【作者】 柴新义；

【导师】 李增智；

【作者基本信息】 安徽农业大学 ， 森林保护学， 2004， 硕士

【摘要】 球孢白僵菌（Beauveria bassiana (Balsamo) Vuillemin）是害虫微生物防治中一种经典的丝孢类昆虫病原真菌，因用途广泛、生产和应用性能优越而倍受关注，国外已有多种商品制剂注册生产。为改变我国目前白僵菌生产技术含量较低的现状，本试验以塑料平板和塑料网格平板作为生产的载体，研究了半固体发酵的培养工艺。同时，还研究了在采取不同的放菌措施下利用球孢白僵菌持续控制松墨天牛(Monochamus alternatus Hope)及其对林间生物群落所产生的影响和应用布氏白僵菌(Beauveria brongniartii (Sacc.) Petch)及金龟子绿僵菌(Metarhizium anisopliae)防治光肩星天牛(Anoplophora glabripennis Motschulsky)，以期为防治提供参考依据。对球孢白僵菌液体种子的生产，实验结果表明：以去皮的马铃薯20%，白砂糖2%作为液体种子的发酵培养基，自然pH，在25oC的培养温度下，发酵36个小时，所得菌体生物量高。对半固体发酵培养基，选择资源丰富的蚕蛹粉做为氮源，自然pH。同时加入合适的粘度剂以增加它在载体上的粘附性，采用浸涂的方法，最后确定的培养基产孢量达4.47亿/cm2,不仅组成成分简单，而且在载体上的粘附性较好。半固体发酵塑料平板培养工艺的试验结果表明：接种量以10%为佳，培养基在载体上粘附的厚度以0.25cm最为合适，培养环境温度25℃，湿度95%最利于该菌的产孢，产孢量可达6.13亿/cm2。从缩短生产周期和获得最大的经济产量综合考验，确定培养9d作为一个生产周期最为经济合理。 利用塑料网格平板生产的孢子粉含量可达1200～2000亿/克(含水率9.05～11.36%)，萌发率在98%以上，用细毛刷子收集塑料网格平板上残留的残渣孢子含量为1.1～2.4亿/克。采用不同的放菌策略防治松墨天牛的实验结果表明：未处理的对照区及两年中只在第一年放菌一次的处理区E死树不仅没有减退，反而分别上升43.1%和67.9%；相比之下，其它各处理区放菌次数均高于实验区E，死树分别减退2.9%～38.6%，其中以放菌频度最高的实验区B(两年中每年各放菌2次)死树减退率最高，达到38.6%。除两年中只在第一年放菌一次的实验区E以外，其他各放菌区与对照区之间经方差分析差异显著，说明林间放菌对松墨天牛的控制发挥了作用。以每年2次放菌控制效果最佳。对林间生物群落调查的结果表明：六个实验区的优势类群基本相同，即为鳞翅目、直翅目、膜翅目、双翅目、鞘翅目和蜘蛛等类群，并且各个目中的种数相等或相近。通过物种多样性相关指数分析表明，林间放菌控制松墨天牛对生物群落尚未产生明显<WP=4>的影响。利用布氏白僵菌、金龟子绿僵菌防治光肩星天牛的实验结果表明：布氏白僵菌、金龟子绿僵菌试验区的校正死亡率分别为21.26%、24.34%；布氏白僵菌、金龟子绿僵菌、对照试验区的天牛平均死亡时间分别为10.38d、17.16d、23.28d。更多还原

【Abstract】 Beauveria bassiana (Balsamo) Vuillemin is a classical entomopathogenic fungus in microbial control of insect pests. It has been attractive for its wide use and superior production property, and has been developed into a variety of commercial formulations of mycoinsecticides in the world. In order to further improve current mass production technology, a set of new technics of semisolid fermentation was developed, which used the plastic board and plastic reseau board as carriers to produce B. bassiana. At the same time, the effect of Monochamus alternatus control with B. bassiana by the different measures and the influence upon the community in the forest were studied and the application of Beauveria brongniartii and Metarhizium anisopliae were tried against Anoplophora glabripennis in order to look for new microbial control agentsLiquid seed culture of B. bassiana was studied at different temperatures. The results showed that biomass was the highest with the 20% peeled potato and 2% white sugar, natural pH, temperature at 25℃, and 36 hours for cultivation. For semisolid fermentation, silkworm powder was chosen as the nitrogen source at natural pH, and an adhesive matter was added into the semisolid medium to make the medium attach fast to the carriers. 4.47×108 conidia per cm2 were yielded by immersing inoculation method. The semisolid medium was composed simple with proper stickiness, suitable for B. bassiana to grow on.Technics of semisolid culture was studied, the results showed that inoculation amount 10%, thickness 0.25cm on the carrier, culture temperature at 25℃ and environmental RH 95% were optimal for the fungus to sporulate. The sporulation reached 6.13×108 conidia per cm2. The optimal harvest time was determined on the ninth day. In the production experiment with plastic reseau board as the carrier, the yield was 1200~2000×108 conidia per gram with water content 9.05～11.36% and germination rate>98%. The residue left on and collected the carriers contained 1.1～2.4×108 conidia per gram.An experiment of M. alternatus control was set with non-woven fabric bands impregnated with B. bassiana in different release frequencies. The results showed that the rates of dead trees increased 43.1% and 67.9%, respectively in the control plot and plot E where only one release of fungal band in the first year of two years, while the rates of others decreased 2.9%～38.6%, respectively. In the high frequency (2 release each year in <WP=6>the 2 years) release plot B the rate was the highest (38.6%). As compared with the control plot, all treatment plots except plot E relieved significantly (p﹤0.05), suggesting that the release of the fungal bands in the plots had effective control of M. alternatus and releasing frequency of twice a year was the best.The results of a biodiversity investigation in forest showed that the dominant groups were Lepidoptera, Orthoptera, Hymenoptera, Diptera, Coleoptera and Araneae similarly in the six different plots. The species numbers were equal or close in these groups.By analyzing indexes of biodiversity, it was suggested that the B. bassiana bands release in pine plantations didn’t have obvious effect on biodiversity of the community. The results of the experiment of A. glabripennis control by the B. brongniartii and M. anisopliae showed that mortalities of A. glabripennis in the B. brongniartii plot and the M. anisopliae plot were 21.26% and 24.34%, respectively. The average lethal time in B. brongniartii plot, M. anisopliae plot and control plot was 10.38d, 17.16d and 23.28d, respectively.更多还原