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Journal of Integrative Agriculture  2025, Vol. 24 Issue (9): 3418-3434    DOI: 10.1016/j.jia.2024.12.034
Horticulture Advanced Online Publication | Current Issue | Archive | Adv Search |
Functional analysis of the xyloglucan endotransglycosylase/hydrolase gene MdXTH2 in apple fruit firmness formation

Li Liu1*, Yifeng Feng2*, Ziqi Han1, Yaxiao Song1, Jianhua Guo1, Jing Yu1, Zidun Wang3, Hui Wang1, Hua Gao1, Yazhou Yang1, Yuanji Wang1#, Zhengyang Zhao1#

1 State Key Laboratory for Crop Stress Resistance and High Efficiency Production, College of Horticulture, Northwest A&F University, Yangling 712100, China

2 College of Horticulture and Forestry, Tarim University, Alaer 843300, China

3 Liaoning Institute of Pomology, Yingkou 115009, China

  Highlights 
MdXTH2 is a key gene influencing apple fruit firmness, identified through transcriptomic and metabolomic analyses.
● Overexpression of MdXTH2 increased fruit firmness, delayed fruit ripening, and inhibited calli growth in apple and tomato.
● MdNAC72 directly binds to the MdXTH2 promoter and represses its transcription, linking fruit firmness with ripening regulation.

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摘要  
苹果果实硬度作为衡量果实内在品质的重要指标之一,不仅影响果实适口性,还决定了果实贮运能力。目前果实发育过程中硬度的形成机制仍不清楚,细胞壁多糖的水解是果实发育过程中硬度下降的主要原因。木葡聚糖内糖基转移/水解酶(XTH)作为解聚细胞壁多糖的关键酶,在果实硬度形成中发挥重要作用。我们通过代谢组和转录组数据联合分析鉴定了一个与果实硬度形成相关的基因MdXTH2,并对MdXTH2在苹果果实硬度形成中的功能及分子机制进行解析。结果表明,果实硬度和细胞壁各组分含量在发育期果实中整体呈下降趋势。细胞壁物质、纤维素和半纤维素含量在不同品种中均与硬度显著正相关,总相关性系数分别为0.871、0.904和0.75,这表明,纤维素和半纤维素含量的降低可能是果实发育过程中硬度下降的普遍原因。过表达MdXTH2显著增加苹果和番茄果实的硬度,并抑制了愈伤组织的生长。MdXTH2基因的上游转录因子MdNAC72可以促进果实成熟相关基因的表达。此外,双荧光素酶、酵母单杂交和凝胶阻滞迁移试验表明,MdNAC72通过与MdXTH2启动子上的NACRS元件结合并下调MdXTH2的转录水平。本研究为果实品质调控机理提供新见解,也为通过遗传改良培育硬度适中的苹果品种提供了理论依据。


Abstract  

Apple fruit firmness is a crucial index for measuring the internal quality of apples, which influences palatability, storage capacity and transportability.  The primary cause of reduced firmness during fruit development is the hydrolysis of cell wall polysaccharides.  Xyloglucan endotransglycosylase/hydrolase (XTH) is a key enzyme involved in the depolymerization of cell wall polysaccharides, but the mechanism of its involvement in the formation of fruit firmness remains unclear.  Here, we identified the gene MdXTH2 by integrating metabolomic and transcriptomic data, and analyzed its function and molecular mechanism in the formation of apple fruit firmness.  The results showed downward trends in both fruit firmness and cell wall components throughout fruit development.  The contents of cell wall material, cellulose, and hemicellulose in various apple varieties exhibited significant positive correlations with firmness, with total correlation coefficients of 0.862, 0.884, and 0.891, respectively.  Overexpression of MdXTH2 significantly increased fruit firmness in apple and tomato, inhibited fruit ripening, and significantly suppressed the growth of calli.  The upstream transcription factor MdNAC72 of the MdXTH2 gene can promote the expression of fruit ripening-related genes.  Furthermore, dual-luciferase, yeast one-hybrid, and electrophoretic mobility shift assay (EMSA) demonstrated that MdNAC72 down-regulates the transcription of MdXTH2 by binding to its promoter.  In summary, the results of this study provide a strategy for examining fruit quality regulation and a theoretical basis for breeding apple varieties with moderate firmness through genetic improvement.


Keywords:  apple       flesh firmness        MdXTH2        transcriptional regulation  
Received: 27 May 2024   Online: 30 December 2024   Accepted: 28 September 2024
Fund: 

This study was supported by the National Natural Science Foundation of China (32302683), the China Postdoctoral Science Foundation (2024M752636), the Earmarked Fund for the China Agriculture Research System (CARS-27), and the Project of Weinan Experimental Demonstration Station of Northwest A&F University, China (2024WNXNZX-1).  

About author:  Li Liu, E-mail: li91761092@163.com, liulily@nwafu.edu.cn; Yifeng Feng, E-mail: f15909073303@163.com; #Correspondence Zhengyang Zhao, E-mail: zhaozy@nwsuaf.edu.cn; Yuanji Wang, E-mail: wangyuanji1019@163.com * These authors contributed equally to this study.

Cite this article: 

Li Liu, Yifeng Feng, Ziqi Han, Yaxiao Song, Jianhua Guo, Jing Yu, Zidun Wang, Hui Wang, Hua Gao, Yazhou Yang, Yuanji Wang, Zhengyang Zhao. 2025. Functional analysis of the xyloglucan endotransglycosylase/hydrolase gene MdXTH2 in apple fruit firmness formation. Journal of Integrative Agriculture, 24(9): 3418-3434.

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