Diversity of facultatively anaerobic microscopic mycelial fungi in soils

The numbers of microscopic fungi isolated from soil samples after anaerobic incubation varied from tens to several hundreds of CFU per one gram of soil; a total of 30 species was found. This group is composed primarily of mitotic fungi of the ascomycete affinity belonging to the orders Hypocreales (Fusarium solani, F. oxysporum, Fusarium sp., Clonostachys grammicospora, C. rosea. Acremonium sp., Gliocladium penicilloides, Trichoderma aureoviride, T. harzianum, T. polysporum, T. viride. T. koningii, Lecanicillum lecanii, and Tolypocladium inflatum) and Eurotiales (Aspergillus terreus, A. niger, and Paecilomyces lilacimus), as well as to the phylum Zygomycota, to the order Mucorales (Actinomucor elegans, Absidia glauca, Mucor circinelloides, M. hiemalis, M. racemosus, Mucor sp., Rhizopus oryzae, Zygorrhynchus moelleri, Z. heterogamus, and Umbelopsis isabellina) and the order Mortierellales (Mortierella sp.). As much as 10-30% of the total amount of fungal mycelium remains viable for a long time (one month) under anaerobic conditions.     

厌氧真菌和甲烷菌共培养的研究进展

厌氧真菌是自然界中降解植物纤维素类物质最高效的微生物之一.近年来,大量厌氧真菌和甲烷菌共培养菌株被分离.共培养中,甲烷菌通过对厌氧真菌代谢产物的利用显著提高厌氧真菌对木质纤维素的降解;厌氧真菌通过为甲烷菌提供能量和营养物质使甲烷菌快速生成大量甲烷.全面深入地了解共培养中两者的互作关系以及共培养降解木质纤维素产甲烷的特性...     

Effect of administration of anaerobic fungi isolated from cattle and wild blue bull ( Boselaphus tragocamelus ) on growth rate and fibre utilization in buffalo calves

Abstract

Fifteen Murrah buffalo calves (age about 10 months, 163 – 176 kg BW) were divided into three groups. Group I (Control) was fed a complete feed mixture consisted of 50% wheat straw and 50% concentrate mixture (contained per kg: maize 330 g, groundnut cake 210 g, mustard cake 120 g, wheat bran 200 g, de-oiled rice bran 110 g, mineral mixture 20 g and common salt 10 g) along with 2 kg green oats per animal and day to meet the vitamin A requirements. Calves of Groups II and III were fed with the Control diet supplemented with Orpinomyces sp. C-14 and Piromyces sp. WNG-12 cultures, respectively. The digestibility of DM was significantly highest with Piromyces sp. WNG-12 in Group III (62.2%) followed by Orpinomyces sp. C-14 in Group II (60.3%), and Control (53.5%). A similar pattern of increase in digestibility of crude protein and cell-wall contents was observed in treatment groups. The digestible energy in terms of percent total digestible nutrients was also significantly enhanced in Groups II (56.6%) and III (59.9%) when compared to Control (49.2%). The rumen fermentation parameters such as pH and NH₃-N were found to be lower, whereas total nitrogen, tricarboxylic acid precipitable-nitrogen, total volatile fatty acids and zoospore counts per millilitre of rumen liquor were significantly higher in fungal administered groups. After administration of fungal cultures, improvements of animal growth rate (i.e. body weight gain) and feed efficiency were also observed.     

ARISA方法研究产甲烷菌共存及去除条件下瘤胃真菌多样性变化

摘要：【目的】建立厌氧真菌多样性分析方法,并研究厌氧真菌与产甲烷菌共培养液在传代过程中厌氧真菌的区系变化及共培养液中去除产甲烷菌条件下厌氧真菌多样性的变化。【方法】根据厌氧真菌ITS1序列长度多态性,设计厌氧真菌特异性引物,然后PCR扩增样品中厌氧真菌ITS1序列,在基因分析仪中分析PCR产物序列长度多态性,分析共培养液在传代过程中及共培养液中去除产甲烷菌后厌氧真菌多样性的变化。【结果】对瘤胃厌氧真菌Caecomyces属YC301菌株、Neocallimastix属菌株（YC501与YC502）的ARI     

