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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。
Insecta fungi vitality is more sensitive to cryogenic storage. Storage method in a wide range of applications, the need for a detailed assessment of the storage effect. Aphidicidae Helicoverpa armigera was stored in the form of primary conidia (2-3 × 105 spores / mL) for cryogenic storage at -80 ℃ for 12 months. The results showed that the daily culture of Sargassy medium containing 0.1% emulsified sesame oil used for culturing the fungus effectively preserved the spores of Aspergillus niger as an ultra-low temperature protective substrate, and the effect was better than common cryoprotectants such as dimethylsulfoxide and glycerol . Spore suspension thawed and cultured to obtain the most biomass, and strains maintained a higher growth rate. More importantly, 4% glucose, 1% peptone and 1% yeast powder, the major components of the culture medium, play a synergistic role in low temperature storage and maintain the high virulence of the bacterium. After one year of storage, the virulence of tested strains against Myzus persicae showed a comparable lethal dose (3.3 spores / mm 2) as reported before storage after multiple dose bioassays. In summary, Cryopreservation at 80 ° C is an acceptable technique for preserving Aspergillus species, whereas Sarsen’s medium is more suitable than Cryptosporidium niger as a cryoprotective substrate than the commonly used 10% glycerol.