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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (9): 2295-2306.doi: 10.3724/SP.J.1006.2025.53016

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Effects of waterlogging stress on root metabolism of maize seedlings

JIANG Huan-Qi1,2(), DUAN Ao1, GUO Chao1, HUANG Xiao-Meng1, AI De-Jun1, LIU Xiao-Xue1, TAN Jing-Yi1, PENG Cheng-Lin2, LI Man-Fei1,*(), DU He-Wei1,*()   

  1. 1College of Life Sciences, Yangtze University, Jingzhou 434025, Hubei, China
    2Plant Protection and Soil Fertilizer Institute, Hubei Academy of Agricultural Sciences / National Soil Quality Hongshan Observation and Experimental Station, Wuhan 430064, Hubei, China
  • Received:2025-03-02 Accepted:2025-06-01 Online:2025-09-12 Published:2025-06-10
  • Contact: *E-mail: mfli_maize@163.com; E-mail : duhewei666@163.com
  • Supported by:
    National Natural Science Foundation of China(32072069);National Natural Science Foundation of China for Young Scientists(32301910);Hubei Provincial Natural Science Foundation Innovation Group Project(2022CFA030);Yangtze University College Students’ Innovation and Entrepreneurship Project(Yz2024251)

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Abstract:

To investigate key metabolites and changes in metabolic pathways in maize (Zea mays L.) seedlings under waterlogging stress, plants were subjected to four stress durations (0, 1, 4, and 7 days). Transcriptome sequencing (RNA-seq) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) were employed to identify differentially accumulated metabolites, followed by KEGG enrichment analysis to explore associated metabolic pathways. A total of 1361 differential metabolites were annotated and quantified. The most pronounced metabolic changes were observed at 4 days, with 414 metabolites significantly altered—372 upregulated and 42 downregulated. The top 20 most differentially accumulated metabolites were all upregulated, including ferulic acid-4-O-glucoside, 2-phenylethanol, 7-methyl-ergochalcone, and S-allyl-L-cysteine. KEGG pathway analysis revealed significant enrichment in flavonoid biosynthesis, carotenoid biosynthesis, phenylpropanoid biosynthesis, plant hormone signaling, ABC transporters, fatty acid degradation, starch and sucrose metabolism, glycine, serine and threonine metabolism, tryptophan metabolism, and glycolysis/gluconeogenesis. Among these, flavonoid biosynthesis was found to be closely associated with waterlogging stress, with key flavonoids such as naringenin and luteolin, and enzymes including chalcone isomerase, flavonoid synthase II, and flavonoid 3',5'-hydroxylase/3'-monooxygenase playing critical roles. These findings provide new insights into the molecular mechanisms underlying maize tolerance to waterlogging and offer a theoretical basis for breeding waterlogging-tolerant maize varieties.

Key words: maize, waterlogging stress, metabolomics, differential metabolites, metabolic pathway

Fig. 1

Analysis of data variability and reproducibility A: samples are shown as points, with PC1 and PC2 indicating the first and second principal components, and the percentage showing the variance explained by each component. B: the plot shows Pearson correlation coefficients between samples, with colors indicating correlation strength and direction: red for strong positive, green for weak, and blue for strong negative correlations, with values annotated in squares."

Fig. 2

Integrated analysis of gene expression and metabolic profiles in maize under waterlogging stress All sig diff: number of significantly different metabolites; Down regulated: number of downregulated metabolites or genes; Up regulated: number of upregulated metabolites or genes. In Fig. D, each circle denotes a comparison group, with overlaps indicating shared differential metabolites and non-overlaps indicating unique ones."

Fig. 3

Phenotypic changes in maize seedlings under waterlogging stress CK: control group; W: waterlogged treatment group, scale bar: 5 cm. * and *** mean significant difference at the 0.05 and 0.001 probability levels, respectively; ns means not significant difference."

Fig. 4

Fold change of metabolites and KEGG enrichment analysis bubble plot A: the x-axis shows the log2FC of differential metabolites, with red indicating upregulation and green indicating downregulation. B: the x-axis shows the pathway enrichment factor (Diff/Background) across omics, the y-axis lists KEGG pathways, color (red→yellow→green) indicates enrichment significance (P-value). Bubble shape denotes omics type (circle represents transcriptomics, triangle represents metabolomics); size reflects differential metabolite/gene counts, larger bubbles indicating higher counts."

Fig. 5

Flavonoid metabolic pathway diagram Boxes represent metabolites, and arrows indicate enzymes. The bar plot displays the relative levels of key metabolites in the control (0 d) versus waterlogging stress (4 d). The heatmap illustrates gene expression changes (0 d vs 4 d) with a green-to-red color gradient, where green indicates downregulation and orange represents upregulation. The metabolites are naringenin chalcone, naringenin, apigenin, dihydrokaempferol, eriodictyol, and luteolin; enzymes include chalcone isomerase, flavone synthase II, flavanone 3-hydroxylase, and flavonoid 3′,5′-hydroxylase/flavonoid 3′-monooxygenase. *** means significant difference at the 0.001 probability level."

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