青岛胶州湾四种类型湿地AM真菌多样性

对青岛胶州湾4种类型湿地（盐田、滩涂、湖泊和河口）中芦苇Phragmites communis、香蒲Typha orientalis和碱蓬Suaeda glauca 根围土壤中丛枝菌根（arbuscular mycorrhizal,AM）真菌进行孢子分离与鉴定,分析湿地生态系统中植物根围AM真菌群落特征。共分离到AM真菌5属10种,包括斗管囊霉属Funneliformis 2种、无梗囊霉属Acaulospora 3种、近明球囊霉属Claroideoglomus 2种、球囊霉属Glomus 1种、巨孢囊霉属Gigaspora 2种,其中,斗管囊霉属Funneliformis及地斗管囊霉Funneliformis geosporum的分离频度和相对多度最高,分别为湿地中AM真菌优势属和优势种。滩涂和河口湿地中植物AM真菌侵染率显著高于湖泊和盐田湿地植物,AM真菌孢子密度以滩涂湿地最高（572个/20mL）,湖泊湿地最低（220个/20mL）;滩涂湿地的种丰度和Shannon-Wiener指数最高,分别为3.8和1.2。从植物种类来看,AM真菌侵染率总体呈现出香蒲>碱蓬>芦苇,AM真菌孢子密度以香蒲最高,芦苇最低,植物种类对AM真菌种丰度和Shannon-Wiener指数影响不显著（P>0.05）。二因素方差分析和典型RDA相关分析表明,寄主植物种类对AM真菌孢子密度有一定影响,但湿地类型对AM真菌多样性的影响更为显著（P<0.05）,胶州湾湿地土壤因子Ca ²⁺、速效P含量与AM真菌孢子密度、物种丰度和多样性指数显著负相关,而速效K、Na ⁺、pH与其显著正相关。结果表明植物种类主要影响AM真菌孢子密度,AM真菌多样性受植物种类和湿地类型综合影响,滩涂湿地较丰富的AM真菌多样性可能与该湿地较良好的理化性质有关。     

甲烷菌对厌氧真菌不同碳源代谢的影响

【目的】探讨碳源和甲烷菌对厌氧真菌碳代谢的影响。【方法】利用体外批次厌氧发酵法,比较厌氧真菌纯培养(Orpinomyces sp.和Neocallimastix sp.)及其与甲烷菌共培养(F1:Orpinomyces sp.+Methanobrevibacter sp.和N3:Neocallimastix sp.+Methanobrevibacter sp.)发酵不同类型碳水化合物代谢产物的差异。【结果】对厌氧真菌和甲烷菌共培养F1和N3的研究显示,F1发酵木薯粉[(26.44±0.22)mmol/L]的乳酸产量是发酵玉米芯[(1.31±0.04)mmol/L]的20.18倍,是N3发酵木薯粉[(1.59±0.03)mmol/L]的16.63倍,玉米芯[(0.79±0.08)mmol/L]的33.47倍。当F1和N3中的厌氧真菌纯培养时,各组乳酸产量均1.90 mmol/L。对F1进一步研究,结果显示发酵体系中木薯粉添加量在0.8%–2.0%之间时,乳酸产量随木薯粉添加量增加而增加。当含量在1.0%–2.4%之间时,随木薯粉添加量增加,甲烷和乙酸产量逐渐降低。比较F1发酵大米粉、木薯粉、玉米粉、小麦粉和土豆粉的发酵结果,发现乳酸产量与底物中支链淀粉的含量成正相关(R2=0.9554)。当F1发酵葡萄糖和麦芽糖时,乳酸产量5.00 mmol/L。当以麦芽糊精为底物时,乳酸产量高达(28.00±0.95)mmol/L。【结论】本文首次报道碳源和甲烷菌能够增强厌氧真菌的乳酸代谢途径并且这种增强存在种属特异性。     

Detection and monitoring of anaerobic rumen fungi using an ARISA method

Aim: To develop an automated ribosomal intergenic spacer region analysis (ARISA) method for the detection of anaerobic rumen fungi and also to demonstrate utility of the technique to monitor colonization and persistence of fungi, and diet‐induced changes in community structure. Methods and Results: The method could discriminate between three genera of anaerobic rumen fungal isolates, representing Orpinomyces, Piromyces and Neocallimastix species. Changes in anaerobic fungal composition were observed between animals fed a high‐fibre diet compared with a grain‐based diet. ARISA analysis of rumen samples from animals on grain showed a decrease in fungal diversity with a dominance of Orpinomyces and Piromyces spp. Clustering analysis of ARISA profile patterns grouped animals based on diet. A single strain of Orpinomyces was dosed into a cow and was detectable within the rumen fungal population for several weeks afterwards. Conclusions: The ARISA technique was capable of discriminating between pure cultures at the genus level. Diet composition has a significant influence on the diversity of anaerobic fungi in the rumen and the method can be used to monitor introduced strains. Significance and Impact of the Study: Through the use of ARISA analysis, a better understanding of the effect of diets on rumen anaerobic fungi populations is provided.     

