Abstract
BACKGROUND: Several yeast species have potential applications in biotechnology and the identification of such yeast species is of great interest. The first step in the identification of yeasts is the establishment of an effective isolation method. Thus, we compared the efficacy of different yeast media in the isolation of yeast associated with Aloe vera and Aloe saponaria. METHODS AND RESULTS: In this study, we spread homogenized A. vera and A. saponaria leaves onto 4 different yeast selective media containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose. We observed high selectivity for yeast and many colonies on media. We isolated 67 yeast strains from A. vera and 42 yeast strains from A. saponaria. We used phylogenetic analysis to identify the yeast isolates based on ITS region sequencing and performed sequence analysis on representative isolates from each agar plate. Further, we compared the sequences obtained with reference sequences. The yeast species isolated from A. vera were as follows: 56 isolates of Meyerozyma, 9 isolates of Cryptococcus, and 1 isolate each of Rhodotorula and Sporobolomyces. Those isolated from A. saponaria were as follows: 41 isolates of Rhodosporidium and 1 isolate of Sporobolomyces. CONCLUSION(S): All the isolates obtained using large agar plate containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose were identified as yeast. Therefore, we concluded that this method is useful for selective screening of yeast species.
효모의 유용 기능을 탐색하기 위해서는 효모만을 스크리닝 하는 방법의 정립이 요구되는데 이를 위해서 알로에에 정착하는 효모 군집을 분석하였다. 본 연구에서는 알로에 베라(Aloe vera)와 알로에 사포나리아 (Aloe saponaria)의 잎을 파쇄하여 세균을 억제하기 위해서 항생제 chloramphenicol과 streptomycin과 곰팡이 생장을 억제하기 위해서 Triton X-100과 L-sorbose를 포함한 4가지 선택 배지(DOB with CS, GPY, YM, SCG)에 도말 하였고, A. vera에서 총 67균주, A. saponaria에서 총 42균주를 분리해 내었다. 분리된 균주를 ITS 1, 4 primer를 사용하여 sequence의 계통분석을 실시한 결과, A. vera에서는 Meyerozyma 가 56균주, Rhodotorula 가 1균주, Sporobolomyces 가 1균주, Cryptococcus 가 9균주, A. saponaria에서는 Rhodosporidium 가 41균주, Sporobolomyces가 1균주로 분포 되었다. 계통분석한 모든 분리 균주가 효모로 밝혀졌으며, 특히 대형 플레이트를 도입하여 신속하고 간편하게 대량의 효모를 발굴할 수 있음을 보였다.