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. 2025 Jan 17;26(2):786.
doi: 10.3390/ijms26020786.

Analysis of the Transcriptome Provides Insights into the Photosynthate of Maize Response to Salt Stress by 5-Aminolevulinic Acid

Ying Jiang  1 , Min Li  1 , Yumei Qian  1 , Hao Rong  1 , Tao Xie  1 , Shanshan Wang  1 , Hong Zhao  1 , Liangli Yang  1 , Qingyun Wang  2 , Yanyong Cao  3
Affiliations

Analysis of the Transcriptome Provides Insights into the Photosynthate of Maize Response to Salt Stress by 5-Aminolevulinic Acid

Ying Jiang et al. Int J Mol Sci. .

Abstract

Salt stress is a significant environmental factor that impedes maize growth and yield. Exogenous 5-aminolevulinic acid (ALA) has been shown to mitigate the detrimental effects of various environmental stresses on plants. However, its regulatory role in the photosynthesis mechanisms of maize seedlings under salt stress remains poorly understood. Transcriptome sequencing and physiological index measurements were conducted on the leaves of the "Zhengdan 958" cultivar subjected to three different treatments. Differential expression analysis revealed 4634 differentially expressed genes (DEGs), including key transcription factor (TF) families such as NAC, MYB, WRKY, and MYB-related, across two comparisons (SS_vs_CK and ALA_SS_vs_SS). Significant enrichment was observed in the metabolic pathways related to porphyrin metabolism, photosynthesis-antenna proteins, photosynthesis, and carbon fixation in photosynthetic organisms. ALA treatment modulated the expression of photosynthesis-related genes, increased photosynthetic pigment content, and enhanced the activities of superoxide dismutase (SOD) and catalase (CAT), thereby mitigating the excessive accumulation of reactive oxygen species (ROS). Furthermore, ALA increased starch content under salt stress. These findings establish a foundational understanding of the molecular mechanisms through which ALA regulates photosynthesis under salt stress in maize seedlings. Collectively, exogenous ALA enhances maize's salt tolerance by regulating photosynthesis-related pathways.

Keywords: 5-aminolevulinic acid; Zea mays L.; photosynthesis; salt stress; transcriptome.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Effect of ALA on (A) phenotype and (B) microstructure of maize leaves under salt stress.
Figure 2
Figure 2
Analysis of transcriptome sequencing data. (A) Venn plot of the number of expressed genes in three sample groups. (B) Principal component analysis (PCA) based on gene expression profiles of three sample groups. (C) Heatmap of the gene expression correlation for all samples. Green box represents the CK treatment group. Blue box represents the SS treatment group. Red box represents the ALA_SS treatment group. (D) Histograms of the number of up- and down-regulated DEGs in two comparisons (SS_vs_CK and ALA_SS_vs_SS). (E) The Venn diagram of DEGs in two comparisons.
Figure 3
Figure 3
Functional annotation and enrichment analysis of shared DEGs conducted by GO and KEGG. (A) A histogram of the GO annotation analysis. (B) A histogram of the KEGG annotation analysis. (C) A bubble diagram of the GO enrichment analysis. (D) A bubble diagram of the KEGG enrichment analysis.
Figure 4
Figure 4
Analysis of transcription factors. (A) The stacked bar chart of the top 10 TF families in two comparisons. (B) The top 10 TF families’ number of co-regulated DEGs in two comparisons.
Figure 5
Figure 5
DEGs involved in porphyrin metabolism and photosynthesis-antenna proteins in two comparisons. The log2 fold change of FPKM represents the gene expression level in two comparisons. (A) Pathway of selected DEGs involved in porphyrin metabolism. The schematic pathways were modified according to KEGG enrichment. Solid arrows represent direct reactions. Dashed arrows represent indirect reactions. The red and green boxes express up- and down-regulated genes in two comparisons. (B) Statistics of Chl a content under different treatments. Different lowercase letters indicate a significant difference (p < 0.05). (C) Statistics of Chl b content under different treatments. (D) Statistics of total Chl content under different treatments. (E) Statistics of Car content under different treatments. (F) Heatmap of selected DEGs involved in photosynthesis-antenna proteins.
Figure 6
Figure 6
DEGs involved in photosynthesis in two comparisons. The log2 fold change of FPKM represents the gene expression level in two comparisons. (A) Pathway of selected DEGs involved in photosynthesis. The schematic pathways were modified according to KEGG enrichment. Solid arrows represent direct reactions. Dashed arrows represent indirect reactions. (B) Heatmap of selected DEGs involved in photosynthesis. (C) Statistics of O2•– content under different treatments. Different lowercase letters indicate a significant difference (p < 0.05). (D) Statistics of H2O2 content under different treatments. (E) Statistics of SOD activity under different treatments. (F) Statistics of CAT activity under different treatments.
Figure 7
Figure 7
DEGs involved in carbon fixation in photosynthetic organisms in two comparisons. The log2 fold change of FPKM represents the gene expression level in two comparisons. (A) Pathway of selected DEGs involved in carbon fixation in photosynthetic organisms. The schematic pathways were modified according to KEGG enrichment. Solid arrows represent direct reactions. Dashed arrows represent indirect reactions. The red and green boxes express up- and down-regulated genes in two comparisons. (B) Statistics of starch content under different treatments. Different lowercase letters indicate a significant difference (p < 0.05).
Figure 8
Figure 8
Comparisons of log2 fold change of 8 selected DEGs in RNA-Seq and qRT-PCR results.
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