温度及碳源对纤维素分解菌群分解活性与稳定性的影响

为探索秸秆纤维素分解菌群筛选过程中,温度及碱处理小麦秸秆对菌群纤维素分解活性及菌群结构的影响,利用高温秸秆堆肥为筛选菌源,以碱处理小麦秸秆和未经碱处理小麦秸秆为碳源,分别在50和60℃条件下进行限制性筛选,最终获得18组具有纤维素分解活性的菌群。选择其中4组代表性菌群进行连续继代培养,监测相关性质,并利用PCR-DGGE技术结合主成分分析(PCA)方法对菌群结构进行分析。结果表明,从高温堆肥环境筛选秸秆纤维素分解菌群,培养温度及秸秆碳源均影响菌群的筛选效果。以碱处理小麦秸秆为碳源的菌群在分解秸秆过程中能够保持较好的菌群结构稳定性;60℃的温度条件和碱处理小麦秸秆的碳源条件更有利于获得高活性的纤维素分解菌群,并在此条件下成功筛选到菌群WDC2。该菌群分解碱处理小麦秸秆的纤维素内切酶活性(CMCase)达到1.01 U/mL,分解率最高为60.8%。     

一组纤维素-林丹分解菌复合系的稳定性研究

本试验通过对一组高效降解纤维素林丹的复合微生物菌系的性质、功能和菌种组成进行研究,以确定该菌系的稳定性.结果表明,该复合系能在较大的pH范围内保持高的纤维素林丹分解活性,不同培养代在培养过程中的pH均呈现先降后升的趋势;不同代次的复合系在培养第3 d时对滤纸的分解率均达到了90%以上,在第7 d时对林丹的降解率均达到50%以上,且差异很小,说明该复合菌系产酶及分解纤维素和林丹的能力也已经非常稳定;平板培养基培养证明该复合菌系全部由细菌组成,用变性梯度凝胶电泳(DGGE)法比较来源于不同培养代的16S rDNA条带,分析菌种组成的变化,结果显示,不同培养代的菌种组成的主要条带没有变化,在DGGE图上出现的菌种能够稳定存在.     

玉米秸秆低温高效降解复合菌系GF-20的菌种组成及降解稳定性

【目的】探索不同温度及pH条件对玉米秸秆低温高效降解复合菌系的玉米秸秆分解稳定性及菌群结构稳定性的影响,完善菌系培养方法,促进其应用开发利用。【方法】以低温高效降解复合菌系GF-20为研究对象,在10℃条件下连续继代培养45代、不同温度和pH条件下连续继代培养15代,分别获得多组不同代数（F）、不同温度（T）和pH（P）条件下的菌系。测定各复合菌系发酵液pH、玉米秸秆降解率及纤维素酶活,评价复合菌系的玉米秸秆分解稳定性;利用PCR-DGGE技术结合主成分分析法对菌群组成结构的稳定性进行研究。【结果】在10℃条件下经连续继代培养40代和在温度4-30℃、pH 6.0-9.0条件下继代培养获得的不同复合菌系发酵液pH均随发酵时间的延长趋近中性;玉米秸秆降解率在27.59%-32.53%,除F40显著高于F5外,其余无显著差异;纤维素酶活性呈高代菌系大于低代菌系,温度4-10℃和pH 6.0-9.0条件下,对复合菌系产酶有促进作用,纤维素酶活为1.34-1.84 IU·mL^-1;复合菌系的纤维素酶在较低的温度和较宽的pH范围内具有良好的温度和pH稳定性,酶促反应温度15-30℃和pH 4.0-9.0内仍保持80%以上的纤维素酶活力;复合菌系F5-F45、T4-T30和P6.0-P9.0的DGGE条带差异不显著,表明菌系的菌种组成稳定;而在偏酸（pH=4、5）和偏碱性（pH=10）条件下继代培养,复合菌系秸秆降解率和纤维素酶活均显著降低,菌种组成发生变化,进而影响性质与功能稳定性。PCR-DGGE共检测到18个条带,其中关键菌株分别为Bacillus licheniformis、Azonexus hydrophilusd、Azospira oryzae、Arobacter cloacae、Cellvibrio mixtus、Bacillus tequilensis、Clostridium populeti subsp. Mixtus和Clostridium xylanolyticum。【结论】复合菌系GF-20在温度4-30℃、pH 6.0-9.0条件下经过多代继代培养,仍然保持了较高的玉米秸秆分解活性和菌种组成稳定性,具有良好的应用前景。     

