转录组和代谢组联合解析‘泽州芹菜’露地越冬的机制

doi:10.13560/j.cnki.biotech.bull.1985.2025-1038

摘要/Abstract

摘要：

目的解析‘泽州芹菜’露地越冬的分子调控机制，为珍贵种质资源利用与作物抗寒分子育种提供理论支撑。方法以幼苗期‘多利王芹菜’（CRY）为对照，分析不同品种以及‘泽州芹菜’幼苗期（ZRY）和开花期（ZR）不同时期根部的差异代谢物和差异表达基因。结果代谢组学分析发现，ZRY vs CRY差异显著代谢物有57种；ZR vs ZRY差异显著代谢物有187种，进一步筛选抗寒差异代谢物，共发现9种，包括苯丙素类和聚酮类、脂质和类脂分子、有机含氧化合物等，且均上调。转录组学分析发现，ZRY vs CRY筛选到2 988个差异表达基因，其中1 001个上调表达，1 987个下调表达；ZR vs ZRY共筛选到8 869个差异表达基因，其中有4 153个上调表达，4 716个下调表达，这些差异表达基因主要富集在植物激素信号转导等通路。‘泽州芹菜’露地越冬后，差异代谢物水杨酸含量增加，分析水杨酸合成路径，发现Ag2G02930在水杨酸的合成中发挥重要作用。结论 ‘泽州芹菜’通过积累水杨酸等关键代谢物，并激活植物激素信号转导通路抵御低温胁迫，水杨酸合成相关基因Ag2G02930对其露地越冬抗寒性起核心调控作用。

关键词: 泽州芹菜, 露地越冬, 代谢组, 转录组, 水杨酸

Abstract:

Objective To elucidate the molecular regulatory mechanisms underlying the overwintering of ‘Zezhou’ celery in open fields, providing theoretical support for utilization of valuable germplasm resource and molecular breeding for cold tolerance in crops. Method Using ‘Duoliwang’ celery (CRY) at the seedling stage as the control, differentially accumulated metabolites and differentially expressed genes in the roots were analyzed between different varieties and different stages (seedling stage ZRY and flowering stage ZR) of ‘Zezhou’ celery. Result Metabolomic analysis revealed 57 significantly differentially accumulated metabolites in ZRY vs. CRY, and 187 in ZR vs. ZRY. Further screening identified 9 cold-tolerance-related differential metabolites, including phenylpropanoids and polyketides, lipids and lipid-like molecules, and organo-oxygen compounds, all of which showed increased accumulation. Transcriptomic analysis identified 2 988 differentially expressed genes in ZRY vs. CRY, with 1 001 upregulated and 1 987 downregulated. In ZR vs. ZRY, 8 869 differentially expressed genes were identified, comprising 4 153 upregulated and 4 716 downregulated genes. These genes were primarily enriched in pathways such as plant hormone signal transduction. Following overwintering in open fields, the content of the differential metabolite salicylic acid increased in ‘Zezhou’ celery. Analysis of the salicylic acid biosynthesis pathway indicated that the gene Ag2G02930 plays a significant role in its synthesis. Conclusion ‘Zezhou Celery’ resists cold stress by accumulating key metabolites such as salicylic acid and activating plant hormone signal transduction pathways. The salicylic acid synthesis-related gene Ag2G02930 plays a core regulatory role in its resistance to cold during overwintering in the open field.

Key words: Zezhou celery, overwintering in open-field, metabolome, transcriptome, salicylic acid

申瑞雪, 郭子卿, 庞星辰, 李志远, 丁炜, 董琦, 李森, 宋红霞. 转录组和代谢组联合解析‘泽州芹菜’露地越冬的机制[J]. 生物技术通报, 2026, 42(8): 251-262.

SHEN Rui-xue, GUO Zi-qing, PANG Xing-chen, LI Zhi-yuan, DING Wei, DONG Qi, Li Sen, SONG Hong-xia. Combined Transcriptome and Metabolome Analysis Elucidates the Mechanism of Open-field Overwintering in ‘Zezhou’ Celery[J]. Biotechnology Bulletin, 2026, 42(8): 251-262.

