Phylogenetic congruence between subtropical trees and their associated fungi

Recent studies have detected phylogenetic signals in pathogen–host networks for both soil‐borne and leaf‐infecting fungi, suggesting that pathogenic fungi may track or coevolve with their preferred hosts. However, a phylogenetically concordant relationship between multiple hosts and multiple fungi in has rarely been investigated. Using next‐generation high‐throughput DNA sequencing techniques, we analyzed fungal taxa associated with diseased leaves, rotten seeds, and infected seedlings of subtropical trees. We compared the topologies of the phylogenetic trees of the soil and foliar fungi based on the internal transcribed spacer (ITS) region with the phylogeny of host tree species based on matK, rbcL, atpB, and 5.8S genes. We identified 37 foliar and 103 soil pathogenic fungi belonging to the Ascomycota and Basidiomycota phyla and detected significantly nonrandom host–fungus combinations, which clustered on both the fungus phylogeny and the host phylogeny. The explicit evidence of congruent phylogenies between tree hosts and their potential fungal pathogens suggests either diffuse coevolution among the plant–fungal interaction networks or that the distribution of fungal species tracked spatially associated hosts with phylogenetically conserved traits and habitat preferences. Phylogenetic conservatism in plant–fungal interactions within a local community promotes host and parasite specificity, which is integral to the important role of fungi in promoting species coexistence and maintaining biodiversity of forest communities.     

辽宁省9种兰科植物根内与根际土壤中真菌群落结构的差异

兰科植物的生存及生长高度依赖其根中的共生真菌, 其中的菌根真菌更是对兰科植物的种子萌发与后续生长有着非常重要的作用, 研究兰科植物根中的真菌, 尤其是菌根真菌, 对兰科植物的保护有重要作用。该研究利用第二代测序技术, 对中国辽宁省境内的9种属于极小种群的兰科植物的根、根际土和根围土中的真菌群落和菌根真菌组成进行了研究。结果显示, 兰科植物根中的真菌群落和根际土、根围土中的真菌群落具有显著差异。兰科植物根中的总操作分类单元(OTU)数目远小于根际土和根围土中的总OTU数目。同时, 兰科植物根中菌根真菌的种类和丰度与根际土、根围土中菌根真菌的种类与丰度没有明显联系。FunGuild分析结果显示, 丛枝菌根真菌在根际土与根围土中的丰度非常高, 但在兰科植物的根中却数量极少。这些结果表明, 兰科植物根中的真菌群落与土壤中的真菌群落在一定程度上是相互独立的。     

四川黄龙沟优势兰科植物菌根真菌多样性及其季节变化

在自然条件下,兰科菌根真菌对兰花的种子萌发和植株生长都是必不可少的。为了解高原兰科植物菌根真菌的多样性状况及其季节性变化规律,选取了四川黄龙沟的两种生境中生长的8种优势兰科植物,分别于植株的萌芽期(4月份)、生长期(7月份)和果期(9月份)采集营养根进行菌根真菌的多样性研究。其中,黄花杓兰(Cypripedium flavum)、少花鹤顶兰(Phaiusdelavayi)、二叶匍茎兰(Galearis diantha)和广布小蝶兰(Ponerorchis chusua)分布在开阔生境;筒距兰(Tipularia szechuanica)、小花舌唇兰(Platanthera minutiflora)、珊瑚兰(Corallorhiza trifida)和尖唇鸟巢兰(Neottia acuminate)则分布在密林生境。通过对分离所得的50个菌株进行形态观察和ITS序列测定相结合的鉴定,共获得菌根真菌41种。对担子菌和子囊菌分别进行的系统发育树构建结果显示,子囊菌为优势种类(35种),以柔膜菌目(Helotiales)、炭角菌目(Xylariales)和肉座菌目(Hypocreales)内的种类为主,担子菌则以胶膜菌(Tulasnellaceaesp.)为主。在8种兰科植物中,二叶匐茎兰表现出极高的专一性,其菌根真菌均属于Hypocrea。其余兰科植物的菌根真菌分别属于不同的科,专一性相对较低。物种丰富度和Simpson多样性指数分析结果表明,密林生境的兰科植物的菌根真菌多样性在各生长季节基本高于开阔生境。此外,两种生境的优势兰科植物的菌根真菌物种多样性随生长季节转变所呈现的变化规律是相似的:萌发期和生长期的多样性均较高,峰值出现在生长期,到果期时则大幅下降。这与高原兰科植物的生长特性及营养供求规律基本相符。     

