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以产孢量、菌落生长速率、菌丝干重等为指标,设计单因素试验和正交试验,对重庆地区柑桔粉虱上的渐狭蜡蚧菌TN002菌株的培养条件进行研究。结果表明,在马铃薯葡萄糖琼脂培养基上,10~35℃范围内TN002菌株均可生长,25℃条件下产孢量和菌落生长速率最大。pH值(4.5~8.0)对TN002菌株生长无显著影响,pH值5.5时产孢量最大。与无机氮相比,有机氮更利于TN002菌株生长和产孢,以酵母浸粉为氮源时菌落生长速率最大,以蛋白胨为氮源时产孢量最大。在不同碳源中,蔗糖和葡萄糖比较利于TN002菌株生长和产孢。在摇瓶培养的各液体培养基中,以葡萄糖酵母浸粉液体培养基的产孢量和菌丝干重较大。摇瓶培养时间为60h时产孢量和菌丝干重较大。TN002菌株产孢发酵培养优选方案为500mL锥形瓶中葡萄糖酵母浸粉液体培养基装量100mL、接种量4%(孢子悬液浓度4×107个孢子/mL)、摇床转速190r/min。
Taking sporulation quantity, colony growth rate and mycelium dry weight as indexes, the single factor experiment and orthogonal experiment were designed to study the culture conditions of T. waxytum strain TN002 in Citrus whitefly in Chongqing. The results showed that on the potato dextrose agar medium, the TN002 strain could grow in the range of 10-35 ℃, the sporulation amount and colony growth rate were the highest at 25 ℃. The pH value (4.5 ~ 8.0) had no significant effect on the growth of TN002 strain, and the sporulation amount was the highest at pH 5.5. Compared with inorganic nitrogen, organic nitrogen was more conducive to the TN002 strain growth and sporulation, yeast extract powder as the nitrogen source when the colony growth rate of the largest peptone nitrogen source spores maximum. In different carbon sources, sucrose and glucose are more conducive to TN002 strain growth and sporulation. In each liquid medium cultured in shake flasks, the amount of sporulation and the dry weight of mycelium of glucose leaching liquid culture medium were large. Shake flask culture time 60h spores and mycelial dry weight greater. The optimal culture conditions of strain TN002 for sporulation were as follows: 100 mL of inoculated liquid of glucose yeast in 500 mL Erlenmeyer flask with inoculum size of 4% (spore suspension concentration of 4 × 107 spores / mL) and shaking speed of 190 r / min.