不同种类冷冻保护剂对海带配子体及孢子的毒性评价

Toxicity evaluation of different kinds of cryoprotectants on gametophytes and spores of Sacchatina japonica

为筛选用于海带超低温保存的冷冻保护剂,采用10%二甲基亚砜(DMSO)、10%甘油、10%蔗糖、10%乙二醇、10%DMSO+10%甘油、10%DMSO+8%葡萄糖、10%DMSO+10%蔗糖、10%DMSO+10%甘油+8%葡萄糖、10%DMSO+10%山梨醇等9种单一或复合冷冻保护剂,在0~4℃条件下分别对海带配子体和孢子进行处理,对配子体处理30 min、60 min、24 h,对孢子处理15、30、60 min,通过统计配子体细胞的存活率以及孢子萌发成配子体的比例,对不同种类的冷冻保护剂进行毒性评价。结果表明:9种冷冻保护剂均对海带配子体和孢子有一定的毒性作用,并且毒性随着处理时间的延长而增强。10%DMSO对海带配子体细胞的毒性最小,处理60 min后配子体细胞的存活率为100%;其次是10%乙二醇,处理30 min后配子体的存活率为100%;含10%甘油的单一或复合冷冻保护剂对配子体细胞毒性较大,处理后配子体的成活率显著低于DMSO和乙二醇组(P<0.05)。单一冷冻保护剂对海带孢子的毒性显著低于复合冷冻保护剂,除10%甘油组处理30、60 min外,其他单一冷冻保护剂试验组的配子体发生率均在60%~80%。

In order to screen the cryoprotectants of ultra-low temperature preservation of kelp,nine single or compound cryoprotectants including 10%Dimethyl sulfoxide(DMSO),10%glycerol,10%sucrose,10%ethylene glycol,10%DMSO+10%glycerol,10%DMSO+8%glucose,10%DMSO+10%sucrose,10%DMSO+10%glycerol+8%glucose,and 10%DMSO+10%sorbitol were used to treat the gametophytes and spores of kelp for 30 min,60 min,24 h and 15 min,30 min,60 min,respectively,at 0-4℃.The toxicity of different cryoprotectants was evaluated by calculating the survival rate of gametophyte cells and the proportion of spores germination.The results showed that all the 9 cryoprotectants had certain toxic effects on gametophytes and spores of S.japonica,and the toxicity increased with the extension of treatment time.The toxicity of 10%DMSO to gametophytic cells was minimal,and the survival rate of was 100%after 60 min treatment,followed by 10%ethylene glycol.The survival rate of gametophyte treated with single or compound cryoprotectant containing 10%glycerol was significantly lower than that of DMSO and ethylene glycol groups(P<0.05).The toxicity of single cryoprotectant was significantly lower than that of compound cryoprotectant,the incidence of gametophyte treated with single cryoprotectant was 60%-80%except 10%glycerol group treated for 30 min,60 min.