云南松外生菌根真菌分离培养研究

就滇中一带云南松林进行了多次调查,共采集、鉴定标本130余号;从中选取13科, 19属, 33种真菌进行了菌种分离、培养的研究。选用并修改4种培养基进行了外生菌根真菌分离、培养的比较,结果共有11科, 17属, 25种真菌分离获得成功,成功率高达75.8%, 4种培养基的分离成功率依次为36.4%、63.6%、33.3%和57.6%。其中,修改PDA培养基和松针、玉米培养基较适宜于云南松外生菌根真菌的分离、纯化和培养。     

西藏昆诺阿藜种子可培养内生真菌多样性

探究西藏不同地区昆诺阿藜种子内生真菌的物种组成、多样性等群落结构特征,丰富昆诺阿藜种子相关微生物的可利用资源,对于微生物资源利用和昆诺阿藜病害的生物防控具有重要意义。本研究收集西藏不同海拔3个地区(日喀则、拉萨和林芝)的昆诺阿藜种子,利用传统分离方式进行了真菌的培养。从46份昆诺阿藜种子中共分离出947株真菌,经ITS序列分析结合形态观察鉴定为1门9目12科26属77种。所有的真菌都归类为子囊菌门Ascomycota,菌株数量相对丰度最高的3个属为链格孢属Alternaria (40.2%)、镰刀菌属Fusarium (17.4%)和茎点霉属Phoma (13.9%)。按照多样性指数,日喀则地区的昆诺阿藜内生可培养真菌的多样性明显高于拉萨和林芝两个地区。     

小麦种子内携真菌多样性研究

为揭示小麦种子所携带真菌的种类和多样性,丰富小麦相关微生物的可利用资源,我们对来自俄罗斯、澳大利亚、哈萨克斯坦、加拿大以及中国的57份小麦种子样品进行内携真菌分离并对其中的21份种子利用高通量测序进行多样性分析.结果显示,分离获得564株真菌,经ITS序列分析结合形态观察鉴定为31属69种,其中菌株数量相对多度最高的5...     

一株南海指海绵共附生真菌多样性及其抗菌活性

本研究通过采用6种不同的培养基对南海指海绵上的共附生真菌进行平板涂布法分离培养,结合ITS-rDNA基因序列特征和形态学特征进行鉴定来研究海绵共附生真菌多样性。共分离培养出55株(21种)真菌菌株,优势菌属为木霉菌属Trichoderma sp.(31株),其中的哈茨木霉Trichoderma harzianum 22株作为优势菌种,占真菌菌株总数的40%。其余真菌则属于枝孢菌属Cladosporium、迈耶氏酵母属Meyerozyma、茎点霉属Phoma、红酵母属Rhodotorula、青霉菌属Penicillium、曲霉属Aspergillus等10个属,还有两株未鉴定到种的子囊菌。选取金黄色葡萄球菌Staphylococcus aureus等7种病原指示菌,采用琼脂扩散法对21种真菌的代表性菌株进行抗菌活性筛选,其中有3种真菌至少对一种指示菌表现出抑菌效果,占菌株种类总数的11%。     

辽宁地区药用植物根际土壤真菌多样性的研究

作者于2007和2008年的5-10月间在辽宁省的丹东、凤城、清原、桓仁等药材产区采集土壤样品200余份,通过采用稀释平板法和土壤颗粒平板法进行土壤真菌的分离和培养,共分离和鉴定出27属54种真菌,其中接合菌2属3种、子囊菌1属1种、无性型真菌24属50种。分离到的部分真菌种类是重要的药用植物病原菌,如Cylindrocarpon destructans、Fusarium oxysporum和F.solani。研究发现,Penicillium paxilli、P.expansum、Trichoderma atroviride及T.viride的分离频率最高,是辽宁省4个调查地区药用植物根际土壤真菌的优势菌群。     

