摘要乙基多杀菌素是由刺糖多孢菌生产发酵产生的生物杀虫剂多杀菌素的K和J组分,再经过一步化学合成得到的。由于兼具生物农药的安全性和化学合成农药的速效性特点,现已广泛应用于农业生产中,是多杀菌素的换代产品。【目的】通过利用UV诱变微生物的基因发生突变并进行突变菌株的筛选。得到提高多杀菌素组分K和组分J的产量的突变株【方法】根据对乙基多杀菌素生产原始菌株的传代培养而得到单孢子悬浮液,并在具有甘氨酸的抗性平板上筛选,待菌种长出后,再进一步的传代培养,然后便用实验室30W的紫外灯管进行照射而发生诱变,再把诱变过的菌种进一步的传代培养,当菌种长得饱满之后,再进行4天的种瓶培养和7天的发酵摇瓶培养,之后便将其离心,再用液相色谱进一步检测其诱变后产生乙基多杀菌素的含量。【结果】由本实验筛选得到甘氨酸最小抑制浓度为20g/l;在含有甘氨酸最小抑制浓度的菌株中进行UV诱变后便得到UV诱变的最佳时间段为60s;UV诱变后得到菌落的形态有草帽型、褶皱型、光秃型。经液相检测得到诱变前组分总的峰面积为129439;诱变后总的峰面积248401。【结论】在甘氨酸的浓度为20g/l、UV照射时间段为60s下,经过UV诱变后菌落形态为草帽型的菌株的诱变效果较好,这为以后乙基多杀菌素生产菌的诱变研究方向提供一些基础。90056
Abstract Spinetoram is biologically produced by Saccharopolyspora spinosa strain producing a fermentation insecticide spinosyn J and K components, and then through the step of chemical synthesis。 Due to have both the safety characteristics of biological pesticides and the quick-acting characteristics of chemical synthesis of pesticides, it has been widely used in agricultural production and is a substitute product of spinosad。 [Objective]Mutations were made by mutagenesis of mutant strains by using UV mutagenesis mutations in microorganisms' genes and dressing by screening the mutant。 Thereby obtaining a mutant strain for increasing the yield of the spinosyn component K and the component J。 [Method]A monospore suspension was obtained according to subculturing of the original strain for the production of the original strain of spinetoram and screened on a resistant plate with glycine, Until the bacteria grow, and then further subculture, the culture was further subcultured and then use the laboratory 30W UV lamp irradiation and mutagenesis, and then mutated bacteria to further passaged culture。 When the bacteria grows full, then another 4 days of the bottle culture and 7 days of fermentation shake flask culture。 After the bacterial fluid were centrifuged。 and then liquid chromatography to further detect the mutagenesis of spinetoram production。 [Results]The minimum inhibitory concentration of glycine was 20g/l by this experiment。 The optimum times for UV mutagenesis after UV mutagenesis in a strain containing minimal inhibitory concentration of glycine was 60s。 After the UV mutagenesis。 the form of colony straw have hate type, fold type, bare type。 [Conclusion] In the case of glycine concentration of 20g/l。 UV irradiation time period of 60s。 After mutagenesis, the mutagenic effect of the colonies was better。 This provides some basis for the future research of mutagenesis of spinetoram-producing bacteria。源Q于W优E尔A论S文R网wwW.yOueRw.com 原文+QQ75201,8766
毕业论文关键词:乙基多杀菌素;多杀菌素; UV诱变;甘氨酸;
Key words:spinetoram;spinoad;UV mutagenesis;glycine;
目录
1引言 5
1。1 多杀菌素概述 5
1。2 乙基多杀菌素的概述 5
1。2。1乙基多杀菌素的来源