黄栌根内生真菌分离鉴定及拮抗真菌筛选

采用常规分离法从不同地区健康黄栌根系中共分离得到33株内生真菌,其优势类群主要包括无孢菌群、腐质霉属、青霉属等,而且不同地区黄栌的根系内生真菌种类和数量有一定差异。内生真菌对黄栌枯萎病病原——大丽轮枝孢的拮抗试验结果表明:团炭角菌、黑乌霉属、假丝酵母属的菌株抑制作用较强,其它内生真菌菌株对大丽轮枝孢均有不同程度的抑制作用。     

长白山分解纤维素的瓶霉属和外瓶霉属真菌

瓶霉属、外瓶霉属真菌在自然界中广泛分布,是有着重要经济意义的一类真菌。迄今,我国已报道疣状瓶霉（Ｐｈｉａｌｏｐｈｏｒａ ｖｅｒｒｕｃｏｓａ）。裴氏瓶霉（Ｐｈ．ｐｅｄｒｏｓｏｉ）,棘状外瓶霉（Ｅｘｏｐｈｉａｌａ ｓｐｉｎｉｆｅｒａ）、皮炎外瓶霉（Ｅ．ｄｅｒｍａｔｉｔｉｄｉｓ）和甄氏外瓶霉（Ｅ．ｊｅａｎｓｅｌｍｅｉ＝Ｐｈ．ｇｏｕｇｅｒｏｔｉｉ）５种,均是分离自人体的病原真菌。在长白山自然保护区的原始林     

长白山不同植被带大型真菌多样性调查名录Ⅱ亚高山岳桦林带

根据2002年和2007—2009年间在长白山自然保护区岳桦林带所采集的标本,报道了长白山亚高山岳桦林带分布的大型真菌10目29科57属114种,包括中国新记录种5种,吉林省新记录种4种。采集地点包括长白山自然保护区北坡、西坡和南坡的岳桦林带。本名录共引证标本180余份,所有标本均为首次被引证,保藏于吉林农业大学菌物标本馆(HMJAU)。     

从长白山土壤中分离的3种弯颈霉属真菌(英文)

采用土壤稀释平板法从采自长白山自然保护区北坡森林的土壤样品中分离纯化出较多的弯颈霉属(Tolypocladium)真菌菌株,经形态观察鉴定出3个种,即雪白弯颈霉(T.niveum)、柱孢弯颈霉(T.cylindrospo-rum)及地生弯颈霉(T.geodes),其中(T.cylindrosporum)和(T.geodes)为中国新记录种。统计分析结果表明这3种真菌为喜冷真菌,表现出具有海拔分布特点。     

长白山分解纤维素的瓶霉属和外瓶霉属真菌

瓶霉属、外瓶霉属真菌在自然界中广泛分布,是有着重要经济意义的一类真菌。迄今,我国已报道疣状瓶霉（Phialophoraverrucosa）、裴氏瓶霉（Ph.pedrosoi）、棘状外瓶霉（Exophialaspinifera）、皮炎外瓶霉（E.dermatitidis）和甄氏外瓶霉（E.jeanselmei＝Ph．gougerotii）5种,均是分离自人体的病原真菌。在长白山自然保护区的原始林中,用生长锥取样器随机钻取腐朽林木髓心,在实验室进行分离培养。共鉴定出瓶霉属真菌2种,外瓶霉属真菌3种,其中美州瓶霉（Ph.americana、烂木瓶霉（Ph．richardsiae）和鲑外瓶霉（E.salmonis）为国内新记录种。     

云南元江干热河谷五种优势植物的内生真菌多样性

从蔓草虫豆（Atylosia scarabaeoides）、余甘子（Phyllanthus emblica）和黄花稔（Sida acuta）等5种云南元江干热河谷植物的525个组织块中,共分离得到内生真菌371株,内生真菌的分离频率在0.61～0.92之间,且所有植物叶内生真菌的分离频率都明显高于茎（P<0.05）。经形态学鉴定,内生真菌分属于拟茎点霉属（Phomopsis sp.）、离蠕孢属（Bipolaris sp.）和交链孢属（Alternaria sp.）等32个分类单元。拟茎点霉属为干热河谷植物优势内生真菌属,从所有被调查植物的茎叶中都分离得到该属真菌,且相对分离频率高达12.90%～50.54%。内生真菌群落组成的多样性和相似性分析结果表明,云南元江干热河谷植物内生真菌多样性偏低、宿主专一性较小。     

