中国大陆地区丛枝菌根真菌菌种资源的分离鉴定与形态学特征

【目的】分离收集保藏中国大陆各个地区不同生态环境的丛枝菌根真菌菌种资源,为丛枝菌根的研究提供资源、奠定基础。【方法】以高粱为宿主植物,采用诱导培养、单孢培养和扩繁培养分离土壤样品中的丛枝菌根真菌菌种并鉴定。【结果】从我国大陆的45个地区50余种宿主植物根区土壤中分离到丛枝菌根真菌135株,隶属于23个种;对各个菌株的形态特征进行了描述。【结论】我国蕴藏着丰富的丛枝菌根真菌菌种资源,文中描述的菌种资源是目前从我国大陆地区获得的种类和数量最多、覆盖范围最广的AM真菌菌种资源。     

球囊菌门丛枝菌根真菌最新分类系统菌种名录

球囊菌门Glomeromycota是菌物界(Kingdom of Fungi)晚近新增加的一个门,下设1纲4目11科27属约300种丛枝菌根(AM)真菌,均是植物根系重要的共生真菌。同其他真菌分类研究过程相似,随着新种的不断发现、分类技术的进步与研究的深入,AM真菌分类系统及其菌种学名经历着持续不断的变更。然而,这些变动给AM真菌研究者造成了一定的困扰和混乱。本文在AM真菌系统发育分类重建的基础上,结合当前国际上AM真菌分类的最新进展,规范、正确并一致描述全球已知的AM真菌菌种拉丁文和中文学名,以纠正错误和统一AM真菌中文学名;同时标注了中国新记录种、新种和种质资源保藏种名录,以期促进中国AM真菌分类、资源多样性、群落结构及其相关研究。     

丛枝菌根真菌种质资源库

“中国丛枝菌根真菌种质资源库”（Bank of Glomales in China英文缩写BGC）是国家自然科学基金（30070028）资助,北京市农林科学院植物营养与资源研究所承担,中国农业大学植物营养系协作,2003年建成的拥有我国自主知识产权的丛枝菌根真菌资源库。     

保藏条件对丛枝菌根真菌Glomus mosseae侵染的影响

以高粱为宿主植物,接种丛枝菌根真菌Glomus mosseae,测定了经不同保藏温度(18~20,4,-20,-80℃)处理,在不同保藏时间(3,6,12,18,24,30个月)下丛枝菌根真菌G.mosseae对根系的侵染率。结果表明:丛枝菌根真菌G.mosseae在18~20,4,-20℃温度下保藏12~18个月时,需进行转接,用高粱为宿主植物进行更新复壮培养,以保持菌种活力。     

丛枝菌根结构与功能研究进展

丛枝菌根(arbuscular mycorrhiza,AM)是陆地生态系统中分布最广泛、最重要的互惠共生体之一,对提高植物抗逆性、修复污染生境、保持生态系统稳定与可持续生产力的作用显著.AM结构特征是判断菌根形成的主要指标,与其功能密切相关.本文总结了AM丛枝结构、泡囊结构、菌丝结构和侵入点结构等发育特征;分析了A型丛枝结构、P型丛枝结构、泡囊结构和根外菌丝结构与促进寄主植物养分吸收和生长、提高植物抗旱性、耐涝性、耐盐性、抗高温、拮抗病原物、提高植物抗病性、抗重金属毒性、分解有毒有机物、修复污染与退化土壤等功能的关系,及其所发挥的重要作用;探讨了影响AM结构与功能的因子,以及基于AM不同结构所发挥功能的作用机制.旨在为系统研究AM真菌发育特征、AM真菌效能机制,以及评价和筛选AM真菌高效菌种提供依据.     

AM真菌遗传多样性研究进展

丛枝菌根（arbuscular mycorrhiza,AM）真菌是一类专性共生多核生物,至今尚未获得纯培养,与植物根系共生后才能完成其生活史。该类真菌无性繁殖,具有独特的遗传特性,属于真菌界球囊菌门（Glomeromycota）,共有200余种。研究发现AM真菌种群间以及种群内,甚至单一孢子内都存在大量基因变异,表明该类真菌具有丰富的遗传多样性。本文总结了近年来有关AM真菌遗传多样性方面的研究进展,并讨论了存在的问题。     

目前AM真菌培养特性方面研究的基本概况

从盆钵培养,双相培养和纯培养3个层次讨论了当前探索丛枝菌根（AM）真菌培养特性的研究现状,并探讨了当前培养AM真菌的动向与展望。     

Arbuscular mycorrhizal fungi protect a subtropical tree species exposed to simulated acid rain by accelerating photosynthetic ability,antioxidant enzymes and osmolyte accumulation

