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Variation in Bufadienolide Composition of Parotoid Gland Secretion From Three Taxa of Japanese Toads

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Abstract

Toads of the genus Bufo synthesize and accumulate bufadienolides (BDs) in their parotoid glands. BDs are cardiotonic steroids that play an important role in defense against the toads’ predators. Three bufonid taxa occur in mainland Japan, Bufo japonicus formosus, B. j. japonicus, and B. torrenticola. The chemical structures of BDs isolated from B. j. formosus were studied several decades ago, but there is no further information on the toxic components of Japanese toads and their metabolism. In this study, we analyzed BDs of toads from throughout Japan and compared the BD profiles by liquid chromatography/mass spectrometry (LC/MS) and hierarchical cluster analysis (HCA). We observed BDs in three taxa of Japanese toads, and identified five of the most common BDs by nuclear magnetic resonance (NMR) analyses. Of the five BDs, only bufalin was detected in all individuals. HCA of individual BD profiles divided the three taxa into five primary clusters and several subclusters. This result indicates that BD profiles differ both among and within the taxa. The clustering pattern of BDs is generally concordant with a phylogenetic tree reconstructed from the mitochondrial cytochrome b gene of Japanese toads. Our results suggest that the BDs of Japanese toads have diversified not in response to specific selective pressures, but simply due to population structuring over evolutionary time.

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Acknowledgments

We thank many colleagues for collecting toads in Japan. (JSPS) and National Natural Science Foundation of China (NSFC 31411140033) and from.

Funding

This research was supported by grants from Japan-China Joint Research Project (2014–2016) from the Japan Society for the Promotion of Science (JSPS) and National Natural Science Foundation of China (NSFC 31411140033) and from JSPS KAKENHI Grant Numbers JP26440213, JP17H03719, and JP18KK0205. Additional funding was provided by Utah State University.

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  1. Ryu Nakata

    Present address: Department of Bioscience and Biotechnology, Kyoto University of Advanced Science, 1-1 Nanjo Otani, Sogabe, Kameoka, Kyoto, 621-8555, Japan

Authors and Affiliations

  1. Division of Applied Life Science, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo, Kyoto, Kyoto, 606-8502, Japan

    Takato Inoue, Ryu Nakata, Naoko Yoshinaga & Naoki Mori

  2. Department of Biology, Utah State University, Logan, UT, 84322-5305, USA

    Alan H. Savitzky

  3. Department of Zoology, Graduate School of Science, Kyoto University, Kitashirakwa, Sakyo, Kyoto, Kyoto, 606-8502, Japan

    Akira Mori

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Contributions

Conceptualization: Inoue T, Nakata R, Savitzky AH, Yoshinaga N, Mori A, Mori N.

Data curation: Inoue T, Nakata R, Savitzky AH, Yoshinaga N, Mori A, Mori N.

Formal analysis: Inoue T, Nakata R, Yoshinaga N, Mori N.

Funding acquisition: Mori A.

Investigation: Inoue T, Nakata R, Savitzky AH, Mori A, Mori N.

Methodology: Inoue T, Nakata R, Yoshinaga N, Mori A, Mori N.

Project administration: Inoue T, Nakata R, Mori A, Mori N.

Resources: Inoue T, Mori A.

Supervision: Inoue T, Nakata R, Savitzky AH, Yoshinaga N, Mori A, Mori N.

Validation: Inoue T, Nakata R, Yoshinaga N, Mori A, Mori N.

Visualization: Inoue T, Nakata R, Savitzky AH, Yoshinaga N, Mori A, Mori N.

Writing – original draft: Inoue T, Nakata R.

Writing – review & editing: Inoue T, Nakata R, Savitzky AH, Mori A, Mori N.

All authors approved the manuscript to be published and agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

Corresponding author

Correspondence to Naoki Mori.

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Inoue, T., Nakata, R., Savitzky, A.H. et al. Variation in Bufadienolide Composition of Parotoid Gland Secretion From Three Taxa of Japanese Toads. J Chem Ecol 46, 997–1009 (2020). https://doi.org/10.1007/s10886-020-01217-y

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