兰科植物菌根真菌研究新见解

兰科(Orchidaceae)在地球生命系统演化中占有十分重要的地位,几乎全部兰科植物均处于不同程度的濒危状态,研究兰科菌根真菌对于保护珍稀濒危兰科植物具有重要意义。该文在对菌根真菌相关的概念及研究方法进行综述的基础上,对兰科菌根真菌的主要类群、特异性及其与兰科植物稀有性之间的关系,以及兰科菌根真菌与兰科植物之间的营养关系和进化关系进行了总结。兰科菌根真菌的研究方法可以归纳为经典研究方法、早期分子生物学方法、rDNA片段高通量测序法、宏基因组学方法。兰科菌根真菌类群主要隶属于担子菌门(Basidiomycota)、子囊菌门(Ascomycota)和毛霉门(Mucoromycota)。根据兰科菌根真菌特异性与否,首次明确了兰科菌根真菌定植关系可分为三大类:特异性定植、广泛性定植和特异-广泛兼性定植。根据营养关系特点,首次将兰科植物与菌根真菌之间的营养关系划分为三大类:兰花单向利好型、典型共生型、分工合作型。兰科菌根真菌特异性与兰科植物稀有性之间的关系呈现出两面性,而兰科菌根真菌与兰科植物之间是否存在协同进化关系还需要更多的研究才能阐明。此外,该文还对兰科菌根真菌领域今后的研究提出了一些思路。     

辽宁省八种兰科植物根内生真菌多样性

兰科植物的生存强烈依赖菌根真菌,研究兰科菌根真菌对兰科植物的保护有重要作用。本研究以辽宁省的无柱兰(Amitostigma gracile)、二叶舌唇兰(Platanthera chlorantha)、小斑叶兰(Goodyera repens)、蜻蜓兰(Tulotis fuscescens)、山兰(Oreorchis patens)、羊耳蒜(Liparis japonica)、长苞头蕊兰(Cephalanthera longibracteata)和绶草(Spiranthes sinensis) 8种属于极小种群的野生兰科植物为例,利用第二代测序技术对其根内生真菌多样性进行了研究。结果表明:无柱兰、二叶舌唇兰、小斑叶兰和山兰都偏好与角担菌科(Ceratobasidiaceae)真菌共生,羊耳蒜偏好与胶膜菌科(Tulasnellaceae)真菌共生;长苞头蕊兰主要与革菌科(Thelephoraceae)和蜡壳耳科(Sebacinaceae)真菌共生;绶草不仅能与丝核菌属(Rhizoctonia)真菌共生,还能与蜡壳耳科真菌共生;同种兰科植物的不同植株在同一生境下所选择的菌根真菌有差异,而同一地点的不同兰科植物的菌根真菌群落各不相同。由此可见,兰科植物根中菌根真菌的组成并非完全受植物自身控制,但主要菌根真菌仍取决于兰科植物的选择;这8种兰科植物根中的真菌绝大多数为非菌根真菌,而菌根真菌的丰度通常很低,这可能是辽宁地区兰科植物稀少的原因之一。     

菌根真菌与兰科植物氮营养关系的研究进展

兰科植物是典型的菌根植物。兰菌根是兰科植物根与真菌形成的菌根共生体。兰菌根真菌的营养来源影响宿主植物的生活方式和营养水平。氮是植物生长的主要限制因子。兰科植物具有富集氮的特征, 其组织和器官的氮含量通常高于同生境中的其他植物。该文综述了兰菌根真菌类别、兰科植物氮营养特征和兰菌根的氮转移机制等的研究进展, 以期为兰科植物资源的保护、再生及可持续利用的相关研究提供参考和借鉴。     

兰科植物与菌根真菌的营养关系

兰科植物与真菌具有天然的菌根共生关系。兰科植物种子细小,无胚乳,自然条件下只有与适宜的真菌共生才能萌发;兰科植物作为有花植物中最大的科之一,有光合自养、混合营养及完全真菌异养等营养类型。近年研究表明,不同营养类型的兰科植物其菌根真菌常具有一定的差异性,表现出不同的营养作用关系。本文对不同营养类型兰科植物与菌根真菌的营养作用关系的研究进展进行综述,并对兰科植物菌根营养机理、营养关系的变化等进行讨论,以期为兰科菌根营养学研究及菌根技术应用于兰科植物快繁和保育工作等提供参考。     

