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Comparative transcriptomic screen identifies expression of key genes involved in pattern-triggered immunity induced by salicylic acid in strawberry

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Abstract

The destructive illness powdery mildew, caused by Podosphaera aphanis, reduces strawberry yield. However, the mechanism through which exogenous salicylic acid can enhance resistance to P. aphanis remains unknown. Therefore, this study aimed to investigate the underlying mechanism of SA-induced resistance in strawberry leaves against P. aphanis using a comparative RNA-seq approach. Upon observing the symptoms, it was found that SA partially contributed to strawberry resistance against P. aphanis by increasing H2O2 levels during the initial stages. To mitigate the effects of H2O2, SA-treated leaves significantly boosted the activities of enzymes such as superoxide dismutases (SOD), catalases (CAT), and peroxidases (POD). RNA profiling identified several differentially expressed genes (DEGs) associated with the reactive oxygen species (ROS)-redox pathway. Moreover, SA-induced leaves fortified the cell wall to prevent P. aphanis penetration. Using weighted gene coexpression network analysis (WGCNA), three modules (red, green, and yellow) were constructed, showing a strong correlation with enhanced resistance. Predictably, cyclic nucleotide-gated channels (CNGC) were identified as potential players in Ca2+ signaling linked to the PTI response. Additionally, the presence of WRKY33 and defense-related genes such as PR10 were associated with enhanced resistance. It is plausible that strawberries exposed to SA trigger ROS and Ca2+ signaling, along with significantly elevated WRKY33 expression, to facilitate the production of PR10 and camalexin, which help protect against P. aphanis.

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Data Availability

The data analyzed in this study have been included in the article and its supplementary information. The RNA-seq data has been deposited to the Genome Sequence Archive in BIG Data Center under accession numbers CRA001964.

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Acknowledgements

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Funding

JF reports financial support was provided by Fundamental Research Program of Shanxi Province (Number 20210302124305). CXZ reports financial support was provided by National Natural Science Foundation of China (Number 31870571).

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Authors and Affiliations

  1. College of Biological Sciences and Technology, Taiyuan Normal University, Shanxi, 030619, China

    Jun Feng & Xiao-Ying Wu

  2. College of Biological Sciences and Technology, Beijing Forestry University, Beijing, 100083, China

    Yu Xiu & Cai-Xia Zheng

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  1. Jun Feng
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  2. Xiao-Ying Wu
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  3. Yu Xiu
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  4. Cai-Xia Zheng
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Contributions

JF designed the experiment, and conducted the laboratory experiments along with YX; JF and XYW analyzed the data and wrote the manuscript, with valuable assistance from YX and CXZ.

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Correspondence to Jun Feng or Cai-Xia Zheng.

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Communicated by Inhwa Yeam.

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Supplementary Information

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13580_2023_548_MOESM1_ESM.pdf

Analysis of enriched Gene Ontology (GO) and KEGG enrichment pathways. (A) Top 20 enriched GO biological processes in the CK group; (B) Top 20 enriched GO biological processes in the SA group; (C) Top 20 KEGG enrichment pathways in the CK group; (D) Top 20 KEGG enrichment pathways in the SA group.

13580_2023_548_MOESM2_ESM.pdf

The number of genes associated with ROS that are differentially expressed. (B) expression of FaRBOHD genes in strawberry after two pretreatments under P. aphanis stress ; and (C) phylogenetic analysis of RBOH proteins from Arabidopsis and strawberry, with the red arrows indicating candidate proteins identified in this study.

13580_2023_548_MOESM3_ESM.pdf

Strawberry DEGs were studied concerning enzymatic antioxidants. (A) phylogenetic analysis of SOD proteins from Arabidopsis and strawberry; (B) fundamental SOD genes expression in strawberry after two pretreatments under P. aphanis stress. The red words represent candidate proteins discovered in this study.

13580_2023_548_MOESM4_ESM.pdf

Strawberry DEGs were studied about enzymatic antioxidants. (A) phylogenetic analysis of CAT proteins from Arabidopsis and strawberry; (B) phylogenetic analysis of POD proteins from Arabidopsis and strawberry; (C) key CAT genes expression in strawberry after two pretreatments under P. aphanis stress; (D) expression of critical POD genes in strawberry after two pretreatments under P. aphanis stress. The red words represent candidate proteins discovered in this study.

13580_2023_548_MOESM5_ESM.pdf

The number of DEGs involved in plant defense. (A) VENN analysis between the CK group and SA group; (B) a heatmap of common DEGs associated with plant defense

The heatmap of common DEGs related to plant pathogen interaction. Green indicates 16 common DEGs.

13580_2023_548_MOESM7_ESM.pdf

Weighted gene co-expression network of strawberry leaves. (A) Clustering dendrogram of all genes, with dissimilarity based on the topological overlap, together with assigned module colors. (B) Correlation between modules. (C) Module-trait associations. Each row corresponds to a module characteristic gene, and each column corresponds to a trait. Each cell contains a corresponding correlation coefficient and p-value. (D–F) D. moniliforme transcriptome co-expression network diagram: (D) red module; (E) green module; (F) yellow module.

13580_2023_548_MOESM8_ESM.pdf

The relative transcript levels of 5 genes in infected (with P. aphanis) and uninfected Fragaria tissues at 0, and 3 dpi (days post infection). Relative expression of phenylpropanoid intermediates genes. Application of two levels of preharvest wounding on leaves and its effects in strawberry fruit when wounding occurred 7 and 14 days before harvest. (A) Major allergen Pru ar 1-like (FaRP10); (B) Major allergen Pru ar 1-like (FaRP10); (C) Phenylalanine ammonia lyase (FaPAL); (D) NAC domain-containing protein 2 (FaNAC2); and (E) WRKY transcription factor 33 (FaWRKY33). Data presented are means standard error of three replicates.

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Feng, J., Wu, XY., Xiu, Y. et al. Comparative transcriptomic screen identifies expression of key genes involved in pattern-triggered immunity induced by salicylic acid in strawberry. Hortic. Environ. Biotechnol. 64, 835–848 (2023). https://doi.org/10.1007/s13580-023-00548-5

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