供给天麻种子萌发营养的真菌——紫萁小菇

从天麻种子发芽的原球茎中,共分离到12种可供给天麻种子萌发营养的真菌,其中最优良的01号菌株,诱导培养出子实体,经鉴定为紫萁小菇(Mycena osmundicola Lange),属国内新纪录。用01号菌株及紫萁小菇培养的染菌树叶,伴播天麻种子都可发芽,经酯酶同工酶凝胶电泳分析,二者酶带条数、迁移率(RF值)基本相同,证明鉴定结果可靠(标本保存在医科院药植所真菌研究室)。     

东北地区保护地土壤真菌多样性研究I

20 0 3年4月至2 0 0 4年7月在辽宁、吉林、黑龙江三省的2 0多个市县共采集不同年限的保护地土样15 0多份,采用稀释平板法在孟加拉红培养基上分离近千个真菌菌株,通过纯化培养和形态鉴定,将各菌株鉴定到种。本文报道了部分可培养的子囊菌和有丝分裂孢子真菌的研究结果,其中子囊菌5属6种,包括两个中国新记录种(Lophotrichusbrevirostratus和Microascuscinereus)和一个国内记录但未正式发表的子囊菌(Emericel lopsisterricola) ;有丝分裂孢子真菌7属10种。     

温室黄瓜叶部微生物区系

采用稀释分离法和消毒叶片研磨液培养法对温室黄瓜叶围和内生微生物进行了分离,共分离到248个菌株,初步鉴定出13个属的叶围真菌,其中链格孢属(Alternaria)和青霉属(Penicillium)真菌为优势类群;鉴定出4个属的内生真菌,其中曲霉属(Aspergillus)真菌为优势类群;10个属的叶围细菌,其中芽孢杆菌属(Bacillus)和黄单胞菌属(Xanthomonas)细菌为优势类群;6个属的内生细菌,其中芽孢杆菌属和假单胞菌属(Pseudomonas)细菌为优势类群;6个属的叶围酵母菌,其中隐球酵母属(Cryptococcus)为优势类群;已鉴定出2个属的叶围放线菌,分别为链霉菌属(Streptomyces)和小多孢菌属(Micropolpspora).未分离到内生酵母菌和放线菌.     

新疆特色植物恰玛古块根内生真菌的分离与鉴定

植物体的内生真菌与其自身生长及生物防御均有密切关系,因此对健康恰玛古块根组织进行内生真菌分离与纯化,利用形态学结合分子鉴定方法对内生真菌群落多样性进行了初步研究。从恰玛古块根中共分离到可培养内生真菌29株,鉴定结果分属于8个属的11个种,其中支顶孢属(Acremonium sp.)和青霉属(Penicillium sp.)为优势属,分别占总菌株数的34.48%和27.59%。为国内外首次对新疆特色药用植物恰玛古内生真菌进行分离研究,并对其分类地位进行了确定。     

从麦草浆污泥中分离产木聚糖酶的木质纤维降解真菌

采用涂布平板法在马丁氏培养基平板上对麦草浆污泥作真菌分离研究,共分离纯化获得27株真菌,其中青霉属(Penicillium)、曲霉属(Aspergillus)、麦轴梗霉属(Tritirachium)根霉属(Rhizopus)和毛霉属(Mucor)为其优势菌群;在麦芽提取物-OPP培养基平板上分离得到10株真菌,其中拟指突孢曲霉属(Emercellopsis)为其优势菌群。通过对麦草浆污泥连续富集培养得到真菌10株,经初步鉴定为曲霉属(Aspergillus)和青霉属(Penicillium)。对富集培养分离的菌种进行木聚糖酶产酶特性研究,结果表明:最优菌株为Aspergillussp.MC-JAⅡ,其木聚糖酶酶活性最高值达到2060 U/mL,发生在产酶培养的第84 h,相应纤维素酶活为99.5 U/mL。为国内首次报道麦草浆污泥中的降解木质纤维真菌的分离、鉴定和产酶研究。     

