ナラ枯れ原因菌 Raffaelea quercivora 侵入に応答するミズナラの抽出成分に関する研究

Title

ナラ枯れ原因菌 Raffaelea quercivora 侵入に応答するミズナ

_{ラの抽出成分に関する研究( 本文(Fulltext) )}

Author(s)

今井, 香代子

Report No.(Doctoral

Degree)

博士(農学) 甲第622号

Issue Date

2014-03-13

Type

博士論文

Version

ETD

URL

http://hdl.handle.net/20.500.12099/49102

※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。

ナラ枯れ原因菌

Raffaelea quercivora

侵入に応答する

ミズナラの抽出成分に関する研究

2013年

岐阜大学大学院連合農学研究科

生物資源科学

（岐阜大学）

今 井 香 代 子

ナラ枯れ原因菌

Raffaelea quercivora

侵入に応答する

ミズナラの抽出成分に関する研究

1

⥴ゝ ᆅ⌫㝣ᆅࡢ୕ศࡢ୍ࢆ༨ࡵࡿ᳃ᯘࡣ㔜せ࡞ᆅ⌫⎔ቃ㈨※࡛࠶ࡿࠋ᳃ᯘࡣ┠࡟ぢ࠼࡞ ࠸ᚤ⏕≀࠿ࡽᡃࠎே㛫ࡢࡼ࠺࡞኱ᆺື≀ࡲ࡛ 㸪ከ✀ከᵝࡢ⏕≀ࡢ⏕ᜥᆅࡸ㔜せ࡞㣗ᩱ ࢆ୚࠼࡚࠸ࡿࠋ≉࡟㉥㐨௜㏆࡟ᗈࡀࡿ⇕ᖏ㞵ᯘ࡟ࡣ 700 ୓✀࡜ࡶ 2000 ୓✀࡜ࡶゝࢃ ࢀࡿᆅ⌫࡟⏕ᜥࡍࡿ⏕≀✀ࡢ⣙༙ᩘࡀ⏕ᜥࡋ࡚࠾ ࡾ㸪᳃ᯘࡀ⏕≀ከᵝᛶࡢ⥔ᣢ࡟㔜せ ୙ྍḞ࡛࠶ࡿࡇ࡜ࡣゝ࠺ࡲ࡛ࡶ࡞࠸ࠋ᳃ᯘࡢᣢࡘ㔜せ࡞ᙺ๭ࡣ⏕≀ከᵝᛶࡢࡳ࡛ࡣ࡞ ࠸ࠋ㇏࠿࡞᳃ᯘᅵተࡣ㸪ᚤ⏕≀ศゎ࡟ࡼࡗ࡚✵㝽ࡢከ࠸ᅵተࢆసࡾᅵተỈศࡸᆅୗỈ ࢆ㇏࠿࡟ࡋ㸪ᅵ◁ᔂࢀࡸᆅ⾲ᔂቯ࡞࡝ࢆ㜵ࡄࠋࡲࡓ 㸪⏕≀࿧྾࡟ࡼࡗ࡚ቑຍࡍࡿ஧㓟 ໬Ⅳ⣲ࢆගྜᡂ࡟ࡼࡗ࡚ᅛᐃ໬ࡋ㸪ᆅ⌫ ᗘࢆಖࡕ㸪✜ࡸ࠿࡞Ẽ㇟⎔ቃࢆసࡿᙺ๭ࢆ ᢸࡗ࡚࠸ࡿࠋ᳃ᯘࡢ㔜せ࡞ᙺ๭ࡀᗈࡃྉࡤࢀ࡚࠸ࡿ⌧ᅾ࡛ࡶ㸪ᆅ⌫つᶍࡢ᳃ᯘ◚ቯࡣ ๻ⓗ࡞ࢫࣆ࣮ࢻ࡛ᗈࡲࡾ㸪ࡑࡢࡦࡎࡳࡣࡉࡲࡊࡲ࡞࡜ࡇࢁ࡛Ⓨ⌧ࡋ࡚࠸ࡿࠋ ᳃ᯘ◚ቯࡢཎᅉࡢ࡯࡜ࢇ࡝ࡣ 㸪ே㛫࡟ࡼࡿ↓సⅭ࡞᳃ᯘఆ᥇࡛࠶ࡿࠋ㈋ᅔᆅᇦ࡛ࡢ ↝⏿㎰ᴗ㸪୙㐺ษ࡞ၟᴗఆ᥇ࡸ᳃ᯘ௨እࡢ฼⏝┠ⓗ࡟ࡼࡿ ᅵᆅࡢ㌿⏝࡞࡝㸪ࡑࡢཎᅉ ࡣࡉࡲࡊࡲ࡛࠶ࡿࠋࡓࡋ࠿࡟ே㛫࡟ࡼࡿ᳃ᯘ◚ቯࡣⴭࡋ࠸ࡶ ࡢࡀ࠶ࡾ㸪⮬↛ᅇ᚟ຊࢆ ୖᅇࡿ㏿ᗘ࡛ᔂቯࡉࡏ࡚࠸ࡿࠋࡋ࠿ࡋ᳃ᯘ◚ቯࡢཎᅉࡣ㸪ே㛫࡟ࡼࡿ┤᥋ⓗ࡞◚ቯ ࡔ ࡅ࡛ࡣ࡞࠸ࠋᒣⅆ஦ࡸᒣᔂࢀ㸪⑓⹸ᐖ࡞࡝ࡶཎᅉࡢ㸯ࡘ࡛࠶ࡿࠋ⸄Ⅳࢆ࢚ࢿࣝࢠ࣮※ ࡜ࡋ࡚฼⏝ࡋ࡚࠸ࡓ᫬௦ࡣ 㸪ࡓ࡜࠼⑓⹸ᐖࡀⓎ⏕ࡋ࡚ࡶ⿕ᐖ ࢆཷࡅࡓᶞᮌࢆே㛫ࡀᒣ ᯘ࠿ࡽᣢࡕฟࡋ㸪⇞↝฼⏝ࡍࡿࡇ࡜࡛⿕ᐖࡀ㐣๫࡟ᗈࡀࡽࡎ㸪ࡲࡓ⑓⹸ᐖࡶ㥑㝖ࡍࡿ ࡇ࡜ࡀ࡛ࡁࡓࠋࡘࡲࡾே㛫ࡀ⏕ែ⣔ࡢ୍ဨ࡜ࡋ࡚ ⏕ែ⣔⥔ᣢࡢ㔜せ࡞ᙺ๭ࢆᢸࡗ࡚ ࡁ ࡓࠋࡋ࠿ࡋ⇞ᩱ㠉࿨࡜࠸ࢃࢀࡿ᫬௦௨㝆࡟࡞ࡿ ࡜ே㛫ࡣᒣᯘ࡟ධࡽ࡞ࡃ࡞ࡾ㸪⿕ᐖࢆ

2

ཷࡅࡓᮌࡣ᳃ᯘෆ࡟ᨺ⨨ࡉࢀࡓࠋ⑓⹸ᐖࡣ㥑㝖ࡉࢀࡿࡇ࡜࡞ࡃ⦾Ṫࢆ⥆ࡅ㸪ே㛫ࡀ㥑 㝖ࢆヨࡳࡼ࠺࡜ࡋࡓ᫬࡟ࡣࡍ࡛࡟ᗈ⠊ᅖ࡟ࡑࡢ⿕ᐖࢆᣑ኱ࡉࡏ㸪㥑㝖ࡀ㏣࠸ࡘ࠿࡞࠸ ≧ែ࡟ࡲ࡛῝้໬ࡋ ࡚࠸ࡿࡢࡀ⌧≧࡛࠶ࡿ 1㸧_ࠋ ୡ⏺ⓗ࡟⮬↛ಖㆤ㸪᳃ᯘಖㆤࡀ㔜せどࡉࢀ 㸪ᣢ⥆ྍ⬟࡞᳃ᯘ⤒Ⴀࡀᅜ㝿ⓗ࡟ồࡵࡽ ࢀ࡚࠸ࡿ௒᪥㸪᳃ᯘ⿕ᐖ㜵㝖ࡣ⏕≀ከᵝᛶࡢಖ඲࡜୧❧ࡍࡿࡇ࡜ࡀ୙ྍḞ࡛࠶ࡿࠋ⸆ ๣࡟ࡼࡿ⹸ᐖ㜵㝖ࡣ 㸪㜵㝖ᑐ㇟࡜ࡍࡿ⏕≀ࡢࡳࢆ㑅ᢥⓗ࡟ つไ࡛ࡁࡿ⸆๣ࡸ௚ࡢ⏕ែ ⣔࡟ᙳ㡪ࢆཬࡰࡉ࡞࠸⸆๣ࡢ฼⏝ᢏ⾡ࡢ㛤Ⓨࡀᚲせ࡛࠶ࡾ 㸪ࡉࡲࡊࡲ࡞◊✲ࡀ㐍ࡵࡽ ࢀ࡚࠸ࡿࠋᶞᮌࡢᑑ࿨ࡣ௚ࡢ⏕≀࡜ࡣ␗࡞ࡾ 㸪㠀ᖖ࡟㛗࠸ࠋࡑࢀࡺ࠼㛗࠸ᖺ᭶ࢆ࠿ࡅ ࡚ࡉࡲࡊࡲ࡞㜵ᚚ≀㉁ࢆ஧ḟ௦ㅰ≀࡜ࡋ࡚ྜᡂࡋ㸪ᶞ⓶ࡸⴥ㸪ᮦ࡟㈓ⶶࡋ࡚࠸ࡿࠋࡇ ࢀࡽ㜵ᚚ≀㉁ࡣ⳦㢮ࡸ᪻⹸࡟ᑐࡋ࡚ᚷ㑊ࡸẘᛶຠᯝࢆⓎ᥹ࡋ㸪ᶞᮌࡸᮌᮦࡢ⪏ஂᛶࢆ ྥୖࡉࡏ࡚࠸ࡿ 2,3,4㸧_{ࠋࡇࡢࡼ࠺࡞㜵ᚚ≀㉁ࡣࡑࡢᶞᮌࡀᮙࡕࡓᚋࡣᚤ⏕≀࡞࡝࡟ࡼࡗ} ࡚ศゎࡉࢀ㸪⏕ែ⣔࡟ఱࡽᙳ㡪ࢆ୚࠼ࡿࡇ࡜࡞ࡃ ⮬↛⏺࡟ᡠࡗ࡚ࡺࡃࠋࡇࡢࡼ࠺࡞ኳ ↛≀⏤᮶ࡢᡂศࡣ᪻⹸࡟ᑐࡋ࡚ㄏᘬཬࡧᚷ㑊ຠᯝࢆ♧ࡍ 5,6,7㸧_{ࡇ࡜࠿ࡽ㸪ᶞᮌᡂศࢆࡶ} ࡕ࠸ࡓㄏᘬ㸪ᚷ㑊๣ࡢ㛤Ⓨࡀ⾜ࢃࢀ࡚࠸ࡿࠋᐇ㝿 㸪᪥ᮏࡢ࣐ࢶᯤࢀ⿕ᐖ࡟࠾࠸࡚ 㸪ᶞ ᮌᡂศ࡛࠶ࡿȘ-ࣆࢿࣥࡸ࢚ࢱࣀ࣮ࣝࡢΰྜᾮࢆ౑⏝ࡋࡓ㜵㝖ἲ 6,8㸧_{ࡀ☜❧ࡉࢀ࡚࠸ࡿࠋ} ࡋ࠿ࡋ㸪ࡇࡢࡼ࠺࡞ኳ↛≀࡟ࡼࡿ㜵㝖᪉ἲࡣᮍࡔ☜❧ࡉࢀ࡚࠸࡞࠸ࡶࡢࡀከ࠸ ࡢࡀ⌧ ≧࡛࠶ࡿࠋ ᪥ᮏ࡟࠾ࡅࡿ᳃ᯘ⿕ᐖࡢ 1 ࡘ࡟ࣈࢼ⛉ᶞᮌࡢ㞟ᅋᯤᦆ⿕ᐖࡀ࠶ࡿࠋࣈࢼ⛉ᶞᮌࡣ᪥ ᮏࡢཎ⏕ᯘࡢ୺せᵓᡂᶞ✀࡛࠶ࡾ㸪ୡ⏺ⓗ࡟ぢ࡚ࡶࣈࢼཎ⏕ᯘࡀ⮬↛ࡢ≧ែ࡛ṧࡗ࡚ ࠸ࡿࡢࡣ᪥ᮏࡔࡅ࡛࠶ࡿࠋࡋ࠿ࡋ㸪᪥ᮏᾏഃࢆ୰ᚰ࡟ᗈࡀࡿࣈࢼ⛉ᶞᮌࡢ㞟ᅋᯤᦆ⿕

3

ᐖࡣᣑ኱ഴྥ࡟࠶ࡿࡇ࡜࠿ࡽၥ㢟どࡉࢀ࡚࠸ࡿࠋࡇࡢ⿕ᐖࡣ㸪㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ࡛࠶

ࡿ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ㸦Platypus quercivorus MURAYAMA㸧ࡀᶞᮌ࡟✸ධࡋ㸪ᶞయ

ෆ࡛ࡑࡢඹ⏕⳦࡛࠶ࡿ Raffaelea quercivora ࡢఏ᧛࡟ࡼࡾᶞᮌ㎶ᮦ㒊ࡀ〓ኚࡋ㸪ࡑࢀ࡟ క࠺ࢳ࣮ࣟࢫᙧᡂ 9,10,11㸧_{࡞࡝࡟ࡼࡗ࡚ᶞᮌࡢ㏻Ỉ㜼ᐖࡀ⏕ࡌ 㸪⤖ᯝⓗ࡟ᶞᮌࡣᯤṚࡍ} ࡿࠋࡇࡢ⿕ᐖ࡟ࡘ࠸࡚᪻⹸Ꮫ 12,13,14,15㸧_{㸪⳦㢮Ꮫ 16,17,18,19,20,21}㸧_{࡞࡝ࡉࡲࡊࡲ࡞どⅬ࠿ࡽ} ࡢ◊✲ࡀࡍࡍࡵࡽࢀ࡚࠸ࡿࡀ 㸪ࡇࡢ⿕ᐖ࡟㛵ࡋ࡚໬Ꮫⓗ 㸪⏕໬Ꮫⓗ࡞どⅬ࠿ࡽࡢ◊✲ ࡣ࡯࡜ࢇ࡝࡞ࡉࢀ࡚࠸࡞࠸ࠋࡑࡇ࡛ᮏ◊✲࡛ࡣ R. quercivora ࡟ᑐࡋ࡚ឤཷᛶࡢ㧗࠸࣑ ࢬࢼࣛࢆ⏝࠸㸪ኚⰍ㒊࡜೺඲㒊ࡢᢳฟᡂศࢆẚ㍑ࡋ㸪R. quercivora ࡢㄏᑟࡍࡿ㓝⣲࡟ ࡘ࠸࡚ࣔࢹࣝ໬ྜ≀ࢆ⏝࠸᳨࡚ウࡋࠊࡉࡽ࡟ྠ㓝⣲࡟ࡼࡿ ᢳฟᡂศࡢኚ໬ࢆ☜ㄆࡍࡿ ௚ࠊ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ✸ධ࡞ࡽࡧ࡟ R. quercivora ࡢឤᰁ࡟ࡼࡗ࡚ᯤṚࡋࡓ࣑ࢬࢼ ࣛ࡟ྵࡲࢀࡿ໬ྜ≀ࢆ༢㞳ྠᐃࡍࡿࡇ࡜࡛ࠊ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ࠿ࡽ࣑ࢬࢼࣛࢆಖ ㆤࡍࡿࡓࡵࡢᇶ♏ⓗ▱ぢࢆᥦ౪ࡍࡿࡇ࡜ࢆ┠ᣦࡍࠋ

4

ཧ⪃ᩥ⊩ 1. ᯇᾆ㑥᫛㸦1997㸧᳃ᯘ⿕ᐖ㜵㝖࡜⏕≀ከᵝᛶಖ඲㸬 ᳃ᯘ⥲◊㛵す◊✲ 46㸸1 2. Ᏻ⏣ᚁᕷ㸦2002㸧ᶞᮌࡢ㢦㸫ᶞᮌᢳฟᡂศ࡜ࡑࡢ฼⏝㸫㸬ᾏ㟷♫ ⁠㈡┴ 196-201 3. ㇂⏣㈅ගඞ㸦1996㸧ᮌࡢࡩࡋࡂ࡞ຊ㸬ᩥ◊ฟ∧ ᮾி 4-53 4. ᶓ⏣ಙ୕㸪ගỌ ᚭ㸦2002㸧ᮌ㉁ࡢᙧᡂ ࣂ࢖࣐࢜ࢫ⛉Ꮫ࡬ࡢᣍᚅ㸬ᾏ㟷♫ ⁠ ㈡┴ 307-310

5. Huber, D. P. W. and Borden, J. H. (2001) Angiosperm bark volatiles disrupt response of Douglas-fir beetle, Dendroctonus pseudotsugae, to attractant-baited traps. J. Chem. Eco. 27 : 217-233

6. Sadik, G., Islam, R., Rahman, M. M., Khondkar, P., Rashid, M. A. and Sarker, S.D. (2003) Antimicrobial and cytotoxic constituents of Loranthus globosus. Fitoterapia,

74 : 308–311

7. Yamasaki, T., Saito, M. and Sakoguchi, H. 㸦 1997 㸧 (-)germacrene D:Masking Substances of Attractants for the Cerambycid Beetle, Monochamus alternatus (HOPE). Appl. Entmol. Zool. 32: 423-429

8. Vattem, D.A., Lin, Y.-T., Labbe, R.G. and Shetty, K. (2003) Antimicrobial activity against select food-borne pathogens by phenolic antioxidants enriched in cranberry pomace by solid-state bioprocessing using the food grade fungus Rhizopus oligosporus. Proc. Biochem. in press.

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fungus of a new wilt disease vectored by the ambrosia beetle Platypus quercivorus. J. Wood. Sci. 47: 425-42

10. 㯮⏣៞Ꮚ㸪㧗⏿⩏ၨ㸦2002㸧ࢼࣛ㢮㞟ᅋᯤᦆᶵᵓࡢゎ᫂㸬᳃ᯘ⥲◊ᡤሗ  11㸸 8-9

11. 㯮⏣៞Ꮚ㸪ᒣ⏣฼༤㸦1996㸧ࢼࣛ㢮㞟ᅋᯤᦆ࡟ぢࡽࢀࡿ㎶ᮦࡢኚⰍ࡜㏻Ỉᶵ⬟

ࡢపୗ㸬᪥ᯘㄅ 78㸸84-88

12. Igeta, Y., Esaki, K., Kato, K. and Kamata, N.㸦2003㸧Influence of light condition on the stand-level distribution and movement of the ambrosia beetle Platypus quercivorus (Coleoptera: Platypodidae) Appl. Entomol. Zool. 38:167-175

13. 㙊⏣┤ே (2002) ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ⏕ែ㸬 ᳃ᯘ⛉Ꮫ 35: 26-34 14. Kobayashi, M., Ueda, A. and Takahata, Y. (2001) Inducing Infect ion of Oak logs by a

Pathogenic Fungus Carries by Platypus quercvorus (MURAYAMA) (Coleoptera : Platypododae). J. For. Res. 6 : 153-136

15. Ueda, A. and Kobayashi, M. (2001) Aggregation of Platypus quercivorus (Murayama) (Coleoptera : Platyposisae) on Oak Log Bored by Males of the Species. J. For. Res.