Optimisation of DNA extraction for AFLP analysis of mycorrhizal fungi of terrestrial orchids caladeniinae and drakaeinae

Published DNA extraction methods present a number of problems when applied to mycorrhizal fungi of native Australian terrestrial orchids. Grinding with liquid nitrogen shears the DNA, and other pulverisation methods yield too little DNA. We found that freezing the fungal sample with liquid nitrogen, with no grinding, followed by the Qiagen DNeasy extraction procedure produced good yields of high-molecular-weight DNA. The DNA was then used for amplified fragment length polymorphism (AFLP) fingerprinting. Good fingerprints were produced by restriction withEcoRI/MseI enzymes, the use of preamplification primer mix II (for small genomes), and a 2-base extensionMseI primer (m-cc) with 3-base extensionEcoRI primers in the selective amplification. This protocol may be of general utility for other fungi with similarly fragile DNA.     

外生菌根菌与森林树木的相互关系

生态系统的每个过程都伴随着各种微生物的活动,其中最重要的功能群之一是菌根真菌(菌根菌)。一般认为,菌根菌是自然界多数植物生存最基本的组成部分,陆地上约90％以上的高等植物都具有菌根菌。这些菌类的菌丝体与植物根系结合形成菌根,使植物生长成为可能,使不同种类植物的根系联在一起。根据菌根菌入侵植物根系的方式及菌根的形态特征,菌根可分为外生菌根、内生菌根和内外生菌根3组共7种类型。外生菌根主要出现在松科、桦木科、壳斗科等树种的森林生态系统中,在根系表面形成菌丝鞘,部分菌丝进入根系皮层细胞间隙形成哈氏网表面。菌根菌剂在森林经营中得到广泛地应用。外生菌根菌对森林树木的作用可归纳为：1)促进造林或育苗成活与生长;2)提高森林生态系统中植物的多样性、稳定性和生产力;3)对森林生态系统的综合效应,主要表现在增加植物一土壤联结,改善土壤结构,促进土壤微生物,增强植物器官的功能;4)抗拮植物根部病害病原菌等。树木与菌根菌相互关系研究主要包括：1)菌根共生的机理;2)菌根菌在退化森林生态系统恢复与改造中的作用;3)菌根菌的分布格局与森林生态系统服务功能的关系;4)菌根菌对森林生态系统的综合效应,如菌根菌与森林植物群落结构、物种多样性以及森林系统稳定性和生产力的研究。     

菌根真菌对白及种子萌发和幼苗生根的作用

自然条件下,兰科植物需要依赖菌根真菌获得营养才能萌发。本研究对白及根和原球茎中分离的4株菌根真菌（WQ17-33、WQ17-43、JST-3和SL15-7）进行分子鉴定,并评价光照和黑暗条件下不同菌株促白及种子萌发和幼苗生根的效果。结果表明,4个菌株分别隶属于鬼伞属Coprinus、胶膜菌属Tulasnella、腊壳菌属Sebacina和Serendipita。在种子萌发前期（未形成叶子）进行暗培养较光照对菌株JST-13和SL15-7处理组原球茎阶段萌发具有显著的促进作用。不同菌株共生萌发效果不同,菌株SL15-7较其他处理原球茎和幼苗发育阶段的萌发率高。菌株JST-13和SL15-7处理组形成的幼苗较其他处理组强壮,定殖的菌丝团也较多,其幼苗生根效果也较对照组好。该研究表明白及可与多种不同类群的菌根真菌菌株形成共生关系,这些真菌在促进白及种子萌发和生根能力方面存在差异。     

Increased photosynthetic acclimation in alfalfa associated with arbuscular mycorrhizal fungi (AMF) and cultivated in greenhouse under elevated CO2