从长白山森林土壤中分离的茎点霉属(Phoma)真菌新记录种(英文)

长白山自然保护区是目前整个欧亚大陆生态系统保存较完整的森林生态系统之一,自然环境复杂多变,该保护区蕴藏着极其丰富的动物、植物及微生物资源。本研究从长白山自然保护区北坡森林土壤中分离获得1株茎点霉属(Phoma)真菌,经形态观察鉴定为淡黄茎点霉(P.flavescens),为我国首次报道。     

长白山森林土壤真菌区系研究

本研究对长白山自然保护区不同海拔、林型、土壤剖面的森林土壤真菌进行分离鉴定,从而为建立森林真菌资源库提供种质资源,并为森林生物资源管理提供科学依据。在该地区6个不同林型中,采用“之”形路线及土壤剖面取样法多点采集土壤样品,按照标准方法初步分离鉴定出真菌28个属,其中包括1个中国新纪录属,即球壳属(Sphaerodes)。研究结果表明,土壤真菌区系与森林(植被)类型、土壤剖面层次密切相关:美人松落叶松林、红松阔叶林和红松云冷杉林土壤真菌的种类明显多于鱼鳞云冷杉林、岳桦林和高山苔原带;土壤不同剖面层次上的真菌种群数量高低依次为落叶层>腐殖质层>土壤层;同一林型下不同土壤层面上优势菌属的组成也具有一定的差异,且不同层面上也存在着其特有的优势菌属。     

除虫菊内生真菌类群与分布的初步研究

为了解除虫菊内生真菌的种类和分布情况,采用常规分离方法,从除虫菊(Pyrethrum cinerariifolium Trev.)根、茎、叶、花中分离获得内生真菌共计128株,经形态观察,鉴定出12个属:镰孢霉属(Fusarium),毛壳菌属(Chaetomium),茎点霉属(Phoma),链格孢属(Alternaria),内多隔孢属(Endophragmiella),拟盘多毛孢属(Pestalotiopsis),腐殖霉属(Humicola),黑孢霉属(Nigrospora),韦氏孢属(Wiesneriomyces),色串孢属(Torula),阜孢霉属(Papularia),丝核菌属(Rhizoctonia),其中镰孢菌属、链格孢属、腐殖霉属为优势种群。除虫菊不同部位内生真菌的数量、分布、种群及其组成存在差异。     

烟草内生真菌多样性和种群结构

从云南“世界烟草品种园”10种烟草的根、茎、叶中分离得到199株内生真菌,根据rDNA-ITS系统发育分析鉴定为17属25种,其中格孢腔菌目Pleosporales内生真菌的种类和数量最多;茎点霉属Phoma、链格孢属Alternaria和镰孢菌属Fusarium为主要优势属,相对频率分别为25.1%、24.6%和11.6%,优势度Y值分别为0.251、0.172和0.104。烟草不同组织内生真菌的种群结构存在显著差异,分离自根的内生真菌的主要优势属为Fusarium和Phoma,Y值分别为0.235和0.123;分离自茎的内生真菌的主要优势属为Phoma和Alternaria,Y值分别为0.186和0.155;分离自叶片的内生真菌的主要优势属为Alternaria,Y值为0.286。Phoma从烟草根茎叶中均可分离得到,而Alternaria只分布在地上部茎叶中,Fusarium只分布在根茎中,表明这3个优势属真菌对根茎叶组织的专化性不同。     

Higher host plant specialization of root‐associated endophytes than mycorrhizal fungi along an arctic elevational gradient

