菌糠高效降解菌剂的研发及其在堆肥中的应用

发布时间：2018-02-04 21:34

本文关键词： 菌糠堆肥木质纤维素纤维素降解菌菌剂腐熟度微生物群落　出处：《浙江大学》2017年硕士论文　论文类型：学位论文

【摘要】：随着食用菌产量的日趋增加,菌糠产生量不断增加,随之也带来了因菌糠不合理利用造成的环境污染。菌糠堆肥化处理转化为有机肥,是使其资源化利用的有效途径之一,也是符合我国国情的最佳处理方法。目前,堆肥化处理存在堆肥发酵慢、腐熟时间长、堆肥质量低等问题。基于此,本文系统研究了菌糠高效降解菌剂的研发及其在堆肥中的应用,以期促进堆肥发酵、缩短腐熟时间、提高堆肥质量。主要研究结论如下:1)筛选鉴定了 4株高效纤维素降解细菌、2株商效纤维素降解放线菌以及2株高效纤维素降解真菌。通过富集培养、划线分离,经滤纸崩解实验和菌株CMCase、FPase酶活测定实验,共筛选出4株高效纤维素降解细菌(GX-5、GX-7、GX-13和GX-16)、2株高效纤维素降解放线菌(GF-1和GF-2)以及2株高效纤维素降解真菌(GZ-1和ZZ-2)。对筛选出的菌株进行了形态学观察、生理生化特性和16SrRNA/ITS序列鉴定,结果为:GX-5为芽孢杆菌属(Paenibacillus),GX-7、GX-13和GX-16均为短芽孢杆菌属(Brevibacillus);GF-1为链霉菌属(Streptomyces),GF-2为小双孢菌属(Microbispora);GZ-1 为樟绒枝霉(Malbranchea cinnamomea),ZZ-2 为小囊菌属(Micraascus)。2)优化了高效纤维素降解菌的产酶条件。优化了菌株产酶的培养条件,主要包括培养基初始pH、接种体积、转速和温度对菌株产酶的影响;也优化了菌株产酶的培养基质,主要包括碳源、氮源和碳氮比对菌株产酶的影响。经产酶条件优化,获得了菌株的最适产酶条件,菌株的CMCase和FPase较优化前都得到了很大提高。其中,菌株GX-5的CMCase和FPase较优化前分别提高了 37.2%、49.0%;菌株GX-7的CMCase和FPase较优化前分别提高了 36.2%、44.7%;菌株GX-13的CMCase和FPase较优化前分别提高了 34.2%、42.6%;菌株GX-16的CMCase和FPase较优化前分别提高了32.6%、44.5%;菌株GF-1的CMCase和FPase较优化前分别提高了 63.9%、96.4%;菌株GF-2的CMCase和FPase较优化前分别提高了 58.5%、82.1%;菌株GZ-1的CMCase和FPase较优化前分别提高了 35.9%、28.5%;菌株ZZ-2的CMCase和FPase较优化前分别提高了 24.9%、26.5%。3)研制了一种菌糠高效降解菌剂,并将其成功运用到菌糠实际堆肥中。选用本实验室自行筛选、驯化和构建的具有高效降解木质纤维素能力的微生物菌剂HJ、菌剂JK和纯菌GX-5,以及企业原有有机肥作为菌剂组的不同处理,以不接种菌剂的处理作为对照进行菌糠堆肥,研究不同菌剂对菌糠堆肥的促腐效果和菌糠高效降解菌剂的高效性。在整个堆肥过程中,重点检测了气味与色度、温度、含水率、酸碱度、电导率、木质纤维素、总氮和腐殖质的变化,并通过种子发芽试验对堆肥的毒性和腐熟度进行了评估,主要结果为:相较于CK,菌剂HJ可明显促进堆肥发酵,其纤维素、半纤维素和木质素的降解率分别提高了 5.1%、6.5%和4.6%,并可将腐熟时间缩短6天。综合各项堆肥腐熟度指标的检测结果,表明菌剂的添加可促进堆肥发酵,缩短腐熟时间并提高堆肥质量。其中,菌剂促腐效果:菌剂HJ菌剂JK纯菌GX-5=企业原有有机肥CK。4)探索了堆肥微生物群落演替规律。不同堆肥处理系统微生物群落存在一定的群落演替。随着堆肥进程的发展,细菌群落中的丛毛单胞菌属(Comamonas)等逐渐减少,而芽孢杆菌属(Bacillus)、类芽孢杆菌属(Paenibacillus)等逐渐成为优势菌群;真菌群落中的假丝酵母(Candida)等逐渐减少,而嗜热真菌属(Thermomyces)等逐渐成为优势菌群。随着堆肥进程的发展,堆肥微生物群落呈现出了明显的演替规律。
[Abstract]:With the increasing of the yield of edible fungus, mushroom bran production continues to increase, it also brings with WMLE unreasonable utilization caused by environmental pollution. WMLE composting into organic fertilizer, which is one of the effective ways of resource utilization, the best approach is consistent with China's national conditions. At present. Composting composting maturity there is slow, long time, the problem of compost low quality. Based on this, this paper studies research WMLE degrading bacteria and its application in composting, in order to promote composting, shorten the composting time, improve the quality of compost. The main conclusions are as follows: 1) screening and identification 4 cellulose degrading bacteria, 2 strains of highly effective cellulose degrading actinomycete and 2 cellulose degrading fungi. By enrichment culture, line separation, the disintegration of filter paper experiment and strain CMCase, FPase enzyme activity were measured Selected 4 cellulose degrading bacteria (GX-5, GX-7, GX-13 and GX-16), 2 strains of high efficient cellulose degrading actinomycetes (GF-1 and GF-2) and 2 strains of high efficient cellulose degrading fungi (GZ-1 and ZZ-2). The selected strains by morphological observation, physiological and biochemical characteristics and 16SrRNA /ITS sequence identification results GX-5: as Bacillus (Paenibacillus), GX-7, GX-13 and GX-16 are short bacillus (Brevibacillus); GF-1 (Streptomyces), Streptomyces GF-2 Microbispora (Microbispora); GZ-1 Zhang Thamnidium (Malbranchea cinnamomea) cashmere, ZZ-2 