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Expression of alcohol acyltransferase is a potential determinant of fruit volatile ester variations in Capsicum

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The transcript level of alcohol acyltransferase 1 (AAT1) may be the main factor influencing the variations in volatile esters that characterizing the fruity/exotic aroma of pepper fruit.

Abstract

Volatile esters are key components for characterizing the fruity/exotic aroma of pepper (Capsicum spp.) fruit. In general, the volatile ester content in the fruit is the consequence of a delicate balance between their synthesis by alcohol acyltransferases (AATs) and degradation by carboxylesterases (CXEs). However, the precise role of these families of enzymes with regard to volatile ester content remains unexplored in Capsicum. In this study, we found that the volatile ester content was relatively low in C. annuum and much higher in C. chinense, particularly in pungent varieties. Additionally, fruits collected from multiple non-pungent C. chinense varieties, which harbor loss-of-function mutations in capsaicinoid biosynthetic genes, acyltransferase (Pun1), putative aminotransferase (pAMT), or putative ketoacyl-ACP reductase (CaKR1) were analyzed. The volatile ester contents of non-pungent C. chinense varieties (pamt/pamt) were equivalent to those of pungent varieties, but their levels were significantly lower in non-pungent NMCA30036 (pun12/pun12) and C. chinense (Cakr1/Cakr1) varieties. Multiple AAT-like sequences were identified from the pepper genome sequences, whereas only one CXE-like sequence was identified. Among these, AAT1, AAT2, and CXE1 were isolated from fruits of C. chinense and C. annuum. Gene expression analysis revealed that the AAT1 transcript level is a potential determinant of fruit volatile ester variations in Capsicum. Furthermore, enzymatic assays demonstrated that AAT1 is responsible for the biosynthesis of volatile esters in pepper fruit. Identification of a key gene for aroma biosynthesis in pepper fruit will provide a theoretical basis for the development of molecular tools for flavor improvement.

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Acknowledgements

The authors thank Dr. Paul Bosland (New Mexico State University) for providing NMCA30036, Caribbean Agricultural Research and Development Institute (CARDI) for providing C. chinense varieties (Faria, Moruga Red, Joyce, Phyllis, and West Indies Red), Dr. Norio Yamamoto (National Museum of Ethnology) for providing CaKR1 mutants (No.3327, No.3341, No.4026, and No.4028), and the National Agriculture and Food Research Organization (NARO) Genebank for providing C. annuum varieties. We thank Shinya Kanzaki (Kindai University) for useful discussion. We thank Jennifer Smith, PhD, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript. We thank the anonymous reviewers for the constructive comments and suggestion which helped us to improve the manuscript.

Funding

This study was supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers 25850018, 16K07605, 18H03446, 22H03827 and Kindai University Faculty of Agriculture Fund for Scientific Research.

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Authors and Affiliations

  1. Graduate School of Agriculture, Kindai University, Nara, 3327-204, Japan

    Sota Koeda, Tomona Noda, Shinkai Hachisu & Hiroto Yamamoto

  2. Faculty of Agriculture, Kindai University, Nara, 3327-204, Japan

    Sota Koeda, Akiha Kubo, Yasuto Tanaka, Sayaka Ozaki, Machiko Kinoshita & Kouki Ohno

  3. Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan

    Yoshiyuki Tanaka

  4. Japan Society for Scientific Aromatherapy, Tokyo, 164-0003, Japan

    Kenichi Tomi

  5. College of Bioresource Sciences, Nihon University, Kanagawa, 252-0880, Japan

    Yusuke Kamiyoshihara

  6. Graduate School of Bioresource Sciences, Nihon University, Kanagawa, 252-0880, Japan

    Yusuke Kamiyoshihara

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  1. Sota Koeda
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  2. Tomona Noda
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  3. Shinkai Hachisu
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  4. Akiha Kubo
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  5. Yasuto Tanaka
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  12. Yusuke Kamiyoshihara
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Contributions

SK conceived the idea of the study. SK and TN designed experiments, interpreted the results, and wrote the manuscript. TN, SH, AK, Yasuto T, HY, SO, MK, and KO analyzed volatile compounds by GC. KT analyzed volatile compounds by GC–MS. Yoshiyuki T analyzed capsaicinoid. TN isolated and analyzed the gene expression of AAT and CXE genes. YK analyzed the function of the recombinant AAT protein. All authors read and approved the final manuscript.

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Correspondence to Sota Koeda.

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Koeda, S., Noda, T., Hachisu, S. et al. Expression of alcohol acyltransferase is a potential determinant of fruit volatile ester variations in Capsicum. Plant Cell Rep 42, 1745–1756 (2023). https://doi.org/10.1007/s00299-023-03064-z

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