Diversity and Antimicrobial Activity of Culturable Fungi Isolated from Six Species of the South China Sea Gorgonians

Fungi in gorgonians are now known to cause gorgonian diseases, but little attention has been paid to the nature of fungal communities associated with gorgonians. The diversity of culturable fungi associated with six species of healthy South China Sea gorgonians were investigated using a culture-dependent method followed by analysis of fungal internal transcribed spacer sequences. A total of 121 fungal isolates were recovered and identified using the Basic Local Alignment Search Tool search program. These belonged to 41 fungal species from 20 genera. Of these, 30 species and 12 genera are new reports for gorgonians, and the genera Aspergillus and Penicillium were the most diverse and common in the six gorgonian species. Comparison of the fungal communities in the six gorgonian species, together with results from previous relevant studies, indicated that different gorgonian species and the same gorgonian species living in different geographic locations had different fungal communities. The gorgonian Dichotella gemmacea harbored the most fungal species and isolates, while Echinogorgia aurantiaca had the least fungal diversity. Among the six media used for fungal isolation, potato glucose agar yielded the highest isolates (27 isolates), while glucose peptone starch agar had the best recoverability of fungal species (15 species). The antimicrobial activity of the 121 fungal isolates was tested against three marine bacteria and two marine gorgonian pathogenic fungi. A relatively high proportion (38?%) of fungal isolates displayed distinct antibacterial and antifungal activity, suggesting that the gorgonian-associated fungi may aid their hosts in protection against pathogens. This is the first report comparing the diversity of fungal communities among the South China Sea gorgonians. It contributes to our knowledge of gorgonian-associated fungi and further increases the pool of fungi available for natural bioactive product screening.     

Diversity and distribution of cultured endolichenic fungi in the Ny-Ålesund Region,Svalbard (High Arctic)

Endolichenic fungi within 17 lichen species in the area near Ny-Ålesund (Svalbard, High Arctic) were studied by a culture-based method. The 247 fungal isolates were obtained from 2712 lichen thallus segments. The colonization rate of endolichenic fungi ranged from 1.6 to 26.5 %, respectively. These isolates were identified to 40 fungal taxa, including 35 Ascomycota (10 orders), 4 Basidiomycota (3 orders), and 1 unidentified fungus. Thelebolales was the most abundant order, while Sordariales were the most diverse order. The common fungal taxa shared by more than 3 lichen species were Thelebolus microsporus (93 isolates), Coniochaeta hoffmannii (7 isolates), Sarocladium kiliense (33 isolates), Coniochaeta sp. 1 (5 isolates), Coniochaeta sp. 4 (28 isolates), and Coniochaeta sp. 2 (5 isolates). Low Sorenson’s similarity coefficients were observed among different lichen species, indicating that host-related factor may shape the endolichenic fungal communities in this region. In addition, no endolichenic fungal taxa were previously found in the Antarctica and Austrian Alps, suggesting endolichenic fungal communities in this region might be also shaped by the Arctic climate. The results demonstrate the existence of specific cultured endolichenic fungal species, which may be suitable objects for further study of their possible functional roles in the lichen thalli.     

分离自冬虫夏草可培养真菌的多样性研究

冬虫夏草是生长于青藏高原的一种名贵中药材。天然冬虫夏草及其微环境中生活着多种真菌。作者使用常规分离培养方法对冬虫夏草的真菌区系进行研究。从天然冬虫夏草的子座、菌核和菌膜3个部位共分离到572个真菌菌株,并根据形态特征将大部分菌株鉴定到37个不同的属。这些菌株经SSCP(single-strand conformation polymorphism)分析后,再根据nrDNAITS序列的相似性(以97%为阈值)共区分出92种不同的分类单元(operational taxonomic unit,OTU)。其中,属于子囊菌的菌株数及OTU数均比接合菌和担子菌多。从菌膜分离的菌株数及OTU数都明显多于子座和菌核。分离自子座的优势真菌是产黄青霉Penicillium chrysogenum,而分离自菌核和菌膜的优势真菌均为玫红假裸囊菌Pseudogymnoascus roseus。尚未最终鉴定的部分真菌可能为新的真菌物种。     

