Biodegradation of Gaseous Chlorobenzene by White-rot Fungus Phanerochaete chrysosporium

Objective To evaluate the effect of white rot fungus Phanerochaete chrysosporium on removal of gaseous chlorobenzene. Methods Fungal mycelium mixed with a liquid medium was placed into airtight bottles. A certain amount of chlorobenzene was injected into the headspace of the bottles under different conditions. At a certain interval, the concentrations in the headspace were analyzed to evaluate the degradation of chlorobenzene by P. chrysosporium. Results The degradation effects of P. chrysosporium on chlorobenzene under different conditions were investigated. The difference in the optimum temperature for the growth of the fungi and chlorobenzene degradation was observed. The data indicated that a lower temperature （28℃） would promote the degradation of chlorobenzene than the optimum temperature for the growth of the fungi （37℃）. A low nitrogen source concentration （30 mg N/L） had a better effect on degrading chlorobenzene than a high nitrogen source concentration （higher than 100 mg N/L）. A high initial concentration （over 1100 mg/m3） of chlorobenzene showed an inhibiting effect on degradation by P chrysosporium. A maximum removal efficiency of 95% was achieved at the initial concentration of 550 mg/m3. Conclusion P. chrysosporium has a rather good ability to remove gaseous chlorobeuzene. A low nitrogen source concentration and a low temperature promote the removal of chlorobenzene by P. chrysosporium. However, a high initial chlorobenzene concentration can inhibit chlorobenzene degradation.

Biotreatment of Wastewater from Soda-pretreatment Process of Corn Stover Using White-rot Fungus Z-6

Alkaline pretreatment of straw materials prior to enzyme hydrolysis is a key step for bioconversion of lignocellulose to bioethanol and chemicals.Wastewater from the alkaline pretreatment process must be treated before discharge to minimize its environmental impact.In this study,biotreatment of the wastewater from soda-pretreatment process of corn stover was investigated using fungus Z-6,and some indexes such as color,chemical oxygen demand(COD),and lignin content of wastewater before and after biotreatment were determined to assess the effect of the biotreatment.Results showed that fungus treatment could remove color up to approximately 72%after 2 d,and decrease COD and lignin content by about 63%and 60%,respectively after 3 d.The wastewater was fractionated using dynamic ultrafiltration method,and the changes in lignin contents of the effluent fractions with different molecular weights before and after biotreatment were analyzed.Some compounds produced by the fungus during treatment were identified using gas chromatography-mass(GC-MS)spectrometer,which revealed that depolymerization of lignin occurred during the biotreatment process.

Effects of White-Rot Fungi Composting on Wood Powder-Chicken Manure Mixture

[ Objective] To explore the possibility of treating livestock manure with white-rot fungi. [Method] The wood powder-chicken manure mixture was respectively composted by three kinds of white-rot fungi, the weight of lignin, cellulose, protein, fat, ash and crude polysaccharide as well as the mycelial growth was determined during this process. [ Result] The mixture was better composted with white-rot fungi than the control. The best effect was achieved in P. ostreatus composting group. In this group, the rate of weight reduction, lignin degradation and cellulose degradation were respectively 15.68% （6.79 times as great as that of the control group）, 39.92% （6.54 times as great as that of the control group） and 32.26% （2.77 times as great as that of the control group）. The weight of protein and fat were increased by 31.68% and 146.58%, respectively. The content of crude polysaccharide was 2.43%. No crude polysaccharide was detected, and the weight of protein and fat decreased by 21.96% and 70.99%, respectively. [ Conclusion] It is feasible to compost livestock and poultry manure with white-rot fungi.

Differentiation of Fungal Destructive Behaviour of Wood by the White-Rot Fungus Fomes fomentarius by MALDI-TOF Mass Spectrometry

There are many methods to identify and recognize the molecular and behavioural differences between organisms.One of the methods for the detection and identification of unknown organisms as well as intermolecular and intramolecular structural differences is MALDI-TOF mass spectrometry.Therefore,differentiation of Fomes fomentarius decay capabilities on the chemical properties of the wood cell wall of the tree species Quercus castaneifolia,Juglans regia,and Carpinus betulus were used to determine and characterize the destructive behaviour of F.fomentarius decay by MALDI-TOF mass spectrometry.The results showed that the fungus had more significant destructive behaviour on J.regia than the other species.For this evidence,completely removal of xylan hemicellulose fragment+Na+at peak 1227 Da and severe digestion of fragment of glucomannan hemicellulose at peak 1477–1480 Da that it seems that signs of soft-rot patterns were obtained from the decayed sample of J.regia,while these were incremental and unchanged for C.betulus and Q.castaneifolia,respectively.However,C.betulus had different peaks of atomic mass than J.regia and Q.castaneifolia wood,respectively.These results showed that this technique could be useful for separating and identifying unknown compounds of the wood cell wall attacked by fungi relying on their biological behaviour.