丛枝菌根真菌对其宿主光合能力、抗氧化酶和渗透物质积累的促进作用 及其抗酸雨机制的探讨 酸雨在中国南方发生频繁,对亚热带树种生长具有明显抑制作用。以往研究表明,丛枝菌根真菌(AM真菌)可以缓解酸雨对宿主植物的胁迫效应。榉树(Zelkova serrata)为中国南方主要经济树种之一,其如何与共生AM真菌协同、增强其抗酸雨胁迫的能力是本项研究所要探讨的关键科学问题。通过温室控制实验,将榉树幼苗随机接受4个水平的AM真菌接种处理(接种灭菌菌种;单独接种Rhizophagus intraradices;单独接种Diversispora versiformis;接种这两种菌种的混合菌种)和3个pH水平(pH2.5、pH4.0和pH5.6)的硫酸型酸雨和硝酸型酸雨处理组成的12个处理组合,同时测定其生长、光合性能、抗氧化酶、渗透调节和土壤酶的响应格局。研究发现酸雨处理显著降低了非菌根榉树幼苗的总干重、总叶绿素含量、叶片净光合速率和可溶性蛋白的含量;接种AM真菌,特别是接种混合菌种,显著提高了强酸胁迫下榉树幼苗的总干重、光合性能、丙二醛、过氧化物酶、超氧化物歧化酶、可溶性蛋白和根系酸性磷酸酶活性。此外,菌根效应依赖于AM真菌的种类和酸胁迫的梯度。本研究 结果表明,AM真菌对榉树幼苗抗酸胁迫的调控作用主要源于调节宿主植株光合能力、抗氧化酶和渗透物质的积累。榉树与其共生AM真菌在应对酸胁迫上协同机制的解析为该树种在中国南方酸雨区的栽培提供理论基础、具有重要的实践指导意义。     

丛枝霸王（Zygophyllum dumosum）根际AM真菌生态学研究

AM真菌是一类广泛分布的土壤真菌,与宿主植物形成共生结构后,对于植物生长和植被恢复有多种有益的生理学和生态学作用。1999年11月至2000年10月,通过每月分别从0－10cm和10－20cm土层采集根际土样,对以色列荒漠地区丛枝霸王（Zygophyllum dumosum Boiss）根际AM真菌进行了系统的生态学研究。AM真菌的分布和定殖与季节变化和采样浓度密切相关。菌根真菌的最高定殖率并不伴随有最大的孢子密度,最高的定殖率发生在1999年11月,而最大的孢子密度则出现在2000年9月。10－20cm土层中的菌根真菌定殖率和孢子密度明显高于0－10cm土层。土壤温度与所有菌根结构的定殖率呈正相关,土壤有机质含量与泡囊和丛枝定殖率呈正相关,而土壤总可溶性氮对泡囊和丛枝定殖有显著正效应,对孢子密度有显著负效应。结果建议,孢子密度、泡囊和丛枝定殖程度可作为检测荒漠土壤状况的生态指标。研究应用于我国特别是西部荒漠地区的植被建立和恢复,可望发挥重任作用：（1）AM真菌能与绝大多数高奶系形成共生联合体,促进根系对土壤矿质营养和水分的吸收,提高植物对干旱、高温、盐碱、根部病害等的抗性,提高逆境条件下植物的成活率,深入研究荒漠生态系统中AM真菌动态分布,以及筛选优势AM菌种和人工接种,进行菌根化育苗,为植被建立和恢复提供优质苗木;（2）菌根的丛枝定殖时间短,主要发生在幼根,泡囊定殖时间长,主要发生在老根,而AM真菌的生长发育和繁殖所需的碳水化合物来自植物根系的分泌活动,所以,通过检测不同时期菌根各种结构的定殖程度和孢子的丰富度,可以获得宿主植物根系的生长状况,进而对土壤环境作出科学的评估。     

西双版纳地区龙脑香科植物AM真菌的初步研究

 对云南省西双版纳地区17种龙脑香科树种根系丛枝菌根（Arbuscular mycorrhiza, AM）真菌的定居情况进行了调查,并对根围土壤中AM真菌进行了分离和鉴定。结果表明,调查根样均有不同程度的菌根感染,感染率最高可达40%,调查揭示了西双版纳地区龙脑香科植物在自然条件下可形成丛枝菌根。初步从龙脑香科植物根际土壤中分离、鉴定出32种AM真菌,隶属于无梗囊霉属（Acaulospora）、球囊霉属（Glomus）、原囊霉属（Achaeospora）、拟球囊霉属（Paraglomus）和盾巨孢囊霉属（Scutellospora）,其中,无梗囊霉属和球囊霉属真菌为西双版纳地区龙脑香科植物AM真菌优势类群。     

Can intra‐specific genetic variation in arbuscular mycorrhizal fungi (Glomus etunicatum) affect a mesophyll‐feeding herbivore (Tupiocoris notatus Distant)?