烟草根际土壤真菌多样性的研究

烟草土传病害发生普遍,危害严重。为深入了解烟草根际土壤真菌区系与土传病害间的相互关系,选取黑龙江、山东、广西和云南四省区的代表性烟田,开展了烟草根际土壤真菌多样性的研究。对采自上述四省区12个县市的224份烟草根际土壤样品进行了真菌的分离和培养,共分离获得1,278株真菌,通过形态学特征鉴定出真菌25属86种。其中接合菌门4属6种,子囊菌门1属1种,无性型真菌20属79种。通过分析发现,烟草根际土壤中的优势真菌种群及其菌株数量所占分离真菌总菌株数量的比例分别为：青霉属Penicillium 16.74%,木霉属Trichoderma 14.32%,镰孢菌属Fusarium 13.54%和曲霉属Aspergillus 11.50%。在12个采样地点中,山东诸城的多样性指数（H￠=2.0666）和均匀度指数（J=0.6898）最高,云南玉溪的丰富度指数（R=4.0687）最高。不同采样地点的烟草根际土壤真菌种群的相似性水平也存在差异,其中云南永胜和大理的烟草根际土壤真菌种群的相似性系数最高（Cj=0.9375）,山东沂水和云南玉溪间的相似性最低（Cj=0.5217）。     

Characterization of novel <Emphasis Type="Italic">Trichoderma</Emphasis> spp. isolates as a search for effective biocontrollers of fungal diseases of economically important crops in Argentina

Monoconidial cultures of 33 isolates of Trichoderma from Buenos Aires Province, Argentina were characterized on the basis of twenty eight morphological, physiological and biochemical features. All of them were screened for proteinase, endochitinase and β-1,3 glucanase activity. Universally primed PCR (UP-PCR) and inter-simple sequence repeat (ISSR) techniques were used to examine the genetic variability among isolates, which resulted in 127 bands for the total number of isolates. These results were subjected to numerical analysis revealing 20 haplotypes grouped in five clusters. The ability of Trichoderma isolates to antogonize soil-borne fungal plant pathogens using a dual culture assay was done against five fungal species: Alternaria sp., Bipolaris sorokiniana, Fusarium graminearum, F. solani, and Pyricularia oryzae. The highest inhibition values (85% RI) were obtained against B. sorokiniana and P. oryzae. Three isolates of T. harzianum named as FCCT2, FCCT3 and FCCT9 were capable of causing a high growth inhibition on four of the fungal species assayed, which was in agreement with their higher extracellular hydrolytic activity. Our results suggest that these isolates have the potential to be effective agents for biocontrol of cereal and tomato fungal pathogens.     

Evidence for specificity of cultivable bacteria associated with arbuscular mycorrhizal fungal spores

Bacteria associated with arbuscular mycorrhizal (AM) fungal spores may play functional roles in interactions between AM fungi, plant hosts and defence against plant pathogens. To study AM fungal spore-associated bacteria (AMB) with regard to diversity, source effects (AM fungal species, plant host) and antagonistic properties, we isolated AMB from surface-decontaminated spores of Glomus intraradices and Glomus mosseae extracted from field rhizospheres of Festuca ovina and Leucanthemum vulgare. Analysis of 385 AMB was carried out by fatty acid methyl ester (FAME) profile analysis, and some also identified using 16S rRNA gene sequence analysis. The AMB were tested for capacity to inhibit growth in vitro of Rhizoctonia solani and production of fluorescent siderophores. Half of the AMB isolates could be identified to species (similarity index 0.6) within 16 genera and 36 species. AMB were most abundant in the genera Arthrobacter and Pseudomonas and in a cluster of unidentified isolates related to Stenotrophomonas. The AMB composition was affected by AM fungal species and to some extent by plant species. The occurrence of antagonistic isolates depended on AM fungal species, but not plant host, and originated from G. intraradices spores. AM fungal spores appear to host certain sets of AMB, of which some can contribute to resistance by AM fungi against plant pathogens.     

