灵芝漆酶对直接蓝86的催化脱色性能

利用灵芝菌Ganoderma lucidum U-281漆酶对直接蓝86进行酶促氧化脱色,并对其降解机理进行了探讨。结果表明,染料-漆酶共反应体系在20～50℃及pH小于5.0范围内,直接蓝86均可脱色50%以上;漆酶对直接蓝86具有宽泛的浓度适应性,对300 mg/L的该染料仍具有耐受性。最优脱色工艺参数为温度40℃、pH 5.0、染料初始浓度200 mg/L、漆酶用量1 U/mL。在优化条件下,直接蓝863 h的脱色率达到54.54%,48 h脱色率达到91.54%。紫外-可见吸收光谱分析表明,漆酶的酶促氧化导致染料的分子结构产生了变化,是造成直接蓝86脱色的主要发生机制。     

漆酶对活性黑KN-B和直接大红染料的脱色性能

利用白腐真菌漆酶对活性黑KN-B和直接大红2种偶氮染料进行脱色实验。考察反应时间、加酶量、pH值、染料浓度、温度对脱色率的影响,研究了ABTS介体以及金属离子存在下的脱色效果,并分析了漆酶脱色的动力学性能以及其对偶氮染料的降解规律。结果表明,活性黑KN-B和直接大红脱色适宜条件为：反应时间为30 min,加酶量8 U/mL,pH=7,染料浓度分别为50 mg/L和80 mg/L,温度40~45℃。ABTS介体对酶促偶氮染料脱色没有明显促进作用。Fe²⁺对漆酶脱色有较强的抑制作用;Cu²⁺对漆酶催化活性黑KN-B促进作用较大,对直接大红影响较小。漆酶对2种染料的脱色反应符合米氏方程,其催化活性黑KN-B和直接大红染料的K_m值分别为114.81 mg/L,317.5 mg/L,v_max值分别为6.57 mg/（L·min）和26.0 mg/（L·min）。     

Prevalence of the Crayfish Plague Pathogen Aphanomyces astaci in Invasive American Crayfishes in the Czech Republic

Abstract:  In Central Europe invasive North American crayfishes are carriers of the oomycete Aphanomyces astaci, which causes crayfish plague. This lethal disease currently represents one of the major threats to native European crayfishes. We used molecular methods—species-specific amplification and sequencing of the pathogen DNA—to investigate the prevalence of individuals latently infected with A. astaci in 28 populations of two invasive American crayfish species (6 of the signal crayfish [ Pacifastacus leniusculus ] and 22 of the spiny-cheek crayfish [ Orconectes limosus ]) in the Czech Republic. The pathogen occurred in 17 investigated populations. We recorded a high variation in positive reactions, ranging from 0% to 100%, in populations of O. limosus . In P. leniusculus, however, only one individual out of 124 tested positive for the pathogen. There was a clear relationship between the water body type and pathogen prevalence in O. limosus . Infection ratios in isolated standing waters were usually low, whereas in running waters, pathogen prevalence often exceeded 50%. Other evaluated characteristics of potential plague pathogen carriers (size, sex, and the presence of melanized spots in the cuticle) seemed to be unrelated to infection. Our data suggest that in contrast to other European countries, O. limosus seems to be the primary reservoir of crayfish plague in the Czech Republic. Although all populations of alien American crayfishes may be potential sources of infections and should be managed as such, knowledge on the prevalence of the plague pathogen at various localities may allow managers to focus conservation efforts on the most directly endangered populations of native crayfishes.     