6 : 173-179

16. ఀ⸨㐍୍㑻㸦2000㸧ࢼࣛ㢮㸪ࢩ࢖࣭࢝ࢩ㢮ᯤṚ⿕ᐖࡢ⌧≧㸬ᒣᯘ 㸳㸳 㸸60-68

17. ఀ⸨㐍୍㑻㸦2000㸧ࣈࢼ⛉ᶞᮌࡢ㞟ᅋᯤṚ㸫⳦㢮࡜᪻⹸ࡢඹ⏕⳦ࡢ࡞ࡒ㸫㸬᪥ ᮏ⳦Ꮫ఍す᪥ᮏᨭ㒊఍ሗ 10㸸16-22㸬

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㐃ࡍࡿ⳦㢮㸬᪥ᯘㄅ  80㸸170-175

19. ᮌᮧ໷Ꮚ㸪ᯇ⏣㝧௓㸪❑㔝㧗ᚨ㸪ఀ⸨㐍୍㑻㸦 2003㸧Raffaelea quercivora ⳦ᰴ

㛫ࡢᙧែᏛⓗ≉ᚩ㸬  ୰᳃◊ 51㸸199-200

20. Kubono, T. and Ito, S. (2002) Raffaerea quercivora sp. nov. associated with mass mortality of Japanese oak, and the ambrosia beetle  (Platypus quercivorus). Mycoscience 43㸸255-260

21. ගỌ ᚭ㸪Ọᯇ࿴ᡂ㸪す஭Ꮥᩥ㸪኱㈡⚈἞㸦1999㸧ᢸᏊ⳦㢮ࡢ⳦⣒ᡂ㛗࡟ཬࡰࡍ ᶞ⓶࣏ࣜࣇ࢙ࣀ࣮ࣝࡢຠᯝ㸬᪥⳦ሗ 40㸸91-96

7

ᗎㄽ ୡ⏺࡟ᗈࡀࡿ᳃ᯘ⿕ᐖ ᳃ ᯘࡢ ⾶㏥ 㸪ᯤ Ṛࡢ ཎᅉ ࡣ㸪 ᅜࡸ ᆅᇦ 㸪⿕ ᐖࡢ ᵝᏊ ࡞࡝ ࡟ࡼ ࡗ࡚ ␗࡞ ࡗ࡚ ࠸ࡿ ࡀ 㸪 ୍ ⯡ ࡟ ࡣ ࡑ ࡢ ᡂ 㛗 㐣 ⛬ ࡟ ࠾ ࡅ ࡿ ࡉ ࡲ ࡊ ࡲ ࡞ ⎔ ቃ ࢫ ࢺ ࣞ ࢫ ࡟ ࡼ ࡗ ࡚ ᘬ ࡁ ㉳ ࡇ ࡉ ࢀ ࡿ ࡇ ࡜ࡀㄆ㆑ࡉࢀ࡚࠸ࡿࠋ౛࠼ࡤ 㸪࢟ࢡ࢖࣒ࢩࡸ࣐ࢶࣀࢨ࢖ࢭࣥࢳࣗ࢘࡞࡝ ᪻⹸㢮ࡸࢩ࢝ ࡞࡝ࡢ㫽⋇㢮࡟ࡼࡿᦤ㣗⿕ᐖ 㸪࡞ࡽࡓࡅ⑓࡟௦⾲ࡉࢀࡿᅵተ⑓ᐖ࡜࠸ࡗࡓ⏕≀ⓗࢫࢺ ࣞࢫ㸪㓟ᛶ㞵ࡸ኱Ẽởᰁ 㸪ᅵተởᰁ≀≀㉁㸪Ỉศ᮲௳ࡸ ᗘ᮲௳ ࡜࠸ࡗࡓ㠀⏕≀ⓗࢫ ࢺࣞࢫ࡞࡝㸪ࡉࡲࡊࡲ࡞せᅉࡀ」ྜⓗ࡟స⏝ࡋ࡚Ⓨ⏕ࡍࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿ 1)_ࠋ ୡ ⏺ ⓗ ࡟ ၥ 㢟 ど ࡉ ࢀ ࡚ ࠸ ࡿ ᳃ ᯘ ⾶ ㏥ ࡢ ୍ ࡘ ࡟ ࣈ ࢼ ⛉ ᶞ ᮌ ࡢ ⾶ ㏥ ᯤ Ṛ ⿕ ᐖ ࡀ ࠶ ࡿ ࠋ 600 ✀࡟ࡶ࠾ࡼࡪࣈࢼ⛉ᶞᮌࡣୡ⏺ࡢ⇕ᖏ࠿ࡽ ᖏᆅᇦ࡟ᗈࡃศᕸࡋ࡚࠸ࡿᶞ✀࡛࠶ ࡾ㸪ࡑࢀࡽࡢᮦࡣ㓇ᶡࡸᘓ⠏⏝ᮦ࡜ࡋ࡚ࡢ฼⏝౯್ࡀ㧗ࡃ 㸪ࡲࡓᗈⴥᶞᯘࡢ୺せᵓᡂ ᶞ✀࡛࠶ࡿࡇ࡜࠿ࡽ㸪ࡇࢀࡽࡢᶞ✀ࡢ⾶㏥㸪ᯤṚၥ㢟ࡣࡼࡾ㔜せどࡉࢀ࡚࠸ࡿࠋ࢔࣓ ࡛ࣜ࢝Ⓨ⏕ࡋ࡚࠸ࡿࢼࣛ㸪࢝ࢩ㢮ⴎ෼⑓㸦Oak wilt㸧ࡣ 1930 ᖺ௦࡟Ⓨ⏕ࡋ㸪⌧ᅾࡶࡑ ࡢ ⿕ ᐖ ࡣ ၥ 㢟 ࡜ ࡞ ࡗ ࡚ ࠸ ࡿ ࠋ ࡇ ࡢ ⿕ ᐖ ࡣ 㸪 ࢟ ࢡ ࢖ ࣒ ࢩ ࡟ ࡼ ࡗ ࡚ 㐠 ᦙ ࡉ ࢀ ࡿ ⑓ ཎ ⳦ Ceratocystis fagacearm ࡟ࡼࡗ࡚ᘬࡁ㉳ࡇࡉࢀࡿࡇ࡜ࡀ᫂ࡽ࠿࡟ࡉ ࢀ࡚࠸ࡿࠋ୍᪉㸪ࣚ ࣮ࣟࢵࣃ༡㒊ࡢᅜࠎ࡛ࡣ Pytophthora ᒓ⳦㸪≉࡟ P. cinnamomi ࡟ࡼࡗ࡚ࢼࣛ㢮ࡢ⾶㏥ ᯤṚࡀࡦࡁ࠾ࡇࡉࢀ࡚࠸ࡿ࡞࡝ 㸪ከࡃࡢ᳃ᯘ⾶㏥⿕ᐖࡀሗ࿌ࡉࢀ࡚࠸ࡿ 2, 3)_ࠋ

8

᪥ᮏ࡟࠾ࡅࡿࣈࢼ⛉ᶞᮌࡢᯤṚ⿕ᐖ  ᪥ᮏࡢࢼࣛ㢮࡟ࡣ⣙ 50 ✀ࡢ⑓ᐖࡀグ㘓ࡉࢀ࡚࠸ࡿࠋࡋ࠿ࡋ࡞ࡽࡓࡅ⑓㸦Armillaria spp.㸧࡞࡝ࡢᅵተ⑓ᐖࢆ㝖ࡁ㸪ᡂᮌࢆᯤṚࡉࡏࡿࡼ࠺࡞⑓ᐖࡣ࡞ࡃ㸪ࡲࡓ⑓ᐖ࡟ࡼࡗ ࡚ࢼࣛ㢮ࡀ㞟ᅋⓗ࡟ᯤṚࡋࡓグ㘓ࡶ࡯࡜ࢇ࡝ṧࡉࢀ࡚࠸࡞࠸ࠋ ࡋ࠿ࡋ1980ᖺ௦௨㝆㸪ᮏᕞ᪥ᮏᾏഃࢆ୰ᚰ࡟ࢼࣛ㢮㸦ࢥࢼࣛ㸪࣑ࢬࢼࣛ࡞࡝㸧ࡀ㞟ᅋ ࡛ᯤṚࡍࡿ⿕ᐖ㸦෗┿1㸧ࡀⓎ⏕ࡋ㸪⌧ᅾ࡛ࡶࡑࡢ⿕ᐖࡣᣑ኱ഴྥ࡟࠶ࡿࡇ࡜࠿ࡽၥ 㢟どࡉࢀ࡚࠸ࡿ4㸧_{ࠋࡲࡓᐑᓮ┴ࡸ㮵ඣᓥ┴ࡢ஑ᕞᆅ᪉ࡸ ⣖ఀ༙ᓥࡢ୕㔜┴㸪࿴ḷᒣ┴㸪} ዉⰋ┴ࡢ┴ቃ࡛ࡣࢩ࢖࣭࢝ࢩ㢮㸦࣐ࢸࣂࢩ࢖ 㸪࢘ࣛࢪࣟ࢞ࢩ㸧࡟ᯤṚ⿕ᐖࡀⓎ⏕ࡋ࡚ ࠸ࡿࠋࡇࢀࡽࡢ⿕ᐖᆅ࡛ඹ㏻ࡍࡿ⏕≀ⓗせᅉ ࡜ࡋ࡚㸪ࡍ࡭࡚ࡢᯤṚᮌ࠿ࡽ㣴⳦ᛶ࢟ࢡ ࢖࣒ࢩࡢ1✀࡛࠶ࡿ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ㸦Platypus quercivorus Murayama㸧ࡀ✸ධࡋ

࡚࠸ࡿࡇ࡜ࡀ☜ㄆࡉࢀࡓ5㸧_{ࠋ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධࢆཷࡅࡓᶞᮌࡣ㎶ᮦ㒊࡟Ꮝ} 㐨࡟ࡑࡗࡓ〓ⰍࡢኚⰍ㒊ࢆᙧᡂ 㸦෗┿2㸧ࡋ㸪≉࡟ࢼࣛ㢮࡛ࡣ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ ࡢ✸ධᚋ㸪ᛴ⃭࡟ⴎ෼㸪ᯤṚࡍࡿࠋ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධ࡟ࡼࡗ࡚ᙧᡂࡉࢀࡓ ኚⰍ㒊࡛ࡣ㸪ࢳ࣮ࣟࢫ࡞࡝ࡀ⏕ᡂࡍࡿࡇ࡜࡛ ㏻Ỉࡀ㜼ᐖࡉࢀᶞᮌࡀᯤṚ࡟⮳ࡿ6,7,8㸧_ࠋ ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ✸ධ ᮌ ㎶ ᮦ ࡟ࡳࡽ ࢀ ࡿ ኚ Ⰽ 㒊 㸪 ࢝ ࢩ ࣀ ࢼ࢞ ࢟ ࢡ ࢖ ࣒ࢩࡢ య ⾲ 㸪 ⬊Ꮚ㈓ⶶჾᐁ㸪࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢᗂ⹸ࡢయ⾲࠿ࡽඃ༨ⓗ࡟ศ㞳ࡉࢀࡓRaffaelea

quercivora Kubono et Shin-Ito sp.nov.9㸧_{ࡣ㸪ⱑᮌ࡟ᑐࡍࡿ᥋✀ヨ㦂 ࡟࠾࠸࡚㎶ᮦ㒊ࢆኚ}

ⰍࡉࡏᯤṚ࡟⮳ࡽࡏࡿࡇ࡜10㸧_{ࡼࡾ㸪R. quercivoraࡢឤᰁ࡟ࡼࡗ࡚㎶ᮦ㒊ࡀ〓ኚࡋ㸪ᶞ}

10

⿕ᐖᆅᇦ ࢼࣛ㢮ࡢᯤᦆ⿕ᐖࡣ 㸪1934 ᖺ࡟ᐑᓮ┴ࡸ㮵ඣᓥ┴࡛࢖ࢳ࢖࢞ࢩࡸ࢘ࣛࢪࣟ࢞ࢩ 㸪࢔ ࢝࢞ࢩ࡞࡝ࡢᯤṚࡀ☜ㄆࡉࢀࡓࠋࡑࡢᚋ㸪㧗▱┴㸦1950 ᖺ㸪1952 ᖺ㸧㸪රᗜ┴㸦1952 㹼1956 ᖺ㸧㸪ᒣᙧ┴㸦1958㹼1960 ᖺ㸪1979 ᖺ㸧࡞࡝࡛⿕ᐖࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋᒣᙧ┴ ࡛ࡣ 1900 ᖺ௦ึ㢌࡟⹸ࡢ✸ධ࡟ࡼࡗ࡚࣑ࢬࢼࣛ㸪ࢥࢼࣛࡀ඲⁛ࡋࡓ࡜࠸࠺ሗ࿌ࡶ࠶ ࡾ㸪㠀ᖖ࡟ྂࡃ࠿ࡽ⿕ᐖࡀᏑᅾࡋ࡚࠸ࡓ 㸦⾲ 1㸧ࠋ1980 ᖺ௨㝆ࡢ⿕ᐖ࡟ࡘ࠸࡚ᐇែㄪ ᰝࡀ⾜ࢃࢀࡓ⤖ᯝ㸪ᮏᕞ᪥ᮏᾏഃ࡛ࡣᒣᙧ┴ 㸪᪂₲┴㸪⚟ᓥ┴㸪▼ᕝ┴㸪⚟஭┴㸪ᒱ 㜧┴㸪⁠㈡┴㸪ி㒔ᗓ㸪රᗜ┴㸪㫽ྲྀ┴㸪ᓥ᰿┴ࡢ 1 ᗓ 10 ┴࡛⿕ᐖࡀሗ࿌ࡉࢀ㸪ࡲ ࡓ஑ᕞᆅ᪉࡛ࡶ㮵ඣᓥ┴ࡸᐑᓮ┴࡟࠾࠸࡚⿕ᐖࡀሗ࿌ࡉࢀࡓࠋ ࡉࡽ࡟ 1999 ᖺ࡟ࡣ⣖ ఀ༙ᓥ㸦୕㔜┴㸪࿴ḷᒣ┴㸪ዉⰋ┴ࡢ┴ቃ㸧࡟࠾࠸࡚ࡶ⿕ᐖࡀⓎ⏕ࡋ 㸪⌧ᅾ 1 ᗓ 15 ┴㸦ᅗ 1㸧࡟ᗈࡀࡗ࡚࠸ࡿ 4㸧_ࠋ ⿕ᐖᶞ✀ ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡣ㸪17 ⛉ 27 ᒓ 45 ✀ࡢᶞᮌ࡟ຍᐖࡍࡿࡇ࡜ࡀ ▱ࡽࢀ࡚࠸ ࡿ ࠋ ஑ᕞࡸ⣖ఀ༙ᓥ࡛ࡣ 㸪࢔࢝࢞ࢩ㸪࢔ࣛ࢝ࢩ㸪࢖ࢳ࢖࢞ࢩ㸪࢘ࣛࢪࣟ࢞ࢩ㸪ࢶࢡࣂࢿ࢞ ࢩ࡞࡝ࡢࢥࢼࣛᒓ㸦Quercus㸧ࡢ࢝ࢩ㢮ࡸࢶࣈࣛࢪ࢖㸪ࢫࢲࢪ࢖࡞࡝ࡢࢩ࢖ᒓ 㸦Castanopsis㸧㸪࣐ࢸࣂࢩ࢖ᒓ㸦Pasania㸧㸪ࢱࣈࣀ࢟ᒓ㸦Persea㸧࡞࡝ࡢࣈࢼ⛉㸦Fagaceae㸧 ᖖ⥳ᶞࡀຍᐖࡉࢀࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ୍᪉㸪᪥ᮏᾏഃࡢᆅᇦ࡛ࡣࢥࢼࣛ 㸪࣑ࢬ ࢼࣛ㸪ࢡࢾࢠ࡞࡝ࡢࢥࢼࣛᒓ 㸪ࢡࣜᒓ㸦Castanea㸧㸪ࣈࢼᒓ㸦Fagus㸧࡞࡝ࡢࣈࢼ⛉ࡢ ⴠⴥᶞࡀຍᐖࡉࢀࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦⾲ 2㸧ࠋ≉࡟╔┠ࡍ࡭ࡁⅬࡣ㸪᪥ᮏᾏഃᆅ

11

ᇦ࡛ࡣࢼࣛ㢮㸪≉࡟࣑ࢬࢼࣛࡸࢥࢼࣛ࡜࠸ࡗࡓࣈࢼ⛉ⴠⴥᶞࡀ㞟ᅋⓗ࡟኱㔞 ᯤṚࡋ࡚

࠸ࡿ୍᪉࡛㸪஑ᕞᆅ᪉࡞࡝ࡢࣈࢼ⛉ᖖ⥳ᶞ࡛ࡣ 㸪ᯤṚ⿕ᐖࡣሗ࿌ࡉࢀ࡚࠸ࡿࡀ᪥ᮏᾏ ഃࡢࡼ࠺࡞㞟ᅋⓗ࡞ᯤṚ⿕ᐖ࡛ࡣ࡞ࡃ㸪࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ࡟ຍᐖࡉࢀ࡚ࡶᯤṚࡋ

12

࢟ࢡ࢖࣒ࢩ ࢟ࢡ࢖࣒ࢩ࡜ࡣ㸪ࡑࡢྡࡢ㏻ࡾࠕᮌ႞࠸⹸࡛ࠖ࠶ࡿࠋᶞ⓶ୗࡶࡋࡃࡣ㎶ᮦෆ࡟ࢺࣥ ࢿࣝ㸦Ꮝ㐨㸧ࢆ࠶ࡅ㸪ࡑࡢ୰࡛⏕άࡍࡿ✸Ꮝ⹸ࡢ⥲⛠࡛࠶ࡿࠋ࢟ࢡ࢖࣒ࢩࡣ㠧⩟┠ࢰ ࣒࢘ࢩୖ⛉࡟ᒓࡍࡿ⏥⹸࡛࠶ࡾ 㸪య㛗ࡣ 0.4̺25mm㸦ከࡃࡣయ㛗 2㸫5mm㸧࡛㸪యⰍ ࡣ࡯࡜ࢇ࡝ࡢ✀ࡀ㯮Ⰽ࠿ࡽ〓Ⰽࢆ♧ࡍࠋ࢟ࢡ࢖࣒ࢩ⛉ࡣ⌧ᅾ 215 ᒓ 7000 ✀㢮௨ୖࡀ ሗ࿌ࡉࢀ࡚࠾ࡾ㸪᪥ᮏ࡟ࡣ⣙ 300 ✀ࡀศᕸࡋ࡚࠸ࡿࠋࡇࢀࡽ࢟ࢡ࢖࣒ࢩࡣ㸪㣵࡟ࡼࡾ ᶞ⓶ୗ࢟ࢡ࢖࣒ࢩ࡜㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ࡟኱ูࡍࡿࡇ࡜ࡀ࡛ࡁࡿࠋᶞ⓶ୗ✸Ꮝ࢟ࢡ࢖࣒ ࢩ㸦Bark beetle㸧ࡣࡑࡢྡࡢ㏻ࡾ㸪ᶞ⓶ୗ㸪୺࡟ෆᶞ⓶ୗ࡟Ꮝ㐨ࢆ᥀ࡾ㸪ෆᶞ⓶⤌⧊ ࢆ㣵࡟ࡋ࡚⦾Ṫࡍࡿ࢟ࢡ࢖࣒ࢩ㢮࡛࠶ࡾ 㸪᪥ᮏ࡛ࡣ࣐࢝ࣛࢶࣖࢶࣂ࢟ࢡ࢖࣒ࢩࡸࣄࣂ ࣀ࢟ࢡ࢖࣒ࢩ㸪࣐ࢩࣀࢥ࢟ࢡ࢖࣒ࢩ㸪࢟࢖ࣟࢥ࢟ࢡ࢖࣒ࢩ࡞࡝ࡀ▱ࡽࢀ࡚࠸ࡿࠋ୍᪉㸪 㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ㸦Ambrosia beetle㸧ࡣᶞ⓶ୗ✸Ꮝ࢟ࢡ࢖࣒ࢩ࡜ࡣ␗࡞ࡾ㸪㎶ᮦෆ࡟ Ꮝ㐨ࢆ᥀ࡾ⏕⫱ࡍࡿࡀ㸪ᮦࢆ㣵࡟ࡣࡋ࡚࠸࡞࠸ࠋ㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ㢮ࡢ㞤➼ࡢ⫼⬚㒊

