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Functional divergence of two closely related carboxylesterases in pepper (Capsicum annuum L.)

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Abstract

Carboxylesterases (CXEs) catalyze the hydrolysis of carboxylic acid esters into the corresponding acids and alcohols. Previously, a carboxylesterase gene was isolated from pepper (Capsicum annuum L.) and designated as PepEST. This gene is highly expressed during the incompatible interaction between ripe fruits of pepper and an anthracnose fungus (Colletotrichum gloeosporioides), and the PepEST protein exhibits an antifungal activity by suppressing fungal growth with the inhibition of appressorium formation. In this study, a PepEST-homologous gene was newly isolated from the pepper genome, which is designated as PepEST2 (the original PepEST gene is re-named here as PepEST1). Compared with PepEST1 whose expression is only induced by inoculation of the fungus, PepEST2 is not induced by fungus infection, rather expressed mainly in roots. Subcellular localization analysis using tobacco protoplasts showed that both PepEST1 and PepEST2 proteins are localized in the cytoplasm, and enzymatic activity assays showed similar Vmax and Km values of PepEST2 to those of PepEST1. Antifungal activity assays against C. gloeosporioides demonstrated that not only PepEST1 but also PepEST2 was able to suppress the fungal growth by inhibiting the appressorium formation. In addition, the antifungal activity of PepEST2 was higher than that of PepEST1. Collectively, this study suggests that two closely related members of pepper CXEs are functionally diversified due to the differential expression patterns, and also provides PepEST2 as an alternative gene to confer disease resistance to plants.

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Acknowledgements

This work was supported by the New Breeding Technologies Development Program (Project No. PJ016543 to JIK) through the Rural Development Administration (RDA), Republic of Korea, and in part by the National Research Foundation of Korea (NRF) grants funded by the Korea government (MSIT) (grant no. 2021R1A2C1012562 and 2022R1A5A1031361 to JIK).

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Author notes

  1. Ha-Young Kang

    Present address: Tomocube Inc., Daejeon, 34109, Republic of Korea

  2. Young Soon Kim

    Present address: Plant Cell Research Institute of BIO-FD&C Co., Ltd., Incheon, 21990, Republic of Korea

  3. Yun-Jeong Han and Ha-Young Kang have contributed equally.

Authors and Affiliations

  1. Kumho Life Science Laboratory, Chonnam National University, Gwangju, 61186, Republic of Korea

    Yun-Jeong Han, Young Soon Kim & Jeong-Il Kim

  2. Department of Biotechnology, Chonnam National University, Gwangju, 61186, Republic of Korea

    Ha-Young Kang & Jeong-Il Kim

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Correspondence to Jeong-Il Kim.

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Han, YJ., Kang, HY., Kim, Y.S. et al. Functional divergence of two closely related carboxylesterases in pepper (Capsicum annuum L.). Plant Biotechnol Rep 17, 499–507 (2023). https://doi.org/10.1007/s11816-023-00849-2

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