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Fungus Symbionts Colonizing the Galleries of the Ambrosia Beetle Platypus quercivorus

  • Invertebrate Microbiology
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Abstract

Isolations were made to determine the fungal symbionts colonizing Platypus quercivorus beetle galleries of dead or dying Quercus laurifolia, Castanopsis cuspidata, Quercus serrata, Quercus crispula, and Quercus robur. For these studies, logs from oak wilt-killed trees were collected from Kyoto Prefecture, Japan. Fungi were isolated from the: (1) entrances of beetle galleries, (2) vertical galleries, (3) lateral galleries, and (4) the larval cradle of P. quercivorus in each host tree. Among the fungus colonies which appeared on YM agar plates, 1,219 were isolated as the representative isolates for fungus species inhabiting in the galleries based on their cultural characteristics. The validity of the visual classification of the fungus colonies was checked and if necessary properly corrected using microsatellite-primed PCR fingerprints. The nucleotide sequence of the D1/D2 region of the large subunit nuclear rRNA gene detected 38 fungus species (104 strains) of which three species, i.e., Candida sp. 3, Candida kashinagacola (both yeasts), and the filamentous fungus Raffaelea quercivora were isolated from all the tree species. The two yeasts were most prevalent in the interior of galleries, regardless of host tree species, suggesting their close association with the beetle. A culture-independent method, terminal restriction fragment length polymorphism (T-RFLP) analysis was also used to characterize the fungus flora of beetle galleries. T-RFLP patterns showed that yeast species belonging to the genus Ambrosiozyma frequently occurred on the gallery walls along with the two Candida species. Ours is the first report showing the specific fungi inhabiting the galleries of a platypodid ambrosia beetle.

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Acknowledgments

This research was supported in part by a Grant-in-Aid for Scientific Research (A) from the Ministry of Education, Sports, Culture, Science and Technology of Japan (no. 18208015 to KF, 2006–2008) and a grant from the Japan Society for the Promotion of Science (no. 21·1976 to RE, 2009–2010). We are grateful to Dr. J. Sutherland for advice on preparing the manuscript and correcting the English. We thank Dr. M. Kobayashi (Kyoto Pref. Univ.), the technical staffs at Kamigamo Experimental Station of Kyoto University, Mr. H. Qi (Kyoto Univ.), and Mr. T. Hagus (Kyoto Univ.), and other members of our labortory for their precious assistance during wood sample collection. We also thank Dr. M. Sakamoto (RIKEN), and Dr. Y. Benno (RIKEN) for their technical advice on T-RFLP analysis. Too, we thank Dr. K.-D. An (RIKEN) for his assistance on optimizing the PCR condition especially for R. quercivora. We deeply appreciate anonymous reviewers for their helpful suggestions on our manuscript.

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Authors and Affiliations

  1. Laboratory of Environmental Mycoscience, Division of Environmental Science and Technology, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, 606–8502, Japan

    Rikiya Endoh, Yuko Takeuchi & Kazuyoshi Futai

  2. Microbe Division/Japan Collection of Microorganisms (JCM), RIKEN BioResource Center, Wako, Saitama, 351–0198, Japan

    Rikiya Endoh, Motofumi Suzuki & Gen Okada

Authors
  1. Rikiya Endoh
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  2. Motofumi Suzuki
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  3. Gen Okada
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  4. Yuko Takeuchi
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  5. Kazuyoshi Futai
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Corresponding author

Correspondence to Rikiya Endoh.

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Table 1

Sources of fungus isolation (DOC 35 kb)

Table 2

Additional fungus isolates of which the D1/D2 region of the LSU rDNA sequences were determined and sporulating condition of the species isolated in this study (DOC 122 kb)

Table 3

Beetle galleries of Platypus quercivorus used for the T-RFLP analysis (DOC 43 kb)

Table 4

Error rate on the cultural characteristic-based identifications among the first representatives (DOC 70 kb)

Table 5

Percent ratio of fungus colony counts and frequency as determined by a plating method. (DOC 211 kb)

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Endoh, R., Suzuki, M., Okada, G. et al. Fungus Symbionts Colonizing the Galleries of the Ambrosia Beetle Platypus quercivorus . Microb Ecol 62, 106–120 (2011). https://doi.org/10.1007/s00248-011-9838-3

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