࡟⿄≧ࡢ⳦ࡢ࠺ Mycangia ࢆᣢࡕ㸪ࡑࡢ୰࡟㓝ẕ࡞࡝ࡢඹ⏕⳦㸦ambrosia fungi㸧ࢆ

㈓ⶶࡋ࡚࠸ࡿࠋ㣴⳦ᛶ࢟ࢡ࢖࣒ࢩࡣ᪂ࡓ࡞ᮌ࡟✸ධࡍࡿ࡜ 㸪⫼⬚㒊ࡢ⳦ࡢ࠺࡟㈓ⶶࡋ ࡚࠸ࡿ⳦㢮ࢆᮦෆ࡛⦾Ṫࡉࡏ 㸪⳦⣒ࡸ⬊Ꮚࢆ㣗ࡍࠋ㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ㢮ࡢぶ⹸ࡣ 㸪Ꮝ 㐨ෆ࡛ඹ⏕⳦ࡢᇵ㣴⟶⌮ 㸪Ꮝ㐨ෆࡢΎᤲ㸪᥮Ẽ㸪‵ᗘㄪᩚ㸪㞵Ỉࡢὶධ㜵Ṇ㸪ࢲࢽ㢮 ࡢ౵ධ㜵Ṇ࡞࡝ࡢసᴗࢆ⾜࠸ 㸪ᗂ⹸ࡀᡂ⹸࡟࡞ࡿࡲ࡛௜ࡁῧࡗ࡚ୡヰࢆࡍࡿࡢࡀ≉ᚩ ࡛࠶ࡿࠋࡲࡓ㸪㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ࡟ࡼࡿᐖࡣᮦ࡟┤᥋Ꮝ㐨ࡀసࡽࢀࡿࡔࡅ࡛࡞ࡃ 㸪ඹ ⏕⳦࡟ࡼࡗ࡚Ꮝ㐨ཬࡧᮦࡀ〓Ⰽࡲࡓࡣ㯮Ⰽ࡟ኚⰍࡋ㸪ᡂ⹸ࡢ⬺ฟᚋࡢᏍ㐨࡟ࡣྛ✀ࡢ ⭉ᮙ⳦ࡀ⦾Ṫࡍࡿࡇ࡜࡛ᮦࡀෆ㒊࠿ࡽ㏿ࡸ࠿࡟ศゎࡉࢀࡿࠋ⮬↛⏺࡟࠾࠸࡚ࡣ 㸪᳃ᯘ

13

ෆࡢಽᮌ➼ࡢศゎࢆ᪩ࡵࡿᙺ๭ࢆᢸࡗ࡚࠸ࡿࡀ 㸪ᮦࢆ฼⏝ࡍࡿே㛫࡟࡜ࡗ࡚㠀ᖖ࡟གྷ ௓࡞ၥ㢟࡛࠶ࡿࠋ ࢟ࢡ࢖࣒ࢩ㢮ࡢ≉ᚩⓗ࡞⾜ື࡜ࡋ࡚㸪ྠࡌᶞ✀ࡢᶞᮌࡀఱᮏࡶ⏕ᜥࡋ࡚࠸ࡿ࡟ࡶ࠿ ࠿ࢃࡽࡎ 1 ᮏࡢᶞᮌ࡟㞟୰ࡋ࡚✸ධࡍࡿ⌧㇟ࡀ࠶ࡿࠋࡇࢀࡣ㸪ඛ㥑⪅࡜࡞ࡿ⹸ࡀ࢝࢖ ࣟࣔࣥ࡜࿧ࡤࢀࡿᶞᮌ⏤᮶ࡢㄏᘬ≀㉁࡞࡝࡟ࡼࡗ࡚ᐤ୺㑅ᢥࡀ⾜ࢃࢀ 㸪㞟ྜࣇ࢙ࣟࣔ ࣥࢆᨺฟࡍࡿࡇ࡜࡛ከᩘࡢ㞤㞝ࢆ㞟୰ⓗ࡟ㄏᘬࡋ㸪࣐ࢫ࢔ࢱࢵࢡࡀ⏕ࡌࡿࠋࡲࡓ㸪࢟ ࢡ࢖࣒ࢩ㢮࡟㣗ᐖࡉࢀࡓᶞᮌࡢᖿ࡞࡝࡟ࡣ୍ᵝ࡟✰ࡀ࠶ࡅࡽࢀ 㸪ࣇࣛࢫ㸦Frass㸪෗┿ 3㸧࡜࿧ࡤࢀࡿᮌᒌ࡜⣅ࡢΰྜ≀ࡀ኱㔞࡟᤼ฟࡉࢀ࡚࠸ࡿࡢࡶ≉ᚩⓗ࡞⌧㇟ࡢ㸯ࡘ࡛ ࠶ࡿ 42㸧_ࠋ ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ  ࢼࣛ㢮㞟ᅋᯤᦆ⿕ᐖࡢཎᅉ࡛࠶ࡿ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡣ 㸪㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ࡟ᒓ ࡍయ㛗 5mm ⛬ᗘࡢ⏥⹸࡛࠶ࡿ㸦෗┿ 4㸧ࠋ㞤⫼⬚㠃࡟࣐࢖࢝ࣥࢠ࢔ࢆ᭷ࡋ㸪ඹ⏕⳦࡜ ࡋ࡚ R. quercivora ࡸ㓝ẕ࡞࡝ࢆ㈓ⶶࡋ࡚࠸ࡿࠋ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢᒓࡍࡿࢼ࢞࢟ ࢡ࢖࣒ࢩ⛉ࡣࡍ࡭࡚ࡀ㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ࡛࠶ ࡾ㸪ࡑࡢከࡃࡀ⾶ᙅᮌࡸఆಽᮌ࡞࡝࡟ຍ ᐖࡍࡿ஧ḟᛶࡢᐖ⹸࡛࠶ࡿ࡜ࡉࢀ࡚ࡁࡓࠋࡋ࠿ࡋ㸪⇕ᖏᆅ᪉࡛ࡣ⏕❧ᮌ࡟ࡶ✸ධࡍࡿ ୍ḟᛶᐖ⹸✀ࡶ▱ࡽࢀ࡚࠸ࡿࠋࣇ࢕ࢪ࣮࡛ࡣ㸪࣐࢝࣎ࢽ࣮㸦Swietenia macrophylla㸧⏕ ❧ᮌࡀࢯࢺࣁࢼ࢞࢟ࢡ࢖࣒ࢩ㸦Crossotarsus externe-dentatus㸧ࡸࣇ࢕ࢪ࣮ࢼ࢞࢟ࢡ࢖࣒ ࢩ㸦Platypus gerstaeckeri㸧࡟ࡼࡗ࡚✸ධࡉࢀࡿࡇ࡜ࡀ㸪࣮࢞ࢼ࡛ࡣ࣡࣡㸦Triplochiton

14

ࡇࢀࡽࡢࢼ࢞࢟ࢡ࢖࣒ࢩ࡟ࡼࡿ⿕ᐖࡣ✸ධࡢࡳ࡛ᶞᮌࡀᯤṚ࡟⮳ࡿࡇ࡜ࡣ࡞ࡃ 㸪࢝ࢩ ࣀ ࢼ ࢞ ࢟ ࢡ ࢖ ࣒ ࢩ ࡟ ࡼ ࡿ 㞟 ᅋ ᯤ Ṛ ⿕ ᐖ ࡣ ࡇ ࢀ ࡽ ࢼ ࢞ ࢟ ࢡ ࢖ ࣒ ࢩ 㢮 ࡢ ⏕ ❧ ᮌ ⿕ ᐖ ࡜ ࡣ ඲ࡃ␗࡞ࡗ࡚࠸ࡿࠋ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡣ 㸪௚ࡢ࢟ࢡ࢖࣒ࢩ࡜ྠᵝ࡟㞝ࡀඛ㥑⹸࡜ ࡋ࡚ᐤ୺㑅ᢥࢆ⾜࠸ 㸪㞟ྜࣇ࢙ࣟࣔࣥ࡟ࡼࡗ࡚ከᩘࡢ㞤㞝ࢆㄏᘬࡋ 㸪࣐ࢫ࢔ࢱࢵࢡࡀ ⏕ࡌࡿࠋࡋ࠿ࡋ㸪๓ᖺᗘࡲ࡛࡟ࡍ࡛࡟✸ධṔࢆᣢࡘᶞᮌ ࡟ᑐࡋ࡚ࡣ㸪㞝ࡀ✸ධࡋ࡚ࡶ ࡍࡄ࡟✰࠿ࡽ⬺ฟࡍࡿࠋࡋࡓࡀࡗ࡚ 㸪࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ㞝ᡂ⹸ࡀ࢝ࢩࣀࢼ࢞࢟ࢡ ࢖ ࣒ ࢩ ✸ Ꮝ ᒚ Ṕ ࢆ ᣢ ࡘ ᶞ ᮌ ࢆ ୙ 㐺 ษ ࡜ ุ ᩿ ࡍ ࡿ ࡼ ࠺ ࡞ ኚ ໬ ࡀ ᶞ ᮌ ෆ ࡸ ኚ Ⰽ ቯ Ṛ 㒊 ࡛ ㉳ࡇࡗ࡚࠸ࡿࡇ࡜ࡀ᥎ ࡉࢀ࡚࠸ࡿ 18㸧_ࠋ ࢼࣛ㢮㞟ᅋᯤᦆ⿕ᐖ࡟࠾ࡅࡿ㜵ᚚ ᪉ἲ ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ࡟ࡼࡿࢼࣛ㢮㞟ᅋᯤᦆ⿕ᐖࢆ㜵ࡄࡓࡵ࡟⌧ᅾ 㸪᪩ᛴ࡞㜵㝖ᢏ ⾡ࡢ㛤Ⓨࡀồࡵࡽࢀ࡚࠸ࡿࠋ㜵ᚚ᪉ἲ࡜ࡋ࡚ 㸪⸆๣࡟ࡼࡿ R. quercivora ࡸ࢝ࢩࣀࢼ࢞ ࢟ࢡ࢖࣒ࢩࡢẅ⳦㸪ẅ⹸ᢏ⾡ࡀ⪃࠼ฟࡉࢀ࡚ࡁࡓࠋ1941 ᖺ࡟ࡣ㸪ᯘෆ࡟ᨺ⨨ࡋࡓ㣵ᮌ ୸ኴ࡟࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࢆ✸ධࡉࡏ 㸪✸ධᚋࡢ୸ኴࢆ㏿ࡸ࠿࡟↝༷ࡍࡿ᪉ἲࡀ⏝ ࠸ࡽࢀࡓࠋࡋ࠿ࡋ㸪㏆ᖺ࡛ࡣ⇞ᩱ࡜ࡋ࡚⸄Ⅳࢆ⏝࠸ࡿᚲせࡀ࡞ࡃ 㸪㣵ᮌࡢฎ⌮ࡀᅔ㞴 ࡛࠶ࡿࡇ࡜㸪ᛴᓧ࡞ᩳ㠃࠿ࡽ✸ධࢆཷࡅࡓ㣵ᮌࢆࡍࡳࡸ࠿࡟ᦙฟࡋ࡞ࡅࢀࡤ࡞ࡽ࡞࠸ ࡞࡝ࡢၥ㢟Ⅼࡀ࠶ࡾ 㸪ᐇ⏝ⓗ࡛ࡣ࡞࠿ࡗࡓࠋ ࣐ࢶࣀࢨ࢖ࢭࣥࢳࣗ࢘࡟ࡼࡿᯇᯤࢀ⿕ᐖ ࡛ࡣ㸪ᯤṚᮌࡢఆಽ㥑㝖ἲࡀ⏝࠸ࡽࢀ࡚࠸ ࡿࠋࡇࡢ᪉ἲࡣ㸪ᯤṚᮌࢆఆಽࡋ㸪⋢ษࡗࡓ୸ኴ࡟࣮࢝ࣂ࣒⣔ࡃࢇ⵨๣ࢆᩓᕸࡋ 㸪ࣅ

15

ࢽ࣮ࣝࢸ࣮ࣉ࡛⿕そࡍࡿᡭἲ࡛࠶ ࡿࡀ㸪ࢼࣛ㢮ࡣᛴᩳ㠃࡟⏕ᜥࡋ࡚࠸ࡿࡇ࡜࠿ࡽ 㸪ᯤ Ṛᮌࢆఆಽ㸪ࡃࢇ⵨๣ฎ⌮ࡍࡿࡇ࡜ࡣ㠀ᖖ࡟༴㝤࡛࠶ࡿࡇ࡜ࡀ᥎ ࡉࢀࡓࠋ⌧ᅾ⾜ࢃ ࢀ࡚࠸ࡿ㥑㝖᪉ἲ࡜ࡋ࡚㸪⿕ᐖᮌ࡟ࢻ࡛ࣜࣝ✰ࢆ㛤ࡅ 㸪┤᥋ᶞᮌ࡟࣮࢝ࣂ࣒⣔ࡃࢇ⵨ ๣ࢆὀධࡍࡿ᪉ἲࡀ࠶ࡿ 41㸧_{ࠋ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡣᆅ⾲㏆ࡃ࡟㞟୰ⓗ࡟✸ධࡍࡿࡇ} ࡜࠿ࡽ㸪࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩཬࡧ R. quercivora ࡢຠ⋡ⓗ࡞ẅ⹸㸪ẅ⳦ຠᯝࡀᚓࡽࢀ ࡚ ࠸ࡿ ࠋࡋ ࠿ࡋ 㸪❧ ᮌ࡟ ┤᥋ ⸆๣ ࢆὀ ධࡍ ࡿࡇ ࡜࡟ ࡼࡿ ⎔ቃ ㈇Ⲵ ࡞࡝ ࡀᠱ ᛕࡉ ࢀࡿ ࠋ ୍᪉㸪࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධࢆཷࡅࡓᶞᮌࢆᯖᮌ࡟ 㸪ࢩ࢖ࢱࢣ࡞࡝ࡢ⭉ᮙ⳦ࢆ ⏕⫱ࡉࡏࡿࡇ࡜࡛R. quercivora ࡢ⏕⫱ࢆጉᐖࡍࡿ࡜࠸ࡗࡓ᪉ἲ࡞࡝ࡶヨࡳࡽࢀ࡚࠸ࡿ 13,41㸧_ࠋ

16

ᶞᮌࡢᣢࡘ᢬ᢠᛶ ᶞᮌ࡟ࡣ㸪ࢭ࣮ࣝࣟࢫ㸪࡬࣑ࢭ࣮ࣝࣟࢫ㸪ࣜࢢࢽࣥ࡟ᣲࡆࡽࢀࡿ ୺せᡂศࡢ࡯࠿࡟㸪 ᩘ 㸣 ࡢ ᢳ ฟ ᡂ ศ ࡜ ࿧ ࡤ ࢀ ࡿ ᭷ ᶵ ⁐ ፹ ࡸ ࢔ ࣝ ࢝ ࣜ ࡞ ࡝ ࡛ ᢳ ฟ ࡍ ࡿ ࡇ ࡜ ࡢ ࡛ ࡁ ࡿ ᚤ 㔞 ᡂ ศࡀྵࡲࢀ࡚࠸ࡿࠋࡇࢀࡽࡣᮌᮦࡢእほ 㸪Ⰽㄪ㸪ໝ࠸㸪ࡑࡢ௚ࡢᛶ㉁࡜῝࠸㛵ಀࢆ᭷ ࡋ㸪໬Ꮫⓗ࡟ᶞ✀ࡢಶᛶࢆ⾲⌧ࡍࡿ㔜せ࡞ᡂศ࡛࠶ࡿࠋࡲࡓ⛣ື࡛ࡁ࡞࠸ᶞᮌ࡟࡜ࡗ ࡚ࡣ㸪እᩛ࠿ࡽࡢᨷᧁࡸ่⃭࡟ᑐࡋ࡚㑊㞴ࡍࡿࡇ࡜ࡀฟ᮶࡞࠸ࡓࡵ࡟ࡉࡲࡊࡲ࡞໬ྜ ≀ࢆྜᡂࡋ㸪⮬ࡽࡢ㌟ࢆᏲࡗ࡚࠸ࡿࠋᢳฟᡂศࡣ㸪኱ࡲ࠿࡟᥹Ⓨᛶࡢᡂศ࡜㞴 ᥹Ⓨᛶ ࡢᡂศ࡟ศࡅࡿࡇ࡜ࡀฟ᮶ࡿࠋ᥹Ⓨᡂศ࡟ศ㢮ࡉࢀࡿࢸࣝ࣌ࣥ㢮ཬࡧ⬡⫫ᒓ࢔ࣝࢥ࣮ ࣝ㢮࡞࡝ࡢ⢭Ἔᡂศࡣ㸪ࣇ࢕ࢺࣥࢳࢵࢻ㸦᳜≀࡟ࡼࡗ࡚⏕⏘ࡉࢀࡿ௚ࡢ᳜≀ࡸᚤ⏕≀ ࡞࡝࡟స⏝ࡍࡿ≀㉁㸧࡜ࡋ࡚▱ࡽࢀ࡚࠸ࡿࡶࡢࡶ࠶ࡿࠋࣇ࢕ࢺࣥࢳࢵࢻ࡟ࡣ 㸪㟷ⴥ࢔ ࣝ ࢥ ࣮ ࣝ ࡞ ࡝ ࡟ ௦ ⾲ ࡉ ࢀ ࡿ ࡼ ࠺ ࡞ ࿘ ㎶ ᶞ ᮌ ࡟ እ ᩛ ࡢ Ꮡ ᅾ ࢆ ▱ ࡽ ࡏ ࡿ ࢔ ࣞ ࣟ ࣃ ࢩ ࣮ ຠ ᯝࡸ㸪ࢡࢫࣀ࢟ࡢⴥ࡟ከࡃྵࡲࢀࡿࢩࣙ࢘ࣀ࢘ ࡞࡝ࡢ⹸⋇ࢆᚷ㑊ࡉࡏࡿຠᯝ㸪ࢡ࣑ࣝ ࡢ ⴥ ࡟ ࡩ ࡃ ࡲ ࢀ ࡿ ࣘ ࢢ ࣟ ࣥ ࡢ ࡼ ࠺ ࡟ ࿘ ㎶ ᳜ ≀ ࡢ ᡂ 㛗 ࢆ ᢚ ไ ࡉ ࡏ ࡿ ࣇ ࢓ ࢖ ࢺ ࢺ ࢟ ࢩ ࣥ ຠ ᯝ ࡸ 㟷 ᳃ ࣄ ࣂ ࡟ ከ ࡃ ྵ ࡲ ࢀ ࡿ ࣄ ࣀ ࢟ ࢳ ࢜ ࣮ ࣝ ࡸ ࣄ ࣀ ࢟ ࡢ ࢝ ࢪ ࣀ ࣮ ࣝ ࡟ ぢ ࡽ ࢀ ࡿ ᢠ ⳦ᛶࡢຠᯝࡀ࠶ࡿ 53,54㸧_ࠋ ୍᪉㸪㞴᥹Ⓨᛶᡂศ࡟ࡣࢱࣥ ࢽࣥ㢮ࡸࣇࣛ࣎ࣀ࣮ࣝ㢮࡜࠸ࡗࡓ࣏ࣜࣇ࢙ࣀ࣮ࣝ㢮࡞ ࡝ࡀศ㢮ࡉࢀࡿࠋࡇࢀࡽ࣏ࣜࣇ࢙ࣀ࣮ࣝࡣ ᶞ⓶㸪ⴥཬࡧᮦෆ࡟⵳࠼ࡽࢀ࡚࠾ࡾ 㸪ᚲせ ࡟ᛂࡌ࡚ᢠ⳦ᛶࡣ㜼ᐖຠᯝࢆⓎ᥹ ࡋ㸪⏕య㜵ᚚ≀㉁࡜ࡋ࡚⏕ᡂࡉࢀࡿࠋⴥෆࡢࢱࣥࢽ ࣥ ࡣ ࡑ ࡢ ཰ ᩡ ᛶ ࢆ ά ⏝ ࡋ ࡚ ࣈ ࢼ ࢔ ࢜ ࢩ ࣕ ࢳ ࣍ ࢥ ࡢ ࡼ ࠺ ࡞ 㣗 ⴥ ᐖ ⹸ ࡢ ᾘ ໬ ୙ Ⰻ ࢆ ㉳ ࡇ ࡋ㸪ࢡࣜ㸦Castanea crenata㸧ࡣᮦෆ࡟࢚ࣛࢪࢱࣥࢽࣥࢆ⵳✚ࡉࡏ࡚᰿࠿ࡽ㐍ධࡍࡿᚤ