Diversity and metal tolerance of nematode-trapping fungi in Pb-polluted soils

The diversity of nematode-trapping fungi (NTF) in two lead (Pb) mines in Yunnan Province, China was investigated in 2004. In total, 20 species belonging to five genera were identified from 500 samples collected at the Lanping and the Huize mines. Pb concentrations ranged from 216 approximately 7,150 mg/kg for the former and 132 approximately 13,380 mg/kg for the latter, respectively. The fungi were divided into five groups based on different trapping mechanisms. The trapping-net producer group contained the largest number of species, with nine. Two predators, Dactylellina ellipsosporum and Arthrobotrys oligospora, were found at frequencies of 32.85% and 15.41%, respectively. The diversity indexes of NTF were positively correlated with Pb pollution levels in both the Lanping Mine (r=0.66) and the Huize Mine (r=0.72), suggesting that the distribution of NTF was not negatively affected by Pb contamination. For most strains of a given species, there was no significant difference (P>0.01) in the Pb tolerance between the strains isolated from habitats with low or high Pb concentrations. However, Pb toxicity exerted adverse effects on trap formation and predacious capability of fungi. We discuss the possible metal tolerance mechanisms and their relationships to the survival strategy of NTF in Pb-polluted environments.     

构树可培养内生真菌的多样性初探

构树Broussonetia papyrifera为桑科重要的资源植物。为调查逆境构树内生真菌的多样性,我们对铅锌矿区构树根、茎、叶和果实各部位进行内生真菌分离鉴定,从中分离得到187株内生真菌,根据形态学与分子生物学方法鉴定为20属,其中链格孢属Alternaria（50.27%）、镰孢属Fusarium（11.76%）为优势属。不同培养基的内生真菌分离率结果表明,马铃薯葡萄糖琼脂培养基（PDA）分离出的菌株数最多,占总数的73.96%;构树不同部位的菌株分离数目显示,叶中分离出的菌株较多,占38.02%。在属的分类水平上,根、茎、叶、果实不同部位的真菌数量也存在明显差异。Shannon-Wiener多样性指数和Simpson多样性指数分析结果提示不同样地的构树内生真菌的多样性也存在差异。Chao1指数和ACE指数也提示构树内生真菌丰富度与样地和组织有关。     

贵州马比木内生真菌的多样性研究

为了解贵州省马比木内生真菌的多样性,通过采集贵阳修文、遵义播州、铜仁万山3个县区的不同季节的健康马比木植物不同组织部位样本进行内生真菌的分离,分析分离菌ITS rDNA序列,采用分子系统学方法,对3个地区的不同季节马比木植物不同组织部位的内生真菌进行鉴定、归类,并进行多样性评价。结果表明,从1 444个马比木组织块中分离出1 037株内生真菌,分别隶属于30个属,其中间壳座属Diaporthe为优势属,分离率与分离频率分别为40.24%与57.58%;夏季内生真菌的多样性指数最高,4个季节的相似度在0.40-0.71之间,贵阳的相似性指数最高,三地的相似性指数在0.42-0.55之间,果实的多样性指数最高,各个组织部位的相似性指数在0.00-0.54之间。表面植株生长环境及部位对内生真菌的组成和多样性均有影响。     