图/表 10

表1 ZRY vs CRY、ZR vs ZRY抗寒差异代谢物

Table 1 ZRY vs CRY, ZR vs ZRY cold resistance differential metabolites

比较的样品对 The sample pairs for comparison	代谢物名称 Metabolite name	类型 Class	Up/down
ZRY vs CRY	5-乙酰水杨酸 5-Acetylsalicylic acid	苯丙素类和聚酮类 Phenylpropanoids and polyketides	Up
	矢车菊-3-芸香苷 Cyanidin-3-rutinoside	苯丙素类和聚酮类 Phenylpropanoids and polyketides	Up
	γ-亚麻酸 Gamma-linolenic acid	脂质和类脂分子 Lipids and lipid-like molecules	Up
ZR vs ZRY	水杨酸 Salicylic acid	苯环型化合物 Benzenoids	Up
	酪醇 Tyrosol	苯环型化合物 Benzenoids	Up
	反式玉米素9-O-葡萄糖苷 trans-zeatin 9-O-glucoside	/	Up
	景天庚酮糖-7-磷酸 s7p	有机含氧化合物 Organic oxygen compounds	Up
	D-（-）-甘露醇 D-（-）-Mannitol	有机含氧化合物 Organic oxygen compounds	Up
	D-脯氨酸 D-Proline	有机酸及其衍生物 Organic acids and derivatives	Up

图1 正离子（pos）和负离子（neg）模式下样本的PCA分析

Fig. 1 PCA analysis of samples in positive ion (pos) and negative ion (neg) modes

图2 正离子（pos）和负离子（neg）模式下样本差异代谢物聚类热图

Fig. 2 Cluster heat map of differential metabolites in positive ion (pos) and negative ion (neg) modes

图3 样本间相关性热图

Fig. 3 Correlation heat map between samples

图4 ZRY vs CRY和ZR vs ZRY差异表达基因维恩图（A、B）以及火山图（C、D）

Fig. 4 Differential expression gene Venn diagram (A, B) and volcano map (C, D) of ZRY vs CRY and ZR vs ZRY

图5 ZRY vs CRY（A、C）、ZR vs ZRY（B、D）差异表达基因的GO富集分析和KEGG通路分析横坐标为注释到KEGG通路上的差异基因数与差异基因总数的比值，纵坐标为KEGG通路，点的大小代表注释到KEGG通路上基因数的多少，颜色从红到紫代表富集显著性逐渐增加

Fig. 5 GO enrichment analysis and KEGG pathway analysis of differentially expressed genes in ZRY vs CRY (A, C) and ZR vs ZRY (B, D)The horizontal axis indicates the ratio of the number of differentially expressed genes annotated to the KEGG pathway to the total number of differentially expressed genes. The vertical axis indicates the KEGG pathways. The size of the points indicates the number of genes annotated to the corresponding KEGG pathway, while the color gradient from red to purple reflects increasingly significant enrichment

图6 ZR vs ZRY苯丙氨酸代谢途径中差异表达基因与差异代谢物苯丙氨酸代谢途径中蓝色代表下调，红色代表上调。转录组热图表示基因在不同样品中的表达水平（FPKM），代谢组热图表示代谢物在不同样品中的积累量

Fig. 6 Differentially expressed genes and differentially accumulated metabolites in the phenylpropanoid biosynthesis pathway in ZR vs ZRYIn the phenylalanine metabolic pathway, blue indicates down-regulated and red indicates up-regulated. The transcriptome heat map indicates the expressions of the gene in different samples (FPKM), and the metabolome heat map indicates the accumulation of metabolites in different samples

图7 苯丙氨酸代谢途径中差异表达基因与差异代谢物的相关性分析（A）以及水杨酸合成路径和相关差异表达基因热图（B）在相关性分析中，红色表示正相关，蓝色表示负相关；在热图中，红色表示上调表达，蓝色表示下调表达

Fig. 7 Correlation analysis between differentially expressed genes and differentially accumulated metabolites in the phenylpropanoid biosynthesis pathway in (A), and salicylic acid synthesis pathway and related differentially expressed gene heat map (B)In correlation analysis, red indicates positive correlation and blue indicates negative correlation. In the heat map, red indicates up-regulated expression and blue indicates down-regulated expression. A: ZR vs ZRY; B: ZRY vs CRY, ZR vs ZRY

图8 水杨酸信号转导途径及相关差异基因表达热图在水杨酸信号转导途径中方框表示酶，圆表示代谢物，红色代表上调，绿色代表下调，黄色代表上调和下调；在差异基因表达热图中，蓝色代表下调，红色代表上调

Fig. 8 Salicylic acid signal transduction pathway and related differential gene expression heat mapIn the salicylic acid signaling pathway, boxes indicate enzymes, circles indicate metabolites, red indicates up-regulation, green indicates down-regulation, and yellow indicates both up- and down-regulation. In the heatmap of differentially expressed genes, blue indicates down-regulation, and red indicates up-regulation

图9 芹菜根系中差异表达基因RT-qPCR验证

Fig. 9 RT-qPCR validation of differentially expressed genes in celery (Apium graveolens L.) roots

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