Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Isolation and Characterization of Six Microorganisms from the Digestive Tract of the Cricket Gryllus bimaculatus

쌍별귀뚜라미(Gryllus bimaculatus) 소화기관에서 분리한 6종류의 특성규명

  • Kwon, Kisang (Department of Biomedical Laboratory Science, College of Nursing & Health, Kyungwoon University) ;
  • Lee, Eun Ryeong (Department of Biomedical Laboratory Science, College of Nursing & Health, Kyungwoon University) ;
  • Yoo, Bo-Kyung (Department of Anatomy and Cell Biology, College of Medicine, Chungnam National University) ;
  • Ko, Young Hwa (Department of Anatomy and Cell Biology, College of Medicine, Chungnam National University) ;
  • Shin, Hyojung (Department of Anatomy and Cell Biology, College of Medicine, Chungnam National University) ;
  • Choi, Ji-Young (Applied Entomology Division, National Academy of Agricultural Science, RDA) ;
  • Kwon, O-Yu (Department of Anatomy and Cell Biology, College of Medicine, Chungnam National University)
  • Received : 2017.04.28
  • Accepted : 2017.06.15
  • Published : 2017.09.30
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Abstract

We describe the isolation and characterization of six different intestinal microorganisms from the digestive tract of the cricket Gryllus bimaculatus. Based on 16S rRNA gene sequences, we obtained six isolates belonging to four different genera: Staphylococcus, Bacillus, Citrobacter, and Proteus. All the isolates were resistant to ampicillin. Ampicillin is an irreversible inhibitor of the enzymeetranspeptidase, which is needed to make bacterial cell walls. None of the isolates were resistant to kanamycin, which binds to the 30S subunit of the bacterial ribosome and then inhibits total protein synthesis. Gram staining was conducted, in addition to morphological classification under a microscope. Four grampositive isolates and two gram-negative isolates were detected. The gram-positive isolates were GL1 (round shaped, 2 am in diameter), GL2 (rod shaped, $2.5{\mu}m$ in length), GL3 (rod shaped, $2{\mu}m$ in length), and GL4 (round shaped, $1.5{\mu}m$ in diameter). The gram-negative isolates were GL5 (rod shaped, $2{\mu}m$ in length) and GL6 (rod-shaped, $2.5{\mu}m$ in length). Notably, two of the isolates, GL2 and GL4, secreted specific extracellular proteins. These were determined by MALDI-TOF-MS spectral analysis to be a 87 kDa collagenase, 56 kDa hypothetical protein, and 200 kDa hypothetical protein. The six isolates in this study could be used for various biotechnological applications and pest management, both in the field and in greenhouse systems. In addition, it would be interesting to determine the relationship between these isolates and their host.

우리는 처음으로 쌍별귀뚜라미 소화기관에서 6종류의 미생물을 분리하고 특성을 규명하였다. 이들은 16S rDNA을 기준으로 분류한 결과 4종류(Staphylococcus, Bacillus, Citrobacter, Proteus)에 속하였다. 분리된 6종류의 미생물은 공통적으로 ampicillin에 저항성을 보이지만 kanamycin 저항성은 보이지 않았다. 이들을 Gram염색하여 미생물의 형태적 특징을 확인하였다. Gram-positive한 rod-shaped GL2와 round-shaped GL4는 다른 분리 균 보다 많은 양의 세포외분비물을 만들었다, 이들을 MALDI-TOF-MS spectral analysis결과 87-kDa collagenase, 56-kDa & 200-kDa hypothetical protein이였다. 새롭게 분리된 6종류의 미생물은 귀뚜라미의 생리에 미치는 영향과 이들의 생물공학적 혹은 해충 방제에 이용될 수 있는 연구가 기대된다.

Keywords

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