Transcriptome Analysis of White-Rot Fungi in Response to Lignocellulose or Lignocellulose-Derived Material Using RNA Sequencing Technology

White-rot fungi are the only organisms that can completely degrade all components of lignocellulosic biomass, including the recalcitrant lignin polymer. Lignin degradation is important for the industrial application of lignocellulosic biomass as a raw material for producing value-added chemicals and materials. Therefore, elucidating the lignin degradation mechanism in white-rot fungi will help researchers develop efficient and eco-friendly methods enabling the production of value-added chemicals from lignocellulosic biomass. A transcriptome analysis is an effective way to compare gene expression patterns between different samples under diverse conditions and can provide insights into biological processes. The democratization of next-generation sequencing technology, especially RNA-sequencing, has made transcriptome sequencing and analysis a common research approach for many laboratories. In this review, we focus on the transcriptome profiles of two well-characterized white-rot fungi (Phanerochaete chrysosporium and Dichomitus squalens) in response to various lignocellulosic materials. The application of RNA-seq technology combining with other techniques remains the best approach for investigating fungal secretomes and elucidating the mechanisms of fungal responses to lignocellulose.

不同条件下白腐真菌多糖调控合成的铁(氢)氧化物的特征

对白腐真菌胞外多糖在铁(氢)氧化物的生物形成过程中的作用展开具体研究,使用傅里叶红外光谱仪研究了不同多糖浓度、陈化时间、Fe^(3+)初始浓度、矿化体系pH和陈化温度条件下白腐真菌胞外多糖调控形成的铁(氢)氧化物的特征,探究了在不同条件下白腐真菌生物矿化的铁(氢)氧化物的晶型组成和不同相之间的转化规律。结果表明:适当增大多糖含量可促进铁(氢)氧化物在多糖分子中成核,且陈化过程中铁(氢)氧化物的晶型和结晶程度均会发生变化;Fe^(3+)初始浓度的升高有助于提高铁(氢)氧化物羟基结合态的稳定程度;酸性条件下有利于α-FeOOH的形成,而在碱性条件下则有利于α-Fe_(2)O_(3)的形成;温度的适当升高有利于Fe—O结晶度的提高,温度的进一步升高有利于铁(氢)氧化物转化成更稳定的相。

絮凝沉淀与生物曝气滤池结合处理杨木心材生物化学机械法制浆废水

本研究利用白腐菌Trametes sp.48424预处理杨木心材,结合絮凝沉淀与生物曝气滤池(BAF),处理化学机械法制浆过程中产生的综合废水,研究废水回用对生物化学机械法制浆及BAF的影响,探究处理后废水回用的可行性。结果表明,采用白腐菌预处理技术,能够抵消添加2%(相对于原料绝干质量)的NaOH所产生的CODCr。絮凝沉淀处理后,CODCr的最高去除率为49.9%。BAF处理后COD_(Cr)最高去除率可达91.8%,出水CODCr浓度降至39.8 mg/L,达到制浆废水回用和排放的标准。废水回用会降低BAF的整体效率,且处理后的废水回用于制浆系统,对浆料的成纸性能也会产生一定的影响;回用1次后出水CODCr浓度升至76.3 mg/L,故处理后的废水只能回用1次。

3种病害海带的颜色差异及生化组分分析

为分析不同病害海带叶片颜色和生化组分的差异及二者间的关系,分别对患点状白烂病、绿烂病和卷曲病3种病害海带的颜色指标(明度、红绿色度、黄蓝色度、褐度、色泽饱和度和色调角)和生化组分含量(含水率、叶绿素a、类胡萝卜素、可溶性蛋白和褐藻酸盐)进行了差异分析及相关性分析探索。结果表明:3种病害海带的总色差值ΔE为7.41~15.24,明度、红绿色度、黄蓝色度和色泽饱和度等颜色指标差异显著(P<0.05),其中红绿色度的变异系数最高,为48.18%。绿烂病海带红绿色度最低,明度、黄蓝色度和色泽饱和度最高;卷曲病海带与之相反,红绿色度最高,明度、黄蓝色度和色泽饱和度最低;点状白烂病海带各颜色指标介于二者之间。3种病害海带的叶绿素a、类胡萝卜素和可溶性蛋白含量差异显著(P<0.05),其中可溶性蛋白含量的变异系数最高,为22.08%。点状白烂病海带的叶绿素a含量最高,可溶性蛋白含量最低,绿烂病海带的类胡萝卜素含量最低,卷曲病海带的类胡萝卜素含量最高。3种病害海带基于颜色指标和生化组分含量的综合判别率分别为78.90%和100.00%。绿烂病海带的红绿色度与叶绿素a、类胡萝卜素含量呈显著正相关(r=0.99,r=0.99),3种病害海带的明度、黄蓝色度和色泽饱和度等颜色指标与可溶性蛋白含量存在较强的相关关系(|r|=0.94~0.99)。