17

⏕≀࡟ᑐࡋ࡚ᢠ⳦ᛶࢆ♧ࡋ㸪ࡑࡢ⤖ᯝࢡࣜᮦࡣ⪏ஂᛶࡢࡍࡄࢀࡓᮦ࡜ࡋ࡚ά⏝ࡉࢀ࡚ ࠸ࡿ 51)_{ࠋ஬ಸᏊࡢⴥࡣ㸪࢔ࣈ࣒ࣛࢩࡢ⏘༸่⃭࡟ࡼࡗ࡚⹸ࡇࡪෆ࡟ࢱࣥࢽࣥ㓟ࢆከ㔞} ࡟⏕ᡂࡉࢀ㸪࣮ࣘ࢝ࣜࡢ๧ᐃയ࡟ࡣ౵ධࡍࡿ⳦㢮࡟ᑐࡋ࡚ࢱࣥࢽࣥ㢮ࢆከࡃྜᡂࡉࡏ 㸪 ⭉ᮙ⳦࡞࡝ࡢ⳦㢮ࡢ㐍ධࢆ㜵 ࡄ 3㸧_{࡞࡝ࡢ㜵ᚚ཯ᛂࡶ࠶ࡿࠋࡲࡓ㸪ࢱࣥࢽࣥࡢࡼ࠺࡞ 㧗} ศᏊ㔞࣏ࣜࣇ࢙ࣀ࣮ࣝ ࡔࡅ࡛ࡣ࡞ࡃ㸪పศᏊ㔞࣏ࣜࣇ࢙ࣀ࣮ࣝ㢮ࡣ㧗࠸ᢠ㓟໬ᛶࡸᢠ ⳦ᛶࢆ᭷ࡍࡿ໬ྜ≀ࡀከࡃ㸪ᶞᮌෆ࡛ࡣ㔜せ࡞⏕య㜵ᚚ≀㉁࡛࠶ࡿࠋࡇࡢࡼ࠺࡞ᶞᮌ ࡢ㜵ᚚ≀㉁ࡢከࡃࡣ 㸪ே㛫࡟ᑐࡋ࡚ࡶ᭷ຠ࠿ࡘ㠀ᖖ࡟㧗࠸ᢠ⳦ᛶ㸪ᢠ㓟໬ᛶࢆⓎ⌧ࡉ ࡏࡿࡇ࡜࠿ࡽ⌧ᅾὀ┠ࢆ㞟ࡵ࡚࠸ࡿ㸦ᅗ 2㸧ࠋ

18

ࣈࢼ⛉ᶞᮌ࡟ྵࡲࢀࡿᡂศ ࣈࢼ⛉ᶞᮌࡣⴠⴥᛶཬࡧᖖ⥳ᛶ㧗ᮌ࡛࠶ࡾ 㸪ள⇕ᖏ࠿ࡽ ᖏ࡟ศᕸࡍࡿ㸴ᒓ 600 ✀ ௨ୖࡀ࠶ࡿࠋ᪥ᮏ ࡛ࡣࢩ࢖ᒓ㸦Castanopsis㸧㸪ࢡࣜᒓ㸦Castanea㸧㸪ࣈࢼᒓ㸦 Fagus㸧㸪 ࢥࢼࣛᒓ㸦Quercus㸧㸪࣐ࢸࣂࢩ࢖ᒓ㸦Pasania㸧࡞࡝ࡀ⏕⫱ࡍࡿࠋࡇࢀࡽࡢከࡃࡣ㸪࣡ ࢖ ࣥࡸ ࢘࢕ ࢫ࢟ ࣮ᶡ 㸪໬ ⢝ရ 㸪་ ⸆ရ 㸪ࢱ ࣥࢽ ࣥ〇 㐀࡞ ࡝࡟ ฼⏝ ࡉࢀ ࡿᶞ ᮌ࡛ ࠶ࡾ 㸪 ࢱࣥࢽࣥ㸪ࢸࣝ࣌ࣥ㸪ࢫ࣋ࣜࣥ㸪㓄⢾య࡞࡝ࡢᶞ⓶ࡸᮌ㒊ࡢ໬Ꮫᡂศ 㸪ศ㢮㸪ࡑࡢ⏕ ⌮άᛶ࡟ࡘ࠸࡚◊✲ࡀࡉ࠿ࢇ࡟⾜ࢃࢀࡓࠋ ࣈࢼ⛉ᶞᮌⴥ㒊ࡢᡂศ࡜ࡋ࡚㸪࣮ࣚࣟࢵࣃࣈࢼ㸦F. sylvatica㸧ࡢ᥹Ⓨᡂศ࡜ࡋ࡚㸦Z㸧

-3-hexenol ࡞࡝ࡢࢸࣝ࣌ࣥ㢮ࡸ Sabinen ࡀ㸪Q. laurifolia ࡸࣞࢵࢻ࣮࢜ࢡ㸦Q. rubra㸧ⴥ 㒊࠿ࡽᾘ⅖ຠᯝࡢ࠶ࡿᩘ✀ࡢࢣࣥࣇ࢙࣮ࣟࣝ㓄⢾యࡸࢧ࣏ࢽࣥ࡞࡝ࡀ༢㞳ࡉࢀ㸪་⸆

ရ࡜ࡋ࡚฼⏝ࡉࢀ࡚࠸ࡿ 52㸧_ࠋ

୍᪉㸪ࣈࢼ⛉ᶞᮌࡢከࡃࡣᮦෆࡢ୺せᡂศ࡜ࡋ࡚࢚ࣛࢪࢱࣥࢽࣥࢆ᭷ࡋ࡚࠸ࡿࡇ࡜

ࡀ▱ࡽࢀ࡚࠸ࡿࠋCastalagin ࡸ Vescalagin ࡣ㸪࣮ࣚࣟࢵࣃࢢࣜ㸦Castanea sativa㸧㸪ࢡࣜ

㸦Castanea crenata㸧㸪ࢭࢩ࣮࣮ࣝ࢜ࢡ㸦Q. petraea㸧㸪࣮ࣚࣟࢵࣃࢼࣛ㸦Q. robur㸧㸪ࢥ

ࣝࢡ࢞ࢩ㸦Q. suber㸧➼㸪ᗈࡃศᕸࡋ࡚࠸ࡿࠋࡑࡢ௚㸪valoneic acid dilactone㸪Castalagin

ࡸVescalagin ࡢ㔜ྜయ㸪Castacrenin 㢮㸪Castanopsin㸪grandinin㸪roburin 㢮㸪pedunculagin㸪

catechin 㢮㸪taxifolin 㓄⢾య㸪⦰ྜᆺࢱࣥࢽ࡛ࣥ࠶ࡿ hamamelitannin ࡞࡝ࡀࣈࢼ⛉ᶞᮌ

ࡢᮦ㸪ᶞ⓶࠿ࡽ༢㞳㸪ྠᐃࡉࢀ࡚࠸ࡿ 6,27,52)_{㸦ᅗ 3㸧ࠋࢼࣛ㢮࡟ྵࡲࡿࡇࢀࡽࡢ࢚ࣛࢪ}

ࢱ ࣥ ࢽ ࣥ 㢮 ࡣ 㸪 ࣡ ࢖ ࣥ ࡸ ࢘ ࢕ ࢫ ࢟ ࣮ ࡢ 〇 㐀 ࡟ 㔜 せ ࡞ ᙺ ๭ ࢆ ᢸ ࡗ ࡚ ࠾ ࡾ 㸪 ࣡ ࢖ ࣥ ࠿ ࡽ ✀ ࠎ ࡢ ࢚ ࣛ ࢪ ࢱ ࣥ ࢽ ࣥ 㢮 ࡸ ࡑ ࡢ ࢔ ࢢ ࣜ ࢥ ࣥ ࡛ ࠶ ࡿ ࢚ ࣛ ࢢ 㓟 ࡸ ࢞ ࣜ ࢵ ࢡ 㓟 ࡀ ᳨ ฟ ࡉ ࢀ

19

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ཧ⪃ᩥ⊩ 1. ఀ⸨㐍୍㑻 (2002) ⌧ᅾၥ㢟࡜࡞ࡗ࡚࠸ࡿࣈࢼ⛉ᶞᮌࡢ⾶㏥㸪ᯤṚ㸬 ᳃ᯘ⛉Ꮫ 35: 4-9 2. ఀ⸨㐍୍㑻㸦2000㸧ࣈࢼ⛉ᶞᮌࡢ㞟ᅋᯤṚ㸫⳦㢮࡜᪻⹸ࡢඹ⏕⳦ࡢ࡞ࡒ㸫㸬᪥ᮏ ⳦Ꮫ఍す᪥ᮏᨭ㒊఍ሗ 10㸸16-22㸬 3. ఀ⸨㐍୍㑻㸪❑㔝㧗ᚨ㸪బᶫ᠇⏕㸪ᒣ⏣฼༤㸦1998㸧ࢼࣛ㢮㞟ᅋᯤᦆ⿕ᐖ࡟ 㛵㐃 ࡍࡿ⳦㢮㸬᪥ᯘㄅ 80㸸170-175 4. ఀ⸨㐍୍㑻㸪ᒣ⏣฼༤㸦1998㸧᪥ᮏᾏഃ࡟Ⓨ⏕ࡍࡿࢼࣛ㢮㞟ᅋᯤṚ⿕ᐖࡢศᕸ㸬  ᳃ᯘ㜵␿ 47㸸222-229 5. ఀ⸨㐍୍㑻㸦2000㸧ࣈࢼ⛉ᶞᮌࡢ㞟ᅋᯤṚ㸫⳦㢮࡜᪻⹸ࡢඹ⏕⳦ࡢ࡞ࡒ㸫㸬᪥ᮏ ⳦Ꮫ఍す᪥ᮏᨭ㒊఍ሗ 10㸸16-22㸬

6. Kuroda, K. (2001) Responses of Quercus sapwood to infection with the pathogenic fungus of a new wilt disease vectored by the ambrosia beetle Platypus quercivorus. J. Wood. Sci. 47: 425-42

7. 㯮⏣៞Ꮚ㸪㧗⏿⩏ၨ㸦2002㸧ࢼࣛ㢮㞟ᅋᯤᦆᶵᵓࡢゎ᫂㸬᳃ᯘ⥲◊ᡤሗ  11㸸8-9 8. 㯮⏣៞Ꮚ㸪ᒣ⏣฼༤㸦1996㸧ࢼࣛ㢮㞟ᅋᯤᦆ࡟ぢࡽࢀࡿ㎶ᮦࡢኚⰍ࡜㏻Ỉᶵ⬟ࡢ

పୗ㸬᪥ᯘㄅ 78㸸84-88

9. Kubono, T. and Ito, S. (2002) Raffaerea quercivora sp. nov. associated with mass mortality of Japanese oak, and the ambrosia beetle  (Platypus quercivorus). Mycoscience 43㸸255-260

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10. ගỌ ᚭ㸪Ọᯇ࿴ᡂ㸪す஭Ꮥᩥ㸪኱㈡⚈἞㸦1999㸧ᢸᏊ⳦㢮ࡢ⳦⣒ᡂ㛗࡟ཬࡰࡍᶞ ⓶࣏ࣜࣇ࢙ࣀ࣮ࣝࡢຠᯝ㸬᪥⳦ሗ 40㸸91-96

22

➨ 1 ❶ 1-1 ⥴ゝ ᪥ᮏ࡟࠾ࡅࡿ᳃ᯘ⿕ᐖࡢ 1 ࡘ࡟ࣈࢼ⛉ᶞᮌࡢ㞟ᅋᯤᦆ⿕ᐖࡀ࠶ࡿࠋࣈࢼ⛉ᶞᮌࡣ᪥ᮏ ࡢཎ⏕ᯘࡢ୺せᵓᡂᶞ✀࡛࠶ࡾ㸪ୡ⏺ⓗ࡟ぢ࡚ࡶࣈࢼཎ⏕ᯘࡀ⮬↛ࡢ≧ែ࡛ṧࡗ࡚࠸ ࡿࡢࡣ᪥ᮏࡔࡅ࡛࠶ࡿࠋࡋ࠿ࡋ㸪᪥ᮏᾏഃࢆ୰ᚰ࡟ᗈࡀࡿࣈࢼ⛉ᶞᮌࡢ㞟ᅋᯤᦆ⿕ᐖ ࡣᣑ኱ഴྥ࡟࠶ࡿࡇ࡜࠿ࡽၥ㢟どࡉࢀ࡚࠸ࡿࠋࡇࡢ⿕ᐖࡣ㸪㣴⳦ᛶ࢟ࢡ࢖࣒ࢩ࡛࠶ࡿ

࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ㸦Platypus quercivorus MURAYAMA㸧ࡀᶞᮌ࡟✸ධࡋ㸪ᶞయෆ

࡛ࡑࡢඹ⏕⳦࡛࠶ࡿ Raffaelea quercivora ࡢఏ᧛࡟ࡼࡾᶞᮌ㎶ᮦ㒊ࡀ〓ኚࡋ㸪ࡑࢀ࡟క ࠺ࢳ࣮ࣟࢫᙧᡂ 1,2,3㸧_{࡞࡝࡟ࡼࡗ࡚ᶞᮌࡢ㏻Ỉ㜼ᐖࡀ⏕ࡌ㸪⤖ᯝⓗ࡟ᶞᮌࡣᯤṚࡍࡿࠋ} ࡇࡢ⿕ᐖ࡟ࡘ࠸࡚᪻⹸Ꮫ 4,5,6㸧_㸪⳦㢮Ꮫ 7,8,9,10,11,12,13㸧_{࡞࡝ࡉࡲࡊࡲ࡞どⅬ࠿ࡽࡢ◊✲ࡀ} ࡍࡍࡵࡽࢀ࡚࠸ࡿࡀ㸪ࡇࡢ⿕ᐖ࡟㛵ࡋ࡚໬Ꮫⓗ㸪⏕໬Ꮫⓗ࡞どⅬ࠿ࡽࡢ◊✲ࡣ࡯࡜ࢇ ࡝࡞ࡉࢀ࡚࠸࡞࠸ࠋࡑࡇ࡛ ೺඲࠶ࡿ࠸ࡣ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ ✸ධ⿕ᐖࢆཷࡅࡓ࣑ ࢬࢼࣛᮦ࡟ࡘ࠸࡚㸪㎶ᮦ࠿ࡽᢳฟ≀ࢆᚓ࡚ 㸪ࡇࢀࡽ࡟ࡘ࠸࡚ᛶ≧ศᯒࢆ⾜ࡗࡓࠋ

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1-2 ᐇ㦂᪉ἲ 1-2-1 ౪ヨᩱ 1-2-1-1 ೺඲࡞࣑ࢬࢼࣛ㎶ᮦヨᩱ ⿕ᐖࢆཷࡅ࡚࠸࡞࠸೺඲࡞࣑ࢬࢼࣛ㸦⣙ 40 ᖺ⏕㸧ࡣ 2007 ᖺ 11 ᭶࡟ᒱ㜧኱Ꮫ₇⩦ᯘ ࡼ ࡾ ධ ᡭ ࡋ ࡓࠋ ᶞ ⓶ ࢆ๤ ࡂ ྲྀ ࡾ 㸪㎶ᮦ ࡜ ᚰ ᮦ 㒊 ࡟ศ ࡅ ࡓ ࠋ㎶ ᮦ 㒊 ࡣ ⢊○ᶵ(Operating

Instructions Ultra Centrifugal Mill Type ZM200, Retsch)ࢆ⏝࠸࡚ᚤ⢊໬ࡋࡓࠋ㎶ᮦ⢊ᮎ 580.6g ࢆ 70%࢔ࢭࢺࣥỈ㸦ィ 35.9L㸧࡛ᢳฟࡋ㸪ᢳฟᾮࡣ࣮ࣟࢱ࣮࢚ࣜࣂ࣏࣮ࣞࢱ࣮ ࢆ⏝࠸࡚⃰⦰ࡋ㸪〓Ⰽࡢ⢓ᛶ≀ᚓࡓ(NS-e)ࠋNS-e ࡟ࡘ࠸࡚ HPLC ศᯒࢆ⾜ࡗࡓࠋ 1-2-1-2 ⿕ᐖࢆཷࡅࡓ࣑ࢬࢼࣛ㎶ᮦヨᩱ ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධࢆཷࡅᯤṚ⿕ᐖࢆཷࡅࡓ࣑ࢬࢼࣛᮦ㸦⣙ 40 ᖺ⏕㸧ࡣ 2007 ᖺ 11 ᭶࡟ᐩᒣ┴ᯘᴗᢏ⾡ࢭࣥࢱ࣮࡟࡚ఆ᥇ࡉࢀ㸪ᥦ౪ࢆཷࡅࡓࠋ-30Υ࡛෾⤖ಖᏑࡉ ࢀ࡚࠸ࡓ෇┙ࢆ୍ᬌ㸪37Υ࡟ಖࡗࡓ஝⇱ᶵ࡟ධࢀ࡚஝⇱ࡉࡏ࡚࠿ࡽᶞ⓶㸪㎶ᮦ㸪ᚰᮦ ࡟ศࡅࡓࠋ㎶ᮦࡢእഃ 1/2 ࡣኚⰍࡀ㐍ࢇ࡛࠸࡞࠿ࡗࡓࡢ࡛㸪ෆഃ 1/2 ࢆኚⰍ⿕ᐖࢆཷ

ࡅࡓ࣑ࢬࢼࣛ㎶ᮦヨᩱ࡜ࡋࡓࠋ⢊○ᶵ(Operating Instructions Ultra Centrifugal Mill Type

ZM200, Retsch)࡟࡚ᚤ⢊໬ᚋ㸪725.6g ࡢ⢊ᮎࢆ 70%࢔ࢭࢺࣥỈ㸦11.0L㸧࡟࡚ᢳฟࡋࡓࠋ ᢳฟᾮࡣ࣮ࣟࢱ࣮࢚ࣜࣂ࣏࣮ࣞࢱ࣮ࢆ⏝࠸࡚⃰⦰ࡋ㸪ࡑࡢ୍㒊ࢆ࣑ࢬࢼࣛ⿕ᐖᮌ㎶ᮦ