Abstract.  1. Arbuscular mycorrhizal fungal (AMF) infection can have negative, positive or neutral effects on insect herbivore populations, but patterns are difficult to predict.
2. Intra-specific genetic variation in nutrient uptake ability between fungal isolates may also have indirect effects on insect herbivores due to changes in plant quality. In preliminary studies mirid ( Tupiocoris notatus ) populations were significantly reduced on tobacco ( Nicotiana rustica ) colonised by AMF but it was unknown if same-species fungal isolates differed in their effect.
3. An experiment was performed as a first test of the effect of intra-specific genetic variation in the mycorrhizal fungus Glomus etunicatum on mirid nymphal population structure, dynamics, and growth rate.
4. Mirid nymphal populations were lower on mycorrhizal fungal-infected plants. Population size, however, did not differ between the mycorrhizal isolates. While no statistical difference in population between isolates was found, one isolate consistently had 1.7–2.4 times lower mirid populations compared with the controls, indicating that the magnitude of effect is different between mycorrhizal isolates.
5. The significantly negative effect of AMF on mirid populations likely resulted from AMF-induced changes in plant quality (e.g. increased defence). This study lends further support to recent demonstrations that below-ground symbionts significantly influence above-ground processes. In addition, mycorrhizal fungi can affect insect population structure, which may have consequences for future herbivory.     

A 60-year journey of mycorrhizal research in China: Past,present and future directions

The significance of mycorrhizas (fungal roots in 90% of land plants) in plant nutrient acquisition and growth, element biogeochemical cycling and maintaining of terrestrial ecosystem structures has been globally established for more than 120 years. Great progress in mycorrhizal research in the past 60 years (1950–2009, 1981–2009 in particular) has also been made across China, particularly in the mainland, Hong Kong and Taiwan. For instance, a total of 20 new and ∼120 records of arbuscular mycorrhizal (AM) fungal species, 30 new and ∼800 records of ectomycorrhizal (EM) fungal species, a dozen of new and ∼100 records of orchid mycorrhizal (OM) fungal species have been isolated by morphological observation and/or molecular identification in China since the 1950s. Great accomplishment has also been made in the following area, including fungal species richness and genetic structure, relationships between species composition and plant taxa, effects of mycorrhizal fungi on plant nutrient uptake and growth, resistances to pathogens and interactions with other soil microorganisms, potential of mycorrhizal fungi in phytoremediation and/or land reclamation, alterations of enzymatic activities in mycorrhizal plants, and elevated CO₂ and O₃ on root colonization and species diversity. Unfortunately, the international community cannot easily appreciate almost all Chinese mycorrhizal studies since the vast majority of them have been published in Chinese and/or in China-based journals. The aim of this review is to make a comprehensive exposure of the past and present China’s major mycorrhizal research to the whole world, and then to suggest potential directions for the enhancement of future mycorrhizal research within and/or between the Chinese and international mycorrhizal community.     

Identity and Specificity of Rhizoctonia-Like Fungi from Different Populations of Liparis japonica (Orchidaceae) in Northeast China

Mycorrhizal association is known to be important to orchid species, and a complete understanding of the fungi that form mycorrhizas is required for orchid ecology and conservation. Liparis japonica (Orchidaceae) is a widespread terrestrial photosynthetic orchid in Northeast China. Previously, we found the genetic diversity of this species has been reduced recent years due to habitat destruction and fragmentation, but little was known about the relationship between this orchid species and the mycorrhizal fungi. The Rhizoctonia-like fungi are the commonly accepted mycorrhizal fungi associated with orchids. In this study, the distribution, diversity and specificity of culturable Rhizoctonia-like fungi associated with L. japonica species were investigated from seven populations in Northeast China. Among the 201 endophytic fungal isolates obtained, 86 Rhizoctonia-like fungi were identified based on morphological characters and molecular methods, and the ITS sequences and phylogenetic analysis revealed that all these Rhizoctonia-like fungi fell in the same main clade and were closely related to those of Tulasnella calospora species group. These findings indicated the high mycorrhizal specificity existed in L. japonica species regardless of habitats at least in Northeast China. Our results also supported the wide distribution of this fungal partner, and implied that the decline of L. japonica in Northeast China did not result from high mycorrhizal specificity. Using culture-dependent technology, these mycorrhizal fungal isolates might be important sources for the further utilizing in orchids conservation.     

