白僵菌论文：环境中白僵菌分生孢子检测技术的研究

白僵菌论文：环境中白僵菌分生孢子检测技术的研究 白僵菌论文:环境中白僵菌分生孢子检测技术的研究 【中文摘要】白僵菌(Beauveria)是一种重要的虫生真菌,可以感染松毛虫(Dendrolimus Punctatus Walker)、松小蠹虫(Blasophagus piniperda. L)、天牛(Cerambycidae)、豆毒蛾 (CifunalocuplesWalker)、斜纹夜蛾(Prodenia liturd Fabricius)、棉铃虫(Helicoverpa armigera)、地老虎(Agrotis)等多种农林害虫。白僵菌具有容易培养、杀虫谱广、致病力强以及对人畜无毒、无害等特点,目前作为真菌杀虫剂已被广泛研究,是真菌杀虫剂中应用最多、防治效果最好的一个种类。白僵菌分生孢子接触害虫虫体后,适宜条件下萌发并侵入虫体,死虫尸体 表 关于同志近三年现实表现材料材料类招标技术评分表图表与交易pdf视力表打印pdf用图表说话 pdf 面产生白色分生孢子,随风扩散继续感染其它害虫,形成昆虫流行病。白僵菌分生孢子在空气中大量的散布,可污染桑园或直接扩散到蚕区感染家蚕等经济类昆虫。本实验针对环境中白僵菌分生孢子的扩散进行了监测研究,为有效防治白僵菌分生孢子的污染和危害益虫提供方法。主要的研究内容和结果如下:1.白僵菌分生孢子采集载体的研制微生物的检测方法中使用的采集或培养载体一般都是培养皿,普通的培养皿材质为玻璃,表面光滑、易碎,进行微生物采集时,只能平放。针对检测空气中白僵菌分生孢子的浓度情况,本实验室设计一种新型培养皿,带有独特的挂钩,即可用于空气中悬浮微生物的检测,也可用于沉降微生物的采集。培养皿的材质选为塑料,轻便、不易碎。2.检测培养基的选择室外空气环境复 杂,微生物种类繁多,需要一种即适合白僵菌生长又能有效抑制一些非目标真菌类的选择性培养基,以检测环境中白僵菌分生孢子的扩散和污染情况。白僵菌易于培养,营养条件要求低。本实验使用一种营养贫瘠的碱性培养基,配方如下:黄豆芽200g,蔗糖20g,链霉素0.30g孟加拉红0.33g琼脂18g,水1000mL,调节PH值为8.0,该培养基具有制作简单,成本低等特点。结果表明,使用该培养基即能有效的检测出环境中的白僵菌分生孢子又能够抑制一些非目标真菌的生长,易于观察菌落特征。能够应用于白僵菌分生孢子污染和扩散的检测。3.吸附垫型培养基的制备吸附垫型培养基是由添加了聚丙烯酸钠的培养液和含20%丙纶、80%棉的吸水性纸片通过吸附和干燥制成,可较长时期的保存和使用。该吸附垫型培养基可与特制配套培养皿联用,检测时只需加入一定量的无菌水即可。进行白僵菌分生孢子采集和检测时省去了配制培养基、清洗、消毒等大量辅助性工作,具有成本低廉、使用方便、操作简单、保水性能好等优点。4.对白僵菌分生孢子检测力的试验室内构建一个长2.08米,宽2.2米,高1.85米,共8.5m3的检测空间。称取不同浓度的白僵菌分生孢子粉进行检测,得出能检测到的最低孢子浓度。之后对家蚕进行检测,得出感染家蚕的最低孢子浓度。结果表明:空气中检测出的最低孢子浓度为0.00035g/m3(7.9×105个/m3)。感染家蚕的最低浓度也为0.00035g/m3(7.9×105/m3),感染率为3.3%。感染率为43.67,100%的孢子浓度分别是0.012g/m3(2.65×106个/m3)、0.027g/m3(6.0×107个/m3)。5.养蚕 场所及周边环境的检测试验白僵菌分生孢子的广泛撒播扩散致使环 境中的孢子浓度不断升高,监测环境中孢子浓度状况是有效避免白僵 菌分生孢子污染的前提。本实验室发明的挂钩培养皿可方便的进行室 外检测,解决了普通培养皿只能平放、不能悬挂的问题。 【英文摘要】Beauveria bassiana is an important entomogenous fungus which has been widely used as biological control agents against pests of agricultural and forestry such as the Dendrolimuss, Blasophagus, Monochamus, Cifunalocuples Walker, Spodopteralitura, Helicoverpa armigera and Agrotis. It has the characteristics that easy to cultivate, broad host range, high pathogenicity and safely to human beings and animals, which as the fungus insecticide has the most widely used and the best control effect and been studied extensively. Ordinary, after the Beauveria conidia exposed to pest body, it will sprout and intrude into the insect body under suitable conditions, then produce large numbers of conidia after the death of the insects and spread by the winds to infect other insects, formation an epidemic of insects. However, large of conidia suspended in the air can pollution the mulberry or direct spread to