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1-2-2 HPLC ศᯒ

NS-e㸪DS-e ࡣࡑࢀࡒࢀ඘ศ࡟⃰⦰ᚋ㸪ᑡ㔞ࡢ࣓ࢱࣀ࣮ࣝ࡟⁐ゎࡋ㸪㧗㏿ᾮయࢡ࣐ࣟࢺ

ࢢࣛࣇ࢕࣮㸦HPLC㸧࡟࡚ศᯒࡋࡓࠋ

ᶵჾ୍ぴ

Pump : SHIMADZU LC-10AD (0.05 % TFA in water) SHIMADZU LC-10AT (Methanol)

Detector : SHIMADZU SPD-M10Avp Column ocen : SHIMADZU CTO-10A

Communication bus module : SHIMADZU CBM-10A Degasser : SHIMADZU DGU-12A

Column 㸹 Deverosil ODS HG-5Φ4.6x250mm Flow rate 㸹1ml/min

Column temp 㸹40 Υ

Detector wave length 㸹200-600nm 㸦SHIMAZU SPD-M10A VP㸧 Elution 㸹MeOH / 0.01%TFA aq. = 5 / 95 Ѝ 100 / 0 㸦45min㸧

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1-2-3 ᢳฟ≀ࡢᛶ≧ศᯒ

NS-e ࡜ DS-e ࡢᛶ≧ศᯒࡣࠊࡇࢀࡲ࡛࡟ሗ࿌ࡉࢀ࡚࠸ࡿ᪉ἲ࡛⾜ࡗࡓ 14,15)_{ࠋࡑࢀࡽࡢ}

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1-2-3-1 ඲ࣇ࢙ࣀ࣮ࣝ㔞ࡢᐃ㔞㸦Folin- Ciocalteu ἲ㸧 ᮏἲࡣ㸪඲ࣇ࢙ࣀ࣮ࣝ㔞ࡢᐃ㔞ἲ࡛㸪ࣇ࢙ࣀ࣮ࣝ㢮ࡀཷࡅࡿ㓟໬཯ᛂࢆ฼⏝ࡋ࡚ࣇ ࢙ࣀ࣮ࣝヨ⸆୰ࡢࣜࣥࣔࣜࣈࢹࣥ㓟ࢆ㑏ඖࡋ㸪ࡑࢀ࡟ࡼࡗ࡚⏕ࡌࡿࣜ ࣥࣔࣈࢹࣥࣈࣝ ࣮ࡢ࿊Ⰽࢆ ᐃࡍࡿ᪉ἲ࡛࠶ࡿ 16)_ࠋ ᪉ἲ 40㸣࢚ࢱࣀ࣮ࣝỈ࡛ㄪ〇ࡋࡓヨᩱ⁐ᾮ㸦⣙ 0.1mg / ml㸧1ml ࢆ࣮࣍ࣝࣆ࣌ࢵࢺ࡛ 24ml ᐜ〓Ⰽヨ㦂⟶࡟᥇ྲྀࡋ㸪⬺࢖࢜ࣥỈ 14ml ࢆῧຍࡋࡓࠋ1㹌ࣇ࢙ࣀ࣮ࣝヨ⸆㸦࿴ග ⣧⸆ᕤᴗᰴᘧ఍♫〇㸧1ml ࢆῧຍࡋ㸪ࢱࢵࢳ࣑࢟ࢧ࣮࡛᧠ᢾࡋࡓࠋ 3 ศ㛫ᨺ⨨ᚋ㸪㣬 ࿴Ⅳ㓟ࢼࢺ࣒ࣜ࢘⁐ᾮ㸦୍⣭  ࿴ග⣧⸆ᕤᴗᰴᘧ఍♫〇㸧2ml ࢆῧຍࡋ㸪⬺࢖࢜ࣥỈ ࡛ 20ml ࡟ᐜ㔞つṇࡋ࡚ࡼࡃ᧠ᢾࡋࡓࠋᐊ ࡛ 10 ศẖ࡟᧠ᢾࡋ㸪1 ᫬㛫ᚋ࡟⬺࢖࢜ࣥ Ỉࢆࣜࣇ࢓ࣞࣥࢫ࡜ࡋ࡚Ἴ㛗 750nm ࡟࠾ࡅࡿ྾ගᗘࢆศගගᗘィ㸦 Jasco 〇 V-520㸧 ࡛ ᐃࡋࡓࠋ㸦+㸧-࢝ࢸ࢟ࣥ㸦ᮾி໬ᡂᕤᴗᰴᘧ఍♫〇㸧࣓ࢱࣀ࣮ࣝ⁐ᾮࢆ⏝࠸࡚㸪྾ ගᗘ࡜⃰ᗘࡢ㛵ಀࢆ⾲ࡍ᳨㔞⥺ࢆసᡂࡋࡓࠋḟᘧࢆ⏝࠸࡚ᮍ▱ヨᩱ⁐ᾮࡢ྾ගᗘࢆ᳨ 㔞⥺࡟ᙜ࡚ࡣࡵ࡚㸦+㸧-࢝ࢸ࢟ࣥᙜ㔞ࢆồࡵ㸪ୗᘧ࡟ᚑ࠸඲ࣇ࢙ࣀ࣮ࣝ㔞ࢆ⟬ฟࡋࡓࠋ P = 100 ™ Y / S P㸹඲ࣇ࢙ࣀ࣮ࣝ㔞 ( % ) Y㸹᳨㔞⥺ࡼࡾ⟬ฟࡋࡓ࢝ࢸ࢟ࣥᙜ㔞 (mg / ml)

27

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1-2-3-2 ࣇࣛࣂࣀ࣮ࣝ㔞ࡢᐃ㔞㸦ࣂࢽࣜࣥሷ㓟ἲ㸧 ᮏ ἲࡣ ࣇࣛ ࣂࣀ ࣮ࣝ 㢮ࡢ ᐃ㔞 ἲ࡛ 㸪ヲ ࡋ࠸ ཯ᛂ ᶵᵓ ࡟ࡘ ࠸࡚ ࡣ౫ ↛୙ ࡛᫂ ࠶ࡿ ࡀ 㸪 㓟ᛶ⁐ᾮ୰࡛ࣂࢽࣜࣥࡀࣉࣟࢺࢿ࣮ࢩࣙࣥࡉࢀࡿࡇ࡜࡟ࡼࡾ㸪ồ㟁Ꮚ࢖࢜ࣥࡀࣇࣛࣂ ࣀ࣮ࣝ㢮ࡢ㟁Ꮚᐦᗘࡢ㧗࠸㸿⎔ࡢ㹁8 ఩ཬࡧ㹁6 ఩࡜཯ᛂࡍࡿࡇ࡜࡟ࡼࡾ㸪㉥Ⰽ࿊Ⰽ ≀㉁ࢆ⏕ᡂࡍࡿࡶࡢ࡜⪃࠼ࡽࢀ࡚࠸ࡿ 17)_{ࠋ᭱㏆ࡢ◊✲࡛㸪ࣉࣟࣇ࢕ࢭࢳࢽࢪ࡛ࣥࡣⓎ} Ⰽ య ࡢ ᴟ ኱ ྾ ཰ Ἴ 㛗 ཬ ࡧ ྾ ග ᗘ ࡀ ࢝ ࢸ ࢟ ࣥ ࢆ ࣔ ࣀ ࣐ ࣮ ࡜ ࡍ ࡿ ࣉ ࣟ ࢩ ࢔ ࢽ ࢪ ࣥ ࡜ ࠿ ࡞ ࡾ␗࡞ࡿࡓࡵ㸪ࢱࣥࢽࣥࡢᵓ㐀࡟ࡼࡗ࡚ࡣᮏᡭἲࡢྲྀࡾᢅ࠸࡟ὀពࢆせࡍࡿࡇ࡜ࡀᣦ ᦬ࡉࢀ࡚࠸ࡿ 18)_{ࠋᮏ◊✲࡛ࡣ㸪ࣉࣟࢩ࢔ࢽࢪࣥᆺࡢࣇࣛࣂࣀ࣮ࣝ 㢮ࡢᐃ㔞ࡢࡓࡵ࡟㸦+㸧} -࢝ࢸ࢟ࣥࢆᇶ‽≀㉁࡜ࡋ㸪᭱኱྾཰Ἴ㛗㸦500nm㸧࡟࠾ࡅࡿ྾ගᗘࢆ⏝࠸㸪ࣇࣛࣂࣀ ࣮ࣝ㢮ࢆᐃ㔞ࡋࡓࠋ ᪉ἲ 40㸣࢚ࢱࣀ࣮ࣝỈ࡛ㄪ〇ࡋࡓヨᩱ⁐ᾮ㸦 0.2̺0.3mg / ml㸧1ml ࢆ࣮࣍ࣝࣆ࣌ࢵࢺ࡛ 25ml ᐜ〓Ⰽヨ㦂⟶࡟᥇ྲྀࡋ㸪4㸣ࣂࢽࣜࣥ㸦≉⣭ ࢼ࢝ࣛ࢖ࢸࢡࢫᰴᘧ఍♫〇㸧-࣓ࢱ ࣀ࣮ࣝ⁐ᾮ 6ml ࡜⃰ሷ㓟 3ml ࢆ࣮࣍ࣝࣆ࣌ࢵࢺ࡛ῧຍࡋ㸪ࢱࢵࢳ࣑࢟ࢧ࣮࡛᧠ᢾࡋࡓࠋ ᐊ ࡛ 5 ศẖ࡟᧠ᢾࡋ㸪15 ศᚋ࡟࣓ࢱࣀ࣮ࣝࢆࣜࣇ࢓ࣞࣥࢫ࡜ࡋ࡚Ἴ㛗 500nm ࡟࠾ ࡅࡿ྾ගᗘࢆศගගᗘィ㸦Jasco 〇 V-520㸧࡛ ᐃࡋࡓࠋ㸦+㸧-࢝ࢸ࢟ࣥ㸦ᮾி໬ᡂᕤ ᴗᰴᘧ఍♫〇㸧࣓ࢱࣀ࣮ࣝ⁐ᾮࢆヨᩱ࡟⏝࠸࡚㸪྾ගᗘ࡜⃰ᗘࡢ㛵ಀࢆ⾲ࡍ᳨㔞⥺ࢆ సᡂࡋࡓࠋḟᘧࢆ⏝࠸࡚ᮍ▱ヨᩱ⁐ᾮࡢ྾ගᗘࢆ᳨㔞⥺࡟ᙜ࡚ࡣࡵ࡚㸦+㸧-࢝ࢸ࢟ࣥ ᙜ㔞ࢆồࡵ㸪ୗᘧ࡟ᚑ࠸඲ࣇ࢙ࣀ࣮ࣝ㔞ࢆ⟬ฟࡋࡓࠋࡲࡓ㸪4㸣ࣂࢽࣜࣥ-࣓ࢱࣀ࣮ࣝ

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⁐ᾮࡣ㸪 ᐃẖ࡟᪂ࡋࡃㄪ〇ࡋࡓࠋ 㹄 = 100 ™ 㹗 / 㹑 P㸹඲ࣇࣛࣂࣀ࣮ࣝ㔞 ( % ) Y㸹᳨㔞⥺ࡼࡾ⟬ฟࡋࡓ࢝ࢸ࢟ࣥᙜ㔞 (mg / ml) S㸹⏝࠸ࡓヨᩱ⃰ᗘ (mg / ml)

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1-2-3-3 ࢱࣥࣃࢡ㉁྾╔ヨ㦂 ࢱࣥࣃࢡ㉁཰ᩡస⏝ࢆ᭷ࡍࡿ໬ྜ≀⩌ࢆࢱࣥࢽࣥ࡜⥲⛠ࡍࡿࡇ࡜࠿ࡽ㸪ᢳฟ≀୰ࡢ ࢱ ࣥ ࢽ ࣥ ࡢ Ꮡ ᅾ ࢆ ☜ ㄆ ࡍ ࡿ ᪉ ἲ ࡜ ࡋ ࡚ ࢱ ࣥ ࣃ ࢡ ྾ ╔ ⬟ ࢆ   ᐃ ࡍ ࡿ ᪉ ἲ ࡀ ᗈ ࡃ ⏝ ࠸ ࡽ ࢀ࡚࠸ࡿ 19,20)_ࠋ ᪉ἲ  0.1㹋㓑㓟⦆⾪ᾮ*1_{࡛ 1mg / ml ࡟ㄪ〇ࡋࡓ∵⾑Ύ࢔ࣝࣈ࣑ࣥ㸦BSA㸧200ȣl ࡜ 40㸣} ࢚ࢱࣀ࣮ࣝỈ࡛ㄪ〇ࡋࡓヨᩱ⁐ᾮ㸦5mg / ml㸧200ȣl ࢆΰྜࡋ㸪1 ᫬㛫ᐊ ࡛྾╔཯ ᛂࡉࡏࡓࠋ཯ᛂᚋ㐲ᚰศ㞳ࡋ㸪࢟ࣕࢵࣉ௜ࡁヨ㦂⟶࡟ୖ⃈ࡳ 200ȣl ࢆධࢀ㸪Ỉ㓟໬ࣂ ࣒ࣜ࢘㸦୍⣭ ࿴ග⣧⸆ᕤᴗᰴᘧ఍♫〇㸧100mg ࢆῧຍࡋ㸪ຍᅽ≧ែ࡟࡚ 120Υ㸪10 ศ㛫ຍ⇕ࡋࡓࠋỈᾎ࡛෭༷ᚋ㸪㓑㓟ࢼࢺ࣒ࣜ࢘⦆⾪ᾮ 2ml ࢆຍ࠼㸪Ỉ㓟໬ࣂ࣒ࣜ࢘ࢆ ᏶඲࡟⁐ゎࡋ㸦pH 5.1s0.1㸧㸪ḟ࠸࡛ࢽࣥࣄࢻࣜࣥᾮ*2 _{3ml ࢆຍ࠼㸪108Υ࡛ 5 ศຍ⇕} ࡋࡓࠋ෭༷ᚋ㸪Ἴ㛗 570nm ࡟࠾ࡅࡿ྾ගᗘࢆศගගᗘィ㸦Jasco 〇 V-520㸧࡛ ᐃࡋ ࡓࠋBSA ⃰ᗘ࡜྾ගᗘࡢ㛵ಀࢆ⾲ࡍ᳨㔞⥺ࢆసᡂࡋ㸪྾╔ヨ㦂ᚋࡢୖ⃈ࡳᾮࡢ྾ගᗘ ࢆ᳨㔞⥺࡟ᙜ࡚ࡣࡵ㸪㐟㞳 BSA ⃰ᗘࢆồࡵ㸪྾╔⋡ࢆ⟬ฟࡋࡓࠋ *1 _{㓑㓟ࢼࢺ࣒ࣜ࢘⦆⾪ᾮ㸸㓑㓟ࢼࢺ࣒ࣜ࢘୕Ỉሷ㸦࢔࣑ࣀ㓟⮬ືศᯒ⏝  ࿴ග⣧⸆} ᕤᴗᰴᘧ఍♫〇㸧36g ࢆ⬺࢖࢜ࣥỈ࡟⁐ゎࡋ㸪ị㓑㓟 18.5ml ࢆຍ࠼ࡓࠋ᭦࡟ࢩ࢔ࣥ໬ ࢼࢺ࣒ࣜ࢘㸦≉⣭ ࿴ග⣧⸆ᕤᴗᰴᘧ఍♫〇㸧⁐ᾮ 2ml ࢆຍ࠼㸪⬺࢖࢜ࣥỈ࡛ 200ml ࡟ᐜ㔞つṇࡋࡓࠋ

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*2 _{ࢽࣥࣄࢻࣜࣥᾮ㸸⢭␃ࡋࡓ࣓ࢳࣝࢭࣝࢯࣝࣈ㸦࿴ග⣧⸆ᕤᴗᰴᘧ఍♫〇㸧࡟ࢽࣥ}

ࣄࢻࣜࣥ㸦࢔࣑ࣀ㓟⮬ືศᯒ⏝  ࿴ග⣧⸆ᕤᴗᰴᘧ఍♫〇㸧ࢆ 0.5㸣㸦㹵 / 㹴㸧࡟࡞

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1-2-3-4 㓟ຍỈศゎ ᮏἲࡣ㸪㧗 㸪ᙉ㓟᮲௳ୗ࡛㓟ຍỈศゎࢆ㉳ࡇࡋ㸪㓄⢾య࡞࡝ࡢ࢚ࢫࢸࣝ⤖ྜࢆᙉ ไⓗ࡟ศゎࡉࡏ㸪㐟㞳ࡋࡓ࢔ࢢࣜࢥࣥࢆ ᐃࡍࡿࡇ࡜࡟ࡼࡾ㸪ຍỈศゎᆺࢱࣥࢽࣥࡢ Ꮡᅾࡸࡑࡢศゎ≀ࢆ☜ㄆࡍࡿ᪉ἲ࡜ࡋ࡚⏝࠸ࡽࢀࡿ 21㸧_ࠋ ᪉ἲ ヨᩱ 5mg ࡜ 3N ◲㓟 5ml ࢆ࣮࣍ࣝࣆ࣌ࢵࢺ࡛࢟ࣕࢵࣉ௜ࡁヨ㦂⟶࡟ῧຍࡋ㸪ࢱࢵࢳ ࣑࢟ࢧ࣮࡛᧠ᢾᚋ㸪110Υ㸪10 ᫬㛫ຍ⇕ࡋࡓࠋỈᾎ࡟࡚෭༷ᚋ㸪㸴N Ỉ㓟໬ࢼࢺࣜ࢘ ࣒Ỉ⁐ᾮ 2.5ml ࢆῧຍࡋ㸪ヨᩱࢆ᏶඲࡟⁐ゎࡉࡏࡓᚋ㸪Ỉ࡛ 8ml ࡟ᐜ㔞つṇࡋࡓࠋࡑ ࡢ⁐ᾮࢆ 100ȣl ᥇ྲྀࡋ㸪◲㓟ࢆᩘ⁲⁲ୗࡋ㸪⁐ᾮࢆ㓟ᛶ࡟ࡋࡓࠋ❅⣲࡛Ỉࢆ᏶඲࡟␃ ཤࡋࡓᚋ㸪100ȣl ࡢ࣓ࢱࣀ࣮࡛ࣝ⁐ゎࡋሷࢆỿẊࡉࡏࡓࠋ㓟ຍỈศゎ࡟࡚⏕ᡂࡋࡓ࢔ ࢢࣜࢥࣥࡣ HPLC ࡟࡚ศᯒࡋࡓࠋ HPLC ศᯒ᮲௳ Column㸹Cosmosil 5CMS WATERS 4.6mm × 250mm Flow rate㸹1ml / min

Column temp㸹35Υ

Detector wave length㸹SHIMADZU SPD-M10A VP㸪200㹼600nm Solvent㸹 Initial MeOH / 0.01%TFA 㸻 5 / 95㸪

33

34

1-3 ⤖ᯝ࡜⪃ᐹ 1-3-1 ೺඲࣑ࢬࢼࣛ㎶ᮦ࡜ᯤṚ⿕ᐖ࣑ࢬࢼࣛ㎶ᮦ࡟࠾ࡅࡿ໬Ꮫศᯒ ೺඲࣑ࢬࢼࣛ㎶ᮦࠊᯤṚ⿕ᐖࢆཷࡅࡓ࣑ࢬࢼࣛ㎶ᮦࡑࢀࡒࢀ࠿ࡽᚓࡓᢳฟ≀ࡣࠊඹ࡟ 〓Ⰽ࡛ࠊ⢓ᛶࡀ㧗࠿ࡗࡓࠋᚓࡽࢀࡓᢳฟ≀ࡢ཰⋡ࢆ Table1-1 ࡟♧ࡋࡓࠋ೺඲࣑ࢬࢼࣛ ㎶ᮦᢳฟ≀㸦NS-e㸧㸪ᯤṚ⿕ᐖࢆཷࡅࡓ࣑ࢬࢼࣛ㎶ᮦᢳฟ≀ (DS-e)ࡢ୧᪉࡟ࡘ࠸࡚ࠊ඲ యࡢ཰⋡࡟㢧ⴭ࡞ᕪࡣぢࡽࢀ࡞࠿ࡗࡓࠋ

