贵州浓香型白酒大曲中霉菌的18S rDNA系统发育分析

从贵州某名酒厂大曲中分离到104株丝状真菌(霉菌),通过形态特征和生理特性检测等传统鉴定方法,初步把形态和生理生化特征一致的霉菌归类为14个大类群;并采用18S rDNA序列分析,进一步把菌株归类为MJ-I至MJ-V五个类群,随后构建系统发育树.结果表明,样品中的霉菌以曲霉属(Aspergillus)为主,占总分离菌数...     

一株甲醛降解真菌Aspergillus spp.H4的分离鉴定

甲醛是一种广泛使用和很重要的化工原材料,但它同时也是对大多数生物有机体有高毒性的物质。通过添加甲醛的选择性培养基从淤泥里分离了一株甲醛耐受真菌,将其接种于加孟加拉红的PDA培养基、察氏培养基、麦芽汁培养基平板培养,观察它的培养特征和孢子结构,结合DNA提取、PCR扩增、产物测序、GenBank比对等分子鉴定手段,结果表明:其培养特征、显微特征和黄曲霉(Aspergillus flavus)相似,18SrDNA序列与黄曲霉(Aspergillus flavus)同源率达99.8%。我们把它命名为Aspergillus flavus H4。该真菌最适生长pH值为pH5.5,在培养的144h内ρ(甲醛)从1241mg·L-1下降到4mg·L-1。在培养的96h内ρ(甲醛)下降迅速,A值上升缓慢,当ρ(甲醛)下降到10mg·L-1时,也就是120h时,A值迅速上升,ρ(甲醛)下降缓慢。结论是这株Aspergillus flavus H4是甲醛耐受(降解)真菌。     

Alteration in Rhizosphere Soil Properties of Afforested <Emphasis Type="Italic">Rhamnus lycioides</Emphasis> Seedlings in Short-Term Response to Mycorrhizal Inoculation with <Emphasis Type="Italic">Glomus intraradices</Emphasis> and Organic Amendment

The reestablisment of autochthonous plant species is an essential strategy for recovering degraded areas under semiarid conditions. A field experiment was carried out to assess the short-term effect of two reafforestation methods involving mycorrhizal inoculation and compost addition on soil quality parameters and Rhamnus lycioides seedling growth. The nutrient content (NPK) and enzymatic activities (dehydrogenase, urease, protease-BAA, acid phosphatase and β-glucosidase) increased and bulk density decreased in the rhizosphere soil with the organic amendment. Biomass C of rhizosphere soil increased by at least 240% with respect to the control soil after mycorrhizal inoculation and the combination of compost addition + mycorrhizal inoculation. Both mycorrhizal inoculation and composted organic residue addition increased R. lycioides seedling growth in the same proportion. In the short term, we conclude that the application of both reafforestation methods not only enhances the establishment of R. lycioides seedlings, but also improves soil quality.     

Multivariate characterization of elements accumulated in Wolfiporia extensa mushroom from Yunnan province of China

ABSTRACT

Dried sclerotia of Wolfiporia extensa have been used as medicine in Asia from Eastern Han Dynasty, and also used as traditional snack called “fulingjiabing” in Beijing, China. In this paper, 18 macro and trace elements (Ag, As, Ba, Cd, Co, Cr, Cs, Cu, Fe, Li, Mn, Ni, Pb, Rb, Se, Sr, V, and Zn) in both flesh and peel of Wolfiporia extensa from seven sites of Yunnan province in China were determined by inductively coupled plasma mass spectrometer. The average recovery rates of certified reference materials for GBW10015 (spinach leaves) ranged from 90.5 to 113%, for GBW10028 (citrus leaves) from 92.8 to 106%, and for GBW07603 (bush branch and leaves) from 83.3 to 114.6%. Generally speaking, the concentration of all elements determined was at common level. The results of this survey indicate that mineral compositions in peel were higher than in flesh. In peel, the contents of investigated trace metals in mushroom samples were found to be in the range of 1,660–13,400 µg·g^?1 dry matter (dm) for Fe and 29.6–710 µg·g^?1 dm for Mn. The mean contents of Cr, Cu, Rb, V, and Zn in peel were between 10 and 20 µg·g^?1 dm, followed by As, Co, Li, Ni, Pb, Se, and Sr with mean contents between 1 and 10 µg·g^?1 dm, while Ag, Cd, and Cs had mean contents of <1 µg·g^?1 dm. In flesh, the concentration of Fe was in the range of 54–900 µg·g^?1 dm, and it was 1.5–49 µg·g^?1 dm for Mn, followed by Ba, Cu, Rb, and Zn in the range of 1 to 10 µg·g^?1 dm, while for Ag, As, Cd, Co, Cr, Cs, Li, Ni, Pb, Se, Sr, and V it was <1 µg·g^?1 dm. The concentration of toxic elements, such as As, Cd, and Pb, in both flesh and peel was below the permissible limits of World Health Organization. However, As and Pb contents in peel were higher than the limits permitted in the Chinese Pharmacopoeia. The results of principal component analysis showed that the flesh of Wolfiporia extensa from all the seven sites of the Yunnan province tend to cluster together, most probably because the origin of mineral elements in both flesh and peel is wood substrate (old and dead pine trees).     

