蚜科专化菌努利虫疠霉菌种超低温储存  

Cryopreservation of aphid-specific pathogen Pandora nouryi

作　　者：周湘[1] 冯明光[2] 黄志宏[3] 

机构地区：[1]浙江农林大学林业与生物技术学院,浙江临安311300 [2]浙江大学生命科学学院,浙江杭州310058 [3]华侨大学化工学院,福建厦门361021

出　　处：《菌物学报》2012年第2期285-291,共7页Mycosystema

基　　金：华侨大学基本科研业务费专项基金(No.JB-ZR1116);华侨大学高层次人才科研启动(No.09BS402);浙江省教育厅科研项目(No.Y201119481);浙江农林大学科研启动项目(No.2010FR065)

摘　　要：虫霉目真菌的活力对超低温储存较为敏感。储存法在大范围应用前,需对储存效果进行详细评估。将蚜科专化菌努利虫疠霉以初级分生孢子形式(2-3?105个孢子/mL)在?80℃超低温存储12个月。结果显示日常用于培养该真菌的含0.1%乳化芝麻油的萨氏培养基作为超低温保护基质能有效地储存努利虫疠霉孢子,比常见的冷冻保护剂如二甲亚砜和甘油的效果好。孢子悬液经解冻和培养后可获得最多的生物量,而且菌种保持了较高的生长速率。更重要的是,萨氏培养基的主要成分4%葡萄糖、1%蛋白胨和1%酵母粉在低温存储过程中发挥了协同作用,能保持该菌的高毒力性状。存储期一年后,供试菌种对桃蚜若蚜的毒力经多剂量生物测定后显示其半致死剂量(3.3个孢子/mm2)与未储存前所报道的结果相当。总之,?80℃超低温储存是一种可接受的保存虫霉菌种技术,而萨氏培养基比常用的10%甘油更适合作为努利虫疠霉的冷冻保护基质。The viabilities of entomophthoralean fungi are sensitive to cryopreservation procedures.The efficacy of preservation methods for these fastidious fungi should be evaluated in detail before extensive application.In this study,the primary spores（2-3？105conidia/mL） of an aphid obligate pathogen Pandora nouryi was preserved at-80℃ for 12 months.The routine culture medium of Sabouraud dextrose broth plus 0.1% emulsified sesame oil（SDBS） was selected for culture storation,due to more effective in protecting the fungal spores than ordinary cryoprotectants such as dimethylulphoxide and glycerol.The highest biomass and growth rate were obtained after the preserved spore suspensions were thawed and incubated.Moreover,4% glucose,1% peptone and 1% yeast extracts in the liquid culture played cooperative roles in preserving the isolate under ultracold condition,and the virulence retained stable.After one year preservation,the median lethal concentration of the isolate against Myzus persicae nymphs was estimated as 3.3conidia/mm2 by analysis of multiple-dose bioassay data,consistent with that in the previous repeated report.In a word,-80℃ cryopreservation can be accepted as a good preservation technique for entomophthoralean fungi,and Sabouraud dextrose broth is more suitable as cryoprotectantive medium for P.nouryi than 10% glycerol solution.

关 键 词：超低温储存 努利虫疠霉 萨氏培养基 冷冻保护剂 毒力 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治]