松材线虫低温冷冻保存的影响因子  

Factors affecting the cryopreservation of pine wood nematode,Bursaphelenchus xylophilus

作　　者：朱丽华[1] 张文莉[1] 郑俊俊 季锦衣[1] 叶建仁[1] 

机构地区：[1]南京林业大学森林资源与环境学院,江苏省有害生物入侵预防与控制重点实验室,江苏南京210037

出　　处：《中国森林病虫》2013年第4期6-8,共3页Forest Pest and Disease

基　　金：江苏省有害生物入侵预防与控制重点实验室开放基金(PMIS200802);国家林业局推广项目(2011-51)

摘　　要：比较了保护剂平衡时间、解冻温度、线虫浓度及线虫发育期对低温冷冻松材线虫存活的影响。结果表明,平衡时间对冷冻保存有较大影响,以20%甘油为保护剂,在4℃下平衡12 h后转入-80℃超低温冰箱,1 d后线虫存活率为(41.7±4.9)%;冻存存活率与线虫发育期有一定关系,幼虫显著高于成虫,幼虫、雌成虫和雄成虫的存活率分别为(31.5±7.4)%,(7.9%±3.5)%,(6.6±2.8)%。在所选择的解冻温度(30～45℃)和线虫浓度(10 500条/mL～8 4000条/mL)范围内,解冻温度及线虫浓度对存活率影响差异不明显。The effect of equilibrium time,thawing temperature,initial concentration of nematode suspension and development stage of Bursaphelenchus xylophilus on survival of cryopreserved nematodes were evaluated.The results indicated that equilibrium time had significant effect on the survival of cryopreserved B.xylophilus.Significantly more nematodes were motile after cryopreservation in glycerol 20% and equilibrated at 4 ℃ for 12 h and then placing in a-80 ℃ freezer than directly placing them in freezer.The average survival rate of B.xylophilus was（41.7±4.9） %.More juveniles survived deep freezing than adults.The average survival rate of juveniles,females and males were（31.5±7.4）%,（7.9±3.5）%,（6.6±2.8）%,respectively.So,nematodes should be harvested for cryopreservation from just starved plates where juveniles exist predominantly.Thawing temperature（30-45 ℃） and concentration of nematode（10 500/mL-84 000/mL） didn＇t affect the survival ability of cryopreserved nematodes.

关 键 词：松材线虫 冷冻保存 甘油 平衡时间 发育阶段 

分 类 号：S763.16[农业科学—森林保护学]