脱落酸与乙烯协同调控薄皮甜瓜果实软化的机制

doi:10.13560/j.cnki.biotech.bull.1985.2025-0535

摘要/Abstract

摘要：

目的探究外源脱落酸（ABA）及其合成抑制剂NDGA对‘绿美人’薄皮甜瓜采后软化的调控机制，重点阐明ABA通过乙烯信号通路及ASR转录因子的层级调控网络，解析ABA与乙烯协同、独立调控果实软化的时空模式，为靶向保鲜技术的开发提供理论依据。方法以薄皮甜瓜‘绿美人’为材料，外源注射600 μmol/L ABA或50 μmol/L NDGA，测定贮藏期间果实硬度、呼吸速率、乙烯释放量及关键酶活性，结合转录组测序分析差异表达基因（DEGs），并通过qPCR验证ABA和乙烯相关基因的时序表达特征。结果 ABA处理诱导果实提前软化，早期短暂抑制PG活性，激活β-Gal，后期显著提升PG活性、乙烯释放量及呼吸跃变峰值。NDGA有效延缓软化，维持硬度，抑制乙烯释放及PG活性高峰。转录组筛选到4 ABA vs 4 CK的4 014个DEGs和8 ABA vs 8 CK的5 161个DEGs，富集于植物激素信号转导（KO04075）、苯丙烷生物合成（KO00940）及MAPK通路（KO04016）。CmEIN/EIL/ERF与CmNCED/CYP707A形成共表达模块，而CmASR1与乙烯合成基因（LOC103483612）显著正相关。qPCR证实ABA诱导CmASR1/3持续高表达、CmACS1/6和CmACO3早期高表达，晚期通过CmEIN3-like1和CmERF1上调CmACO3，抑制CmCYP707A2。NDGA则激活CmASR2。 结论 ABA可能通过激活ASR家族基因诱导CmEIN3和CmACO基因时序性表达，协同调控乙烯合成与细胞壁代谢，而NDGA通过抑制CmCYP707A2延缓软化。揭示ABA-ASR-乙烯级联通路在薄皮甜瓜采后成熟中的重要作用，为靶向调控果实成熟提供理论依据。

关键词: 薄皮甜瓜, 脱落酸, 乙烯合成, 乙烯信号通路, 采后成熟, 果实软化, ASR基因表达, 转录

Abstract:

Objective To explore the regulatory mechanism of exogenous abscisic acid (ABA) and its synthesis inhibitor NDGA on post-harvest softening of ‘Green Beauty’ oriental melons (Cucumis melo var. Makuwa Makino), focusing on elucidation of ABA regulating network through the ethylene signaling pathway and ASR transcription factor hierarchical, analyzing the spatiotemporal model of ABA and ethylene synergistically or independently regulating fruit softening, thus providing a theoretical basis for the development of targeted preservation technology. Method The oriental melon ‘Green Beauty’ was used as the material, and exogenous injection of 600 μmol/L ABA or 50 μmol/L NDGA was conducted. The fruit hardness, respiration rate, ethylene release amount, and key enzyme activity were determined during storage. Differentially expressed genes (DEGs) were analyzed in combination with transcriptome sequencing, and the timing expression characteristics of ABA and ethylene-related genes were verified by qPCR. Result ABA treatment induced early softening of the fruit, briefly inhibited PG activity in the early stage, activated β-Gal, and significantly improved PG activity, ethylene release and respiratory jump transformation peak value in the later stage. NDGA effectively delayed softening, maintained hardness, and inhibited ethylene release and peak PG activity. The transcriptome was screened into 4 014 DEGs of 4 ABA vs 4 CK and 5 161 DEGs of 8 ABA vs 8 CK, and was enriched in phytohormone signal transduction (KO04075), styrene biosynthesis (KO00940), and MAPK pathway (KO04016). CmEIN/EIL/ERF formed a co-expression module with CmNCED/CYP707A, while CmASR1 was significantly and positively correlated with the ethylene synthesis gene (LOC103483612). qPCR confirmed that ABA induced continuous high expression of CmASR1/3, early high expression of CmACS1/6 and CmACO3, and in the late stage upregulation of CmACO3 through CmEIN3-like1 and CmERF1, inhibiting CmCYP707A2. NDGA activated CmASR2. Conclusion ABA may induce temporal expression of CmEIN3 and CmACO genes by activating ASR family genes, and coordinate the regulation of ethylene synthesis and cell wall metabolism, while NDGA delays softening by inhibiting CmCYP707A2. It reveals the important role of the ABA-ASR-ethylene cascade pathway in post-harvest maturity of oriental melons, and which may provide a theoretical basis for targeted regulation of fruit maturity.