How community‐level specialization differs among groups of organisms, and changes along environmental gradients, is fundamental to understanding the mechanisms influencing ecological communities. In this paper, we investigate the specialization of root‐associated fungi for plant species, asking whether the level of specialization varies with elevation. For this, we applied DNA barcoding based on the ITS region to root samples of five plant species equivalently sampled along an elevational gradient at a high arctic site. To assess whether the level of specialization changed with elevation and whether the observed patterns varied between mycorrhizal and endophytic fungi, we applied a joint species distribution modeling approach. Our results show that host plant specialization is not environmentally constrained in arctic root‐associated fungal communities, since there was no evidence for changing specialization with elevation, even if the composition of root‐associated fungal communities changed substantially. However, the level of specialization for particular plant species differed among fungal groups, root‐associated endophytic fungal communities being highly specialized on particular host species, and mycorrhizal fungi showing almost no signs of specialization. Our results suggest that plant identity affects associated mycorrhizal and endophytic fungi differently, highlighting the need of considering both endophytic and mycorrhizal fungi when studying specialization in root‐associated fungal communities.     

姜黄根茎内生真菌区系分析

目的了解姜黄植物根茎中内生真菌群落组成和生态分布规律。方法采用表面消毒法分别于春、冬季从姜黄植物根茎中分离内生真菌,通过形态特征和基于ITS序列的系统发育分析初步确定其分类地位。结果分离获得51株内生真菌(春季33株、冬季18株),初步鉴定其分别归于5个纲、7个目、8个科、8个属(Fusarium,Gibberel-la,Alternaria,Phomopsis,Diaporthe,Nectria,Botryosphaeria,Mucor),其中镰刀菌属(Fusarium,51.0%)和赤霉属(Gibberel-la,17.6%)为优势菌群;春季分离的内生真菌分属于7个属(分离率为82.5%),而冬季的内生真菌仅归入4个属(分离率为45.0%)。结论姜黄植物中内生真菌具有较丰富的物种和系统发育多样性,在类群组成和分布上存在季节性差异,某些内生真菌(Fusariumsp.)具有宿主偏好性。     

How are plant and fungal communities linked to each other in belowground ecosystems? A massively parallel pyrosequencing analysis of the association specificity of root-associated fungi and their host plants

In natural forests, hundreds of fungal species colonize plant roots. The preference or specificity for partners in these symbiotic relationships is a key to understanding how the community structures of root‐associated fungi and their host plants influence each other. In an oak‐dominated forest in Japan, we investigated the root‐associated fungal community based on a pyrosequencing analysis of the roots of 33 plant species. Of the 387 fungal taxa observed, 153 (39.5%) were identified on at least two plant species. Although many mycorrhizal and root‐endophytic fungi are shared between the plant species, the five most common plant species in the community had specificity in their association with fungal taxa. Likewise, fungi displayed remarkable variation in their association specificity for plants even within the same phylogenetic or ecological groups. For example, some fungi in the ectomycorrhizal family Russulaceae were detected almost exclusively on specific oak (Quercus) species, whereas other Russulaceae fungi were found even on “non‐ectomycorrhizal” plants (e.g., Lyonia and Ilex). Putatively endophytic ascomycetes in the orders Helotiales and Chaetothyriales also displayed variation in their association specificity and many of them were shared among plant species as major symbionts. These results suggest that the entire structure of belowground plant–fungal associations is described neither by the random sharing of hosts/symbionts nor by complete compartmentalization by mycorrhizal type. Rather, the colonization of multiple types of mycorrhizal fungi on the same plant species and the prevalence of diverse root‐endophytic fungi may be important features of belowground linkage between plant and fungal communities.     

锁阳及其寄主白刺内生真菌的生态分布及遗传关系

首次研究了寄生植物锁阳及其寄主白刺内生真菌的分布特征及遗传关系.采用组织块法分离天然白刺、寄生体中锁阳和白刺的内生真菌,利用ITS-rDNA分子序列并结合形态学方法鉴定菌种,研究内生真菌的分离率、定殖率、分离频率、多样性指数、均匀度指数及相似性系数等的差异,以及寄生关系中内生真菌的多样性、遗传关系及分布特征等.结果表明: 本次获得的49株内生真菌隶属于18个分类单元,95.9%为子囊菌,4.1%为担子菌;内生真菌总分离率为15.3%,总定殖率为25.0%;天然白刺中内生真菌Shannon多样性指数最大,为2.13;锁阳花序与锁阳茎的内生真菌相似性系数最大,为0.50;镰孢菌属为白刺的优势菌属,青霉属为锁阳的优势菌属.锁阳与白刺寄生体中真菌类群的差异性分布表明寄生关系对内生真菌群落存在一定影响.     