microascus (Micraascus).2) fermentation conditions of cellulose-degrading fungi were optimized. The optimal culture conditions of producing strains, including medium initial pH, inoculation volume, influence the speed and temperature of producing enzyme; optimize the culture medium producing strains, including carbon source, nitrogen source and carbon The effect of nitrogen on enzyme production strains. The optimization conditions of enzyme parous, obtained the optimum conditions for enzyme production strains, strains CMCase and FPase were optimized and improved. Among them, strain GX-5 CMCase and FPase than before optimization were increased by 37.2%, 49%; GX-7 CMCase and FPase strains than before optimization were increased by 36.2%, 44.7%; GX-13 CMCase and FPase strains than before optimization were increased by 34.2%, 42.6%; GX-16 CMCase and FPase strains than before optimization were increased by 32.6%, 44.5%; GF-1 CMCase and FPase strains than before optimization were increased by 63.9%, 96.4%; GF-2 CMCase and FPase strains than before optimization were increased by 58.5%, 82.1%; GZ-1 CMCase and FPase strains than before optimization were increased by 35.9%, 28.5%; CMCase and FPase strains of ZZ-2 were increased by 24.9% than that before optimization, 26.5%.3) developed a highly efficient degradation of WMLE agents, and put it into The power applied to the mushroom compost. The actual laboratory screening, domestication and construction of high efficient lignocellulose degradation ability of microbial agents HJ, JK and GX-5, the original pure bacteria, organic fertilizer as different treatment agents group, with no inoculated treatment as the control of mushroom compost, study different agents on composting of WMLE decay promoting effect and efficient degradation of WMLE agent efficiency. In the whole composting process, focus on the detection of the odor and color temperature, moisture content, pH, conductivity, lignocellulose, changes of total nitrogen and humus, and the seed germination test on compost maturity and toxicity the degree of evaluation, the main results are as follows: compared with CK, HJ agent can obviously promote the composting, the 5.1% increase in cellulose, hemicellulose and lignin degradation rate were 6.5% and 4.6%, and shorten the time of rotten Short 6 days. The detection results of the compost maturity index, showed that the agent added can promote fermentation, shorten the maturity time and improve the quality of compost. The microbial decay promoting effect: agent HJ agent JK pure GX-5= from original enterprise organic fertilizer CK.4) to explore the regularity in the succession of microbial compost. Different composting microbial community there is a certain system of community succession. With the development of the composting process, the bacterial community in Comamonas (Comamonas), and gradually reduced, Bacillus (Bacillus), paenibucillus (Paenibacillus) has become the dominant bacteria; Candida fungal communities in (Candida) gradually reduced, while thermophilic fungi (Thermomyces) has become the dominant flora. With the development of the process of compost, composting microbial community showed obvious succession.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X172;S141.4

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