Functional diversity of arbuscular mycorrhizal fungal isolates in relation to extraradical mycelial networks

We investigated the functional significance of extraradical mycorrhizal networks produced by geographically different isolates of the arbuscular mycorrhizal fungal (AMF) species Glomus mosseae and Glomus intraradices. A two-dimensional experimental system was used to visualize and quantify intact extraradical mycelium (ERM) spreading from Medicago sativa roots. Growth, phosphorus (P) and nitrogen (N) nutrition were assessed in M. sativa plants grown in microcosms. The AMF isolates were characterized by differences in extent and interconnectedness of ERM. Phenotypic fungal variables, such as total hyphal length, hyphal density, hyphal length per mm of total or colonized root length, were positively correlated with M. sativa growth response variables, such as total shoot biomass and plant P content. The utilization of an experimental system in which size, growth rate, viability and interconnectedness of ERM extending from mycorrhizal roots are easily quantified under realistic conditions allows the simultaneous evaluation of different isolates and provides data with a predictive value for selection of efficient AMF.     

Widespread mycorrhizal specificity correlates to mycorrhizal function in the neotropical, epiphytic orchid Ionopsis utricularioides (Orchidaceae)

Tropical orchids constitute the greater part of orchid diversity, but little is known about their obligate mycorrhizal relationships. The specificity of these interactions and associated fungal distributions could influence orchid distributions and diversity. We investigated the mycorrhizal specificity of the tropical epiphytic orchid Ionopsis utricularioides across an extensive geographical range. DNA ITS sequence variation was surveyed in both plants and mycorrhizal fungi. Phylogeographic relationships were estimated for the mycorrhizal fungi. Orchid functional outcomes were determined through in vitro seed germination and seedling growth with a broad phylogenetic representation of fungi. Most fungal isolates derived from one clade of Ceratobasidium (anamorphs assignable to Ceratorhiza), with 78% within a narrower phylogenetic group, clade B. No correlation was found between the distributions of orchid and fungal genotypes. All fungal isolates significantly enhanced seed germination, while fungi in clade B significantly enhanced seedling growth. These results show that I. utricularioides associates with a phylogenetically narrow, effective fungal clade over a broad distribution. This preference for a widespread mycorrhizae may partly explain the ample distribution and abundance of I. utricularioides and contrasts with local mycorrhizal diversification seen in some nonphotosynthetic orchids. Enhanced orchid function with a particular fungal subclade suggests mycorrhizal specificity can increase orchid fitness.     

Diversity of root-associated fungal endophytes in Rhododendron fortunei in subtropical forests of China

To investigate the diversity of root endophytes in Rhododendron fortunei, fungal strains were isolated from the hair roots of plants from four habitats in subtropical forests of China. In total, 220 slow-growing fungal isolates were isolated from the hair roots of R. fortunei. The isolates were initially grouped into 17 types based on the results of internal transcribed spacer-restriction fragment length polymorphism (ITS-RFLP) analysis. ITS sequences were obtained for representative isolates from each RFLP type and compared phylogenetically with known sequences of ericoid mycorrhizal endophytes and selected ascomycetes or basidiomycetes. Based on phylogenetic analysis of the ITS sequences in GenBank, 15 RFLP types were confirmed as ascomycetes, and two as basidiomycetes; nine of these were shown to be ericoid mycorrhizal endophytes in experimental cultures. The only common endophytes of R. fortunei were identified as Oidiodendron maius at four sites, although the isolation frequency (3–65%) differed sharply according to habitat. Phialocephala fortinii strains were isolated most abundantly from two habitats which related to the more acidic soil and pine mixed forests. A number of less common mycorrhizal RFLP types were isolated from R. fortunei at three, two, or one of the sites. Most of these appeared to have strong affinities for some unidentified root endophytes from Ericaceae hosts in Australian forests. We concluded that the endophyte population isolated from R. fortunei is composed mainly of ascomycete, as well as a few basidiomycete strains. In addition, one basidiomycete strain was confirmed as a putative ericoid mycorrhizal fungus.     

