초록
곤충변원성충(Steinernema carpocapsae Weiser)의 감염태 유충에 대한 액체질소( $-190^{\circ}C$)냉동 저장하는 방법을 개발하기 위하여, 냉동보관 전처리 과정 및 저장기간에 따른 선충생존율과 감염력 변이를 조사하였다. 냉동저장 전에 22% 글리세롤로 선충은 6, 12, 24시간 동안 배양한 후 70% methanol($0^{\circ}C$)안에 10분간 유지하였다. 글리세롤과 methanol에 넣어 둔 샘플을 0,85% 식염수에 넣어 24시간 유지한 후 생존능력을 측정하였다. 글리세롤 배양시간에 따른 선충 생존율의 차이는 없었으나, methanol처리에서는 글리세롤 처리 시간이 길수록 선충 생존율이 높게 나타났다. 냉동저장에 따른 선충의 생존율은 글리세롤 배양시간이 짧았던 처리(6시간)를 제외하고는 5개월 동안 평균 70% 이상으로 높게 나타났다. 냉동저장 되었던 선충의 곤충감염율은 저장기간에 관계없이 90% 이상을 나타내었다. 이상의 결과는 곤충병원선충이 냉동저장에 의해 장기간보존이 가능하다는 것을 제시한다.
Cryopreservation of infective juveniles of entomopathogenic nematode, Steinernema carpocapsae Weiser, was conducted at $-190^{\circ}C$ liquid nitrogen and its, efficacy was analysed on nematode survival and pathogenicity with glycerol pretreatments and storage periods. Infective juveniles were pre-treated before being frozen by incubating the nematodes in 22% glycerol for each of 6, 12, and 24 h, followed by 70% methanol at $0^{\circ}C$ for 10 minutes. Just after glycerol and methanol incubations, subsamp1es of the nematodes were resuspended in 0.85% saline and maintained during 24h for viability determination. Different glycerol incubation periods significantly affected the nematode susceptibility to methanol infiltration. Six hour incubation in glycerol resulted in much less nematode survival than did 12 h or 24 h incubation. About 70% of the infective juveniles frozen at $-190^{\circ}C$ for 5 months, preincubat-ed in glycerol at least for 12h, were able to survive after being resuspended in 30°C saline. They did not also show any change in their pathogenicity during cryopreservation. These results suggest an improved technique for long-term storage of the entomopathogenic nematodes.