玉米秸秆低温高效降解复合菌系GF-20的菌种组成及降解稳定性研究

【目的】探索不同温度及p H条件对玉米秸秆低温高效降解复合菌系的玉米秸秆分解稳定性及菌群结构稳定性的影响,完善菌系培养方法,促进其应用开发利用。【方法】以低温高效降解复合菌系GF-20为研究对象,在10℃条件下连续继代培养45代、不同温度和p H条件下连续继代培养15代,分别获得多组不同代数(F)、不同温度(T)和p H(P)条件下的菌系。测定各复合菌系发酵液p H、玉米秸秆降解率及纤维素酶活,评价复合菌系的玉米秸秆分解稳定性;利用PCR-DGGE技术结合主成分分析法对菌群组成结构的稳定性进行研究。【结果】在10℃条件下经连续继代培养40代和在温度4—30℃、p H 6.0—9.0条件下继代培养获得的不同复合菌系发酵液p H均随发酵时间的延长趋近中性;玉米秸秆降解率在27.59%—32.53%,除F40显著高于F5外,其余无显著差异;纤维素酶活性呈高代菌系大于低代菌系,温度4—10℃和p H 6.0—9.0条件下,对复合菌系产酶有促进作用,纤维素酶活为1.34—1.84 IU·m L~(-1);复合菌系的纤维素酶在较低的温度和较宽的p H范围内具有良好的温度和p H稳定性,酶促反应温度15—30℃和p H 4.0—9.0内仍保持80%以上的纤维素酶活力;复合菌系F5—F45、T4—T30和P6.0—P9.0的DGGE条带差异不显著,表明菌系的菌种组成稳定;而在偏酸(p H=4、5)和偏碱性(p H=10)条件下继代培养,复合菌系秸秆降解率和纤维素酶活均显著降低,菌种组成发生变化,进而影响性质与功能稳定性。PCR-DGGE共检测到18个条带,其中关键菌株分别为Bacillus licheniformis、Azonexus hydrophilusd、Azospira oryzae、Arobacter cloacae、Cellvibrio mixtus subsp.Mixtus、Bacillus tequilensis、Clostridium populeti和Clostridium xylanolyticum。【结论】复合菌系GF-20在温度4—30℃、p H 6.0—9.0条件下经过多代继代培养,仍然保持了较高的玉米秸秆分解活性和菌种组成稳定性,具有良好的应用前景。     

纤维素降解菌的筛选与高效混合菌群的构建

【目的】从土壤中筛选纤维素降解菌,对其进行组合培养获得可高效降解纤维素的混合菌群,为微生物混合培养降解纤维素提供理论基础。【方法】采用刚果红纤维素琼脂平板培养基从土样中初步筛选纤维素降解菌,再以内切酶（CMC）、纤维素全酶（FPA）、外切酶（C₁）和β-葡萄糖苷酶（β-Gase）4种酶活性为指标进行复筛,对复筛获得的高效菌株进行组合培养,筛选高效组合菌群。对复筛后的菌株通过菌落和菌体形态进行初步鉴定。【结果】筛选获得了y3、yi-71、ye-9、er-72和se-93等5株活性较高的纤维素降解菌,对其进行组合培养,得到1个较好组合ye-9/er-72/se-93,其CMC、FPA、C-1和β-Gase 4种酶活性分别为3.18,1.67,1.08和1.12 U/mL,均比单菌株有一定程度提高。初步鉴定ye-9、er-72、se-93均为放线菌。【结论】组合菌群对纤维素的降解效果优于单一菌株。     