NS-e ࡜ DS-e ࡟ࡘ࠸࡚㸪Folin-Denis ἲ㸪Vanillin-HCl ἲ㸪Ninhydrin ἲࡢ᳨㔞⥺ࢆసᡂ ࡋ㸪᳨㔞⥺ࢆࡶ࡜࡟ྛヨᩱࡢ⥲ࣇ࢙ࣀ࣮ࣝ㔞㸪⥲ࣇࣛࣂࣀ࣮ ࣝ㔞㸪ࢱࣥࣃࢡ྾╔⋡ࢆ ồࡵࡓࠋNS-e㸪DS-e ࡢ୧᪉࡟࠾࠸࡚ࠊ⥲ࣇ࢙ࣀ࣮ࣝ㔞ࡣ 45%௨ୖ࡜㠀ᖖ࡟㧗࠸್࡛࠶ ࡗࡓࡀࠊࣇࣛࣂࣀ࣮ࣝ㔞ࡣ ඹ࡟ 1%ᮍ‶࡜ᴟ➃࡟ప࠸್࡜࡞ࡗࡓࠋࡲࡓࠊ F / P㸦ࣇ࢙ ࣀ࣮ࣝ໬ྜ≀୰ࡢࣇࣛࣂࣀ࣮ࣝࡢ┦ᑐ⋡㸧ࡶ 0.02 ௨ୗ࡛࠶ࡗࡓࠋ୍⯡࡟㸪ࣉࣟ࢔ࣥࢺ ࢩ࢔ࢽࢪࣥ㸦⦰ྜᆺࢱࣥࢽࣥ㸧ࢆྵࡴࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿ㔪ⴥᶞᶞ⓶ᢳฟ≀࡛ࡣ F / P ࡀ 0.6㹼0.8 ࢆ♧ࡍࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿ22,23㸧_{ࠋࡇࡢࡇ࡜࠿ࡽ㸪ࡇࢀࡽࡢ࣑ࢬࢼࣛᮦᢳฟ} ≀୰࡟ࡣࣉࣟ࢔ࣥࢺࢩ࢔ࢽࢪࣥ㢮ࡣ࡯࡜ࢇ࡝ྵࡲࢀ࡚࠸࡞࠸ࡇ࡜ࡀண᝿ࡉࢀࡓࠋࡲࡓ㸪 NS-e ࡛ࡣ⥲ࣇ࢙ࣀ࣮ࣝ㔞ࡀ 47.3㸣࡛࠶ࡗࡓࡢ࡟ᑐࡋ࡚ DS-e ࡛ࡣ 85.6㸣࡜ᴟ➃࡟㧗࠸ ್ࢆ♧ࡋࡓࡇ࡜࠿ࡽ㸪ኚⰍ㒊࡛⥲ࣇ࢙ࣀ࣮ࣝ㔞ࡀቑຍࡋ࡚࠸ࡓࡇ࡜ࡀ⪃࠼ࡽࢀࡓࠋ ࡲࡓࠊࢱࣥࣃࢡ྾╔⬟ࢆẚ㍑ࡍࡿ࡜㸪NS-e ࡛ 59.3㸣࡜㧗࠸್ࢆ♧ࡋ㸪ࢱࣥࣃࢡ㉁཰ ᩡᛶࢆ♧ࡍࢱࣥࢽࣥ㢮ࡢᏑᅾࡀ♧၀ࡉࢀࡓࠋࡋ࠿ࡋࣂࢽࣜࣥሷ㓟ἲ࡟ࡼࡿ⥲ࣇࣛࣂࣀ ࣮ࣝ㔞ࡢ ᐃ࡟࠾࠸࡚ࠊࣉࣟ࢔ࣥࢺࢩ࢔ࢽࢪࣥ㢮ࡣྵࡲࢀ࡚࠸࡞࠸ࡇ࡜ࡀ᫂ࡽ࠿ ࡟ࡉ ࢀ ࡚࠸ ࡿࡇ ࡜࠿ ࡽ㸪 ຍỈ ศゎ ᆺࢱ ࣥࢽ ࣥ㢮 ࡀᏑ ᅾࡋ ࡚࠸ ࡿࡇ ࡜ࡀ ᥎  ࡉࢀ ࡓࠋ ࡲࡓ 㸪

35

NS-e ࡢ྾╔⋡࡛࠶ࡿ 59.3㸣࡟ᑐࡋ㸪DS-e ࡛ࡣ 71.6㸣ࢆ♧ࡋࡓࠋDS-e ࡛ࡣ⥲ࣇ࢙ࣀ࣮ ࣝ㔞ࡶከ࠿ࡗࡓࡇ࡜࠿ࡽࠊᮦෆ࡟ྵࡲࢀࡿຍỈศゎᛶࢱࣥࢽࣥࡢ୍㒊ࡀຍỈศゎࡋ࡚ ࢔ࢢࣜࢥࣥࢆ⏕ᡂࡋࠊ࢔ࢢࣜࢥࣥࢆᨺฟࡋࡓศࡔࡅຍỈศゎᛶࢱࣥࢽࣥศᏊࡀᑠࡉࡃ ࡞ࡗࡓࡇ࡜࠿ࡽࠊBSA ࡜྾╔ࡀᐜ᫆࡟࡞ࡿࡇ࡜࡟ࡼࡗ࡚ࠊ྾╔⋡ࡀቑຍࡋࡓࡢ࡛ࡣ࡞ ࠸࠿࡜⪃࠼ࡽࢀࡓࠋ

36

㼟㼍㼙㼜㼘㼑㼟 㼥㼕㼑㼘㼐㻌㻔㻑㻕 㼠㼛㼠㼍㼘㻌㼜㼔㼑㼚㼛㼘㻌㼢㼍㼘㼡㼑㻌㻔㻑㻕 㼠㼛㼠㼍㼘㻌㼒㼘㼍㼢㼍㼚㼛㼘㻌㼢㼍㼘㼡㼑㻌㻔㻑㻕 㻮㻿㻭㻌㼍㼐㼟㼛㼞㼜㼠㼕㼛㼚㻌㼞㼍㼠㼑㻔㻑㻕

㻺㻿㻙㼑 㻟㻚㻡㻝 㻠㻡㻚㻢 㻜㻚㻡 㻢㻡㻚㻤

㻰㻿㻙㼑 㻟㻚㻠㻡 㻢㻤㻚㻢 㼚㻚㼐㻚 㻟㻠㻚㻞

㼔㼍㼞㼠㻌㼣㼛㼛㼐 㻟㻚㻢㻝 㻡㻟㻚㻢 㼚㻚㼐㻚 㻡㻣

Table 1-1 Chemical analysis of the extracts of Quercus crispula

Note :These values were expressed with the percentage based on dry NS-e and DS-e a: NS-e stand for the 70 % aqueous extracts from normal part of Q. crispula sapwood

b: DS-e stand for the 70 % aqueous extracts from infected-colored part of Q. crispula sapwood c: Protein precipitation was calculated by the method described at experimental section

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1-3-2 ᢳฟ≀ࡢ HPLC ศᯒ

NS-e ࡜ DS-e ࡟ࡘ࠸࡚ࠊHPLC ศᯒࢆ⾜ࡗࡓ㸦Fig. 1-1㸧ࠋHPLC ࢡ࣐ࣟࢺࢢ࣒ࣛࢆẚ㍑

ࡋࡓ࡜ࡇࢁࠊNS-e ࡛ࡣࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 30 ศ๓ᚋ࡟ࣈ࣮ࣟࢻ࡞ࣆ࣮ࢡࡀᏑᅾࡋࠊ ࡑࢀࡣ⣙  ศ㛫࡟Ώࡗ᳨࡚ฟࡉࢀࡓࠋࡋ࠿ࡋ࡞ࡀࡽ DS-e ࡛ࡣࡑࡢࣆ࣮ࢡ㧗ࡀ㢧ⴭ ࡟ῶᑡࡋࠊࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 31 ศ㡭࡟ẚ㍑ⓗ኱ࡁ࡞ࣆ࣮ࢡࡀぢࡽࢀࡓࠋࡇࡢ 31 ศ 㡭ࡢࣆ࣮ࢡࡣࠊࡑࡢ UV ࢫ࣌ࢡࢺࣝࡢᙧ≧࡜ࣜࢸࣥࢩࣙࣥࢱ࢖࣒࠿ࡽࠊ࢚ࣛࢢ㓟㸦Ȣ max 㸻252, 365㸪㸧࡜ྠᐃࡋࡓࠋࡲࡓࠊࡑࢀࡒࢀࡢࢡ࣐ࣟࢺࢢ࣒ࣛ࠿ࡽ࢝ࢸ࢟ࣥ㢮࡟㢮 ఝࡋࡓ UV ྾཰ࢆ᭷ࡍࡿ໬ྜ≀㢮ࡸ࢚ࣛࢢ㓟㸪࢞ࣜࢵࢡ㓟࡟㢮ఝ ࡋࡓ UV ྾཰ࢆ♧ࡍ ໬ྜ≀㢮ࡀ᳨ฟࡉࢀࡓࡀ㸪ᶆ‽ヨ⸆࡜ࡢಖᣢ᫬㛫ࡢ୙୍⮴࡟ࡼࡾ㸪ྠᐃࡍࡿࡇ࡜ࡀ࡛ ࡁ࡞࠿ࡗࡓࠋࡲࡓ㸪ᚰᮦᢳฟᡂศ࠿ࡽࡶከ㔞ࡢ࢞ࣜࢵࢡ㓟࠾ࡼࡧ࢚ࣛࢢ㓟ࡀ᳨ฟࡉࢀ ࡓࠋ୍⯡࡟㸪ࢼࣛ㢮࡟ࡣ࢚ࣛࢪࢱࣥࢽࣥࡸ࢞ࣟࢱࣥࢽࣥࡀྵࡲࢀ࡚࠸ࡿࡇ࡜ࡀ▱ࡽࢀ ࡚࠾ࡾ24㸧_{㸪ࡑࢀࡽࡣ࢞ࣜࢵࢡ㓟ࡸ࢚ࣛࢢ㓟࡟㢮ఝࡋࡓUV ࢫ࣌ࢡࢺࣝࢆ♧ࡍࡇ࡜࠿ࡽ㸪} ௒ ᅇ ⏝ ࠸ ࡓ ࣑ ࢬ ࢼ ࣛ ࡟ ࡶ ௚ ࡢ ࢼ ࣛ 㢮 ྠ ᵝ ࡟ ຍ Ỉ ศ ゎ ᆺ ࢱ ࣥ ࢽ ࣥ ࡀ ྵ ࡲ ࢀ ࡚ ࠸ ࡿ ࡇ ࡜ ࡀ᥎ ࡉࢀࡓࠋ

38

㻺㻿㻙㼑㻕

㻰㻿㻙㼑㻕

㻾㼑㼠㼑㼚㼠㼕㼛㼚㻌㼠㼕㼙㼑㻌㻔㼙㼕㼚㻕

㻝㻜

㻞㻜

㻟㻜

㻠㻜

㻡㻜

Fig. 1-1 HPLC chromatograms of extracts from Q. crispula sapwood at 280 nm. NS-e : Extracts from normal sapwood of Q. crispula

39

1-3-3 ᢳฟ≀ࡢ㓟ຍỈศゎ 㓟ຍỈศゎ๓ᚋࡢ NS-e ࡢ HPLC ࢡ࣐ࣟࢺࢢ࣒ࣛࢆ Fig. 1-2 ࡟♧ࡋࡓࠋ㓟ຍỈศゎࡢ ⤖ᯝࠊ࢞ࣜࢵࢡ㓟࡜࢚ࣛࢢ㓟ࡢࣆ࣮ࢡࢆ☜ㄆࡋࡓࠋ࢚ࣛࢢ㓟ࡣຍỈศゎᆺࢱࣥࢽࣥࡢ ୍ࡘ࡛࠶ࡿ࢚ࣛࢪࢱࣥࢽࣥࡢ࢔ࢢࣜࢥ࡛ࣥ࠶ࡿࡇ࡜࠿ࡽ㸪DS-e ࡟ࡣ࢚ࣛࢪࢱࣥࢽࣥᵝ ≀㉁ࢆྵࡴࡇ࡜ࡀ♧၀ࡉࢀࡓࠋ

40

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

㼑㼘㼘㼍㼓㼕㼏 㼍㼏㼕㼐

㼓㼍㼘㼘㼕㼏 㼍㼏㼕㼐

㻾㼑㼠㼑㼚㼠㼕㼛㼚㻌㼠㼕㼙㼑㻌㻔㼙㼕㼚㻕

Fig. 1-2 HPLC chromatograms of hydrolysate of NS-e

41

1-4 ࡲ࡜ࡵ ࢱࣥࢽࣥศᯒࡢ⤖ᯝࠊ࣑ࢬࢼࣛࡢᢳฟᡂศ࡟ࡣ⦰ྜᆺࢱࣥࢽࣥ㢮ࡣ࡯ࡰྵࡲࢀ࡚࠾ࡽ ࡎࠊຍỈศゎᛶࢱࣥࢽࣥ㢮ࡀྵࡲࢀ࡚࠸ࡿࡇ࡜ࡀண᝿ࡉࢀࡓࠋ㓟ຍỈศゎ࡟ࡼࡗ࡚࢚ ࣛࢢ㓟ࡀ⏕ᡂࡋࡓࡇ࡜࡜ࠊQuercus ᒓ࠿ࡽ࢚ࣛࢪࢱࣥࢽࣥ㢮ࡢྠᐃࡢሗ࿌ࡀ࠶ࡿࡇ࡜ ࠿ࡽࠊᮏ◊✲࡛⏝࠸ࡓ࣑ࢬࢼࣛ࡟ࡘ࠸࡚ ࡶࠊ࢚ࣛࢪࢱࣥࢽࣥ㢮ࡀྵࡲࢀ࡚࠸ࡿྍ⬟ᛶ ࡀ࠶ࡿࠋࡲࡓࠊ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධ⿕ᐖࢆཷࡅࡿࡇ࡜࡛ࠊ ࢚ࣛࢪࢱࣥࢽࣥ㢮 ࡣ㢧ⴭ࡟ῶᑡࡋࠊ࢚ࣛࢢ㓟࡜࢞ࣜࢵࢡ㓟ࡀቑຍࡋ࡚࠸ࡓࠋࡇࡢࡇ࡜ࡣࠊ ࡑࡢᡂศᵓᡂ ࡣᚰᮦ࡟࠾ࡅࡿᡂศᵓᡂ࡟ఝ㏻ࡗ࡚࠾ࡾࠊ㎶ᮦ࡟࠾ࡅࡿഇᚰᮦ໬ࡀ㉳ࡇࡗ࡚࠸ࡿࡢ࡛ ࡣ࡞࠸࠿࡜⪃࠼ࡽࢀࡓࠋ

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1-5 ཧ⪃ᩥ⊩

1. Kuroda, K. (2001) Responses of Quercus sapwood to infection with the pathogenic fungus of a new wilt disease vectored by the ambrosia beetle Platypus quercivorus. J. Wood. Sci. 47: 425-42

2. 㯮⏣៞Ꮚ㸪㧗⏿⩏ၨ㸦2002㸧ࢼࣛ㢮㞟ᅋᯤᦆᶵᵓࡢゎ᫂㸬᳃ᯘ⥲◊ᡤሗ  11㸸 8-9

3. 㯮⏣៞Ꮚ㸪ᒣ⏣฼༤㸦1996㸧ࢼࣛ㢮㞟ᅋᯤᦆ࡟ぢࡽࢀࡿ㎶ᮦࡢኚⰍ࡜㏻Ỉᶵ⬟

ࡢపୗ㸬᪥ᯘㄅ 78㸸84-88

4. Igeta, Y., Esaki, K., Kato, K. and Kamata, N.㸦2003㸧Influence of light condition on the stand-level distribution and movement of the ambrosia beetle Platypus quercivorus (Coleoptera: Platypodidae) Appl. Entomol. Zool. 38:167-175

5. 㙊⏣┤ே (2002) ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ⏕ែ㸬 ᳃ᯘ⛉Ꮫ 35: 26-34 6. Kobayashi, M., Ueda, A. and Takahata, Y. (2001) Inducing Infection of Oak logs by a

Pathogenic Fungus Carries by Platypus quercvorus (MURAYAMA) (Coleoptera : Platypododae). J. For. Res. 6 : 153-136

7. Ueda, A. and Kobayashi, M. (2001) Aggregation of Platypus quercivorus (Murayama) (Coleoptera : Platyposisae) on Oak Log Bored by Males of the Species. J. For. Res.

6 : 173-179

8. ఀ⸨㐍୍㑻㸦2000㸧ࢼࣛ㢮㸪ࢩ࢖࣭࢝ࢩ㢮ᯤṚ⿕ᐖࡢ⌧≧㸬ᒣᯘ 㸳㸳 㸸60-68

43

ᮏ⳦Ꮫ఍す᪥ᮏᨭ㒊఍ሗ 10㸸16-22㸬 10. ఀ⸨㐍୍㑻㸪❑㔝㧗ᚨ㸪బᶫ᠇⏕㸪ᒣ⏣฼༤㸦1998㸧ࢼࣛ㢮㞟ᅋᯤᦆ⿕ᐖ࡟㛵 㐃ࡍࡿ⳦㢮㸬᪥ᯘ ㄅ 80㸸170-175 11. ᮌᮧ໷Ꮚ㸪ᯇ⏣㝧௓㸪❑㔝㧗ᚨ㸪ఀ⸨㐍୍㑻㸦 2003㸧Raffaelea quercivora ⳦ᰴ 㛫ࡢᙧែᏛⓗ≉ᚩ㸬  ୰᳃◊ 51㸸199-200

12. Kubono, T. and Ito, S. (2002) Raffaerea quercivora sp. nov. associated with mass mortality of Japanese oak, and the ambrosia beetle  (Platypus quercivorus). Mycoscience 43㸸255-260

13. ගỌ ᚭ㸪Ọᯇ࿴ᡂ㸪す஭Ꮥᩥ㸪኱㈡⚈἞㸦1999㸧ᢸᏊ⳦㢮ࡢ⳦⣒ᡂ㛗࡟ཬࡰࡍ ᶞ⓶࣏ࣜࣇ࢙ࣀ࣮ࣝࡢຠᯝ㸬᪥⳦ሗ 40㸸91-96

14. Takagi K, Shimomura K, Koizumi Y, Mitsunaga T, Abe I (1999) Tyrosinase Inhibitors from the Pericarp of Jatoba (Hymenaea courbaril L.) (in Japanese with English summary). Natural Medicines. 53(1): 15-21

15. Takagi K, Mitsunaga T (2002) A Tyrosinase Inhibitors from the Asam ( Mangifera quadrifida). Natural Medicines. 56(3): 97-103

16. Swain T., and Hillis W.E. 1959. The Phenolic Constitutes of Prunus domestica. II. The analysis of tissues of the Victoria plum tree. J. Sci. Fd. Agri. 10: 63-68

17. Hagernan A.E. 1989. Chemistry of tannin-protein complexation, pp 323-333, in R.W. Hemingway and J.J. Karchesy (eds.). Chemistry and Significance of Condensed Tannins.Plenum Press, New York.