重金属污染土壤接种丛枝菌根真菌对蚕豆毒性的影响

采用盆栽实验的方法,研究了重金属(包括Cu、Zn、Pb和Cd)复合污染和接种丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)Glomus mosseae对蚕豆(Vicia faba)生长及DNA损伤的影响.结果表明,虽然接种菌根真菌对蚕豆生物量的影响并不显著,但是却显著影响植物对重金属的吸收,接种菌根真菌对蚕豆吸收4种重金属元素的作用有差异.采用单细胞凝胶电泳(single cell gel electrophoresis,SCGE)法研究接种菌根真菌对蚕豆叶片的DNA损伤的影响,与重金属吸收的结果相吻合.结果表明,接种处理可显著增加蚕豆叶片的DNA损伤程度,这与接种处理可提高植物的重金属吸收相一致.     

接种白腐真菌堆肥处理含Pb垃圾

选取未污染土壤、家庭厨余、稻草、麸皮加入Pb(NO3)2溶液配成模拟含高浓度Pb垃圾,采用接种白腐菌堆制处理和不接菌堆制处理2种方法分别进行室内模拟堆制.通过监测理化因子(pH、挥发性固体、水溶性有机碳/有机氮、木质素、粗纤维)与生化因子(呼吸量、微生物生物量碳)以及生物毒性分析因子(种子发芽指数、重金属含量)随时间的变化,系统地研究了重金属对垃圾堆肥过程的影响和白腐菌堆肥处理重金属污染垃圾的可行性.结果表明,接种白腐菌处理Pb污染垃圾的堆肥过程能顺利进行,较好地减弱了Pb的迁移性从而降低其生物有效性,降低了堆肥潜在的重金属危害性.此堆肥工艺下,堆肥成品pH为7.9,水溶性C/N达4.01,挥发性固体含量降至36.1%,木质素余量22.4 g,粗纤维余量30.1 g;且堆肥中Pb主要以残留态存在,约为63.38%,水溶交换态Pb含量则降至0%,种子发芽指数高达121%.     

嘉陵江滨岸带不同土地利用类型土壤真菌群落结构与功能多样性

通过明确嘉陵江滨岸带不同土地利用类型土壤真菌群落多样性的差异,为嘉陵江生态环境保护修复提供理论依据.选择嘉陵江中下游滨岸带的人工湿地、天然湿地、林地和农田这4种典型土地利用类型作为研究样地,利用高通量测序技术对土壤真菌群落进行测序,分析不同土地利用类型对土壤真菌群落多样性、结构和功能的影响.结果表明,林地与天然湿地土壤真菌的Chao1指数显著高于其他两种土地利用类型（P<0.05）,林地土壤真菌的Shannon指数显著高于农田和人工湿地两种土地利用类型（P<0.05）;嘉陵江流域滨岸带土壤真菌被划分为15个菌门,在门分类水平上的优势真菌群落（相对丰度>0.01）分别为：子囊菌门（Ascomycota）、担子菌门（Basidiomycota）、油壶菌门（Olpidiomycota）、罗兹菌门（Rozellomycota）、被孢霉门（Mortierellomycota）和壶菌门（Chytridiomycota）;相比之下,罗兹菌门偏好选择林地生境,而油壶菌门和被孢霉门偏好选择农田,担子菌门在人工湿地具有显著优势;天然湿地中的优势功能类群为植物病原菌,人工湿地中的优势功能类群为粪腐菌,农田中优势功能类群为动物病原菌-真菌寄生菌;冗余分析表明,土壤含水率（MC）、全氮（TN）、有机碳（TOC）和碱解氮（AN）是影响真菌群落变化的主要环境因子.由此可见,林地是嘉陵江流域真菌多样性最高和功能类群最均衡的土地利用类型,其次为天然湿地,人为干扰导致嘉陵江滨岸带土壤真菌群落多样性水平降低.     

