The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Genetic Relationships of Internal Transcribed Spacer (ITS) Regions on Entomopathogenic Fungi by RFLP

Entomopathogenic Fungi의 ITS 영역에 대한 RFLP 분석

  • Choi, In-Young (Chonbuk-do Agricultural Research and Extension Service) ;
  • You, Young-Jin (Chonbuk-do Agricultural Research and Extension Service) ;
  • Choi, Joung-Sik (Chonbuk-do Agricultural Research and Extension Service) ;
  • Lee, Wang-Hyu (Department of Agricultural Biology, Chonbuk National University)
  • Published : 2000.10.30
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Abstract

A similarity coefficient were analyzed by RFLP of fourteen species of entomopathogenic fungi, isolated from inhabiting pupa and adult insect at forest. Each rDNA ITS I and ITS II with primers of ITS 1 and ITS 4 was amplified by PCR. The amplified products were conserved to 500 bp were not demarcated between genus and species. Four Paeciliomyces tenuipes, two Beauveria bassiana and six Cordyceps militaris were treated by seven restriction enzymes and confirmed in species except JB3 by electrophoresis band. However, the band of C. scarabaeicola showed the identity with B. bassiana. The result of this experiment indicated that the teleomorph of C. scarabaeicola was the same as that of B. bassiana. CfoI and HpaII of seven restricted enzymes were easily discriminating in the genus between Paecilomyees and Cordyceps. Especially, CfoI was more effective to classify the genera of Paecilomyees, Cordyceps and Beauveria than other restriction enzymes. The band patterns of RFLP of P. tenuipes, C. militaris, C. scarabaeicola and B. bassiana were also analyzed by UPGMA program of NTSYS-pc and showed 100% significance. Thus, the similarity coefficient tended to be lower between genera by RFLP analysis, but was higher between species.

야생에 서식하고 있는 번데기 및 성충에 감염된 기주의 자실체로부터 동충하초속균 3속 14균주를 분리하였고, RFLP에 의한 유전정보를 이용하여 ITS 영역에 대한 유연관계를 분석하였다. 수집된 각 균주의 rDNA ITS I과 II 부위를 primer ITS 1과 ITS 4를 사용하여 PCR에 의해 증폭한 결과 증폭된 산물은 500 bp크기로 잘 보존되어 종간 또는 속간 서로 구분되지 않았다. 따라서 증폭산물을 7종의 제한효소로 절단하여 밴드상을 관찰한 결과 수집된 Paecilomyces tenuipes 4균주와 Beauveria bassiana 2균주, JB3 균주를 제외한 Cordyceps militaris 6균주는 각각 동일 종으로 분류되었다. 한편 C. scarabaeicola 균주는 7종의 제한효소에서 모두 B. bassiana의 밴드와 일치하여 C. scarabaeicola의 불완전세대는 B. bassiana로 간주되었다. 제한 효소 중에 Paecilomyces속과 Cordyceps 속간 구분에 용이한 제한효소는 CfoI, HpaII이었으며, 그중 CfoI는 Paecilomyces, Cordyceps, Beauveria속간 구분에 용이하였다. UPGMA 분석결과 P. tenuipes, C. militaris, C. scarabaeicola와 B. bassiana, JB4 균주 등 4개의 군으로 그룹화되었으며, 그룹간에는 100% 유의수준을 나타냈다. 따라서 rDNA-RFLP 분석에서 속간 유연관계는 적었으나 종간 유연관계는 가까운 결과를 얻었다.

Keywords

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