毛乌素沙地樟子松外生菌根真菌多样性特征

【背景】外生菌根真菌(ectomycorrhizal fungi,ECMF)侵入林木根系形成外生菌根,是樟子松(Pinus sylvestris var. mongholica)适应环境变化和环境胁迫的重要途径。【目的】深入理解沙地樟子松人工林外生菌根真菌群落组成和多样性。【方法】以毛乌素沙地3个林龄(27、33和44 a)沙地樟子松人工林为研究对象,采用rDNA ITS区段PCR扩增测序方法鉴定分析沙地樟子松外生菌根真菌群落组成并计算多样性指数。【结果】毛乌素沙地樟子松共获得56个外生菌根真菌OTU,隶属于2门3纲8目15科21属,优势属为棉革菌属(Tomentella)、地孔菌属(Geopora)和阿太菌属(Amphinema)。27 a人工林外生菌根真菌丰富度指数最高,33 a人工林ECMF的α多样性指数最高,但不同林龄之间不存在显著差异(P>0.05)。不同林龄沙地樟子松ECMF群落组成存在差异,地孔菌属、棉革菌属、Delastria和Mallocybe的动态变化是造成群落组成差异的主要原因。【结论】毛乌素沙地樟子松人工林外生菌根真菌群落组成较为丰富,该研究区的群落组成可能比多样性更能体现外生菌根真菌群落随林龄的动态变化,伴随着林木的生长成熟,部分功能性外生菌根真菌的动态变化可能与沙地樟子松人工林退化存在密切联系,有待进一步研究。     

Complete genome sequence of the facultatively anaerobic, appendaged bacterium Muricauda ruestringensis type strain (B1(T))

Muricauda ruestringensis Bruns et al. 2001 is the type species of the genus Muricauda, which belongs to the family Flavobacteriaceae in the phylum Bacteroidetes. The species is of interest because of its isolated position in the genomically unexplored genus Muricauda, which is located in a part of the tree of life containing not many organisms with sequenced genomes. The genome, which consists of a circular chromosome of 3,842,422 bp length with a total of 3,478 protein-coding and 47 RNA genes, is a part of the Genomic Encyclopedia of Bacteria and Archaea project.     

量天尺根部内生真菌的多样性

为了解不同量天尺（火龙果）品种根部内生真菌菌群组成及多样性,采集GHL-1、GHL-2、GHL-3、ML-1和DL 5个量天尺品种健康根部样品,进行内生真菌分离,采用形态观察和ITS序列分析相结合的方法进行鉴定、归类。共分离得到内生真菌菌株117株,总体分离率为25.71%,分别隶属于13个属,其中Trichoderma、Fusarium、Chaetomium和Phoma为量天尺内生真菌的优势种群,分别占总菌株数的24.79%、35.04%、10.26%和10.26%;不同量天尺品种内生真菌的结构和组成存在一定差异,GHL-2、GHL-3和DL 3个品种中分离频率最高的内生真菌类群为Fusarium,GHL-1和ML-1分离频率最高的类群为Trichoderma;多样性分析结果反映出不同量天尺品种内生真菌菌群的多样性指数、丰富度指数和均匀度指数水平存在差异,其中GHL-2的3项指数均为最高。表明品种差异对内生真菌的组成和多样性均有影响。     

土壤中甲烷厌氧氧化菌多样性的分子检测

甲烷厌氧氧化作用是减少海洋底泥甲烷释放的重要生物地球化学过程,然而在陆地生态系统中甲烷厌氧氧化作用及其功能菌群的生态功能仍然不确定。对甲烷厌氧氧化菌多样性的研究可为减少甲烷排放提供重要科学依据。与传统的分离培养方法比较,分子检测方法是一种更为快速和高效的研究手段,可直接和全面的反映参与甲烷厌氧氧化作用的功能微生物。以DNA分子标记物为研究对象,重点探讨三类主要的分子标记基因,即16S rRNA,mcr A和pmo A,所采用的相关探针和引物信息,同时从定性和定量两个角度综述土壤甲烷厌氧氧化菌的多样性研究的主要进展,最后提出厌氧甲烷氧化菌多样性研究中存在的一些问题和相应的解决思路。     

厌氧真菌纤维降解酶及其应用潜力的研究进展

反刍动物的瘤胃是已知的纤维降解能力最强的天然发酵罐,其发酵粗纤维的能力很大程度上依赖于其中栖息的各类微生物的功能。厌氧真菌作为瘤胃内的一类低丰度菌群,最先定殖到宿主动物摄入的纤维质饲料上,并通过分泌大量高效的碳水化合物活性酶降解粗纤维。然而,由于缺乏足够的基因组信息和有效的厌氧真菌遗传操作系统,目前国内外对厌氧真菌分泌的纤维降解酶及其降解机制的研究进展有限。本文对厌氧真菌的分类及已发表的基因组信息进行概述,介绍了各类纤维降解酶及纤维小体的组成结构和催化机制特点,并对纤维降解酶在生物质能、饲料处理、纺织造纸及食品加工等方面的应用进行总结。研究厌氧真菌纤维降解酶的作用特性,将有助于完善其在瘤胃环境中有效竞争资源并降解粗纤维的知识体系,也将进一步了解其生物技术应用潜力,为工业生产中应用酶制剂提供新思路。     

