Saccharin is generated from probenazole (PBZ) in plants and acts as a plant defense activator. Our study of the mechanism underlying saccharin-induced systemic acquired resistance in Arabidopsis thaliana suggests an antagonistic interaction between salicylic acid (SA)- and jasmonic acid (JA)-signaling as revealed through gene expression analyses. In wild-type plants (Col-0) exposed to saccharin, there was a consistent increase in callose deposition and in expression of SA-marker genes, PR1 and PR2, which coincided with a decrease in expression of JA-marker genes such as VSP2, LOX2 and PDF1.2. Actually, pretreatment of Col-0 with saccharin or PBZ conferred resistance to Pseudomonas syringae pv. tomato DC3000, but not to Pectobacterium carotovorum subsp. carotovorum, Botrytis cinerea, or Colletotrichum higginsianum. Enhanced expression of SA- and JA-marker genes and the augmented deposition of callose were evident after a challenge with virulent DC3000 in saccharin-pretreated plants. Consistently, pretreatment of saccharin and PBZ with SA- and JA-defective mutants led to diminished resistance in NahG-transgenic and npr1 mutant plants, but not in jar1 mutant plants, suggesting that saccharin and PBZ induce resistance in A. thaliana against Pto DC3000 mainly via activation of SA-signaling, leading to suppression of JA/ET-signaling and vice versa. Collectively, an antagonism between SA- and JA-signaling conditioned by saccharin renders resistance to a specific pathogen in Arabidopsis.
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LTP thanks the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) for their financial support during her doctoral course. We acknowledge Dr. Shinji Tsuyumu (Laboratory of Plant Pathology, Faculty of Agriculture, Shizuoka University, Shizuoka, Japan) for generously providing the Pectobacterium carotovorum subsp. carotovorum strain Pc1. This research was supported in part by the Grants-in-Aid for Scientific Research (18K05645) from the Japan Society for Promotion of Science (JSPS).
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Supplementary file1 (PPTX 50517 kb) Effect of saccharin on growth of A. thaliana plants. Col-0 plants were treated with water (as a control) or saccharin ranging from 0.01 to 5 mM by spraying until leaf surface was evenly coated or soil was drenched with 10 ml of solution. Plants were observed for leaf size, shape and color after 7, 11 and 18 days of treatment. This experiment was repeated twice with similar results. Effect of saccharin on growth of Pto DC3000. Liquid and solid KB media were amended with water (control) or saccharin at 0.1, 1 or 10 mM. a Bacterial suspension (10 µl of OD600 = 10-5 or OD600 = 10-4 [right]) was placed onto KB agar disk. b Colonies were counted after 3 days of culture. c Mean ± SD OD600 to estimate bacterial population over 5 days growth in test tubes with shaking at 28C. Three replicates were done for each experiment. Turkey’s test, p > 0.05. Effect of saccharin on expression of SA- and JA/ET-responsive genes and penetration during infection by Colletotrichum higginsianum (Ch). Col-0 plants were pretreated with water or saccharin (1 mM) for 2 days, then inoculated with Ch (5 × 104 spores/ml). a to d Leaves were harvested at 12, 24 and 48 hpi and subjected to qPCR. Water-treated, uninoculated plants were used as the control. Expression value of genes was normalized using EF1-α as an internal standard and expressed relative to average levels in the control. Data are means ± SD from triplicate reactions in each experiment. Letters indicate significant differences between treatments at each time post-inoculation. Tukey’s test, p < 0.05. e Mean (± SD) infection rate (penetration efficiency at 3 dpi calculated as percentage of appressoria that formed primary infection hyphae per total number of appressoria. f Micrographs of large intracellular primary hyphae 3 dpi. ap, appressorium; ph, primary hypha, sh, secondary hypha. Asterisks indicate significant difference from the water treatment. Student’s t-test, **p < 0.01.
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Phuong, L.T., Fitrianti, A.N., Luan, M.T. et al. Antagonism between SA- and JA-signaling conditioned by saccharin in Arabidopsis thaliana renders resistance to a specific pathogen. J Gen Plant Pathol 86, 86–99 (2020). https://doi.org/10.1007/s10327-019-00899-x
- Induced resistance
- Jasmonic acid (JA)
- Probenazole (PBZ)
- Salicylic acid (SA)