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通过对甜菜夜蛾卵超低温冷冻过程中一系列影响因子的研究 ,建立了甜菜夜蛾卵在液氮中冷冻保存的方法。结果表明 ,利用 1 5%次氯酸钠处理卵壳 6min ,再用异丙醇处理 1 5sec和N 己烷处理 30sec可有效去除甜菜夜蛾卵的外卵壳和蜡质层 ,获得 95%以上的渗透化率和 60 %左右的存活率 ;渗透化卵经 2mol L乙二醇初步抗冻 30min和含 1 0 %BSA的 8 5mol L乙二醇脱水 5min ,约有 55%～ 60 %的卵存活和2 4 %的卵孵化 ;将抗冻处理后的卵投入液氮 ( - 1 96℃ )中 2 4h ,在 37℃下解冻 ,结果有 ( 1 6 3± 7 6) %的卵发育至黑头期 ,( 1 6± 1 1 ) %的卵孵化。
Through the study of a series of influencing factors during the cryopreservation of beet armyworm eggs, the method of cryopreservation of beet armyworm eggs in liquid nitrogen was established. The results showed that treatment of eggshell with 1.5% sodium hypochlorite for 6min and then treated with isopropanol for 15sec and N-hexane for 30sec can effectively remove the outer egg shell and wax layer of beet armyworm eggs, and obtain more than 95% And the viability was about 60%. The permeated eggs were dehydrated with 2mol L ethylene glycol for 30min and 85mol L ethylene glycol containing 10% BSA for 5min, and about 55-60% 4% eggs were hatched. The frost-treated eggs were immersed in liquid nitrogen (-196 ℃) for 24 hours and thawed at 37 ℃. As a result, (163 ± 76)% eggs developed to blackheads, (16 ± 1 1)% eggs hatching.