Medicago sativa L. (alfalfa) can exhibit photosynthetic down-regulation when grown in greenhouse conditions under elevated atmospheric CO₂. This forage legume can establish a double symbiosis with nitrogen fixing bacteria and arbuscular mycorrhizal fungi (AMF), which may increase the carbon sink effect of roots. Our aim was to assess whether the association of alfalfa with AMF can avoid, diminish or delay the photosynthetic acclimation observed in previous studies performed with nodulated plants. The results, however, showed that mycorrhizal (M) alfalfa at the end of their vegetative period had lower carbon (C) discrimination than non-mycorrhizal (NM) controls, indicating photosynthetic acclimation under ECO₂ in plants associated with AMF. Decreased C discrimination was due to the acclimation of conductance, since the amount of Rubisco and the expression of genes codifying both large and small subunits of Rubisco were similar or slightly higher in M than in NM plants. Moreover, M alfalfa accumulated a greater amount of soluble sugars in leaves than NM plants, thus favoring a down-regulation effect on photosynthetic rates. The enhanced contents of sugars in leaves coincided with a reduced percentage of arbuscules in roots, suggesting decreased sink of carbohydrates from shoots to roots in M plants. The shorter life cycle of alfalfa associated with AMF in comparison with the NM controls may also be related to the accelerated photosynthetic acclimation in M plants. Further research is needed to clarify to what extent this behavior could be extrapolated to alfalfa cultivated in the field and subjected to periodic cutting of shoots under climatic change scenarios.     

Variation in nutrient-acquisition patterns by mycorrhizal fungi of rare and common orchids explains diversification in a global biodiversity hotspot

Background and Aims

Many terrestrial orchids have an obligate requirement for mycorrhizal associations to provide nutritional support from germination to establishment. This study will investigate the ability of orchid mycorrhizal fungi (OMF) to utilize a variety of nutrient sources in the nutrient-impoverished (low organic) soils of the Southwest Australian Floristic Region (SWAFR) in order to effectively compete, survive and sustain the orchid host.

Methods

Mycorrhizal fungi representing key OMF genera were isolated from three common and widespread species: Pterostylis recurva, Caladenia flava and Diuris corymbosa, and one rare and restricted species: Drakaea elastica. The accessibility of specific nutrients was assessed by comparing growth including dry biomass of OMF in vitro on basal CN MMN liquid media.

Key Results

Each of the OMF accessed and effectively utilized a wide variety of nutrient compounds, including carbon (C) sources, inorganic and organic nitrogen (N) and inorganic and organic phosphorus (P). The nutrient compounds utilized varied between the genera of OMF, most notably sources of N.

Conclusions

These results suggest that OMF can differentiate between niches (micro-niche specialization) in a constrained, highly resource-limited environment such as the SWAFR. Phosphorus is the most limited macronutrient in SWAFR soils and the ability to access phytate by OMF indicates a characterizing functional capacity of OMF from the SWAFR. Furthermore, compared with OMF isolated from the rare D. elastica, OMF associating with the common P. recurva produced far greater biomass over a wider variety of nutritional sources. This suggests a broader tolerance for habitat variation providing more opportunities for the common orchid for recruitment and establishment at a site.     

不同海拔的三种杓兰属植物与菌根真菌群落组成相关性

本研究采集了四川黄龙沟沿海拔梯度3 170-3 400m上4个不同杓兰居群中3种杓兰植物根,利用克隆文库方法获得菌根真菌ITS序列,研究同一栖息地（黄龙沟）不同海拔梯度和不同杓兰对菌根真菌多样性和群落结构的影响。共得到18个可操作分类单元（OTU）,其中14个OTU隶属胶膜菌科Tulasnellaceae,为优势类群（99.6%）;2个OTU隶属腊壳菌科Sebacinaceae,2个OTU隶属亡革菌科Thelephoraceae。随着海拔升高,西藏杓兰菌根真菌多样性减少,而黄花杓兰和无苞杓兰没有明显变化;海拔对3种杓兰菌根真菌群落结构均无显著影响。3种杓兰之间菌根真菌群落结构差异显著且指示物种互不相同,说明在同一栖息地,杓兰对菌根真菌的偏好性显著影响其菌根真菌群落结构。这些研究结果利于了解环境变化对杓兰属植物菌根真菌区系组成的影响,为进一步探索菌根真菌与杓兰属植物的互作机制奠定基础。     