Aureobasidium pullulans UVMU3-1产耐热性胞外漆酶活性物质培养基优化及其对染料脱色作用

对出芽短梗霉无色素突变株Aureobasidium pullulans UVMU3-1产耐热性胞外漆酶活性物质培养基进行优化,并研究其分子量大小、热稳定性及对三苯甲烷类染料孔雀石绿和结晶紫的脱色作用.结果表明:蔗糖、NaNO_3为UVMU3-1产耐热性胞外漆酶活性物质的最佳碳源和氮源,产酶优化培养基配方为:蔗糖66.5 g·L~(-1),NaNO_338.4 g·L~(-1),NaCl 1.08 g·L~(-1),酵母浸粉0.2 g·L~(-1),KH_2PO_41 g·L~(-1),pH=6.0.影响UVMU3-1产耐热性胞外漆酶活性物质的因素依次为:起始pH蔗糖NaClNaNO_3,培养7 d时产生最大酶活(94.81 U·L~(-1)),比对照提高3.85倍.其分子量小于1000 D,具有良好的热稳定性,煮沸30 min后高温酶活仍残留72.50%.粗酶液(7.5 U·L~(-1))与三苯甲烷类代表性染料孔雀石绿和结晶紫75℃反应5 min后,脱色率分别为76.3%、77.5%,反应35 min后脱色率分别达到96.3%、95.3%.     

水稻秸秆经漆酶预处理后产沼气研究

利用秸秆产沼气是秸秆无害化处理解决环境污染问题的可行手段.本研究针对秸秆特殊结构而导致产沼气少、启动慢、利用率低等问题,采用自培养漆酶预处理水稻秸秆,提高了秸秆产气率及利用率.通过研究发现,当漆酶添加量为40 mL、发酵温度为35℃、预处理7d的条件下,秸秆的产气量最高;同时发现,金属离子的添加对沼气产量有很大影响,其中Cu2添加后产气量提高最多.另外,研究发现预处理天数为24 d的秸秆木质素纤维素降解率最高.     

Grazing greatly reduces the temporal stability of soil cellulolytic fungal community in a steppe on the Tibetan Plateau

Excessive livestock grazing degrades grasslands ecosystem stability and sustainability by reducing soil organic matter and plant productivity. However, the effects of grazing on soil cellulolytic fungi, an important indicator of the degradation process for soil organic matter, remain less well understood. Using T-RFLP and sequencing methods, we investigated the effects of grazing on the temporal changes of cellulolytic fungal abundance and community structure in dry steppe soils during the growing months from May to September, on the Tibetan Plateau using T-RFLP and sequencing methods. The results demonstrated that the abundance of soil cellulolytic fungi under grazing treatment changed significantly from month to month, and was positively correlated with dissolved organic carbon (DOC) and soil temperature, but negatively correlated with soil pH. Contrastingly, cellulolytic fungal abundance did not change within the fencing treatment (ungrazed conditions). Cellulolytic fungal community structure changed significantly in the growing months in grazed soils, but did not change in fenced soils. Grazing played a key role in determining the community structure of soil cellulolytic fungi by explaining 8.1% of the variation, while pH and DOC explained 4.1% and 4.0%, respectively. Phylogenetically, the cellulolytic fungi were primarily affiliated with Ascomycota (69.65% in relative abundance) and Basidiomycota (30.35%). Therefore, grazing substantially reduced the stability of soil cellulolytic fungal abundance and community structure, as compared with the fencing treatment. Our finding provides a new insight into the responses of organic matter-decomposing microbes for grassland managements.     

红壤区退化林地表土真菌群落结构对土壤改良措施的响应

真菌群落结构和多样性对于土壤改良效果具有高敏感性.研究南方红壤区侵蚀退化林表土真菌群落对有机肥、生物炭和石灰+微生物肥的响应,以明晰不同改土措施的作用.结果表明：(1)3种土壤改良措施均降低了表土真菌丰富度,其中石灰+微生物肥降低作用最大,3种土壤改良措施对表土真菌多样性也有影响,但影响不显著;(2)表土中优势真菌门为子囊菌门(Ascomycota, 31.29%～46.55%)、担子菌门(Basidiomycota, 30.07%～70.71%),优势真菌属为阿太菌属(Amphinema)和单形古根菌属(Archaeorhizomyces),3种土壤改良措施对表土真菌群落结构的影响不同,有机肥提高了子囊菌门和单形古根菌属的相对丰度,生物炭提高了担子菌门和阿太菌属的相对丰度,而石灰+微生物肥则提高了担子菌门和单形古根菌属的相对丰度;(3)土壤pH是影响表土真菌丰富度的关键因子,而表土真菌群落结构则受pH、全氮和有机碳的影响.研究结果为南方红壤区侵蚀退化林地土壤改良,林下植被生态恢复提供科学指导.     