三种杓兰根相关真菌多样性和生态功能

【背景】除了菌根真菌(Orchid mycorrhizal fungi,OrMF)外,兰科植物根中还有其它内生真菌,称为根相关真菌(Root-associated fungi,RAF)。【目的】采用分离培养的方法获得同一栖息地针叶林和灌木林两种不同生境西藏杓兰、黄花杓兰和无苞杓兰的RAF菌株,研究其真菌谱系、多样性和生态功能结构。【方法】从杓兰根碎屑中分离RAF,通过总DNA提取、PCR扩增及测序得到ITS(Internaltranscribedspacer)序列;进行系统发育和多样性分析,并通过NCBI数据库比对得到相似性最高序列的注释信息来分析RAF生态学特性。【结果】共分离得到278株RAF,25种OTU类型,包括23个子囊菌门OTU,2个毛霉菌门OTU。RAF物种丰富度分析发现西藏杓兰的较黄花杓兰高,不同生境没有显著差异;不同杓兰物种较不同生境的RAF群落分化程度高。生态功能分析显示25个OTU包括共生型、腐生型和致病型3种营养型,以及外生菌根菌群、植物病原菌群、内生真菌群、动物病原菌群、真菌寄生菌群、杜鹃花类菌根群、未定义的腐生菌群和不确定型8种共位群。【结论】阐明不同生境采集的不同杓兰中RAF的分布特点和生态功能,为未来研究RAF与杓兰属植物的共生关系奠定基础。     

独花兰菌根真菌的分离及rDNA ITS序列在其分类鉴定中的应用

独花兰(Changnienia amoena Chien)为我国特有的独属独种兰科植物,仅生长于长江中下游及陕西南部的山地林下及沟谷中,是中国特有兰科物种之一,被列为国家二级保护植物.以浙江天目山自然保护区的野生独花兰中分离获得的菌根真菌为对象,应用传统的形态结构鉴别与rDNA ITS分子生物学手段相结合的研究方法,进行了菌株的分类鉴定研究,确定该菌株为兰科菌根真菌之一的胶膜菌属(Tulasnella)真菌.研究结果对于全面了解兰科植物菌根真菌的特征和有效保护独花兰植物资源均具较大意义和参考价值.     

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。     

濒危药用植物掌裂兰根部内生真菌和根际土真菌多样性分析

【背景】植物内生真菌对宿主植物促生长、抗旱和增强抗病能力等方面有着重大的研究和利用价值,尤其对兰科植物的生长起到重要的作用。【目的】通过对掌裂兰根部内生真菌和根际土真菌多样性进行系统分析,掌握掌裂兰根部内生真菌与根际土真菌群落结构,为进一步探究掌裂兰植物与真菌共生规律提供参考。【方法】采用Illumina MiSeq高通量测序技术分析掌裂兰根部内生真菌和根际土真菌多样性。【结果】掌裂兰根部内生真菌隶属于7门89属,优势菌属为瘤菌根菌属(Epulorhiza)(16.93%)、头梗霉属(Cephaliophora)(10.41%)、酵母属(Saccharomyces)(5.73%)、角担菌属(Ceratobasidium)(5.32%)和镰刀菌属(Fusarium)(5.12%),其中Epulorhiza和Ceratobasidium为兰科植物菌根真菌;根际土真菌隶属于11门269属,优势菌属为镰刀菌属(Fusarium)(8.09%)、丛赤壳属(Neonectria)(6.79%)、Plectosphaerella (3.39%)和被孢霉属(Mortierella)(3.01%)。通过...     