两种盐生植物内生真菌多样性及其功能预测

【背景】植物内生菌是微生物群落中一类非常重要的组成部分,是重要的微生物资源库,在植物促生、抗逆等多个领域有重要的研究和应用价值。【目的】进一步了解干旱荒漠区盐生植物内生真菌的多样性、群落结构和潜在功能特征。【方法】对生长在乌兹别克斯坦西咸海岸边的两种盐生植物毛足假木贼(Anabasis eriopoda abbreviated as AE)和展枝假木贼(A. truncata abbreviated as AT)的内生真菌群落进行扩增子测序分析。【结果】共获得166个ampliconsequencingvariants(ASVs),涉及4门49属,其中Neocamarosporium、Botryosphaeria和Alternaria及其所属高级分类单元是优势类群。多样性和群落组成分析显示两种盐生植物的内生真菌存在较为明显的差异,并包含一些潜在的新分类单元。基于PICRUSt2和FUNGuild的功能预测结果表明这两种盐生植物内生真菌的潜在功能和营养方式多样且表现出宿主差异性。【结论】盐生植物内生真菌具有较高的多样性和潜在的资源价值,有待进一步挖掘和研究。     

Keratinolytic and keratinophilic fungi in the soils of Papua New Guinea

A study was made of soil samples collected during an expedition to the Highlands of Papua New Guinea. Fungi were isolated from the samples by the method of hair baiting (To-Ka-Va). Of the 33 species isolated, about half showed keratinolytic activity. Such activity is previously unreported for Mucor hiemalis f. hiemalis, Myrothecium roridum, Paecilomyces carneus, P. marquandii, Penicillium brevicompactum, Rhinocladiella mansonii and Verticillium lecanii. The species most active keratinolytically were Chrysosporium an. Arthroderma cuniculi, C. an. A. curreyi, C. indicum, Myceliophthora vellerea and Trichophyton ajelloi. The spectrum of fungi with keratinolytic activity isolated from the different sites differed considerably according to the frequency of use by man, heaviest use being correlated with greatest activity. The pH of the soil (varying from 5.8–7.5) had little influence on the type of such fungi isolated.     

Diversity and efficiency of arbuscular mycorrhizal fungi in soils from organic chili (Capsicum frutescens) farms

No previous studies have been conducted on the diversity and population of arbuscular mycorrhizal fungi (AMF) in relation to organically grown chili (Capsicum frutescens L.) in Thailand. This study was carried out to investigate the diversity and status of AMF populations at four organically managed farms in Ubon Ratchathani and Sisaket provinces. The effects of each AMF species on the growth and nutrient uptake of chili grown in sterile, organically managed soil were determined. Fourteen AM fungal taxa belonging to the genera Acaulospora (4 spp.), Entrophospora (1 sp.), Glomus (7 spp.) and Scutellospora (2 spp.) were found. Among these, Glomus was the dominant genus found at all sites, followed by Acaulospora. The spore density and root colonization of AMF on chili did not vary significantly among the sites. The effects of ten selected AMF species on the growth of chili showed that Gl. clarum RA0305 increased the growth, flowering, and fruit production of chili, and also increased the P uptake significantly, compared to non-mycorrhizal plants. This fungus showed the highest potential as a promoter of growth, flowering and yield in organically managed chili production.     

兰科植物黄花白及Bletilla ochracea内生真菌多样性分析

黄花白及Bletilla ochracea是一种地生兰科植物,也是中国传统中药材之一。以黄花白及为实验材料,对其叶和根组织中内生真菌类群组成及多样性进行分析。结果表明,从黄花白及植株的叶片和根组织块(段)中分离到可培养内生真菌140株,根据形态特征将它们鉴定为16个分类单元,其中包括10种子囊菌和6种担子菌。从叶片组织中分离的内生真菌有6种,其中刺盘孢属Colletotrichum真菌为叶片组织中的优势种类;从根组织中分离内生真菌有10种,瘤菌根菌属Epulorhiza和腊壳菌属Sebacina真菌构成了根组织内生真菌的优势类群。根组织可培养内生真菌的多样性(H′=1.968)要高于叶片组织(H′=1.459)。