一组秸秆分解菌群的稳定性及对还田秸秆的促腐效果

探讨一组秸秆还田菌群ADS-3的功能稳定性及初步观察其接种于土壤后对还田秸秆的促腐效果。结果表明：在培养瓶中菌种ADS-3对小麦、水稻和玉米3大农作物秸秆都有稳定高效的分解能力,35℃条件下培养11d,能分解稻秆51．6％、小麦秸秆44．7％、玉米秸秆40．8％;在4、15和35℃条件下,尽管ADS3对小麦秸秆的分解率差异很大,依次为6．8％、21.6％和44．7％;但是用变性梯度凝胶电泳（DGGE）检测其培养液,发现不同温度条件下ADS3的微生物组成稳定。将菌种接种到模拟秸秆还田土壤中,初步试验结果表明ADS-3能显著提高秸秆的腐解速度（P〈0．05）。试验结果为该菌群进一步开发秸秆还田促腐菌剂提供试验参数和依据。     

木屑及农业秸秆对大球盖菇菌丝生长的影响

【目的】通过分析木屑与5种不同农业秸秆的营养成分及其对大球盖菇菌丝生长的影响,为木屑秸秆在大球盖菇上的应用提供依据。【方法】对以木屑和5种秸秆的单一培养基以及复合培养基培养的大球盖菇菌丝的生长情况以及酶活进行了观察记录与测定。【结果】木屑、稻壳、稻草、玉米芯、玉米秸秆、棉籽壳的含氮量范围在0.48%～1.21%;含碳量范围在47.22%～54.35%,玉米芯碳氮比最大,木屑的碳氮比最小;玉米秸秆的纤维素含量最高,玉米芯半纤维素含量最高,木屑木质素含量最高。在木屑与5种农业秸秆单一基质培养基的基础上添加氮源和微量元素可以显著增加大球盖菇菌丝干质量及菌丝密度。木屑、玉米芯、稻草为底物时大球盖菇具有较高的木质纤维素酶活性。在木屑与5种农业秸秆配比的复合培养基上,木屑比例越高,大球盖菇菌丝干质量越大,复合培养基添加氮源和微量元素后则提高菌丝干质量3倍以上。【结论】大球盖菇在复合基质培养基中比在单一基质培养基中生长更好,在氮源和微量元素满足时菌丝生长更好。     

1株高产纤维素酶菌株的筛选鉴定及对稻秆降解的研究

【目的】筛选高效纤维素降解菌,用于稻秆原位还田菌剂的开发。【方法】通过稻秆粉培养基和刚果红-纤维素选择性培养基,从芜湖地区腐殖土中筛选分离出高效纤维素降解菌CX1,测定不同底物条件和不同反应温度下的纤维素酶活性。通过滤纸条崩解试验、土培降解试验、菌株与化学物质协同降解稻秆试验、发酵液对小麦幼苗生长的影响试验,探究菌株CX1腐解稻秆的特性。【结果】通过菌株形态学特征及16S rDNA序列相似性(99%)比对,确定CX1为高温嗜热芽孢杆菌Thermophilic Bacillus sp.。以稻秆粉为底物,50℃条件下菌株CX1纤维素酶活力达13.87 U·mL~(–1),65℃时酶活力仍能达到9.73 U·mL~(–1)。添加菌株CX1培养4 d后滤纸条完全崩解,到15 d时,稻秆纤维素降解率达到52.55%,土培40 d时稻秆相对降解率达到25.38%。预先用质量浓度为0.05 g·mL~(–1)的NaOH溶液浸泡处理的稻秆更利于菌株CX1对稻秆的降解,腐解14 d时稻秆失重率比对照组增加了6.69%。添加经菌株CX1降解稻秆后的发酵液可使小麦的各项生长指标有明显提高,小麦的出苗率、苗高、根鲜质量和苗鲜质量分别提高了9.66%、55.55%、59.71%和118.84%。【结论】菌株CX1对高温具有耐受性,能高效降解纤维素,可在农业生产中促进稻秆原位还田方面发挥积极的作用。     