The chemical composition,antifungal, antioxidant and antimutagenicity properties of bioactive compounds from fungal endophytes associated with Thai orchids

Some plant‐derived bioactive compounds produced by fungal endophytes have been proven to have antimicrobial and antioxidant activities. In this study, endophytic fungi were isolated from 20 orchid samples collected in northern Thailand from 12 genera of orchids. In total, 97 isolates were isolated from the leaves (44.3%), stems (40.2%) and flowers (15.5%) of the orchid samples. The antifungal activity was investigated of the endophytic isolates against the plant pathogenic fungi. The results showed that 13 endophytic isolates provided antifungal activities against Fusarium sp., Colletotrichum sp. and Curvularia sp. The endophyte CK F05‐5, which was isolated from the flower part of Dendrobium lindleyi, was chosen for further testing because it the highest level of antifungal activity against Fusarium sp. The isolate CK F05‐5 was identified as Fusarium oxysporum on the basis of its ITS sequences of 5.8 s rRNA, and phytochemical analysis revealed the presence of coumarins. The ethyl acetate extract of CK F05‐5 was examined for its total phenolic content and antioxidant activity using Folin–Ciocalteu's reagent and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical scavenging assay, respectively. The phenolic content was 160.51 mg of GAE/g of extract, and the free radical scavenging activity was 89.61 µg/ml at the half maximal inhibitory concentration (IC₅₀). The antimutagenic potential of the ethyl acetate extract of CK F05‐5 against Trp‐P‐1 mutagenic substances was determined using the Ames test which revealed that the extract of CK F05‐5 at 10 mg/plate had the highest antimutagenic activity against Trp‐P‐1 (51.2%) and 39.6% for strains TA98 and TA100, respectively. The active compounds present in the acetate extract of CK F05‐5 were examined using GC‐MS analysis, which displayed the presence of gibepyrone A, pyrrolo [1, 2‐a] pyrazine‐1, 4‐dione, hexahydro‐3‐(2‐methylpropyl) and indoleacetic acid as major components. Based on the results, this endophytic fungus contains various bioactive components that have various biological activities. This useful information could help in producing potentially valuable and novel pharmaceutical products.     

菌根食用菌研究进展

与植物根系共生的菌根食用菌（edible mycorrhizal fungi,EMF）,具有较高的营养价值、经济价值和生态价值。总结了菌根食用菌（EMF）种质资源分布与利用、生理与生态学研究、驯化与栽培技术研究最新进展,分析了我国EMF开发利用所面临的挑战与机遇,探讨了EMF研究动向与发展前景。     

Genetic patterns and pollination in Ophrys iricolor and O. mesaritica (Orchidaceae): sympatric evolution by pollinator shift

Ophrys iricolor and O. mesaritica are a pair of morphologically similar, closely related sexually deceptive orchids from the eastern Mediterranean. Ophrys iricolor is known to be pollinated by Andrena morio males and the specific pollinator of Ophrys mesaritica is determined as Andrena nigroaenea. Amplified fragment length polymorphism revealed O. iricolor and O. mesaritica to be genetically intermixed on the whole, although populations of O. iricolor and O. mesaritica in geographical proximity are strongly differentiated, suggesting that specific pollinators locally differentiate these taxa. Based on the available biological data and the system of pollinator attraction operative in Ophrys, we hypothesize that O. mesaritica may have arisen from O. iricolor by pollinator shift and that this is more probable than scenarios invoking hybridization as a result of mispollination by rare, non‐specific flower visitors or specifically attracted insects. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 583–598.     

