The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
권호 표지

Classification of Cordyceps spp. by Morphological Characteristics and Protein Banding Pattern

동충하초(冬蟲夏草)(Cordyceps) 속균의 형태적인 특징과 단백질 Pattern에 의한 계통 분류

  • Sung, Jae-Mo (Department of Agricultural Biology, College of Agriculture Kangwon National University) ;
  • Lee, Hyun-Kyung (Department of Agricultural Biology, College of Agriculture Kangwon National University) ;
  • Yang, Keun-Joo (Department of Agricultural Biology, College of Agriculture Kangwon National University)
  • 성재모 (강원대학교 농과대학 농생물학과) ;
  • 이현경 (강원대학교 농과대학 농생물학과) ;
  • 양근주 (강원대학교 농과대학 농생물학과)
  • Published : 1995.03.30
Download PDF
⟨ Previous Next ⟩

Abstract

Ten species of Cordyceps species were collected throughout Kangwon province including Chuncheon Dongsanmyun KNU forest experiment from June to September, 1993. Collected Cordyceps species were identified as Cordyceps militaris, C. roseostromata, C. kyushuensis, C. scarabaeicola, Phytocordyceps ninchukiospora, C. nutans, Paecilomyces tenuipes, C. sphecocephala, Hymenostilbe odonatae, Torrubiella sp.. C. militaris, type species of Cordyceps species, was mainly formed on pupae of Lepidoptera and found after the rainy season around July. Fruiting body of C. roseostromata was morphologically similar to those of C. militaris, but relatively small in size and they were also found on lawn or pupa of Lepidoptera. Fruiting body of C. scarabaeicola was found on adult Scarabaeidae specifically and collect fruiting bodies of C. kyushuensis were on larva of moth. C. nutans and C. sphecocephala had host specificity on Hemiptera and Hymenoptera, respectively. Each species formed elliptical fertile part attach to the slim and carneous stalk and they were collected the most in specimen number through whole season of the summer. Ascospore of Phytocordyceps ninchukiospora on seed was characterized by two viable, multiseptate, fusiform units linked end-to-end by a long, filiform connective. Paecilomyces tenuipes, imperfect stage of the genus Cordyceps is multi-infective fungi that attack all stages of all groups of insects. Hymenostilbe odonatae attacks only adult Odonata and Torrubiella sp. formed on spider was difficult to collect because it was found the back side of leaf. As results of cultural test PDA medium showed the best mycelial growth. In the experiment of effect of the acidity inside of the media, C. militaris was good on pH 5, C. nutans and Phytocordyceps ninchukiospora were good on pH 6 and Paecilomyces tenuipes was on pH 7 and C. scarabaeicola was on pH 9. All isolates tested showed the best mycelial growth at $20^{\circ}C$. Morphologically similar isolates were used to analyze protein banding pattern among and within species. As a result, C. militaris, C. roseostromata and C. kyushuensis were clustered into close species and C. scarabaeicola and Phytocordyceps ninchukiospora were relatively distant from those species.

강원도 춘성군 강원대 연습림등 장원도일대의 산지에서 1993년 6월부터 9월까지 채집된 Cordyceps 속균의 자실체(子實體)는 Cordyceps militaris, C. roseostromata, C. kyushuensis, C. scarabaeicola, Phytocordyceps ninchukiospora, C. nutans, Paecilomyces tenuipes, C. sphecocephala, Hymenostilbe odonatae, Torrubiella sp. 등 10종이었다. 동충하초속균(冬蟲夏草屬菌)의 Type species이기도 한 C. militaris는 주로 인시목(隣翅目)의 번데기를 기주(寄主)로 한 자실체(子實體)가 채집되었는데 본(本) 종(種)의 발생은 7월경의 장마철이 지나면서 다수 채집되었다. C. roseostromata는 형태적으로 C. mlitaris와 유사하나 크기가 작으며 역시 인시목(隣翅目)의 유충, 번데기 등을 기주(寄主)로 한 자실체(子實體)가 채집되었다. C. scarabaeicola는 풍뎅이 성충만을 침입하여 자실체(子實體)를 형성하였으며 C. kyushuensis는 박각시 나방의 유충만을 기주(寄主)로 하여 자실체(子實體)가 채집되었다. C. nutans와 C. sphecocephala는 각각 노린재와 벌만을 기주(寄主)로 하여 가늘고 질긴 침상의 병(柄)에 타원형의 두부(頭部)를 착생(着生)한 자실체(子實體)를 형성하는데 두 종은 채집지역에 관계없이 곤충기생성균(昆蟲寄生性菌)이 발생하는 6월부터 8월 사이 전 시기에 걸쳐 가장 많이 채집되었다. 식물의 종자를 기주(寄主)로 하여 자실체(子實體)를 형성하는 Phytocordyceps ninchukiospora의 자양포자(子襄胞子)는 다른 Cordyceps 속균과는 달리 양옆 4개의 자양포자(子襄胞子)가 실모양의 구조로서 연결되어 있는 형태를 취하고 있었다. Paecilomyces tenuipes는 모든 곤충의 발달단계 전 시기에 걸쳐 침입하는 다범성 균으로서 분생포자(分生胞子)를 형성하는 동충하초속균의 불완전 세대균이다. 잠자리 성충을 기주로 하여 형성된 Hymenostilbe odonatae는 잠자리만을 특이적으로 침입하며 거미성충을 기주로 형성되는 Torrubiella sp.는 주로 잎의 뒷면에서 형성되므로 채집이 어려웠다. 분리동정된 균주들을 이용하여 배양시험을 한 결과 PDA 배지가 가장 우수한 균사생장을 보여 주었으며 배지내 산도(酸度) 시험(試驗)에서 C. militaris는 pH5에서 C. nutans와 Phytocordyceps ninchukiospora는 PH 6에서 Paerilomyces tenuipes는 pH 7에서 C. scarabaeicola는 pH 9에서 각각 균사생장(菌絲生長)이 왕성하였다. 환경조건으로서의 온도 시험에서는 시험균주에서 공히 $20^{\circ}C$에서 우수한 균사생장율을 보여 주었다. 분리동정된 균주들중 형태적으로 유사한 균주들의 종간 또는 종내 유연관계 구명을 위하여 Protein banding pattern을 분석한 결과 C. militaris, C. roseostromata, C. kyushuensis는 근연종으로 clustering 되었으며 C. scarabaeicola, Phytocordyceps ninchukiospora는 비교적 유연관계가 먼 것으로 나타났다.