Key words: oriental melon, abscisic acid, ethylene synthesis, ethylene signaling pathway, post-harvest ripening, fruit softening, ASR gene expression, transcription

陈常璐, 杨志芳, 曹嵩晓, 李杨青, 叶靖烽, 吕海艳, 陈姗姗, 陈昊. 脱落酸与乙烯协同调控薄皮甜瓜果实软化的机制[J]. 生物技术通报, 2026, 42(3): 302-311.

CHEN Chang-lu, YANG Zhi-fang, CAO Song-xiao, LI Yang-qing, YE Jing-feng, LYU Hai-yan, CHEN Shan-shan, CHEN Hao. Mechanism of Abscisic Acid and Ethylene Collaboratively Regulating the Softening of Oriental Melon Fruits[J]. Biotechnology Bulletin, 2026, 42(3): 302-311.

图/表 7

图1 ABA、NDGA处理对采后薄皮甜瓜果实横切面（A）、硬度（B）、失重率（C）、PG酶活性（D）、β-Gal（E）的影响图中误差线表示标准差。小写字母代表 P<0.05 水平下差异显著，下同

Fig. 1 Effects of ABA and NDGA treatments on cross-sectional section (A), hardness (B), weight loss rate (C), PG enzyme activity (D), and β-Gal (E) of post-harvest oriental melon fruitsThe error bars in the graph indicate the standard deviation. Lowercase letters indicate significant differences at the P<0.05 level, the same below

图2 ABA、NDGA处理对采后薄皮甜瓜果实乙烯释放量（A）、呼吸速率（B）、ACO酶活性（C）的影响

Fig. 2 Effects of ABA and NDGA treatments on ethylene release (A), respiration rate (B) and ACO enzyme activity (C) of post-harvest oriental melon fruits

图3 样品间差异表达基因韦恩图、基因表达火山图和差异基因KEGG通路富集分析A：韦恩图为总DEGs；B：8 ABA与4 ABA的差异表达火山图；ABA代表脱落酸处理组、4和8代表处理后贮藏天数；C：脱落酸处理4 d和对照4 d（4 ABA vs 4 CK）

Fig. 3 Venn diagram, volcano plot of gene expression, and KEGG pathway enrichment analysis of differentially expressed genes between samplesA: Venn diagram is the total DEGs. B: Volcano plot of differential expression of 8ABA vs. 4ABA. ABA refers to abscisic acid treatment group, 4 and 8 refers to storage days after treatment. C: Abscisic acid treatment for 4 d and control for 4 d (4ABA vs 4CK)

图4 4 d和8 d CK及ABA处理中脱落酸、乙烯和软化相关差异表达基因聚类热图

Fig. 4 Cluster heat maps of abscisic acid, ethylene and softening-related differentially expressed genes in 4 d and 8 d CK and ABA treatments

图5 ABA、NDGA处理对采后薄皮甜瓜果实ABA相关基因的时空表达分析

Fig. 5 Spatiotemporal expression analysis of ABA-related genes in post-harvest oriental melon fruit by ABA and NDGA treatments

图6 ABA、NDGA处理对采后薄皮甜瓜果实乙烯相关基因的时空表达分析

Fig. 6 Spatiotemporal expression analysis of ABA and NDGA treatments on ethylene-related genes in post-harvest oriental melon fruit

图7 ABA、NDGA处理对采后薄皮甜瓜果实ABA、乙烯相关基因主成分得分图编号1-45依次代表ABA处理组、NDGA处理组及CK中15个差异表达基因的3次生物学重复样本

Fig. 7 Principal component scores of ABA and ethylene-related genes in post-harvest oriental melon treated with ABA and NDGANumber 1-45 refers to three biological replicates of 15 differentially expressed genes in ABA-treated group, NDGA group, and CK

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