氮添加对不同坡度退化高寒草甸土壤真菌多样性的影响

为明确不同坡度退化高寒草甸土壤真菌多样性对外源氮添加的响应规律,选取三江源区果洛州不同坡度退化程度相近的高寒草甸,进行外源氮添加试验,探讨不同氮添加水平：低等量氮添加（LN,2 g ·m^-2）、中等量氮添加（MN,5 g ·m^-2）和高等量氮添加（HN,10 g ·m^-2）对不同坡度退化草地土壤真菌多样性的影响.结果表明：①土壤中真菌分布类群集中于子囊菌门（Ascomycota）、担子菌门（Basidiomycota）、被孢霉门（Mortierellomycota）和球囊菌门（Glomeromycota）,其中优势菌门为子囊菌门和担子菌门.子囊菌门、根肿黑粉门和油壶菌门等相对丰度在不同坡度间差异显著（P<0.05）.而同一坡度不同氮添加处理下真菌各门丰度变化差异均不显著.②优势属为被孢霉属和Archaeorhizomyces,它们对不同坡度和不同氮添加水平的响应无差异性.③已鉴定95个菌属在不同坡度间存在显著差异（P<0.05）.④一些相对丰度<1%的属在同一坡度不同水平氮添加间存在显著差异（P<0.05）.5 α和β多样性分析整体表现为在不同坡度和不同氮添加水平下土壤真菌群落无差异性.表明退化高寒草甸土壤真菌对外源氮添加的响应不敏感.     

生物炭和秸秆还田对微咸水滴灌棉田土壤真菌群落结构多样性的影响

微咸水灌溉增加土壤盐分,改变土壤环境,进而影响土壤真菌的结构和多样性.在长期微咸水灌溉的基础上,分别添加生物炭和秸秆（采用等碳量设计,分别为3.7 t·hm^-2和6 t·hm^-2）,探究生物炭和秸秆对土壤理化性质和真菌群落结构多样性的影响.结果表明：与不施生物炭和秸秆（对照）相比,生物炭施用显著增加土壤的pH、全碳、速效钾和速效磷含量,但显著降低土壤电导率,降低幅度为20.71%;秸秆处理显著增加土壤的速效钾和速效磷含量,但显著降低土壤容重和电导率,降低幅度为4.17%和64.50%.生物炭和秸秆处理对真菌群落Chao1指数和ACE指数有增加趋势,对Shannon指数和Simpson指数有降低趋势.土壤优势真菌门类为子囊菌门、被孢霉门、担子菌门、壶菌门和球囊菌门;优势真菌属为毛壳菌属、赤霉菌属、镰刀菌属、Idriella和被孢霉属.施加生物炭和秸秆提高子囊菌门、被孢霉门、担子菌门、球囊菌门和毛壳菌属的相对丰度;但降低壶菌门、赤霉菌属和Idriella的相对丰度.LEfSe分析表明,施用生物炭和秸秆还田降低真菌群落潜在生物标志物数量.RDA结果显示,土壤真菌群落结构与EC_1：5和TN显著相关.微咸水灌溉给土壤带来了不利影响,其中EC_1：5和TN是驱动土壤真菌群落结构变化的主要因子,土壤真菌群落通过生物炭和秸秆对土壤的改良作用来适应盐胁迫环境.     

环氧化PVA载体固定漆酶的最佳工艺条件研究

在T=35℃,HAc-NaAc缓冲溶液pH为4的条件下,用环氧化PVA载体对漆酶进行固定化,固定后酶活为游离酶活的95%。用该固定化漆酶处理制浆造纸废水,反复使用6次后,酶活仍保持为初始酶活的45%,且每次COD去除率在40%以上,处理后的废水BOD增大了50%,废水可生化性得到很大提高。