silkworm area infected economic insects. In this article we studied on monitoring about diffusion test of Beauveria bassiana conidia in the environment for provide good method of effective control pollution and harm of Beauveria bassiana conidia to beneficial insects. The main research contents and results are following:1. Invention of Beauveria bassiana conidia collection deviceThe carrier of collection or the culture in the microbial detection method is petri dish, ordinary it is made of glass which smooth and fragile, it can only flat when the microorganisms were collected. In order to detection of Beauveria bassiana conidia in the air, we have designed a novel Petri dish which has a special hook, it can be used for the detection of suspended microorganisms in the air and also be used for settling microbe collection. Material of petri dish is plastics which lightweight and shatterproof.2. Selection of mediumAir environment is complex and has many different kinds of germs, so we need a special culture medium which it is suitable for growth of Beauveria bassiana and can effectively inhibit other fungi, used to detect the diffusion and pollution of Beauveria bassiana conidia in environment. Beauveria bassiana is easy to cultivation and has low nutrition requirement. This experimental use an alkaline medium of nutrient barren which consisted of 200g bean sprouts,20g sucrose,0.30g streptomycin,0.33g 3.4,5,6-Tetrachlorofluorescein,18g agar and 1000mL water, PH8.0. It is very easy to manufacture and not cost much. The results show that, the culture medium is suitable for growth of Beauveria bassiana and easy to observe the characteristics of the colony, it can be applied to detect pollution and diffuse of Beauveria bassiana conidia.3. Preparation of adhesive-pad-culture-mediumAdsorption pad type medium is composed of the culture liquid which adding sodium polyacrylate and absorbent paper which containing 20% polypropylene fiber and 80% cotton, This adhesive-pad-culture-medium can be well matched with the special culture dish, while the only work needs to do before detection is adding a certain amount of sterilized water. It avoids lots of auxiliary work and easy to use in the B. bassiana conidia detection process.4. The detection test of Beauveria bassiana conidiaIn this study, we structure a detection of space that 2.08 meters long,2.2 meters wide and 1.85 meters high, a total of 8.5 cubic meters. Then weighing different concentrations of Beauveria bassiana conidia for