城市菌根真菌多样性、变化机制及功能应用

菌根真菌能够与大多数陆生植物的根系形成菌根共生体,具有改善宿主植物矿质营养、增强抗逆性、改良土壤结构等重要生态功能。城市化过程中气候、土壤、植被、土地利用等因素的改变,对菌根真菌的多样性产生了直接或间接的影响。目前城市菌根真菌的研究多侧重对其空间分布及群落组成的简单描述,缺乏针对城市典型生态现象及生态问题系统性的探讨。分别从城市菌根真菌的多样性变化、影响机制及功能应用等3方面进行了综述,全面揭示城市菌根真菌的研究现状及研究的复杂性,发现当前研究存在多样性评估简单化、研究层次单一化、内在机制现象化及功能应用停滞化等问题,认为今后应建立更为系统、综合、标准的研究体系以深刻而准确地认识与理解城市化对菌根真菌多样性的影响,为城市微生物资源的保存及绿地系统维持提供理论依据。     

丛枝菌根真菌和植物寄生线虫

本文综述了土壤微生物中丛枝菌根真菌和植物寄生线虫的互作关系及其互作机理,并阐述了丛枝菌根真菌在防治植物线虫病害方面的应用前景和实际操作中应注意的技术环节。     

Effects of simultaneous infections of endophytic fungi and arbuscular mycorrhizal fungi on the growth of their shared host grass Achnatherum sibiricum under varying N and P supply

Grasses can be infected by endophytic fungi and arbuscular mycorrhizal (AM) fungi simultaneously. Here, we investigated the interactions of a native grass, Achnatherum sibiricum, with both endophytic and AM fungi (Glomus mosseae, GM and Glomus etunicatum, GE) at different nitrogen (N) and phosphorus (P) levels. The results showed that endophyte infection significantly suppressed the colonization rates and spore density of GE, but had no effect on those of GM. Endophyte infection increased shoot biomass regardless of the nutrient conditions. The effects of AM fungi on host growth were dependent on mycorrhizal species. There was no significant interaction between endophytic fungi and GE on host growth; however, a significant interaction between endophytic fungi and GE occurred in total phenolic concentrations and P uptake. As for GM, a significant interaction among endophytic fungi, AM fungi and nutrient availability occurred in shoot growth. Under sufficient N and P conditions, endophyte infection alleviated the detrimental effects of GM colonization on host growth.     

耐热真菌的分类鉴定及rDNA-ITS系统发育分析

分别从山东、山西、河北、陕西、河南、云南、江西、海南、新疆、内蒙古、甘肃、广州等地实地采集堆肥、培养料、草堆等标本600多份,分离鉴定出耐热真菌28种,其中包括2个中国新记录种,沙生梭孢壳Thielavia arenaria和丰孢木霉Trichoderma saturnisporum,其余26个为中国已知种。在形态分类的基础上,对所分离鉴定的28株耐热真菌及购买的3株耐热真菌,进行rDNA-ITS序列测定,所测序列片段长度为554－895bp。将所测的31个和从GenBank下载的15个耐热真菌的rDNA-ITS序列构建NJ系统发育树,该树较好地显示了耐热真菌的系统进化关系,有力地支持了形态学的观点。     

NO参与AM真菌与烟草共生过程

以烟草（Nicotiana tabacum,品种CF90NF）为材料,利用分光光度法和荧光显微技术结合药理学实验,探讨在AM真菌摩西球囊霉(Glomus mosseae,G.m)与烟草共生过程中一氧化氮(nitric oxide, NO)的作用。结果表明,烟草侧根中含有一定水平的内源NO,苗期接种G.m 10天后,烟草根系NO含量显著增加,侧根中的NO荧光强度也在接种后10天达到最强;一定浓度的NO供体硝普钠(sodium nitroprusside,SNP)能促进G.m对烟草的侵染,而NO的清除剂2-4,4,5,5-苯-四甲基咪唑-1-氧-3-氧化物( 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxidepotassium salt,cPTIO)可明显减弱侧根和菌丝中的NO的荧光强度,降低AM真菌的侵染率,表明NO参与G.m与烟草的共生过程;在G.m与烟草的共生过程中,烟草根系硝酸还原酶(nitrate reductase,NR)活性与Nia-1的表达量明显升高,且NR的抑制剂钨酸钠(sodium tungstate,Na2WO4)可以降低烟草侧根中的荧光强度,但对菌丝中的NO的荧光强度无明显影响。由此推测,来自根系NR途径的NO参与AM真菌与烟草的共生过程,菌丝中可能存在其他来源的NO。     