解淀粉芽胞杆菌SJ1606产脂肽粗提物协同代森锰锌对2种植物病菌的抑制效果

为探明微生物源脂肽粗提物、代森锰锌及二者的复配剂对黄瓜枯萎病菌和葡萄白腐病菌的抑制效果,采用菌丝生长速率法、Horsfall法和Wadley公式法检测了脂肽粗提物和代森锰锌对2种病菌的抑制率和增效比。脂肽粗提物对黄瓜枯萎病菌的EC_(50)为1.93μL·mL^(-1),对葡萄白腐病菌的EC_(50)为5.54μL·mL^(-1),代森锰锌对黄瓜枯萎病菌的EC_(50)为74.37 mg·L^(-1),对葡萄白腐病菌的EC_(50)为48.45 mg·L^(-1)。Horsfall法得出脂肽与代森锰锌混配体积比为8∶2对黄瓜枯萎病菌具显著的增效活性,体积比为1∶9对葡萄白腐病菌具有显著增效活性。通过Wadley公式法得出混配组合对黄瓜枯萎病菌理论EC_(50)为1298.78 mg·L^(-1),实际EC_(50)达472.48 mg·L^(-1),增效比(SR)为2.75;对葡萄白腐病菌理论EC_(50)值为481.41 mg·L^(-1),实际EC_(50)值187.32 mg·L^(-1),增效比(SR)为2.57。结果为2种不同病害的持续防控、化学药剂的减施增效和抗菌脂肽的合理应用提供支撑。

Influence of glucose feeding on the ligninolytic enzyme production of the white-rot fungus Phanerochaete chrysosporium

The present work studied the influence of glucose feeding on the ligninolytic enzyme production of Phanerochaete chrysosporium in a nitrogen-limited(C/N ratio is 56/8.8 mmol/L)medium.Several sets of shaking flask experiments were conducted.The results showed that 2 g/L glucose feeding on the first day of the culture(24 h after the inoculation)stimulated both fungal biomass growth and enzyme production.The manganese peroxidase(MnP)activ-ity was 2.5 times greater than that produced in cultures with-out glucose feeding.Furthermore,the glucose feeding mode in fed-batch culture was also investigated.Compared to cul-tures with glucose feeding every 48 h,cultures with glucose feeding of 1.5 g/L(final concentration)every 24 h produced more enzymes.The peak and total yield of MnP activity were 2.7 and 3 times greater compared to the contrast culture,respectively,and the enzyme was kept stable for 4 days with an activity of over 200 U/L.

铜陵白姜茎基腐的病原鉴定及其噬菌体的分离纯化

目的:鉴定导致铜陵白姜茎基腐病的病原菌科贝肠杆菌(Enterobacter kobei)及其相应的噬菌体,为防治铜陵白姜茎基腐提供科学依据。方法:选用发病的铜陵白姜姜块组织作为试验材料,使用TTC和LB培养基从铜陵白姜腐烂的姜块中分离出科贝肠杆菌SJ16菌株,同时通过半固体培养基从根际土壤中分离得到噬菌体KB-P2。使用2种感染方法(根茎接种法和姜切片接种法),评定该菌株对铜陵白姜的致病性;并利用注射法测定噬菌体的防控效果。结果:共分离出103株细菌分离株,经LB培养基筛选得到10株菌株,其中SJ16菌株经鉴定为科贝肠杆菌(E.kobei)。病原侵染试验表明,该菌株是导致铜陵白姜枯萎的致病菌,并会在后期引起铜陵白姜根茎腐烂。此外,分离纯化得到3种科贝肠杆菌噬菌体,通过双层平板培养方法筛选出裂解能力最强的KB-P2噬菌体。同时,噬菌体接种试验也进一步显示,KB-P2噬菌体具有良好的防控效果。结论:科贝肠杆菌SJ16菌株是一种导致铜陵白姜茎基腐的新病原菌,而KB-P2噬菌体能够有效降低铜陵白姜茎基腐的发病率。该研究为进一步探讨铜陵白姜茎基腐的防治提供了重要依据。