玉米秸秆高效腐解复合菌系CSS-1的选育及其组成分析

【目的】选育在常温(22℃)下对玉米秸秆具有高效稳定腐解功能的微生物复合菌系,明确其菌群组成,以促进选育的复合菌系在中国北方原位还田玉米秸秆中的应用。【方法】采用限制性培养与温度梯度诱导相结合的方法,以山西、山东、北京等省(市)玉米秸秆连续多年还田的土壤样品为菌源,以秸秆失重率、C/N及CMC酶活为筛选指标,获得高效稳定的玉米秸秆腐解复合菌系CSS-1;比较复合菌系不同继代腐解秸秆的失重率、CMC酶活、木聚糖酶活和PCR-DGGE的菌群监测动态变化,评价CSS-1的功能及其组成稳定性;采用克隆文库法,解析复合菌系CSS-1的主要菌群组成。【结果】CSS-1处理玉米秸秆的CMC平均酶活和最高酶活,比目前广泛应用的秸秆腐解菌剂A分别提高了82%和79%;CSS-1处理的秸秆失重率比菌剂A和空白对照分别提高了62.62%和173.65%;CSS-1不同继代腐解秸秆功能及其菌系组成监测表明该复合菌系的稳定性;克隆文库方法显示CSS-1优势菌群由多种具有纤维素与半纤维素降解功能的细菌和真菌构成,其细菌菌群分别归属Enterobacter、Cellulomonas、Streptomyces、Bacillaceae、Pantoe、Cellvibrio6个属,真菌菌群归属Trichoderma、Gibberella2个属。【结论】获得了在中国北方地区常温条件下高效稳定腐解玉米秸秆的复合菌系CSS-1,可研发利用该菌系促进田间原位还田玉米秸秆的快速腐解。     

纤维素分解菌复合系WDC2分解小麦秸秆的特性及菌群多样性

为了加快小麦秸秆资源的资源转化,提高小麦秸秆的生物分解率,从小麦秸秆堆肥中筛选到一组高效的小麦秸秆分解菌复合系WDC2。研究表明,WDC2在PCS培养基中以小麦秸秆作为唯一碳源,60℃培养15 d,对小麦秸秆的分解率达到64.47%,其中纤维素分解44.7%,半纤维素分解13.61%,木质素减少3.85%,pH由初始的8.3迅速下降,60 h下降到6.6,而后逐渐回升接近初始值。羧甲基纤维素酶(CMCase)随小麦秸秆的分解而升高,在15 d达到0.372 U/mL。GC-MS分析WDC2分解小麦秸秆不同阶段的培养液,检测到乙醇、乙酸、乙二醇、丙酸、2-甲基丙酸、丁酸、3-甲基丁酸、2-甲基丁酸、丙三醇共9种挥发性有机酸。其中,除乙醇在分解初期出现后消失外,其他产物在培养的第7～15天的含量都较高。菌群结构分析显示,WDC2菌群中主要含有11种细菌,大部分为不可培养菌株,其中Clostridium thermocellumCTL-6为厌氧培养细菌,是WDC2中具有高效的纤维素分解能力的菌株。     

The Character of Normal Temperature Straw-Rotting Microbial Community

In order to study degradation capability and optimal condition of produced endoglucanase （CMCcase）, the microbial community with efficient cellulose degrading ability in 28℃ was studied. Microbial community came from rotted rice straw which was enriched and domesticated by improved Mandels medium. The standard cellulase activity assays were used to determine cellulase activity, degradation products were analyzed by gas chromatography mass spectrometry （GC/MS）, and denaturing gradient gel electrophoresis （DGGE） was used to identify the composition dynamic of the community. The results showed that the microbial community could degrade 39.6% of rice straw gross weight within 6 d. When culture medium volume was 1/5 and pH = 6 on day 5, the CMCcase activity reached the highest of 14 IU mL-L During the fermentation, more than ten products were detected using GCMS. The microbial changed lots in different fermentation periods which detected by DGGE. Phylogenetic tree derived from 16S rDNA sequence found that the community composed of bacteria, and the closest relative were belong to Clostridium sp., Brevibacillus sp. and Bacterium sp. This microbial community could accelerate rice straw rotting and decomposed products could promote organic matter increase of soil     