Phylogeny of arbuscular mycorrhizal fungi predicts community composition of symbiosis-associated bacteria

Many physicochemical and biotic aspects of the soil environment determine the community composition of bacteria. In this study, we examined the effects of arbuscular mycorrhizal fungi, common symbionts of higher plants, on the composition of bacterial communities after long-term (7-8 years) enrichment culture in the presence of a plant host. We showed that the phylogeny of arbuscular mycorrhizal fungal isolates was a highly significant predictor of bacterial community composition, as assessed by cluster analysis, redundancy analysis and linear discriminant analysis of phospholipid fatty acid patterns. Numerous phospholipid fatty acids differed between the phylogenetic groupings; this pattern also held for fungal-origin phospholipid fatty acids and in a combined bacterial/fungal analysis, suggesting that categorizing phospholipid fatty acids into predominantly bacterial and fungal origin did not affect the overall outcome. The mechanisms underlying this observation could include substrate quality (and quantity) effects, interactions mediated by the host plant (e.g. rhizodeposition) and direct biotic interactions between arbuscular mycorrhizal fungi and bacterial populations. Our results suggest that aspects of arbuscular mycorrhizal fungal functions may be partially explained by the symbiosis-accompanying bacterial communities, a possibility that should be explicitly considered in studies examining the roles of arbuscular mycorrhizal fungal species diversity in soil and ecosystem processes.     

Diversity of culturable ericoid mycorrhizal fungi of Rhododendron decorum in Yunnan, China

The diversity of ericoid mycorrhizal fungi isolated from Rhododendron decorum Franch. in Yunnan, southwestern China, was examined for the first time. In total 300 hair-root samples were collected from 13 R. decorum individuals in two adjacent wild population sites and one cultivated population site. Two hundred eighteen slow-growing isolates were obtained; the ability of some to form ericoid mycorrhiza was tested in vitro. One hundred twenty-five isolates formed hyphal structures morphologically corresponding to ericoid mycorrhiza, and these were determined by morphological and molecular means to belong to 12 fungal species. There were hardly any differences in species among the three sampled populations. The sequences of several isolates were similar to those of Oidiodendron maius and ericoid mycorrhizal fungi from Helotiales, accounting respectively for 18.4% and 24.8% of the total culturable ericoid mycorrhizal fungi assemblage. Dark septate endophytes were detected in the sampled hair roots by microscopy.     

The mycorrhiza of <Emphasis Type="Italic">Schizocodon soldanelloides</Emphasis> var. <Emphasis Type="Italic">magnus</Emphasis> (Diapensiaceae) is regarded as ericoid mycorrhiza from its structure and fungal identities

Structure and fungal identities were examined in the mycorrhizal roots of Schizocodon soldanelloides var. magnus (Diapensiaceae) to determine the mycorrhizal category. Previous studies had suggested the mycorrhizae of Diapensiaceae could be categorized as ericoid, but the mycorrhizal fungi have never been identified. The diameter of the fine lateral roots, in which coiled hyphae were found in epidermal cells, was mostly less than 100 μm. Molecular analyses identified the fungal isolates to be Helotiales and Oidiodendron. From the structure and fungal identities, we confirmed that the mycorrhiza of S. soldanelloides is an ericoid mycorrhiza.     

Mycorrhizas improve nitrogen nutrition of Trifolium repens after 8 yr of selection under elevated atmospheric CO2 partial pressure

Altered environmental conditions may change populations of arbuscular mycorrhizal fungi and thereby affect mycorrhizal functioning. We investigated whether 8 yr of free-air CO2 enrichment has selected fungi that differently influence the nutrition and growth of host plants. In a controlled pot experiment, two sets of seven randomly picked single spore isolates, originating from field plots of elevated (60 Pa) or ambient CO2 partial pressure (pCO2), were inoculated on nodulated Trifolium repens (white clover) plants. Fungal isolates belonged to the Glomus claroideum or Glomus intraradices species complex, and host plants were clonal micropropagates derived from nine genets. Total nitrogen (N) concentration was increased in leaves of plants inoculated with fungal isolates from elevated-pCO2 plots. These isolates took up nearly twice as much N from the soil as isolates from ambient-pCO2 plots and showed much greater stimulation of biological N2 fixation. The morpho-species identity of isolates had a more pronounced effect on N2 fixation and on root length colonized than isolate identity. We conclude that rising atmospheric pCO2 may select for fungal strains that will help their host plants to meet increased N demands.