山楂白纹羽病的诊断与综合防控

白纹羽病是为害山楂等果树根系的重要土传病害之一,严重时造成大树死亡甚至毁园,给山楂生产带来严重损失。总结了山楂白纹羽病的为害症状、病原特征、发病规律及综合防控技术,以期为生产上防控山楂白纹羽病提供参考。

毛栓孔菌XYG422产木质素降解酶的固体发酵工艺研究

为了提高动物粗饲料的消化率,研究以黄芩药渣为基质,采用单因素和正交试验对毛栓孔菌XYG422产木质素降解酶发酵条件进行优化。结果显示,毛栓孔菌XYG422固体发酵的最佳产酶培养基组成为可溶性淀粉∶硫酸铵3∶1、Cu^(2+)浓度3 mmol/L、Mn^(2+)浓度1.8 mmol/L。在此条件下,锰过氧化物酶活力为179.37 nmol/(min·g),漆酶活力为41.68 U/g。研究结果对毛栓孔菌木质素降解酶的生产与应用具有一定的指导意义,可为开发提高纤维消化率的饲料提供借鉴。

迷宫栓孔菌热激蛋白基因的生物信息学与表达分析

为了探究迷宫栓孔菌(Trametes gibbosa)热激蛋白(heat shock proteins,HSPs)家族的功能及结构,对经木屑处理不同时间点的菌丝样品进行cDNA建库,然后根据转录组数据筛选该菌株的所有HSPs基因并进行生物信息学分析;针对HSP100家族进行基因克隆和序列结构分析,并利用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,qRT-PCR)对其在木屑处理下的表达量进行验证。结果如下:在迷宫栓孔菌中共筛选出32个HSPs基因,其编码的蛋白分为5个亚类,分别为HSP100(2个)、HSP90(2个)、HSP70(7个)、HSP60(1个)和小分子热激蛋白[small HSPs(sHSPs),20个],它们在菌体生长调控中具有蛋白翻译后修饰、蛋白质折叠、伴侣蛋白等重要功能。这些HSPs都为疏水蛋白,不同亚类的HSPs理化性质有所差异。HSP100由N-端、核苷酸结合域1(nucleotide-binding domain 1,NBD1)、NBD2、2个NBDs间的接头构成,其中,NBDs具有十分保守的Walker A、Walker B基序及精氨酸指残基。qRT-PCR扩增结果表明,在木屑处理下迷宫栓孔菌HSP100基因表达量有明显上调趋势。综上所述,迷宫栓孔菌中HSPs家族种类多且复杂,在应激情况下HSP100家族承担了重要的蛋白质解聚功能,其序列及结构相对保守。本研究结果为迷宫栓孔菌在胁迫应激方面的研究提供了理论依据。

白腐真菌对邻苯二甲酸二甲酯的降解与机理研究

文章以黄孢原毛平革菌(Phanerochaete chrysosporium)作为白腐真菌的代表,研究其对邻苯二甲酸二甲酯(dimethyl phthalate,DMP)的降解效果与机理。实验结果表明,在最佳条件(pH=5.6,温度为30℃,DMP初始质量浓度为10 mg/L)下,7 d后DMP的去除率为75%。进一步优化反应条件发现:体系中初始葡萄糖质量浓度为5 g/L时,反应结束时DMP降解效率为95%;当反应中加入20μg/L H_(2)O_(2)或27.0 mg/L Mn^(2+),反应7 d后DMP的降解效率显著提升到100%。此外,过氧化物酶在DMP降解过程中起重要作用,实验证明DMP的降解更多地由锰过氧化物酶(manganese peroxidase,MnP)驱动;傅里叶红外光谱证实氨基、羧基、羟基等还原性官能团参与了DMP的去除。文中通过液相色谱-质谱(liquid chromatography-mass spectrometry,LC-MS)联用仪确定DMP的降解产物,并讨论3种可能的降解途径。