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18. Mitsunaga T., Doi T., Kondo Y. and Abe I. 1998. Color development of proanthocyanidins in vanillin-hydrochloric acid reaction. J.Wood. Sci. 44:125-130 19. Harbone, J. B. (1988) The Flavonoids. Chapman and Hall, London, 21-61

20. Takagi, K. and Mitsunaga, T.㸦2002㸧A tyrosinase inhibiter from the wood of Asam (Mangifera quadrifida J.) Nat. Medic. 56㸸97-103

21. ᯇᾆ㑥᫛㸦1997㸧᳃ᯘ⿕ᐖ㜵㝖࡜⏕≀ከᵝᛶಖ඲㸬 ᳃ᯘ⥲◊㛵す◊✲ 46㸸1 22. 㩪ᓥṇᾈ㸪ၿᮏ▱Ꮥ㸦1981㸧㔪ⴥᶞᶞ⓶࣏ࣜࣇ࢙ࣀ࣮ࣝᛶᢳฟᡂศࡢ≉ᚩ࡟ࡘ

࠸࡚㸬ᮌᮦᏛ఍ㄅ  27㸦㸦6㸧:491-497

23. 㧗ᮌၨ஧㸪ୗᮧ೺ḟ㸪ᑠἨ⩏ኵ㸪ගỌ ᚭ㸪㜿㒊 ໏㸦1999㸧Jatoba㸦Hymenaea courbaril L.㸧ᯝ⓶࠿ࡽࡢࢳࣟࢩࢼ࣮ࢮάᛶ㜼ᐖ≀㉁㸬Nat. Medic. 53㸸15-21 24. Ᏻ⏣ᚁᕷ㸦2002㸧ᶞᮌࡢ㢦㸫ᶞᮌᢳฟᡂศ࡜ࡑࡢ฼⏝㸫㸬ᾏ㟷♫ ⁠㈡┴ 196-201

㸦⤖ᯝ㸧

25. K. Imai , T. Mitsunaga , H. Takemoto , T. Yamada , S. Ito , H. Ohashi (2008)

Extractives of Quercus crispula sapwood infected by the pathogenic fungi Raffaelea quercivora (I) : Comparison of sapwood extractives non-infected and infected. J.Wood. Sci. 55:126-132

45

➨ 2 ❶ 2-1 ⥴ゝ ➨୍❶ࡼࡾࠊ೺඲࡞࣑ࢬࢼࣛ㎶ᮦ࡟ࡣຍỈศゎᛶࢱࣥࢽࣥ㢮ࡀከ㔞࡟ྵࡲࢀࡿࡇ࡜ࡀ ࢃ࠿ࡗࡓࠋࡲࡓࠊ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධࢆཷࡅᯤṚࡋࡓ࣑ࢬࢼࣛࡢ㎶ᮦ࡟࠾࠸ ࡚ࡣࠊຍỈศゎᛶࢱࣥࢽࣥ㢮ࡣῶᑡࡋࠊࡑࡢ࢔ࢢࣜࢥ࡛ࣥ࠶ࡿ࢞ࣜࢵࢡ㓟ࡸ࢚ࣛࢪ㓟 ࡢ㔞ࡀቑຍࡍࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋࡇࡢຍỈศゎᛶ཯ᛂࡣ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධ ࡟ᑐࡍࡿ࣑ࢬࢼࣛ⮬㌟ࡢ㜵⾨཯ᛂ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡿ௚ࠊ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ ධ࡟ࡼࡗ࡚ឤᰁࡋࡓࢼࣛ⳦ Raffaerea quercivora ࡀឤᰁࡋࠊᨺฟࡍࡿ㓝⣲ᵝ≀㉁࡟ࡼࡗ ࡚ᘬࡁ㉳ࡇࡉࢀ࡚࠸ࡿྍ⬟ᛶࡶ࠶ࡿࠋࡑࡇ࡛ࠊࢼࣛ⳦ࡀຍỈศゎᛶࢱࣥࢽࣥ㢮ࢆศゎ ࡍࡿ㓝⣲㢮ࢆ᭷ࡋ࡚࠸ࡿ࠿࡝࠺࠿ࢆ᳨ウࡍࡿࡓࡵ࡟ࠊ࢞ࣟ࢝ࢸ࢟ࣥ㢮ࡢΰྜ≀࡛࠶ࡿ ࢱࣥࢽࣥ㓟 1)_{ࢆࣔࢹࣝ໬ྜ≀࡜ࡋ࡚⏝࠸ࠊࢼࣛ⳦ࡢᾮయᇵᆅ࡟ῧຍࡋ࡚ࠊࡑࡢ㔞ⓗኚ} ໬ࢆ HPLC ศᯒ࡟ࡼࡗ࡚ほᐹࡋࡓࠋ

46

2-2 ᐇ㦂᪉ἲ 2-2-1 ౪ヨ⳦ R. quercivora ࡟ศ㢮ࡉࢀࡿ⳦ᰴࡣከᵝ࡞⳦ྀࢆ♧ࡍ 2㸧_{ࠋ௒ᅇࡣ㸪ᒣᙧ┴ࡢ⿕ᐖᮌࡼࡾ} ศ㞳ࡉࢀࡓ R. quercivora ࡢ⳦ᰴ㸦Wa㸫A㸧ࢆ⏝࠸ࡓࠋࡇࡢ⳦ᰴࡣ㸪ᇵᆅࢆ㯤Ⰽ໬ࡉࡏ㸪 ⳦⣒ࡣⓑⰍ࡛࠶ࡿࠋ 2-2-2 ࢱࣥࢽࣥ㓟ῧຍᇵᆅ࡟࠾ࡅࡿࢼࣛ⳦ࡢᇵ㣴࡜ࠊᇵᆅᡂศࡢศᯒ 2-2-2-1 ⳦య㝖ཤᚋࡢࢼࣛ⳦ᇵ㣴ᇵᆅ࡟࠾ࡅࡿࠊࢱࣥࢽࣥ㓟㔞ࡢኚ໬ Ț⥺⁛⳦῭ࡳࢩ࣮ࣕࣞ࡟ࠊ࣮࢜ࢺࢡ࣮ࣞࣈ࡟࡚⁛⳦㸦121Υ㸪15min㸧ࡋࡓ PDA ᇵᆅ ࢆὶࡋࡇࡳࠊ෭༷ࡋ࡚ᅛࡵࡓࠋࡇࡇ࡟ࠊ஦๓࡟ูࡢ PGA ᇵᆅୖ࡟ᇵ㣴ࡋ෭ⶶᗜ࡛ಖ ⟶ࡋ࡚࠸ࡓࢼࣛ⳦ࢆ᳜⳦ࡋࡓࠋࡇࢀࢆ 28Υ࡛ 10 ᪥ᇵ㣴ࡋࠊ๓ᇵ㣴࡜ࡋࡓࠋ 50 ml ᐜ୕ゅࣇࣛࢫࢥ࡟ࣅࢱ࣑ࣥ࡜࣑ࢿࣛࣝ㢮ࢆྵࡴᾮయᇵᆅ㸱 㸧_{ࢆ 10 ml ㄪ〇ࡋࠊ} ⁛⳦ᚋࠊ࠶ࡽ࠿ࡌࡵ๓ᇵ㣴ࡋ࡚࠾࠸ࡓ R. quercivora ࢆ๓ᇵ㣴ᇵᆅࡈ࡜ No. 4 ࡢࢥࣝࢡ ࣮࣮࡛࣎ࣛᡴࡕᢤ࠸࡚᥋✀ࡋࡓࠋ2 ᪥㛫ࡢᇵ㣴ᚋࠊᇵᆅࡢ୍㒊㸦2 ml㸧ࢆ஝⇱ᶵ࡛஝ ⇕⁛⳦ࡋࡓࢧࣥࣉࣝ⎼࡟⛣ࡋࡓࠋ⛣ࡍ㝿࡟ࠊᇵᆅࡣ⁛⳦῭ࡳࣇ࢕ࣝࢱ࣮ࢆ㏻ࡋ࡚࠿ࡽ ࢧࣥࣉࣝ⎼࡟ධࢀࡓࠋࡇࡇ࡟ࠊ200 mg / ml ࡢ࢚ࢱࣀ࣮ࣝ⁐ᾮ࡟ㄪ〇ࡋࡓࢱࣥࢽࣥ㓟ࢆ 0.1 ml / ml ᇵᆅ࡜࡞ࡿࡼ࠺ῧຍࡋࡓࠋῧຍᚋࡣ 28Υ࡟ಖࡗࡓᇵ㣴ᗜ࡟㟼⨨ࡋࠊࢱࣥࢽ ࣥ㓟ῧຍ࠿ࡽ 3 ᪥┠࡟ࡑࡢ୍㒊ࢆ࡜ࡗ࡚ HPLC ヨᩱ࡜ࡋࡓࠋ

47

2-2-2-2 ࢱࣥࢽࣥ㓟ῧຍᇵᆅ࡟࠾ࡅࡿࢼࣛ⳦ᇵ㣴࡜ࠊᇵᆅᡂศࡢ HPLC ศᯒ ࢼࣛ⳦ࡢ๓ᇵ㣴ࠊᮏᇵ㣴 ࡢᇵᆅㄪ〇࠾ࡼࡧ᳜⳦᧯సࡣ2-2-2-1 ࡜ྠᵝ࡟⾜ࡗࡓࠋ2 ᪥ 㛫ࡢᇵ㣴ᚋࠊᇵᆅ࡟ 200 mg / ml ࡢ࢚ࢱࣀ࣮ࣝ⁐ᾮ࡟ㄪ〇ࡋࡓࢱࣥࢽࣥ㓟ࢆ 0.1 ml / ml ᇵᆅ࡜࡞ࡿࡼ࠺ῧຍࡋࡓࠋࡉࡽ࡟ 3 ᪥㛫ᇵ㣴ᚋ㸪୕ゅࣇࣛࢫࢥࡈ࡜㍍ࡃᦂࡍࡗ࡚ࣇࣛ ࢫࢥෆࢆ඘ศ࡟ᨩᢾࡉࡏ࡚࠿ࡽࠊᇵᆅࢆ࢚ࢵ࣌ࣥࢻࣝࣇࢳ࣮ࣗࣈ࡟ 200ȣl ࡜ࡗ࡚㐲ᚰ 㸦12,000 rpm 10 min㸧ࡋࠊୖΎ࡜ỿẊ࡟ศࡅࡓࠋୖΎࡣࡑࡢࡲࡲ HPLC ศᯒ࡟౪ࡋࡓࠋ ỿẊ࡟ࡣ 200ȣl ࡢ⵨␃Ỉࢆຍ࠼࡚࠿ࡽ㉸㡢Ἴ࡛⳦య◚○ࡋࠊࡇࢀࢆ㐲ᚰ㸦12,000 rpm 10 min.㸧ࡋ࡚࠿ࡽୖΎࢆ HPLC ศᯒ࡟౪ࡋࡓࠋ

48

2-3 ⤖ᯝ࡜⪃ᐹ 2-3-1 ࢼࣛ⳦ᇵ㣴ᇵᆅ࡛ฎ⌮ࡋࡓࢱࣥࢽࣥ㓟ࡢኚ໬ ᇵ㣴ᇵᆅ࠾ࡼࡧᇵ㣴୰ࡢᇵᆅ࡟ࢱࣥࢽࣥ㓟ࢆῧຍࡋࠊ3 ᪥ᚋ࡟ᇵᆅࢆᅇ཰ࡋศᯒࡋࡓ HPLC ࢡ࣐ࣟࢺࢢ࣒ࣛࢆ Fig. 3-1 ࡟♧ࡍࠋࡇࡢศᯒ᮲௳࡛ࠊ࢞ࣜࢵࢡ㓟ࡣࣜࢸࣥࢩࣙ ࣥࢱ࢖࣒ 10 ศ㡭࡟ࠊ࢞ࣟࢱࣥࢽࣥ㢮ࡣࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 25 ศ௜㏆࡟ࣈ࣮ࣟࢻ࡞ࣆ ࣮ࢡ⩌࡜ࡋ᳨࡚ฟࡉࢀࡿࠋࡑࢀࡒࢀࡢࢡ࣐ࣟࢺࢢ࣒ࣛ࡟ࡘ࠸࡚࢞ࣜࢵࢡ㓟࡜࢞ࣟࢱࣥ ࢽࣥ㢮ࡢࣆ࣮ࢡ㧗ࢆẚ㍑ࡍࡿ࡜ࠊ࢞ࣟࢱࣥࢽࣥ㢮ࡢῶᑡ ࡣ㢧ⴭ࡛࠶ࡗࡓࠋࡇࡢࡇ࡜࠿ ࡽࠊࢼࣛ⳦ࡀㄏᑟࡍࡿ㓝⣲ᵝ≀㉁࡟ࡣࠊࢱࣥࢼ࣮ࢮᵝάᛶࡀ࠶ࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋ ࡲࡓࠊࡑࡢࢡ࣐ࣟࢺࢢ࣒ࣛࢆᣑ኱ࡍࡿ࡜ࠊ࢞ࣜࢵࢡ㓟࡜࢞ࣟࢱࣥࢽࣥ㢮ࡢ㛫࡟」ᩘᮏ ࡢࣆ࣮ࢡࡀ☜ㄆࡉࢀࡓࠋࡇࢀࡽ࡟ࡘ࠸࡚ࡣࡑࡢ UV ࢫ࣌ࢡࢺࣝ࠿ࡽ໬ྜ≀ࢆྠᐃࡍࡿ ࡇ࡜ࡣ࡛ࡁ࡞࠿ࡗࡓࡀࠊࢼࣛ⳦ࡢ⢒㓝⣲࡟ࡼࡿ࢞ࣟࢱࣥࢽࣥ㢮ࡢ௦ㅰ࡟ࡼࡗ࡚⏕ࡌࡓ ≀㉁࡛࠶ࡿྍ⬟ᛶࡀ࠶ࡿࠋ

49

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

Fig. 3-1 HPLC chromatograms of tannic acid added in culture medium of R. quercivora after fungus body was removed (a) and control (b)

㼍㻕

50

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

Fig. 3-2 HPLC chromatograms of tannic acid added in culture medium of R. quercivora after fungus body was removed (a) and control (b) (zoom up)

㼍㻕

51

2-3-2 ⳦యࢆྵࡴࢼࣛ⳦ᇵ㣴ᇵᆅ࡟࡚ฎ⌮ࡋࡓࢱࣥࢽࣥ㓟ࡢኚ໬ ⳦యࢆྵࡴᇵᆅ࡟ࢱࣥࢽࣥ㓟ࢆῧຍࡋࠊᇵ㣴ᚋࡢᇵᆅ࡜ࠊࡑࡢ㝿࡟ᅇ཰ࡋࡓ⳦యࢆ◚ ○ࡋ࡚ᚓࡽࢀࡓỈ⁐ᾮ࡟ࡘ࠸࡚ࠊHPLC ศᯒࢆ⾜ࡗࡓ㸦Fig. 3-3㸧ࠋࡇࡢศᯒ࠿ࡽࠊᇵ ᆅ࠿ࡽࡣࢃࡎ࠿࡟ࢱࣥࢽࣥ㓟࡟ྵࡲࢀࡿ࢞ࣟࢱࣥࢽࣥ㢮 ࢆ᳨ฟࡋࡓࡀࠊ⳦య◚○⁐ᾮ ࠿ࡽࡣ᳨ฟࡉࢀ࡞࠿ࡗࡓࠋࡲࡓࠊ࢞ࣜࢵࢡ㓟ࡶ᳨ฟࡉࢀ࡞࠿ࡗࡓࠋᇵᆅᡂศࡢศᯒ࡛ ࢞ࣟࢱࣥࢽࣥ㢮ࡀ᳨ฟࡉࢀ࡞࠿ࡗࡓࡇ࡜࠿ࡽࠊࢱࣥࢽࣥ㓟࡟ྵࡲࢀࡿ࢞ࣟࢱࣥࢽࣥ㢮 ࡣࢼࣛ⳦ࡢ⳦యࡀᏑᅾࡍࡿሙྜ࡟ࡼࡾศゎࡀಁ㐍ࡉࢀࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ⳦య◚ ○ࡋ࡚ᚓࡽࢀࡓỈ⁐ᾮ࠿ࡽ࢞ࣟࢱࣥࢽࣥ㢮ࡀ᳨ฟࡉࢀ࡞࠿ࡗࡓࡇ࡜࠿ࡽࡣࠊ࢞ࣟࢱࣥ ࢽࣥ㢮ࡀ⳦య࡟௜╔ࡋ࡚࠾ࡽࡎࡍ࡭࡚ຍỈศゎࡉࢀࡓྍ⬟ᛶ࡜ࠊ⳦య࡟௜╔ࡋ࡚࠸ࡿ ࡀ ࢱ ࣥ ࢽ ࣥ 㢮 ࡣ Ỉ ࡟ 㞴 ⁐ ࡛ ࠶ ࡿ ࡓ ࡵ ࡟ ⳦ య ࠿ ࡽ ⁐ ฟ ࡛ ࡁ ࡞ ࠿ ࡗ ࡓ ྍ ⬟ ᛶ ࡀ ⪃ ࠼ ࡽ ࢀ ࡓࠋࡲࡓࠊ࠸ࡎࢀࡢศᯒ࡟࠾࠸࡚ࡶ࢞ࣜࢵࢡ㓟ࡀ᳨ฟࡉࢀ࡞࠿ࡗࡓⅬ࡟ࡘ࠸࡚ࡣࠊ➨ ஧❶࡛㏙࡭ࡿ⌮⏤࡟ࡼࡾࡑࡢ㔞ࡀῶᑡࡋࡓࡓࡵ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡿࠋ

52

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

Fig. 3-3 HPLC chromatograms of tannic acid added in culture medium of

R. quercivora with fungus body (a), water soluble part of fragmentized fungus

body after cultured with tannic acid (b) and control (c) (zoom up)

㼍㻕

㼎㻕

53

2-4 ࡲ࡜ࡵ ࣔ ࢹ ࣝ ໬ ྜ ≀ ⩌ ࡜ ࡋ ࡚ ຍ Ỉ ศ ゎ ᛶ ࢱ ࣥ ࢽ ࣥ ࡛ ࠶ ࡿ ࢞ ࣟ ࢱ ࣥ ࢽ ࣥ ࡢ ΰ ྜ ≀ ࡛ ࠶ ࡿ ࢱ ࣥ ࢽࣥ㓟ࢆ⏝࠸ࠊࢼࣛ⳦ࢆ㝖ཤࡋࡓᇵ㣴ᇵᆅ࡟ῧຍࡋࡓ࡜ࡇࢁࠊ㢧ⴭ࡞ῶᑡࡀぢࡽࢀࡓࠋ ࡇ ࡢࡇ ࡜࠿ ࡽࠊ ࢼࣛ ⳦ࡀ ᨺฟ ࡍ ࡿ ⢒㓝 ⣲࡟ ࢱࣥ ࢼ࣮ ࢮᵝ άᛶ ࡀ࠶ ࡿࡇ ࡜ࡀ ࢃ࠿ ࡗࡓ ࠋ ࡲࡓࠊ⳦యࢆྵࡴᇵ㣴ᇵᆅ࡟ῧຍࡋࡓሙྜࠊࢱࣥࢽࣥ㓟ࡢࣆ࣮ࢡࡢῶᑡࡣࠊ⳦యࢆྵ ࡲ࡞࠸ሙྜ࡟ẚ࡭࡚ⴭࡋ࠿ࡗࡓࠋࡇࡢࡇ࡜࠿ࡽࠊࢼࣛ⳦ࡢ⢒㓝⣲ࡣᨺฟ㔞ࡼࡾ⳦య⾲ 㠃 ࡬ࡢ ỿ╔ ࡀከ ࡃࠊ ࡼࡗ ࡚ࢼ ࣛ⳦ ࡢ⳦ య⾲ 㠃࡛ ຍỈ ศゎ ࡀࡼ ࡾά Ⓨ࡛ ࠶ࡿ ྍ⬟ ᛶࡸ ࠊ ࢱࣥࢽࣥ㓟ࡀ⳦య࡟྾╔ࡋ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ

54

2-5 ཧ⪃ᩥ⊩

1. K. -T. Chung, Z. Lu and M. W. Chou. 1998. Mechanism of inhibition of tannic acidnext term and related compounds on the growth of intestinal bacteria. Food and Chemical Toxicology. 36: 1053-1060 㸦ࢱࣥࢽࣥ㓟㸧

2. ᮌᮧ໷Ꮚ㸪ᯇ⏣㝧௓㸪❑㔝㧗ᚨ㸪ఀ⸨㐍୍㑻㸦 2003㸧Raffaelea quercivora ⳦ᰴ㛫ࡢ

ᙧែᏛⓗ≉ᚩ㸬 ୰᳃◊ 51㸸199-200 㸦౪ヨ⳦㸧

3. ㉥ᯇ ໏㸪ୖᔱ ὒ㸪⸨஭ ฼㑻㸪ྜྷཎ ୍ᖺ. 1985. ࣜࢢࢽࣥศゎ⬟ࡢᙉຊ࡞ⓑ

Ⰽ⭉ᮙ⳦ࡢࢫࢡ࣮ࣜࢽࣥࢢ. ⣬ࣃᢏㄅ 39:65-74 㸦ᇵᆅ⤌ᡂ㸧

4. K. Imai , T. Mitsunaga , H. Takemoto , T. Yamada, S. Ito, H. Ohashi (2008) Extractives of Quercus crispula sapwood infected by the pathogenic fungi Raffaelea quercivora (I) : Comparison of sapwood extractives non-infected and infected. J.Wood. Sci. 55:126-132

55

➨ 3 ❶ 3-1 ⥴ゝ ➨஧❶࡛㏙࡭ࡓࡼ࠺࡟ࠊࢼࣛ⳦ࡢ⢒㓝⣲ࡣຍỈศゎᛶࢱࣥࢽࣥ㢮࡛࠶ࡿࢱࣥࢽࣥ㓟ࢆ ศゎࡍࡿࡇ࡜ࡀࢃ࠿ࡗࡓࠋࡋ࠿ࡋ࡞ࡀࡽࠊࢱࣥࢽࣥ㓟ࡣࠊຍỈศゎᛶࢱࣥࢽࣥࡢ୰࡛ ࡶ≉࡟ࠊ࢞ࣟࢱࣥࢽࣥࡢΰྜ≀࡛࠶ࡿࠋࡼࡗ࡚ࠊࢱࣥࢽࣥ㓟㔞ࡢῶᑡ࡟ࡘ࠸࡚ࠊᐃ㔞 ⓗ࡞ኚ໬ࢆ㏣࠺ࡇ࡜ࡣᅔ㞴࡛࠶ࡗࡓࠋࡑࡇ࡛ࠊࢱࣥࢽࣥ㓟࠿ࡽ௦⾲ⓗ࢞ࣟࢱࣥࢽࣥࡢ ୍ࡘ࡛࠶ࡿ࣌ࣥࢱ࢞ࣟ࢖ࣝࢢࣝࢥ࣮ࢫ㸦PGG㸧ࢆ༢㞳ࡋࠊࢼࣛ⳦ࡢᇵ㣴ᇵᆅ࡟ῧຍࡍ ࡿࡇ࡜࡛ࠊࡑࡢ㔞ⓗᣲືࢆ㏣࠺ࡇ࡜࡟ࡋࡓࠋ

56

3-2 ᐇ㦂᪉ἲ

3-2-1 PGG ࡢ༢㞳࡜ྠᐃ

࢞ࣟࢱࣥࢽࣥ㢮ࡢΰྜ≀࡛࠶ࡿࢱࣥࢽࣥ㓟㸦ᑠ᐀໬Ꮫ ⸆ရᰴᘧ఍♫㸧ࢆ࣓ࢱࣀ࣮ࣝ࡟

⁐ゎࡋࠊศྲྀ HPLC ࢆ⏝࠸࡚ࠊ௨ୗࡢ᮲௳࡟ࡼࡗ࡚ศ⏬ࡋࡓࠋ

Pump : SHIMADZU LC-10AD (0.05 % TFA in water), SHIMADZU LC-10AT (Methanol) Detector : SHIMADZU SPD-M10Avp

Column ocen : SHIMADZU CTO-10A

Communication bus module : SHIMADZU CBM-10A Degasser : SHIMADZU DGU-12A

Column ; Deverosil ODS HG-5 4.6™250 mm Flow rate ; 2 ml / min.

Column temp. ; 35Υ

Detector wave length ; 280 nm

Solvent; 0.1 % TFA in acetonitrile % TFA in H2O Initial 0 / 100 , Final 30 / 70

Gradient time ; 120 min.

ศ⏬ᚋࠊHPLC ศᯒ࡟ࡼࡗ࡚༢㞳ࡉࢀ࡚࠸ࡿࡇ࡜ࢆ☜ㄆࡋࠊNMR ゎᯒ࡟ࡼࡗ࡚ PGG

57

3-2-2 PGG ῧຍᇵᆅ࡛ࡢࢼࣛ⳦ࡢᇵ㣴࡜ࠊᇵᆅᡂศࡢศᯒ Ț⥺⁛⳦῭ࡳࢩ࣮ࣕࣞ࡟ࠊ࣮࢜ࢺࢡ࣮ࣞࣈ࡟࡚⁛⳦㸦121Υ㸪15min㸧ࡋࡓ PDA ᇵᆅ ࢆὶࡋࡇࡳࠊ෭༷ࡋ࡚ᅛࡵࡓࠋࡇࡇ࡟ࠊ஦๓࡟ูࡢ PGA ᇵᆅୖ࡟ᇵ㣴ࡋ෭ⶶᗜ࡛ಖ ⟶ࡋ࡚࠸ࡓࢼࣛ⳦ࢆ᳜⳦ࡋࡓࠋࡇࢀࢆ 28Υ࡛ 10 ᪥ᇵ㣴ࡋࠊ๓ᇵ㣴࡜ࡋࡓࠋ 50 ml ᐜ୕ゅࣇࣛࢫࢥ࡟ࣅࢱ࣑ࣥ࡜࣑ࢿࣛࣝࢆྵࡴᆅᇵᆅ 1)_{ࢆ10 ml ㄪ〇ࡋࠊ⁛⳦ᚋࠊ} ࠶ࡽ࠿ࡌࡵ๓ᇵ㣴ࡋ࡚࠾࠸ࡓ R. quercivora ࢆ๓ᇵ㣴ᇵᆅࡈ࡜ No. 4 ࡢࢥࣝࢡ࣮࣮࣎ࣛ ࡛ᡴࡕᢤ࠸࡚᥋✀ࡋࡓࠋࡑࡇ࡬ࠊ200 mg / ml 50 % ࣓ࢱࣀ࣮ࣝ⁐ᾮ࡟ㄪ〇ࡋࡓ PGG ࢆ 20 μl ῧຍࡋࡓࠋࣇࣛࢫࢥ࠶ࡓࡾࡢ PGG ⃰ᗘࡣ 4 mg / 10 ml ᇵᆅ ࡛࠶ࡗࡓࠋ28 Υ࡟ ಖࡗࡓᇵ㣴ᗜ࡛ᇵ㣴ࡋ ᇵ㣴࠿ࡽ 1,2,4,6,9,11 ᫬㛫ᚋࠊ࡜ 2 ᪥┠௨㝆ࡣẖ᪥㸪30 μl ࢆࢧ ࣥࣉࣜࣥࢢࡋࠊ-30Υࡢ෭෾ᗜ࡛ಖᏑࡋࡓࠋࡇࢀࢆศᯒ┤๓࡟ゎ෾ࡋࠊ 20μl ࢆ HPLC ศᯒ࡟౪ࡋࡓࠋHPLC ࢡ࣐ࣟࢺࢢ࣒ࣛ࠿ࡽ PGG ࡜ࠊࡑࡢ࢔ࢢࣜࢥ࡛ࣥ࠶ࡿ࢞ࣜࢵࢡ㓟 ࡟┦ᙜࡍࡿࣆ࣮ࢡࡢ㠃✚ࡢ ኚ໬ࢆほᐹࡋࡓࠋࡲࡓࠊPGG ࡜࢞ࣜࢵࢡ㓟࡟ࡘ࠸࡚ࡣࣆ࣮ ࢡ㠃✚࡜⃰ᗘ࡜ࡢ᳨㔞⥺ࢆసᡂࡋࠊᇵᆅ୰࡟ྵࡲࢀࡿ PGG ࡜࢞ࣜࢵࢡ㓟㔞࡟ࡘ࠸࡚ ⟬ฟࡋࡓࠋ

58

3-3 ⤖ᯝ࡜⪃ᐹ 3-3-1 PGG ࡢᵓ㐀ゎᯒ ࢱࣥࢽࣥ㓟࠿ࡽ༢㞳ࡉࢀࡓ PGG ࡣࠊNMR ゎᯒ࡟ࡼࡗ࡚ࡑࡢᵓ㐀ࢆྠᐃࡋࡓ(Fig. 3-1)ࠋ NMR ࢳ࣮ࣕࢺ࠿ࡽࠊ6.8~7.2ppm ࡢ⠊ᅖ࡟ࠊ࢞ࣟ࢖ࣝᇶ 5 ಶศ࡟┦➼ࡍࡿࢩࣥࢢࣞࢵ ࢺࣆ࣮ࢡ 5 ᮏࡀㄆࡵࡽࢀࠊࡲࡓ 4.4~6.3ppm ࡢ⠊ᅖ࡟ぢࡽࢀࡿ 5 ࡘࡢࣆ࣮ࢡࡣࢢࣝࢥ࣮ ࢫṧᇶࡢࡶࡢ࡜୍⮴ࡋࡓࠋࡲࡓࠊࡑࡢࢣ࣑࢝ࣝࢩࣇࢺࡣᩥ⊩ ್ 2)_{࡜ࠊ௨๓࡟ᙜ◊✲ᐊ} ࡛༢㞳ྠᐃࡋࡓ PGG ࡢ NMR ࢹ࣮ࢱ࡜↷ࡽࡋྜࢃࡏࠊPGG ࡛࠶ࡿࡇ࡜ࢆ☜ㄆࡋࡓࠋ

59

Fi g. 3-1 1_H N M R chart of PGG isolated from tannic acid

60

3-3-2 ࢼࣛ⳦ᇵ㣴࡟క࠺ࠊPGG ࠾ࡼࡧ࢞ࣜࢵࢡ㓟㔞ࡢ⤒᫬ⓗኚ໬ R. quercivora ࡢᾮయᇵᆅ࡟ῧຍࡉࢀࡓ PGG ࠾ࡼࡧࡑࡢศゎ≀࡛࠶ࡿ࢞ࣜࢵࢡ㓟㔞ࡣᇵ 㣴㛤ጞ࠿ࡽ 1,2,4,6,9,11 ᫬㛫ᚋࠊ࡜ẖ᪥ࢧࣥࣉࣜࣥࢢࡋࡓࢧࣥࣉࣝࡢ HPLC ศᯒࢆ⾜ࡗ ࡓࠋPGG ࠾ࡼࡧ࢞ࣜࢵࢡ㓟ࡑࢀࡒࢀࡢ᳨㔞⥺ࢆసᡂࡋࠊࣆ࣮ࢡ㠃✚࠿ࡽࡑࢀࡒࢀࡢ㔞 ࢆィ⟬ࡋࡓࠋFig. 3-2 ࡟♧ࡍ࡜࠾ࡾࠊPGG ࡣᇵ㣴 11 ᫬㛫࡛࡯ࡰ༙ῶࡋࠊ2 ᪥┠࡟ࡣ࡯ ࡰㄆࡵࡽࢀ࡞ࡃ࡞ࡗࡓࠋࡲࡓ࢞ࣜࢵࢡ㓟ࡣ PGG ࡜ࡣ㏫࡟ᇵ㣴 1 ᪥┠࠿ࡽᛴቑࡋࠊ5 ᪥ ┠࡟᭱ࡶከ࠿ࡗࡓࠋࡼࡗ࡚ࠊPGG ࡀ R. quercivora ࡢ⏕⫱࡜ඹ࡟ῶᑡࡋࠊศゎ≀࡜ࡋ࡚ ࢞ࣜࢵࢡ㓟ࡀ⏕ᡂࡉࢀࡿࡀࠊ⏕ᡂࡋࡓ࢞ ࣜࢵࢡ㓟㔞ࡶᇵ㣴ࡀ⥆ࡃ࡟ࡘࢀ࡚ῶᑡࡍࡿ࡜ ࠸࠺⤖ᯝ࡟࡞ࡗࡓࠋ࢞ࣜࢵࢡ㓟㔞ࡢῶᑡ࡟ࡘ࠸࡚ࡣࠊ࢞ࣜࢵࢡ㓟ࡀࣛࢵ࣮࢝ࢮ࡟ࡼࡿ

ฎ⌮࡛㔜ྜࡋࠊ㔜ྜ≀࡜ࡋ࡚〓Ⰽࡢ purprogallincarboxylic acid (PGCA)ࢆ⏕ᡂࡍࡿࡇ࡜

࡜ 3)_{ࠊR. quercivora ࡢ⢒㓝⣲ࡀࣛࢵ࣮࢝ࢮάᛶࢆ♧ࡍࡇ࡜} 4)_{࠿ࡽࠊR. quercivora ࡀᨺฟ} ࡋࡓࣛࢵ࣮࢝ࢮᵝ㓝⣲࡟ࡼࡗ࡚࢞ࣜࢵࢡ㓟ࡀ㔜ྜࡋࠊࡑࡢ㔞ࡀῶᑡࡋࡓࡢࡔ࡜⪃࠼ࡽ ࢀࡓࠋࡲࡓࠊࢼࣛ⳦ࡢ⢒㓝⣲άᛶ࡛ࡣࠊࢼࣛ⳦ࡢᇵ㣴ᙜึࡣᙉ࠸ࢱࣥࢼ࣮ࢮάᛶࡀࡳ ࡽࢀࠊࡑࡢᚋࠊ㐜ࢀ࡚ࣛࢵ࣮࢝ࢮάᛶࡀぢࡽࢀࡿࡇ࡜ࡀࢃ࠿ࡗ࡚࠸ࡿࠋࡲࡓࠊ௨๓ࡢ ◊✲ࡼࡾࠊࢼࣛ⳦࡟ᑐࡋ࡚࢞ࣜࢵࢡ㓟ࡀᙉ࠸ᢠ⳦άᛶࢆ♧ࡍࡀࠊ㔜ྜ≀࡛࠶ࡿ PGCA ࡣ࢞ࣜࢵࢡ㓟ࡼࡾᙅ࠸ᢠ⳦άᛶࡋ࠿ࡋࡵࡉ࡞࠸ 4)_{ࠋࡑࡇ࡛ࠊࢼࣛ⳦ࡀࡲࡎᨺฟࡍࡿࢱ} ࣥࢼ࣮ࢮ࡟ࡼࡗ࡚ PGG ࢆ࢞ࣜࢵࢡ㓟࡟ศゎࡋࠊࡑࡢᚋᨺฟࡍࡿࣛࢵ࣮࢝ࢮ࡛࢞ࣜࢵ ࢡ㓟ࢆ㔜ྜࡋࠊࢼࣛ⳦⮬㌟࡟࡜ࡗ࡚ࡼࡾẘᛶࡢప࠸PGCA ࢆ⏕ᡂࡋ࡚࠸ࡿ࡜ᛮࢃࢀࡿࠋ ࡋ࠿ࡋ࡞ࡀࡽࠊ௒ᅇࡢ᳨ウ࡟࠾࠸࡚ࠊPGCA ࡟ヱᙜࡍࡿࣆ࣮ࢡࢆ᳨ฟࡍࡿࡇ࡜ࡣ࡛ࡁ ࡞࠿ࡗࡓࠋࡇࡢⅬ࡟ࡘ࠸࡚ࡣࠊPGCA ࡣࡑࡢ λmax ࡀ 224㸪304㸪398nm3, 4)_{࡛࠶ࡾࠊ௒}

61

ᅇࡢ᳨ฟἼ㛗࡛࠶ࡿ 280nm ࡣ྾཰ࢆ♧ࡉ࡞࠸ࡇ࡜࠿ࡽࠊ᳨ฟࡀᅔ㞴࡛࠶ࡗࡓྍ⬟ᛶࡀ

62

Fig. 3-2 Change of PGG and gallic acid amount added in the culture medium of R.

quercivora.

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63

3-4 ࡲ࡜ࡵ ࢼࣛ⳦ࡢ⢒㓝⣲ฎ⌮࡟ࡼࡗ࡚ࠊPGG ࡣ㏿ࡸ࠿࡟ศゎࡉࢀࠊ࢔ࢢࣜࢥ࡛ࣥ࠶ࡿ࢞ࣜࢵࢡ 㓟ࡀ⏕ᡂࡋࡓࠋࡋ࠿ࡋ࡞ࡀࡽࠊ⏕ᡂࡋࡓ࢞ࣜࢵࢡ㓟ࡶࡲࡓࠊࢼࣛ⳦ᇵ㣴࡟కࡗ࡚ῶᑡ ࡋࡓࠋࡋ࠿ࡋ࡞ࡀࡽࠊ௒ᅇࡢ᳨ฟ࡛ࡣ࢞ࣜࢵࢡ㓟ࡢ㔜ྜ≀࡜ࡋ࡚⪃࠼ࡽࢀࡿ PGCA ࡣ ᳨ฟࡉࢀ࡞࠿ࡗࡓࠋ

64

3-5 ཧ⪃ᩥ⊩

1. ㉥ᯇ ໏㸪ୖᔱ ὒ㸪⸨஭ ฼㑻㸪ྜྷཎ ୍ᖺ. 1985. ࣜࢢࢽࣥศゎ⬟ࡢᙉຊ࡞ⓑ

Ⰽ⭉ᮙ⳦ࡢࢫࢡ࣮ࣜࢽࣥࢢ. ⣬ࣃᢏㄅ 39:65-74 㸦ᇵᆅ⤌ᡂ㸧

2. Richard T, Vitrac X, Merillon JM, Monti JP (2005) Role of peptide primary sequence in polyphenol-protein recognition : An example with neurotensin. Biochimica et Biophysica Acta. 1726:238-243

3. Karl H (1954) Über die Bildung der Purpurogallincarbonsäure durch fermentative

Oxydation der Gallussäure. (zugleich II Mitteilung über gerbstoffartige Substanzen). Archiv der Pharmazie 287:497-503

4. ➟஭⨾࿴ ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩ࡟ࡼࡿ࣑ࢬࢼࣛቯṚኚⰍ࡟㛵ࡍࡿᢳฟᡂศࡢ◊ ✲ ಟኈㄽᩥ

5. K. Imai , T. Mitsunaga , H. Takemoto , T. Yamada, S. Ito, H. Ohashi (2008)

Extractives of Quercus crispula sapwood infected by the pathogenic fungi Raffaelea

quercivora (I) : Comparison of sapwood extractives non-infected and infected. J.Wood. Sci. 55:126-132