外生菌根真菌接种和施磷对油松苗抗盐性的影响

采用3种不同的外生菌根真菌Boletus edulis、Xerocomus chrysenteron和Gomphidius viscidus，在山东东营盐渍土环境胁迫下，研究外生菌根真菌接种和施磷对油松(Pinus tabulaeformis)生长的影响及其可能机理。结果表明，接种和施磷都增加了油松植株的干质量，但施磷不如接种对植物生长的促进作用明显；接种外生菌根后，油松体内的K元素含量显著增加，而Na元素的含量没有明显变化，使得w(K)／w(Na)增加，从而改变了植株体内的离子平衡，减轻离子的毒害作用。而不是通过直接减少植物对Na的吸收来增加植物的抗盐性；同时，接种后外生菌根增加了油松对P的吸收，改善了盐渍土壤导致的植物营养亏缺，促进了植物在盐渍环境下的生长。     

Biodegradation and bioconversion of cellulose wastes using bacterial and fungal cells immobilized in radiopolymerized hydrogels

Annually, great amounts of cellulose wastes, which could be measured in many billions of tons, are produced worldwide as residues from agricultural activities and industrial food processing. Consequently, the use of microorganisms in order to remove, reduce or ameliorate these potential polluting materials is a real environmental challenge, which could be solved by a focused research concerning efficient methods applied in biological degradation processes. In this respect, the scope of this chapter is to present the state of the art concerning the biodegradation of redundant cellulose wastes from agriculture and food processing by continuous enzymatic activities of immobilized bacterial and fungal cells as improved biotechnological tools and, also, to report on our recent research concerning cellulose wastes biocomposting to produce natural organic fertilizers and, respectively, cellulose bioconversion into useful products, such as: ‘single-cell protein’ (SCP) or ‘protein-rich feed’ (PRF). In addition, there are shown some new methods to immobilize microorganisms on polymeric hydrogels such as: poly-acrylamide (PAA), collagen-poly-acrylamide (CPAA), elastin-poly-acrylamide (EPAA), gelatin-poly-acrylamide (GPAA), and poly-hydroxy-ethyl-methacrylate (PHEMA), which were achieved by gamma polymerization techniques. Unlike many other biodegradation processes, these methods were performed to preserve the whole viability of fungal and bacterial cells during long term bioprocesses and their efficiency of metabolic activities. The immobilization methods of viable microorganisms were achieved by cellular adherence mechanisms inside hydrogels used as immobilization matrices which control cellular growth by: reticulation size, porosity degree, hydration rate in different colloidal solutions, organic and inorganic compounds, etc. The preparative procedures applied to immobilize bacterial and fungal viable cells in or on radiopolymerized hydrogels and, also, their use in cellulose wastes biodegradation are discussed in detail. In all such performed experiments were used pure cell cultures of the following cellulolytic microorganisms: Bacillus subtilis and Bacillus licheniformis from bacteria, and Pleurotus ostreatus, Pleurotus florida, and Trichoderma viride from fungi. These species of microorganisms were isolated from natural habitats, then purified by microbiological methods, and finally, tested for their cellulolytic potential. The cellulose biodegradation, induced especially by fungal cultures, used as immobilized cells in continuous systems, was investigated by enzymatic assays and the bioconversion into protein-rich biomass was determined by mycelial protein content, during such long time processes. The specific changes in cellular development of immobilized bacterial and fungal cells in PAA hydrogels emphasize the importance of physical structure and chemical properties of such polymeric matrices used for efficient preservation of their metabolic activity, especially to perform in situ environmental applications involving cellulose biodegradation by using immobilized microorganisms as long-term viable biocatalysts.     

菌根真菌在墨兰和建兰根中的感染特征及生理特性

从产于粤北山区的野生建兰报中分离出4株内生真菌,并对这些真菌在兰根上的感染特征及生理学特性进行了研究。结果表明,菌根真菌多数浸染植株根段离很尖3～18cm的根毛区,而根尖和新长出的根则很少被感染;几乎所有的老根都受过真菌的感染。真菌菌丝从兰报表皮侵入,通过表皮层进入皮层薄壁细胞内部形成菌丝团,而在表皮层并不进入细胞内,也不形成菌丝团。真菌感染极状整后第9天,皮层细胞内的菌丝团开始消解。所分离的菌株能利用大部分简单的有机碳源和氮源,能在营养成分不明确的天然介质中生长良好,但对硝酸盐的利用较差。