Species-specific trade-offs between regrowth and mycorrhizas in the face of defoliation and phosphorus addition

Arbuscular mycorrhizal fungi (AMF) enhance nutrient provision in exchange for carbon. However, a shift from nutrient to carbon limitation, induced by repeated or intense defoliation, can represent a trade-off between plant regrowth and the maintenance of mycorrhiza. The combined effects of AMF, phosphorus and defoliation on growth of Agropyron elongatum (C₃ grass, low mycorrhizal responsiveness) and Brachiaria brizantha (C₄ grass, high mycorrhizal responsiveness) were explored. Each species was subjected to a manipulative experiment with AMF inoculation (non-inoculated, inoculated), soluble P supply (low, high) and defoliation (non-defoliated, 60% defoliated). In A. elongatum, at low P supply, mycorrhizal plants showed increased growth rates following defoliation without substantial changes in AMF colonization. At high P supply instead, we found a clear trade-off between regrowth and the maintenance of mycorrhiza evidenced by growth depression (biomass and tillers) and lower AMF activity (reduction of arbuscules). In contrast, in B. brizantha, defoliation effects on plant regrowth were independent from AMF at any P supply (no trade-off). This indicates that cost-benefit relationship in defoliated plants is highly context-dependent and may vary with mycorrhizal responsiveness of species. This variation of responses can play a decisive role on plant recovery in pastures and natural grasslands subjected to herbivory.     

贵州烟草根围AM真菌多样性的初步研究

从贵州省内烟区不同土壤生态环境下采集烟草根际土样,湿筛离心法分离丛枝菌根（AM）真菌孢子,鉴定出烟草AM真菌4属20种,其中球囊霉属9种,无梗囊霉属7种,巨孢囊霉属3种,盾巨孢囊霉属1种。从土壤样品DNA中扩增AM真菌特异性片段并采用DGGE技术对AM真菌多样性进行分析。测序结果显示烟草根际土壤中菌根真菌主要菌群为球囊霉属,与湿筛离心法的鉴定结果一致。为进一步研究贵州地区AM真菌多样性以及开发应用提供了依据。     

Ericoid mycorrhizal root fungi and their multicopper oxidases from a temperate forest shrub

Ericoid mycorrhizal fungi (ERM) may specialize in capturing nutrients from their host's litter as a strategy for regulating nutrient cycles in terrestrial ecosystems. In spite of their potential significance, we know little about the structure of ERM fungal communities and the genetic basis of their saprotrophic traits (e.g., genes encoding extracellular enzymes). Rhododendron maximum is a model ERM understory shrub that influences the nutrient cycles of montane hardwood forests in the southern Appalachians (North Carolina, USA). We sampled ERM roots of R. maximum from organic and mineral soil horizons and identified root fungi by amplifying and sequencing internal transcribed spacer (ITS) ribosomal DNA (rDNA) collected from cultures and clones. We observed 71 fungal taxa on ERM roots, including known symbionts Rhizoscyphus ericae and Oidiodendron maius, putative symbionts from the Helotiales, Chaetothyriales, and Sebacinales, ectomycorrhizal symbionts, and saprotrophs. Supporting the idea that ERM fungi are adept saprotrophs, richness of root-fungi was greater in organic than in mineral soil horizons. To study the genetic diversity of oxidative enzymes that contribute to decomposition, we amplified and sequenced a portion of genes encoding multicopper oxidases (MCOs) from ERM ascomycetes. Most fungi possessed multiple copies of MCO sequences with strong similarities to known ferroxidases and laccases. Our findings indicate that R. maximum associates with a taxonomically and ecologically diverse fungal community. The study of MCO gene diversity and expression may be useful for understanding how ERM root fungi regulate the cycling of nutrients between the host plant and the soil environment.