Assessment of diversity,distribution and antibacterial activity of endophytic fungi isolated from a medicinal plant <Emphasis Type="Italic">Adenocalymma alliaceum</Emphasis> Miers

A study was conducted for isolation, identification and antibacterial potential of fungal endophytes of Adenocalymma alliaceum Miers., (Bignoniaceae), a medicinal shrub vine plant which has long history for its usages in curing various disorders. A total of 149 isolates of endophytic fungi representing 17 fungal taxa were obtained from 270 segments (90 from each stem, leaf and petiole) of this plant. Hyphomycetes (77.85%) were the most prevalent, followed by Ascomycetes (8.05%) and Coelomycetes (4.03%) respectively. A considerable amount of fungal isolates was kept under (10.07%) Mycelia-Sterilia (MS). Leaf harboured maximum colonization of endophytic fungi (72.22%) which was greater than stem (67.78%) and petiole (25.54%). The Jc similarity index was maximum (0.619) between stem vs leaf followed by leaf vs petiole (0.571) and stem vs petiole (0.428). The dominant endophytic fungi were Alternaria alternata, Aspergillus niger, Stenella agalis, Fusarium oxysporum, Curvularia lunata and Fusarium roseum. Among twelve endophytic fungi tested for antibacterial activity, crude extracts of nine endophytic fungi (75%), showed antibacterial potential against one or more clinical human pathogens. Alternaria alternata, Curvularia lunata, Penicillium sp. and Chaetomium globosum exhibited significant antibacterial activity against 4 of 5 tested pathogens, showing broad spectrum activity. This investigation explains the value of sampling from different tissues of a host plant for the greater species diversity, and additionally, the antibacterial screening of some endophytic fungi from this specific medicinal plant may represent a unique source for many of the useful antibacterial compounds.     

罗汉果内生真菌多样性研究

该文采用传统形态学方法结合r DNA-ITS序列分析,对我国重要药用植物罗汉果中的内生真菌进行了鉴定并研究其多样性。结果表明:采用组织培养法从罗汉果健康植株中共分离得到150株内生真菌,包括罗汉果中雌株的内生真菌96株、雄株的内生真菌54株。122株内生真菌经形态学结合r DNA-ITS序列分析鉴定为9个属,均归属为子囊菌门,包含座囊菌纲(Dothideomycetes)和子囊菌纲(Sordariomycetes)。其中,座囊菌纲(Dothideomycetes)真菌包含3科、3属;子囊菌纲(Sordariomycetes)真菌包含6科、6属。优势属为刺盘孢属(Colletotrichum)和镰刀菌属(Fusarium)。罗汉果雌、雄植株不同组织中内生真菌的定殖率及分离率的变化规律均不相同,雌株中以根中内生真菌的定殖率和分离率最高,叶片中的最低;在雄株中以叶片中的定殖率和分离率最高,根中的最低。不同菌株在雌、雄两种植株的不同组织中的分布均不同,结合内生真菌群落组成的相似性比较结果,表明部分内生真菌对罗汉果雌株和雄株,以及同一植株中的不同组织均具有偏好性。不同组织中内生真菌的多样性指数在0.11~0.69的范围,其中雌株根部的内生真菌多样性指数最高。以上研究结果为后期探究内生真菌与罗汉果互作关系奠定了基础。     