油菜秸秆降解菌群的筛选及降解特性研究

[目的]筛选降解油菜秸秆的最优菌群,并研究其降解特性。[方法]通过刚果红平板法、苯胺兰平板法及纤维素酶、木质素酶产酶试验考察了6个菌群的油菜秸秆降解能力。[结果]腐烂秸秆土菌群生长最快,具有最大的生长量,木质素酶活性最高,纤维素酶活性达到了924 U/ml。腐烂秸秆土菌群固态发酵降解油菜秸秆14 d,对油菜秸秆、纤维素、木质素降解率为71%、82%、53%。前12 d对油菜秸秆、纤维素、木质素的降解量分别占各自总降解量的86.05%、93.35%、93.15%。[结论]腐烂秸秆土菌群为降解油菜秸秆最优菌群,并且在降解的前12 d为高效降解阶段。     

Screening of a Composite Microbial System and Its Characteristics of Wheal Straw Deqradation

To accelerate the decomposition of wheat straw directly returned to soil,we constructed a microbial system (ADS-3) from agricultural soil containing rotting straw residues using a 40-wk limited cultivation.To assess its potential use for accelerating straw decomposing,the decomposing characteristics and the microbial composition of ADS-3 were analyzed.The results indicated that it could degrade wheat straw and filter paper by 63.8 and 80％,respectively,during 15 d of incubation.Straw hemicellulose degraded dramatically 51.2％ during the first 3 d,decreasing up to 73.7％ by the end of incubation.Cellulose showed sustained degradation reaching 53.3％ in 15 d.Peak values of xylanase and cellulase activities appeared at 3 and 11 d,with 1.32 and 0.15 U mL-1,respectively.Estimated pH averaged 6.4-7.6 during the degradation process,which approximated acidity and alkalinity of normal soils.The microbial composition of ADS-3 was stable based on denaturing gradient gel electrophoresis (DGGE) analysis.By using bacterial 16S rRNA and fungal 26S rRNA gene clone library analysis,20 bacterial clones and 7 fungal clones were detected.Closest identified relatives of bacteria represented by Bacillus fusiformis,Cytophaga sp.,uncultured Clostridiales bacterium,Ruminobacillus xylanolyticum,Clostridium hydroxybenzoicum,and uncultured proteobacterium and the fungi were mainly identified as related to Pichia sp.and uncultured fungus.     

Microbial Transformation Process of Straw-Derived C in Two Typical Paddy Soils

【Objective】 The straw degradation rate, microbial community structure changes and functional microbial community composition involved in straw decomposition in soils were researched, and the research results could provide the theoretical foundation for revealing microbial mechanism of the soil organic matter transformation and accumulation. 【Method】 Two typical subtropical paddy soil in China, including Changshu Wushan soil and Yingtan Red paddy soil, were collected as the research materials. We anaerobically incubated the soils with/without ¹³C-enriched rice straw for 38 days. Gaseous samples were regularly collected to investigate mineralization rate of straw in dynamic changes. The soil samples were collected to analyze the dynamic changes of the microbial community composition related to straw decomposition by using ¹³C-PLFA-SIP technology. 【Result】 At the early stage before day 12 of the anaerobic culture, straw degraded slowly, and straw had positive priming effect on soil organic matter (SOM). At the stage of day 12-18, straw degraded rapidly and then the rate tended to be slow after day 18. At the end of incubation, straw mineralization rate was 24% and 33% in Red paddy soil and Wushan soil, respectively. The contribution of straw C to C efflux increased with incubation time, which was 53%-60% and 54%-57% to CO₂ and CH₄ efflux, respectively. The microbial biomass and activity were improved in the soil with straw, and the microbial activity in Wushan soil was higher than that in Red paddy soil. During straw degradation, 16:0 (general bacteria) was the main groups. i16:0, i15:0 (G ⁺ bacteria) and 18:1ω9c (fungi) were also important microbial groups involved in straw degradation. The relative abundance of straw-derived gram-positive (G ⁺) bacteria and actinomycetes increased and gram-positive (G ^-) bacteria decreased with incubation time. The proportions of straw-derived PLFAs were 27%-32% and 18%-24% in Red paddy soil and Wushan soil PLFAs, respectively. The straw utilization efficiency was higher in fungi and general bacteria, while G ^- bacterial and actinomycetes PLFAs were preferentially linked to extant soil organic matter (SOM) mineralization. The microbial community composition was different between Wushan soil and Red paddy soil with rice straw. The straw-derived microbial community composition was similar in two soils, but the SOM-derived microorganisms were differences. 【Conclusion】 The mineralization of straw C lagged behind extant SOM during anaerobic straw degradation. The microbial activity and diversity in soil were important factors influencing the efficiency of straw mineralization. After adding straw in soil, it’s showed differences from the microbial community composition, which were mainly involved in the differences between SOM-derived microorganisms, and SOM was an important factor leading to these differences.     

