几株昆虫病原菌微菌核的诱导培养和条件优化-微生物学专业论文.docxVIP

II II 件下，MS-DE 复苏下降趋势显著增强，但均在也 87％以上，相比之下布氏白僵菌 MS 更耐储藏，31℃储藏 60 天复苏率仍能达到 91.93±1.58％。剂型化对 MS 复苏率有 一定影响，但的确有一定保护作用。室温长期储存实验中，MS-DE 第 4 个月时复苏 率开始急速下降，到第 10 个月最低下降至 40.22％；剂型化 MS 复苏率在第 6 个月才 开始明显下降，10 个月后最低降至 55.19％，可见 MS 本身具备了一定抗逆性，而在 制剂保护下能更长久的维持这种优势。 关键词：微菌核 液体发酵 白僵菌 金龟子绿僵菌 响应面 III III Abstract Fungal insecticides developed with entomopathogenic fungi for soil pest control has showed potential, but propagales tolerant to drought, with long shelf life, are still needed for massproduction. Highly resistant microsclerotia (MS, hereafters) have been discovered to be able to form in submerged culture, showing potential against soil pests. However, the state and art for MS production are still not mature and need to be optimized. A series optimization experiments were run to improve culture condition and provide reliable parameters. Nitrogen sort was determined to be significant affecting factors of MS by change of nutrition conditions of shake culture for Beauveria bassiana, Beuveria brongniarti and Metarhizium anisopliae, 3 isolates each. Different isolates had different optimal nitrogen resource; absence of Tween through a series single factor tests, a highly productive isolate of each species was chosen: Bb2729 for B. bassiana, Bbr17 for B. brongniartii, and Ma55 for M. anisopliae, with MS production at 3.87×103 ml-1, 6.80×103 ml-1 和 4.07×103 ml-1, respectively. Based on a single factor test, proportion of carbon and nitrogen resources was rescreened, with a result that MS production and biomass both were affected by carbon concentration and carbon/nitrogen ratio. On the basis of the single factor test, another optimization was further made by a two-level fractional factor test on non nutrition conditions, with carbon concentration, C/N, rotation speed, amount of conidial suspension inoculum, and load of liquid medium as 5 influencing factors and with MS production and biomass as response to screen significant factors. The result indicated that the significant influen