activation of EjHB1 on EjPRX12 promoter. Furthermore, two highly methylated regions were identified in the promoter of EjXDN1. One of these regions exhibited a negative correlation between methylation level and EjXND1 expression. Additionally, it was shown that hypermethylation of this region weaken the binding affinity of EjXND1 activators to its promoter.

Conclusion: The EjXND1 plays a role in modified Low Temperature Conditioning (mLTC) treatment that alleviates cold-induced lignification in red-fleshed loquat fruit by targeting the EjHB1-EjPRX12 module and EjXND1 is regulated by the dynamic of DNA methylation level in the promoter.

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