Abstract
A so-called priming defense is activated only when a plant suffers from heavy pathogenic invasion, thus avoiding excessive energy and material costs under low pathogen loads. In our previous studies, we demonstrated that elicitors, such as β-aminobutyric acid (BABA), methyl jasmonate (MeJA) or benzothiadiazole (BTH), can induce the priming of a defense response against fungal infections in postharvest peach fruit. Interestingly, the defense priming was accompanied by increased soluble sugar accumulation. In this study, we aimed to elucidate the mechanism underlying the increase in sugar contents in primed peach fruit. We identified a group IIa WRKY transcription factor, PpWRKY40, from Prunus persica. Using yeast one-hybrid assays, electrophoretic mobility shift assays and dual-luciferase reporter assays, we show that PpWRKY40 is a transcriptional activator of the sucrose synthesis-related genes sucrose synthase (PpSS1) and sucrose-phosphate synthase (PpSPS3). Induction of defense priming with BABA, MeJA or BTH upregulated PpWRKY40 expression and promoted the transcript levels of PpSS1 and PpSPS3 and their enzymatic activities in postharvest peach fruit, resulting in the enhancement of sucrose synthesis. Moreover, the activities of sorbitol catabolizing enzymes—S6PDH, NAD+-SDH, NADP+-SDH—increased in the peach fruit treated with the defense priming elicitors, indicating that sorbitol was converted into glucose and fructose. Induction of defense priming also improved the sensory quality and antioxidant levels of fruit. In summary, the three defense priming elicitors improved the soluble sugar accumulation and overall quality in postharvest peach by upregulating the PpWRKY40 gene and increasing sorbitol catabolism.
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Acknowledgements
The present work was funded by the NSFC (No. 31671913) and the Fund for Creative Research Groups in Universities of Chongqing (No. CXQT21036). We appreciate Prof. Daxiang Zhou of Chongqing Three Gorges University for his assistance on CRISPR and mutation experiments.
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KW designed the study and wrote the manuscript. KW, CL, MT, JW, CL and YZ performed the experiments. KW, CL and CL analyzed the data. All authors discussed the results and contributed to the final manuscript.
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Supplementary file 1: Fig. S1. Schematic diagram of the CRISPR-AtWRKY40 vector. Fig. S2. Protein–protein interaction network of Prunus persica WRKY40, in which the line color indicates the type of interaction. Light blue and rose red indicate known interactions from curated databases or experimentally determined interactions, green, red and blue indicate predicted interactions, and grass green, dark blue and purple indicate other interactions from text mining, coexpression and protein homology. Fig. S3. In vivo assays of the Prunus persica WRKY40-SS1/SPS3 interactions were evaluated on the basis of the growth activity of yeast Y1HGold containing PpSS1-AbAi and PpWRKY40-pGADT7 recombinant protein or PpSPS3-AbAi and recombinant PpWRKY40-pGADT7 on SD/-Leu plates containing 200/250/300/350 μg L−1 AbA. Fig. S4. Temporal expression patterns (via 1.2% agarose gel electrophoresis) of PpWRKY40, PpSS1 and PpSPS3 in peach fruit treated with or without elicitors. Table S1. Accession numbers of WRKY transcription factors for Arabidopsis thaliana and Prunus persica derived from the NCBI database. Table S2. Sequences of primers used in this study. Table S3. Changes in O2 and CO2 volume fractions in polyethylene bags from different elicitor treatments during 12 days of storage at 20 °C. Table S4. Potential targets of PpWRKY40 and the confidence score between them obtained from STRING 11.5. Data S1. Three tandem copies of the W-box motif (TTGACT) present in the promoter sequences of PpSS1 and PpSPS3. The TTGACT sites are indicated in red text surrounded by a black box. Data S2. Three tandem copies of the mutated W-box (AAAAAA) in the promoter region of PpSS1 and PpSPS3. The mutated W-box is indicated by a black box and red text. Restriction sites were highlight with underlines. Data S3. Promoter sequences of PpSS1, PpSS2, PpSS4, PpSS5, PpSPS1, PpSPS3, PpSPS4 and PpSPP2, in which the W-box motifs are in bold red text. Data S4. Raw images of Y1H, EMSA, DLR, overexpression lines, mutations and PCR gels.
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Wang, K., Lei, C., Tan, M. et al. Increased soluble sugar accumulation in postharvest peaches in response to different defense priming elicitors. Hortic. Environ. Biotechnol. 64, 115–131 (2023). https://doi.org/10.1007/s13580-022-00464-0
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DOI: https://doi.org/10.1007/s13580-022-00464-0