细胞壁降解酶基因PECTINESTERASE的可变剪接影响葡萄采后软化进程

doi:10.1016/j.jia.2023.11.023

Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (3): 863-875.DOI: 10.1016/j.jia.2023.11.023

细胞壁降解酶基因PECTINESTERASE的可变剪接影响葡萄采后软化进程

收稿日期:2023-04-17 接受日期:2023-10-07 出版日期:2024-03-20 发布日期:2024-03-02

Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape

Hainan Liu^{1, 2*}, Maosong Pei^{1, 2*}, Charles Ampomah-Dwamena³, Yaxin Shang^{1, 2}, Yihe Yu^{1, 2}, Tonglu Wei^{1, 2}, Qiaofang Shi^{1, 2}, Dalong Guo^{1, 2#}

¹College of Horticulture and Plant Protection, Henan University of Science and Technology, Luoyang 471000, China
²Henan Engineering Technology Research Center of Quality Regulation and Control of Horticultural Plants, Luoyang 471000, China
³The New Zealand Institute for Plant & Food Research Limited (PFR), Auckland 1010, New Zealand

Received:2023-04-17 Accepted:2023-10-07 Online:2024-03-20 Published:2024-03-02
About author:Hainan Liu, E-mail: liuhainan0995@163.com; Maosong Pei, E-mail: peimaosong@163.com; #Correspondence Dalong Guo, E-mail: guodalong@haust.edu.cn * These authors contributed equally to this study.
Supported by:
This work was financially supported by the National Natural Science Foundation of China (32202560 and 32302470), the Program for Innovative Research Team (in Science and Technology) in University of Henan Province, China (21IRTSTHN021), the Natural Science Foundation of Henan, China (232300421112), the Program for Science & Technology Innovation Talents in Universities of Henan Province, China (21HASTIT035), and the PhD Research Startup Foundation of Henan University of Science and Technology, China (13480068 and 13480067).

摘要/Abstract

摘要：

果实硬度是鲜食葡萄最重要的品质性状之一，但果实采后软化分子机制尚不完全清楚。为进一步解析采后软化调控机制，我们对采后‘巨峰’葡萄进行叶酸（Folic Acid，FA）处理（以水处理为对照，CK），通过比较各处理组间果实的转录组数据，筛选出差异表达基因（Differentially Expressed Genes，DEGs），并鉴定采后软化相关可变剪接（Alternative splicing，AS）事件及相关差异表达基因。共鉴定出2559个差异表达基因，根据表达模式将其分为四个亚簇，其中亚簇4中的差异表达基因在CK组中高表达。FA和CK处理组果实中特异性AS相关基因数分别为1045个和1042个。GO注释结果表明，CK处理组果实中的AS相关基因主要富集在细胞壁代谢过程中，尤其是细胞壁降解过程。通过比较AS相关基因与亚簇4的差异表达基因，筛选出了8个发生AS事件的差异表达基因，其中包括一个编码细胞壁降解酶的基因（果胶酯酶2、VvPE2、Vitvi15g00704），分析结果显示该基因可能发生了A3SS事件。RT-PCR进一步证实在FA处理组的葡萄果实中存在较高比例的VvPE2的截短转录本变体。本研究运用转录组测序技术分析葡萄采后果实的AS事件，并结合实验验证进一步明确细胞壁降解酶基因的AS事件在葡萄采后果实软化调控中发挥了重要作用。

Abstract:

The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes (DEGs) by comparing the transcriptomes of folic acid (FA)-treated and water-treated (CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced (AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology (GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes, particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2 (VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries. Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.

Key words: Grape , Postharvest softening , Folic Acid , Alternative splicing , Pectinesterase 2 , Alternative 3′ splice site (A3SS)

. 细胞壁降解酶基因PECTINESTERASE的可变剪接影响葡萄采后软化进程[J]. Journal of Integrative Agriculture, 2024, 23(3): 863-875.

Hainan Liu, Maosong Pei, Charles Ampomah-Dwamena, Yaxin Shang, Yihe Yu, Tonglu Wei, Qiaofang Shi, Dalong Guo.

Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape [J]. Journal of Integrative Agriculture, 2024, 23(3): 863-875.

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细胞壁降解酶基因PECTINESTERASE的可变剪接影响葡萄采后软化进程

Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape

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