菲降解体系中细菌多样性的研究

[目的]探索生物多样性在污染物降解中的作用。[方法]以菲为唯一碳源,通过选择培养的方式,从采油废水处理的好氧污泥中富集了菲降解功能微生物类群,并利用PCR结合变性梯度凝胶电泳(DGGE)的方法对它的微生物组成特征进行分析。[结果]混合菌群在菲生物代谢过程中表现出较丰富的细菌多样性,混合菌群的菲降解能力显著高于已报道的纯培养体系。随着培养时间的延长,群落的丰度逐渐增加,群落相似性越来越高。DGGE对于表征一定系统中微生物群落的动态变化有较高的分辨率、方便、快捷,具有良好的应用前景。[结论]群落中多种不同微生物的协同作用可能是目标物高效降解的原因。     

两种典型水稻土中秸秆碳转化的微生物过程

【目的】研究土壤中秸秆腐解速率、腐解过程中微生物群落结构变化和参与秸秆腐解的功能微生物群落组成,为揭示土壤有机质转化和积累的微生物学机制提供理论依据。【方法】以我国亚热带两种典型水稻土——常熟乌栅土和鹰潭红壤性水稻土为研究对象,设置不添加秸秆（CK）和添加 ¹³C标记的水稻秸秆（RS）处理,厌氧恒温培养38 d,在培养过程中定期测定气体释放量,研究秸秆矿化速率的动态变化;采集土壤样品,利用 ¹³C-PLFA-SIP技术分析参与秸秆降解的微生物群落的动态变化。【结果】培养前12 d,秸秆降解缓慢,此时秸秆对土壤有机质（SOM）产生正激发效应;培养12-18 d秸秆快速降解,18 d后趋缓。培养结束时,秸秆碳在红壤性水稻土和乌栅土中的矿化率分别为24%和33%。秸秆碳对CO₂和CH₄贡献率随培养时间的延长而增加,在培养末期分别为53%-60%和54%-57%。添加秸秆可以提高土壤微生物生物量及微生物活性,乌栅土微生物活性高于红壤性水稻土。16:0（一般细菌）是参与秸秆分解主要类群,i16:0和i15:0（G ⁺细菌）和18:1ω9c（真菌）也是参与秸秆分解的重要微生物类群。随培养时间增加,G ⁺细菌和放线菌的相对丰度增加,G ^-细菌呈降低趋势。红壤性水稻土和乌栅土PLFAs中标记利用秸秆碳的PLFAs的比例分别为27%-32%和18%-24%。真菌和一般细菌对秸秆碳的利用效率较高,而土壤原有有机质（SOM）矿化主要与G ^-和放线菌相关联。添加秸秆造成乌栅土和红壤性水稻土两种水稻土微生物群落结构呈现明显差异,但分解利用外源秸秆碳的微生物群落结构相似,而分解利用SOM微生物群落结构有差异。【结论】秸秆厌氧降解过程中秸秆碳的矿化滞后于土壤自身SOM;不同本底微生物活性和多样性是影响秸秆碳矿化速率的重要因素;添加秸秆后不同土壤微生物群落结构的差异主要是参与SOM降解的微生物差异,土壤原SOM是导致这种差异的重要因素。     