detect, get the lowest of spore concentration. Then studies on detection of silkworm, obtain the lowest concentration of Bombyx mori infected. The results show that the minimum spore concentration of air detected is 0.00035g/m3, it is also the minimum concentration of Bombyx mori infected and the infection rate is 3.33%, respectively the infection rate are 43.67% and 100% that the spore concentration is 0.0012g/m3 and 0.027g/m3.5. The detection test of sericulture area and surrounding environmentB. bassiana conidia widely spread in the air, transmitted among insect groups, leads to the increasing spore concentration in environment. The premise to avoid pollution of B. bassiana conidia is to monitor the spore concentration in the air. We invent the dish with hook which is convenient for sericulture area and surrounding environment tests that may provided the way to solve the problem that general petri dish can only be flat, but cannot be hanged. 【关键词】白僵菌 空气 室内 检测技术 【英文关键词】Beauveria Air Indoor Detection technology 【目录】环境中白僵菌分生孢子检测技术的研究 摘要 8-10 Abstract 10-11 第一章 文献综述 12-28 1.1 白僵菌的研究进展 12-23 1.1.1 白僵菌属 的建立及其分类 12-13 1.1.2 白僵菌的分类方法 13-15 1.1.3 白僵菌属种的形态特征 15-20 1.1.4 白僵菌的应用及现状 20-22 1.1.5 白僵菌流行病学的研究 22-23 1.2 空气微生物检测方法与技术进展 23-26 1.2.1 微生物检测技术的方法 24-25 1.2.2 国内外微生物检测的研究现状 25-26 1.3 空气中白僵菌检测的研究进展 26-28 第二章 引言 28-32 2.1 研究背景及意义 28-30 2.2 技术路线 30-32 第三章 采集微生物的培养皿的制作 32-36 3.1 材料与方法 32-33 3.1.1 材料 32 3.1.2 培养皿制作方案 32 3.1.3 培养皿成品 32-33 3.2 结果与分析 33 3.3 讨论 33-36 第四章 选择性培养基的研究 36-38 4.1 材料与方法 36 4.1.1 培养基 36 4.1.2 不同培养基的空气采集实验 36 4.1.3 不同培养基的白僵菌培养实验 36 4.2 结果与分析 36-37 4.2.1 不同培养基的空气采集实验结果 36 4.2.2 不同培养基的白僵菌培养实验结果 36-37 4.3 讨论 37-38 第五章 吸附垫型培养基的制备、保存及检测 38-42 5.1 材料和方法 38-40 5.1.1 材料 38 5.1.2 吸附垫型培养基的制备方法 38 5.1.3 吸附垫型培养基的使用方法 38-39 5.1.4 吸附垫型培养基与普通培养基白僵菌分生孢子的检测研究 39-40 5.1.5 吸附垫型培养基的保存实验 40 5.2 结果与分析 40-41 5.2.1 吸 附垫型培养基与普通培养基的白僵菌分生孢子的检测结果 40-41 5.2.2 吸附垫型培养基的保存实验结果 41 5.3 讨论 41-42 第六章 对白僵菌分生孢子检测力的试验 42-50 6.1 材料和方法 42-46 6.1.1 实验材料 42 6.1.2 对白僵菌分生孢子的检测 42-46 6.1.3 不同白僵菌分生孢子的浓度对家蚕的感染试验 46 6.2 结果与分析 46-49 6.2.1 白僵菌孢子粉的制备 46 6.2.2 空气中悬浮白僵菌分生孢子悬挂检测结果 46-47 6.2.3 空气悬挂平放位点白僵菌分生孢子的检测结果 47-48 6.2.4 地表平放位点的检测结果 48 6.2.5 对家蚕的感染结果 48-49 6.3 讨论 49-50 第七章 养蚕场所及周边环境的检测试验 50-54 7.1 材料和方法 50-52 7.1.1 材料 50 7.1.2 桑园环境的检测 50 7.1.3 松树林的检测 50-51 7.1.4 养蚕农户的检测 51-52 7.2 结果与分析 52 7.2.1 桑园环境检测结果 52 7.2.2 松林环境检测结果 52 7.2.3 养蚕农户检测结果 52 7.3 讨论 52-54 第八章 结论 54-56 8.1 检测培养皿的研制 54 8.2 检测培养基的优化 54 8.3 吸附垫型培养基的研制 54 8.4 对白僵菌分生孢子检测力的试验 54-55 8.5 养蚕场所及周边环境的检测试验 55-56 参考文献 56-64 致谢 64-66 发表论文、专利及参研课题 66