Isolation of fungi from bats of the Amazon basin

A total of 2,886 bats captured in the Amazon Basin of Brazil were processed for the isolation of fungi. From the livers, spleens, and lungs of 155 bats (5.4%), 186 fungal isolates of the genera Candida (123 isolates), Trichosporon (26 isolates), Torulopsis (25 isolates), Kluyveromyces (11 isolates), and Geotrichum (1 isolate) were recovered. Seven known pathogenic species were present: Candida parapsilosis, C. guilliermondii, C. albicans, C. stellatoidea, C. pseudotropicalis, Trichosporon beigelii, and Torulopsis glabrata. Twenty-three culture-positive bats showed identical fungal colonization in multiple organs or mixed colonization in a single organ. The fungal isolation rates for individual bat species varied from 1 fungus per 87 bats to 3 fungi per 13 bats, and the mycoflora diversity for members of an individual fungus-bearing bat species varied from 16 fungi per 40 bats to 7 fungi per 6 bats. Of the 38 fungal species isolated, 36 had not been previously described as in vivo bat isolates. Of the 27 culture-positive bat species, 21 had not been previously described as mammalian hosts for medically or nonmedically important fungi.     

Isolation of fungi from bats of the Amazon basin.

A total of 2,886 bats captured in the Amazon Basin of Brazil were processed for the isolation of fungi. From the livers, spleens, and lungs of 155 bats (5.4%), 186 fungal isolates of the genera Candida (123 isolates), Trichosporon (26 isolates), Torulopsis (25 isolates), Kluyveromyces (11 isolates), and Geotrichum (1 isolate) were recovered. Seven known pathogenic species were present: Candida parapsilosis, C. guilliermondii, C. albicans, C. stellatoidea, C. pseudotropicalis, Trichosporon beigelii, and Torulopsis glabrata. Twenty-three culture-positive bats showed identical fungal colonization in multiple organs or mixed colonization in a single organ. The fungal isolation rates for individual bat species varied from 1 fungus per 87 bats to 3 fungi per 13 bats, and the mycoflora diversity for members of an individual fungus-bearing bat species varied from 16 fungi per 40 bats to 7 fungi per 6 bats. Of the 38 fungal species isolated, 36 had not been previously described as in vivo bat isolates. Of the 27 culture-positive bat species, 21 had not been previously described as mammalian hosts for medically or nonmedically important fungi.     

Random amplified polymorphic DNA markers reveal genetic variation in the symbiotic fungus of leaf-cutting ants

RAPD markers were used to examine the degree of genetic variation within the putatively asexual basidiomycete fungus (Lepiotaceae: provisionally named Leucoagaricus gongylophorus) associated with the leaf-cutting ant species Atta cephalotes. We analyzed fungal isolates from ant nests in two geographically distant sites, two isolates from Panama and five isolates from Trinidad. Ten decamer primers were used to amplify total DNA from these seven fungal isolates, and RAPD banding patterns were compared. Genetic similarity among isolates was determined by pair-wise comparisons of the shared number of DNA bands on an agarose gel. There was considerable genetic variation among isolates of the symbiotic fungus even within sites. Pairs of fungal isolates from the two different sites shared an average of only 36% of the bands in their RAPD profiles, while pairs from the within sites shared an average of 72% of the bands. RAPD markers may be useful for further investigation of the genetic structure of the fungal symbiont within species of leaf-cutting ants.