| Project/Area Number |
17580133
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
林学・森林工学
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| Research Institution | Forestry and Forest Products Research Institute |
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Principal Investigator |
KINUURA Haruo Forestry and Forest Products Research Institute, Kansai Research Center, Senior researcher, 関西支所, 主任研究員 (00353665)
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| Co-Investigator(Kenkyū-buntansha) |
MIYASITA Shunichiro Forestry and Forest Products Research Institute, Kansai Research Center, Senior researcher, 関西支所, 主任研究員 (10353872)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2006: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | ambrosia beetles / symbiotic fungi / Platypus quercivorus / yeasts / アンブロシアビートル / アンブロシアビトール / 養菌性キクイムシ類 |
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| Research Abstract |
We dissected adults of Platypus quercivorus and isolated fungi from the extirpated proventricula. Most of the isolated strains were yeast-like fungi, although filamentous fungi such as Raffaelea quercivora were also isolated. We sequenced the almost complete region of 18S rDNA and the D1/D2 domain of LSU rDNA of the yeast-like isolates. We also sequenced those regions of R. quercivora and compared them with those of yeast-like isolates in order to clarify whether these isolates were R. quercivora. The sequences of the yeast-like isolates were completely different from those of R. quercivora. Furthermore, we searched homologous sequences to the D1/D2 domain of LSU rDNA of the yeast-like isolates in DNA data banks. The phylogenetic analysis based on the determined and searched nucleotide sequence data reveals that some yeast-like strains are related to Candida sp. and others are related to Ambrosiozyma sp.
The ordinary polymerase chain reaction (PCR) assay was performed on DNA in the extirpated proventricula, but little PCR product was yielded or only PCR products from the insects were detected. So PCR was repeated on the PCR reaction solution, and the suitable primer and PCR program were examined. As a result, the PCR product with the same gel mobility as reference DNA of fungi was detected in a few samples. Then we decided to use these PCR products for denaturing gradient gel electrophoresis (DGGE) analysis. The DGGE analysis showed two results: (1) some DGGE bands were detected from proventricula and mycangia, and the number of bands from mycangia was larger than from proventricula, (2) the band with the same mobility as that of yeasts was detected from every sample whereas the band of R. quercivora was inclined not to be detected from proventricula. These results support the hypothesis that the main diets of P. quercivorus are yeasts.
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