XYLEM NAC DOMAIN 1 (EjXND1) relieves cold-induced lignification by negatively regulating the EjHB1-EjPRX12 module in loquat fruit,Journal of Advanced Research

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XYLEM NAC DOMAIN 1 (EjXND1) relieves cold-induced lignification by negatively regulating the EjHB1-EjPRX12 module in loquat fruit
Journal of Advanced Research ( IF 11.4 ) Pub Date : 2024-09-02 , DOI: 10.1016/j.jare.2024.08.032
Zihao Liang ₁ , Yanna Shi ₂ , Yiqing Huang ₁ , Jiao Lu ₁ , Mengxue Zhang ₁ , Xizhi Cao ₁ , Ruoqian Hu ₁ , Dongdong Li ₂ , Wenbo Chen ₂ , Changqing Zhu ₂ , Di Wu ₂ , Kunsong Chen ₂

Affiliation

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.

中文翻译：

木质部 NAC 结构域 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

介绍

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）中发挥作用。此外，它还会积聚在肉质水果中，对水果品质产生负面影响。红肉枇杷对温度敏感，并表现出冷诱导的木质化。已经开发了许多技术，例如低温调节（LTC）治疗，该治疗已被应用于缓解冷损伤的症状。

目标

本研究旨在阐明枇杷果实中低温诱导木质化的调控机制。

方法

通过转录组分析分离靶基因。采用瞬时转基因法分别在烟叶和枇杷果实中分析基因功能，在地钱中稳定过表达。通过体外蛋白质-蛋白质相互作用测定、双荧光素酶测定和 EMSA 实现调节机制研究。

结果

在本研究中，木质部 NAC 结构域转录因子 EjXND1 被鉴定为枇杷果实木质素化的抑制因子。研究表明 EjXND1 可以与表征的木质素激活剂 EjHB1 相互作用，导致 EjPRX12 启动子上 EjHB1 的激活减少。此外，在 EjXDN1 的启动子中鉴定出两个高度甲基化区域。其中一个区域在甲基化水平和 EjXND1 表达之间表现出负相关。此外，研究表明该区域的高甲基化削弱了 EjXND1 激活剂与其启动子的结合亲和力。