木霉菌对白腐真菌所致木材腐朽防效研究

筛选有效抑制白腐真菌生长并降低其对木材腐朽能力的生防菌株,为白腐真菌所致木材腐朽的生物防治提供新思路。以樟子松(Pinus sylvestris)倒木中分离纯化的3株木霉(Trichoderma spp.)作为生防菌株,利用离体对峙培养法测定分离菌株对多种白腐真菌的拮抗效果,结合形态学、分子生物学和系统进化树分析对拮抗效果较好的真菌进行鉴定,并通过检测生防真菌的代谢产物及其与白腐真菌菌丝间的作用关系初步探究其生防机制,最后利用质量损失法检测该生防菌株防治白腐真菌对樟子松木材腐朽的能力。结果表明:筛选得到1株有较好拮抗效果的生防真菌SF2,对峙培养8 d时,该菌株对5种白腐真菌的抑菌率从高到低排序为白囊耙齿菌(Irpex lacteus)>一色齿毛菌(Cerrena unicolor)>粗毛拟革盖菌(Coriolopsis aspera)>彩绒革盖菌(Coriolus versicolor)>裂褶菌(Schizophyllum commune);经过形态观察、ITS1/ITS4 DNA序列扩增比对和系统发育树分析,发现SF2菌株属于长枝木霉(Trichoderma longibrachiatum);生防菌对白腐真菌的抑菌机理显示,菌株SF2可以通过重寄生以及分泌代谢产物对白囊耙齿菌产生抑制;木材接种试验结果表明,接种SF2菌株能显著减少白囊耙齿菌所致樟子松木材质量损失,减少率达46.43%。综上,本研究筛选鉴定的SF2菌株为长枝木霉,其对白腐真菌生长及其所致木材腐朽具有一定的防治效果,本研究为木材白腐病的生物防治奠定理论基础。

白腐真菌Phanerochaete sordida对土壤中吡虫啉的降解研究

为探究白腐真菌对土壤中吡虫啉的降解性能,采用控制变量方法进行了吡虫啉的提取实验和降解实验。结果表明:吡虫啉最优的提取方法为利用甲醇进行超声20 min;在降解实验中,Phanerochaete sordida YK-624对土壤中吡虫啉有良好的降解效果。然而,添加适量碳源,并且增加培养天数都不能提高降解效果,而菌种添加量的增加对降解效果具有积极作用。本项研究初步探索了P.sordida YK-624对土壤中吡虫啉的高效降解条件,对实际吡虫啉污染土壤的白腐真菌修复工作具有重要的理论指导意义。

白腐真菌生长条件及其代谢动力学模型(英文)

To investigate the growth conditions of white-rot fungus and simulate its metabolism kinetic models, the rules how the factors such as biomass, culture fluid, pH value, glucose consumption and exopolysaccharides generation, etc., changed during the batch culture process of white-rot fungi by using an air-lift fermenter, as well as metabolic kinetics of white-rot fungi were studied. Based on Logistic equation, Luedeking-Piret equation and experimental data, the correlation model parameters of mycelia biomass, glucose consumption and exopolysaccharide generation were obtained and found to be change with time in metabolism process. Detailedly, μm=0.071 8 h-1,α= 0.831 8 g/(g·h), β= 0.002 g/(g·h), b1=0.016 3 g/(g·h) and b2=3.023 3 g/(g·h). Hence the mycelial growth kinetic model, exopolysaccharide generation kinetic model and substrate consumption kinetic model which describe fermentation process of white-rot fungi were established. Meanwhile, the experimental data were verified by this model, and a good fitting result with an average relative error less than 10% between the data obtained from experiments and the model was yielded. The results show that these models can predict the growth and metabolic rules of white-rot fungus, the fermentation process of exopolysaccharides and the kinetic mechanism of white-rot fungus accurately.

耐锑白腐真菌的筛选及其吸附特性的研究

随着我国锑相关产业的大规模发展,大量含锑化合物被排入到环境中,对生态环境造成了严重的威胁,如何处理锑污染也成为当今环境科学的研究热点。本研究从几种白腐真菌中筛选出较为耐锑的菌种作为吸附材料,并研究初始浓度、吸附时间及p H条件的改变对锑吸附性能的影响。实验结果表明,最耐锑的白腐真菌是黄孢原毛平革菌(Phanerochaete chrysosporium),在锑初始浓度为50 mg/L、pH=3条件下吸附12 h,对锑的吸附效果最好,吸附量最高达291.76 mg/g。

垃圾堆肥高效复合微生物菌剂的制备

将康氏木霉、白腐菌、变色栓菌与EM菌、固氮菌、解磷菌、解钾菌按一定配比扩大培养制成5组复合微生物菌剂,比较其产脱氢酶、淀粉水解酶和纤维素分解酶能力大小以及在实际堆肥中的应用效果,从中选出一组高效复合微生物菌剂Ⅴ,其配比为:康氏木霉∶白腐菌∶变色栓菌∶EM菌∶固氮菌∶解磷菌∶解钾菌=15∶15∶15∶25∶10∶10。