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Overexpression of salicylic acid carboxyl methyltransferase reduces salicylic acid-mediated pathogen resistance in Arabidopsis thaliana

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Abstract

We cloned a salicylic acid/benzoic acid carboxyl methyltransferase gene, OsBSMT1, from Oryza sativa. A recombinant OsBSMT1 protein obtained by expressing the gene in Escherichia coli exhibited carboxyl methyltransferase activity in reactions with salicylic acid (SA), benzoic acid (BA), and de-S-methyl benzo(1,2,3)thiadiazole-7-carbothioic acid (dSM-BTH), producing methyl salicylate (MeSA), methyl benzoate (MeBA), and methyl dSM-BTH (MeBTH), respectively. Compared to wild-type plants, transgenic Arabidopsis overexpressing OsBSMT1 accumulated considerably higher levels of MeSA and MeBA, some of which were vaporized into the environment. Upon infection with the bacterial pathogen Pseudomonas syringae or the fungal pathogen Golovinomyces orontii, transgenic plants failed to accumulate SA and its glucoside (SAG), becoming more susceptible to disease than wild-type plants. OsBSMT1-overexpressing Arabidopsis showed little induction of PR-1 when treated with SA or G. orontii. Notably, incubation with the transgenic plant was sufficient to trigger PR-1 induction in neighboring wild-type plants. Together, our results indicate that in the absence of SA, MeSA alone cannot induce a defense response, yet it serves as an airborne signal for plant-to-plant communication. We also found that jasmonic acid (JA) induced AtBSMT1, which may contribute to an antagonistic effect on SA signaling pathways by depleting the SA pool in plants.

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Acknowledgements

This work was supported by grants from the Crop Functional Genomics Center (CG2112 to JJC and CG2111 to JK) and the Korea Research Foundation (KRF-2004-005-F00013 to YDC). Financial support, including graduate research assistantships to YJK and CJ, from the Brain Korea 21 project of the Ministry of Education is also acknowledged. Special thanks are due to Professors In Gyu Hwang and Soon Ok Kim (Seoul National University) for the identification of Golovinomyces orontii.

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Authors and Affiliations

  1. School of Agricultural Biotechnology, Seoul National University, Seoul , 151-742, Korea

    Yeon Jong Koo, Myeong Ae Kim, Choonkyun Jung, Joon-Kwan Moon, Jeong-Han Kim, Jong-Joo Cheong & Yang Do Choi

  2. Division of Bioscience, Myongji University, Yongin , 449-728, Korea

    Eun Hye Kim, Sang Ik Song & Ju-Kon Kim

  3. Division of Plant Biosciences, Kyungpook National University, Daegu , 702-701, Korea

    Jong Tae Song

  4. Department of Plant Science, Seoul National University, Seoul , 151-742, Korea

    Hak Soo Seo

  5. School of Biological Sciences, Seoul National University, Seoul , 151-742, Korea

    Jong Seob Lee

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  1. Yeon Jong Koo
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Fig8

Enhanced disease susceptibility of OsBSMT1-overexpressing plants to the avirulent bacterial pathogen Pseudomonas syringae pv. maculicola strain DG6 and their response. (A) Enhanced disease susceptibility of OsBSMT1-overexpressing plants. Wild-type (left) and OsS6 (right) plants were inoculated with P. Syringae DG6. Infected leaves were photographed after 3 days of inoculation. (B) Growth of P. syringae in plants. Wild-type (circles) and OsS6 (triangles) plants were inoculated with P. syringae DG6. On days 2 and 3, P. Syringae grew faster in OsS6 plants than in wild-type plants (P < 0.009, t-test, n = 8). Bars indicate standard deviation. This experiment was repeated two times with similar results. (C) Expression of PR-1 during infection. The fourth or fifth leaves of wild-type and OsS6 plants were infected with P. syringae at OD600 = 0.01 (approximately 5 × 106 cfu/ml) or mock-infected with 10 mM MgSO4. RNA at the indicated time point was isolated, and Northern blot analysis was conducted (GIF 37 kb)

Fig9

SA and BTH insensitivity of OsBSMT1 plants. (A) Northern blot analysis of PR-1 gene expression. Treatment with SA, BTH, or dSM-BTH was performed by spraying wild-type, OsBSMT1, and nahG plants. RNAs were isolated after 1 day of treatment. (B) Metabolism of BTH in plants. In plants, BTH is hydrolyzed to produce dSM-BTH (hydrolyzed BTH) and subsequently conjugated with sugars (Tomlin, 2003). The hydrolyzed BTH is converted to MeBTH by the OsBSMT1 enzyme, as shown in Figure 2. Both BTH and hydrolyzed BTH are known activators of plant disease resistance (Kunz et al., 1997). All experiments were repeated three times with similar results (GIF 47 kb)

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Koo, Y.J., Kim, M.A., Kim, E.H. et al. Overexpression of salicylic acid carboxyl methyltransferase reduces salicylic acid-mediated pathogen resistance in Arabidopsis thaliana . Plant Mol Biol 64, 1–15 (2007). https://doi.org/10.1007/s11103-006-9123-x

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