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生防放线菌153固态发酵条件的优化及其耐热力检测
田晓丽1, 赵红杰1, 唐彩乐1
西北农林科技大学 植物保护学院,陕西省农业分子生物学重点实验室
摘要:
【目的】探讨生防放线菌153的固态发酵条件,并考察其耐热力,为生防放线菌的大规模生产及活体菌剂的研制提供技术支持。【方法】将发酵物的含菌量作为筛选指标,以玉米粉、甘油、蔗糖、葡糖糖和可溶性淀粉为碳源,以小米粉、蛋白胨、酵母浸粉、黄豆粉和尿素为氮源,通过单因素试验获得适宜的碳、氮源,再采用L25(56)正交设计优化固体培养基配方,在此基础上确定固态发酵终点以及所需的接种量和种子液种龄。最后将壳聚糖、非耕作层黄土、高岭土、炉渣、碳酸钙、面粉和几丁质分别与生防放线菌153 菌粉混合,55 ℃水浴至致死中时,测定生防放线菌153的耐热力,筛选适宜的载体。【结果】生防放线菌153固体培养基配方为:80 mg/g麦麸、8 mg/g玉米粉、10 mg/g小米粉和2 mg/g蛋白胨,初始含水量(含0.5 g/L石灰的自来水)为60%,在此条件下培养,培养物的含菌量较对照增加了94.1%。生防放线菌153固态发酵培养终点为216 h,接种量为20%(每100 g固态培养基接种20 mL种子液),种子液种龄为84 h。生防菌153的适宜载体为壳聚糖和非耕作层黄土。【结论】确定了生防放线菌153的固态发酵条件,该条件可以促进生防菌153的生长产孢,并提高菌株的保存稳定性;确定了筛选生放菌153载体的方法,即55 ℃条件下水浴处理136 min。
关键词:  生防放线菌  正交设计  固体发酵培养基  耐热力  活体菌剂
DOI:
分类号:
基金项目:教育部创新团队发展计划项目(200658);国家高等学校学科创新引智计划项目(B07049);西北农林科技大学创新团队项目
Optimization of the fermentation of solid state medium for biocontrol actinomycetes 153 and its heat tolerance ability
Abstract:
【Objective】The research was to investigate the best condition for solid-state fermentations of bioncontrol actinomycetes 153 and its heat tolerance ability.【Method】With corn meal,glycerol,sucrose,D-glucose and starch soluble as the carbon sources,with peptone,mille,yeast extract power,soybean flour and urea as nitrogen sources,appropriate carbon and nitrogen sources were obtained in the single factor experiment.Based on the sporulation quantity of SSF product and the single factor experiment,the medium was optimized by L25(56)orthogonal design.On this basis,the solid-state fermentation was optimized.Chitosan,non-cultivated loess,kaolin,furnace slag,calcium carbonate,flour and chitin were mixed with bio-control actinomycetes 153 bacteria powder respectively.The heat tolerantce ability and carriers of 153 strain were determined at 55 ℃.【Result】The optimum solid state medium of biocontrol actinomycetes 153 was determined as follows:wheat bran 80 mg/g.mille 8 mg/g,corn meal 10 mg/g,peptone 2 mg/g and initial moisture content 60% adjusted by water with 0.5 g/L lime.The spore content was 94.1% higher than that of the control under these conditions.The optimal fermentation effect was inoculating seed culture fluid of 20% seed volume and 84 h seed age duration,and cultivating it for 216 h.At 55 ℃,136 min heat treatment was the standard of 153 thermal resistance;chitosan and Non-cultivated loess were the carriers of 153 strain.【Conclusion】The fermentation conditions for bio-control actinomycetes 153 solid-state were determined,which can not only greatly promote the 153 growths and reproduction,but also increase its storage stability.The conditions were good to its further development and application;At 55 ℃,136 min heat tolerance ability can embody carriers’ protection of 153 long term storage,which was useful in screening large amount of carriers.
Key words:  Biocontrol actinomycete  orthogonal design  solid state fermentation medium  heat tolerance ability  BCA