里氏木霉纤维素酶水解稻草的研究

[目的]探讨碳氮源对里氏木霉发酵产纤维素酶的影响,以及纤维素酶水解稻草的条件。[方法]通过添加不同的碳源和不同浓度的酵母粉,探讨里氏木霉合适的发酵条件;使用不同添加量的纤维素酶对稻草进行酶解;分别利用纤维素酶、纤维素酶和木聚糖酶混合酶对稻草进行酶解反应。[结果]利用乳糖和稻草的复合碳源和12 g/L的酵母粉进行水解时,纤维素酶活性较高。酶解适宜的酶用量为每克稻草底物200 U的滤纸酶。用纤维素酶及木聚糖酶混合酶酶解稻草96 h的酶解得率为65.4%。[结论]该研究可为里氏木霉纤维素酶生产和酶解稻草的应用提供一定的依据。     

降解水稻秸秆细菌-真菌复合菌系的构建与评价

为筛选构建高效降解水稻秸秆的细菌-真菌复合菌系,通过分离筛选、纯化鉴定具备降解水稻秸秆能力的细菌和真菌,选择出降解效率较高的细菌和真菌各两株用于复配。通过全组合将4株菌复配成多样性为1～4的15种复合菌系,采用水稻秸秆降解试验评估复合菌系的降解效果和3种纤维素降解酶的活性,利用线性拟合和多元回归解析复合菌系产酶活力与其降解秸秆能力之间的关系。从腐解的水稻秸秆中筛选获得降解细菌和真菌各4株,其中以细菌分离株雷氏普罗威登斯菌BB18、球形赖氨酸芽孢杆菌JB7和真菌分离株草酸青霉ZA、烟曲霉ZL的秸秆降解能力和产滤纸酶活力较高。将该4株降解效果较高的菌株进行全组合复配,发现复合菌系对水稻秸秆的降解率以及产滤纸酶、纤维素内切酶和木聚糖酶的活性均随菌株多样性的增加而显著提高,且以4株菌株组合的复合菌系BF1对水稻秸秆的降解效果最好,较单菌平均提高了1.6倍,酶活力增加了1～3倍。相关分析结果表明,复合菌系降解水稻秸秆的能力与3种酶的活力呈显著正相关,其中纤维素内切酶和木聚糖酶对复合菌系降解能力的贡献最大。细菌-真菌复合菌系的降解效果与产酶能力具有明显的多样性效应,维生素内切酶和木聚糖酶是驱动细菌-真菌复合菌系高效降解秸秆的关键因子。     

Stability Analysis of a Lignocellulose Degrading Microbial Consortium

An active mesophilic lignocellulose degrading microbial consortium, designated LZF-12, was bred from humus-rich soil by successive subcultivation under facultative aerobic static condition. Batch experiments were performed to investigate the structural and functional stability of lignocellulose degradation of rice straw of 10 g · L-1. The results showed that efficient degradation of rice straw(70%) could be achieved and acetic acid concentration accounted for over 70% of total aqueous products from different generations by microbial consortium LZF-12 within 7 days. Denaturing gradient gel electrophoresis(DGGE) and sequencing of 16 S r DNA sequences amplified from the total consortium DNA representing the presence of sequences were related to those of Clostridium, Clostridium cellulolyticum, Pseudomonas, Acetivibrio and some uncultured bacteria in LZF-12. DGGE pattern profiles from different LZF-12 generations were reproducible, suggesting the relative stabilities of the microbial community structure and succession mechanism in the established consortium.     

一组高温高效纤维素分解菌群的筛选及功能特性研究

从木质纤维材料堆肥样品中驯化筛选出一组高温型纤维素分解菌群N6,对其纤维素分解功能特性进行了研究.结果表明,N6培养体系的pH稳定在6.4～8.0之间,降解纤维的适宜温度为50～ 60℃,最高纤维素酶活(CMCase)为0.83 IU/mL;在50℃下,N6在4d内可使滤纸降解90.0％以上,6d内可使玉米秆粉或稻草粉分别降解75.0％和86.4％,9d内可使玉米秆粉或稻草粉分别降解失重79.0％和90.4％,对木薯渣的降解率也在30.0％以上.可见,N6具有降解天然木质纤维的高活性,是可应用于木质纤维材料生物转化的优良菌群.