Keywords

References

  1. Introductory Mycology Alexopoulos, C.J.;Mims, C.W.
  2. Can. J. Bot. v.46 Studies on the production of perithecial stromata by Cordyceps militaris in artificial culture Basith, M.;Madelin, M.F.
  3. Mycologia v.52 Unusual fruiting of Cordyceps militaris Bigelow, H.E.
  4. Protein method Bollag, D.M.;Edelstein, S.J.
  5. Fungi in Switzerland. Vol. 1 Ascomycetes. Mykologia luzern Breitenbach, J.;Kranzlin, F.
  6. Biological control Insect pest Suppression Coppel, H.C.;Mertins, J.W.
  7. J. Chem. Soc. v.1951 Cordycepin, a metabolic product from cultures of Coydyceps militaris (Linn.) Link Pt. 1. Isolation and characteristics Cunningham, K.G.;Hutchinson, S.A.;Manson, W.;Spring, F.S.
  8. J. Cramer. Kew British Ascomycetes Dennis, R.W.G.
  9. U.S. Dept. Agri. Monogr. v.4 Balansia and the Balansiae in America Diehl, W.W.
  10. A manual of soil fungi Gilman, J.C.
  11. Molecular systematics Hills, D.M.;Moritz, C.
  12. Caner Res. v.21 Inhibition of Ehrlich mouse ascites tumour growth by cordycepin Jaggfer, D.V.;Kredich, N.M.;Guarino, A.J.
  13. Kor. J. Mycol. v.14 Note in unrecorded Fleshy Fungi of Cordyceps in Korea Jang, Yang-Suk;Hong, Soon-Woo
  14. Biochim. Biophysics. Acta. v.80 Effect of cordycepin triphosphate on the incorporation of $(8-^{14}C)$ adenine and $(^{37}P)$ orthophosphate into the acid-soluble ribotides of Ehrlich aseites tumour cells Klenow, H.;Overgaard-Hansen, K.
  15. Sei. Rept. Tokyo Buntika Daigaku v.5 The genus Cordyceps and its allies. Kobayasi, Y.
  16. Trans. Mycol. Soc. Japan v.23 Keys to the taxa of the genera Cordyceps and Torrubiella Kobayasi, Y.
  17. Fundamentals of molecular evolution Li, W.H.;Graur, D.
  18. Physiology of the fungi Lilly, V.G.;Barnett, H.L.
  19. Mycologia v.50 North American entomogenous species of Cordyceps Mains, E.B.
  20. Trans. Brit. Mycol. Soc. v.32 Cordyceps aphodii, a new species, on pasture cockchater grubs Mathiexon, J.
  21. Z. Angew. Entomom. v.55 Cordyceps militaris (Fr.) Link. Becobachtungen und Versuche anlasslich eines Fundes auf Tipula paludosa Meig.(dipt., Tipul.). Muller-Kogler, E
  22. Biochim. Biophys. Acta. v.80 The inhibition of-phosphoribosyl-l-pyrophosphate formation by cordycepin triphosphate in extracts of Ehrlich ascites tumour cells Overgaard-Hansen, K.
  23. Trans. Brit. Mycol. Soc. v.20 Cordyceps militaris and Isaria farinosa Petch, T.
  24. Trans. Brit. Mycol. Soc. Studies on entomogenous fungi Petch, T.
  25. Kor. J. Pl. Pathol. v.3 no.1 Microbial Control: Its Place in Korean Agriculture and Forestry Reed, D.K.
  26. Ascomycete Systematics Renyolds, D.R.
  27. NTSYS-pc, Numerical taxonomy and multivariate analysis system Rohlf, F.J.
  28. Biochim and Biophys. Acta. v.80 The inhibition of purine biosynthesis de noveo in Bacillus subtilis by cordycepin Rottman, F.;Guatino, A.
  29. Mycologia v.3 The Hypocreales of North America-IV. Seaver, F.J.
  30. Kor. J. Mycol. v.21 Studies on distribution and utilization of Cordyceps militaris and C. nutans. Sung, J.M.;Kim, C.H.;Yang, K.J.;Lee, H.K.;Kim, Y.S.
  31. 한국의 버섯 박완희
  32. 韓國動植物圖鑑 高等균류편(버섯류), 28 이지열;홍순우
  33. 漢藥 臨床 應用 神戶中醫學硏究會
  34. 冬蟲夏草 淸水大典
  35. 海軍醫學硏究所 v.10 no.12 冬蟲夏草及人工蟲草菌絲硏究槪況　　 예운운
  36. 冬蟲夏草圖鑑 小林義雄;淸水大典
  37. 昆蟲病理凡論 有賀久雄