云南三个地区马铃薯内生真菌多样性研究

为研究云南马铃薯(Solanum tuberosum)内生真菌的多样性,该文以采自云南省德宏芒市、大理喜洲和临沧双江3个地区的马铃薯植株为研究对象,采用组织块分离法、尖端菌丝挑取法对马铃薯根、茎及块茎中的内生真菌进行分离纯化,并采用形态学鉴定方法和ITS序列分析法对分离得到的内生真菌进行鉴定,对内生真菌的定殖率、分离率及多样性指数进行计算和分析。结果表明:(1)共分离得到内生真菌98株,其中从德宏芒市的样品中获得40株,从大理喜洲的样品中获得27株,从临沧双江的样品中获得31株。(2)经鉴定,分离得到的马铃薯内生真菌共涵盖10目10科13属,大多为子囊菌门和担子菌门,优势菌为镰刀菌属(Fusarium)和青霉属(Penicillium),褶皱裸孢壳(Emericella rugulosa)、接骨木镰刀菌(Fusarium sambucinum)、毛韧革菌(Stereum hirsutum)、Psathyrella sulcatotuberculosa和Epicoccum catenisporum 5种真菌均为首次从马铃薯植株中分离得到。(3)马铃薯块茎内生真菌的定殖率最高,根部内生真菌定殖率最低; 内生真菌的分离率以马铃薯根部为最高,而茎部最低; 不同组织中内生真菌的多样性指数趋势均为根>块茎>茎。从综合来看,云南马铃薯植株中的内生真菌具有较高的多样性,不同地区的马铃薯样品中内生真菌优势菌不同,马铃薯根部具有最丰富的内生真菌种群和最高的分离率,是最适合进行内生真菌分离的材料。该研究结果为后期探究马铃薯内生真菌对病原菌的拮抗作用奠定了基础,也为马铃薯内生真菌多样性研究提供了参考数据。     

云南会泽铅锌矿废弃矿渣堆常见植物内生真菌多样性

从云南会泽铅锌矿废弃矿渣堆上的常见植物硬毛南芥(Arabis hirsuta)、毛萼香茶菜(Rabbosia eriocalyx)和倒挂刺(Rosa longicuspis)等6种植物的690个组织块中共分离得到内生真菌495株,内生真菌的分离频率在0.42—0.93之间,平均为0.72,所有植物茎内生真菌的分离频率都明显高于叶(P<0.05)。经形态学鉴定,内生真菌分属于茎点霉属(Phoma)、交链孢属(Alternaria)和派伦霉属(Peyronellaea)等20个分类单元,其中茎点霉属和派伦霉属为该废弃矿渣堆上常见植物的优势内生真菌属。6种植物内生真菌的多样性指数在1.05—2.29之间,与其它非重金属污染环境植物内生真菌的多样性指数相似,说明在重金属污染地区仍然存在多种重金属耐受的内生真菌种类。6种植物内生真菌的相似性系数(0.455—0.833)表明,会泽铅锌矿区植物内生真菌的宿主专一性较小。     

Effect of different root endophytic fungi on plant community structure in experimental microcosms

Understanding the effects of root‐associated microbes in explaining plant community patterns represents a challenge in community ecology. Although typically overlooked, several lines of evidence point out that nonmycorrhizal, root endophytic fungi in the Ascomycota may have the potential to drive changes in plant community ecology given their ubiquitous presence, wide host ranges, and plant species‐specific fitness effects. Thus, we experimentally manipulated the presence of root endophytic fungal species in microcosms and measured its effects on plant communities. Specifically, we tested whether (1) three different root endophyte species can modify plant community structure; (2) those changes can also modified the way plant respond to different soil types; and (3) the effects are modified when all the fungi are present. As a model system, we used plant and fungal species that naturally co‐occur in a temperate grassland. Further, the soil types used in our experiment reflected a strong gradient in soil texture that has been shown to drive changes in plant and fungal community structure in the field. Results showed that each plant species responded differently to infection, resulting in distinct patterns of plant community structure depending on the identity of the fungus present. Those effects depended on the soil type. For example, large positive effects due to presence of the fungi were able to compensate for less nutrients levels in one soil type. Further, host responses when all three fungi were present were different from the ones observed in single fungal inoculations, suggesting that endophyte–endophyte interactions may be important in structuring plant communities. Overall, these results indicate that plant responses to changes in the species identity of nonmycorrhizal fungal community species and their interactions can modify plant community structure.