兰科菌根的生态学研究进展

兰科植物(Orchidaceae)是典型的菌根植物,自然条件下其种子的成功萌发和生长的早期阶段对菌根真菌有绝对的依赖性,在有些成年兰科植物中菌根真菌仍起着重要的作用。目前大部分兰科植物已为濒危物种,鉴于兰科植物天然的菌根共生关系,开展兰科植物和菌根真菌互作的生态学研究不仅具有极高的科研价值,更有助于兰科植物的物种保护和野生种群的生态恢复。近年研究表明,兰科植物对真菌的选择和二者共生关系的建立与菌根真菌的空间分布和丰度密切相关,然而当前对自然环境中兰科菌根真菌的实际分布还了解甚少,因此文章从生态学角度系统分析兰科植物与菌根真菌的关系,探讨该领域的研究热点,旨在为兰科菌根的生态学研究提供参考。     

华北特有腐生型兰科植物北京无喙兰根际土壤微生物多样性研究

北京无喙兰(Holopogon pekinensis X. Y. Mu & Bing Liu)为华北地区特有珍稀腐生型兰科植物,分布在海拔约1100 m的杂木林内,生境与本区域内其他腐生型兰科植物(常在1600 m以上桦木林中生长)显著不同。本研究针对北京玉渡山和百花山两个北京无喙兰种群的根际土壤样品开展基于高通量测序技术的根际土壤微生物多样性分析,解析北京无喙兰根际土壤微生物群落组成及多样性。测序分析结果显示,共得到4973个细菌OTU(Operational taxonomic unit),发现北京无喙兰根际土壤的优势细菌类群为变形菌门、放线菌门、拟杆菌门等7个门,优势属有MND1、硝化螺菌属(Nitrospira)和Haliangium等。1914个真菌OTU的分析结果表明,根际土壤优势真菌类群为子囊菌门、担子菌门、结合菌门等;优势属有Archaeorhizomyces、蜡壳耳属(Sebacina)和被孢霉属(Mortierella)等;优势真菌多为外生菌根真菌,可能是北京无喙兰潜在的菌根真菌。多样性指数分析显示,北京无喙兰玉渡山种群根际土壤中的真菌和细菌群落的丰富度和均匀度均高于百花山种群,各种群土壤微生物多样性与北京无喙兰所在种群的乔木种类多样性具有一定的相关性。     

Relationships between orchid and fungal biodiversity: Mycorrhizal preferences in Mediterranean orchids

Orchidaceae is one of the most species-rich angiosperm families, and all orchids are fully dependent on fungi for their seed germination and their life cycle. The level of specificity of the association between orchid species and fungi can be related to the number of co-occurring orchid species. To investigate orchid mycorrhizal associations in adult-photosynthetic orchids, 16 Mediterranean orchid species belonging to 4 genera (Anacamptis, Ophrys, Orchis, and Serapias) at 11 different sites were subjected to DNA-based analysis. Eighteen operational taxonomic units representing two fungal families, Tulasnellaceae and Ceratobasidiaceae, were identified. All examined orchid species associated with different mycorrhizal fungi. Interestingly, there was a positive correlation between number of orchid species and number of mycorrhizal. Monospecific populations showed a lower number of fungi, while sympatric populations had a higher number of mycorrhizal fungi. Our results showed that Mediterranean orchid species associated with a higher number of mycorrhizal fungi confirming as photosynthetic orchids are typically generalists toward mycorrhizal fungi. Thus, photosynthetic orchids exhibit low specificity for fungal symbionts showing the potential for opportunistic associations with diverse fungi reducing competition for nutrient. We suggest that these characteristics could confer symbiotic assurance particularly in habitat with resource limitations or prone to stressful conditions.     

Studies of Mycorrhizal Fungi of Chinese Orchids and Their Role in Orchid Conservation in China—A Review

China has over 1,200 species of native orchids in nearly 173 genera. About one fourth of native species are of horticultural merit. Some species are of Chinese medicinal value. In fact, the demand on orchid species with high Chinese medicinal values such as Gastrodia elata, Dendrobium offcinale, along with demands on species of cultural importance, such as those in the genus of Cymbidium, is a major factor causing wild populations to diminish and in some cases, drive wild populations to the brink of extinction. These market demands have also driven studies on the role of mycorrhizal fungi in orchid seed germination, seedling and adult growth, and reproduction. Most of these mycorrhizal studies of Chinese orchids, however, are published in Chinese, some in medical journals, and thus overlooked by the mainstream orchid mycorrhizal publications. Yet some of these studies contained interesting discoveries on the nature of the mycorrhizal relationships between orchids and fungi. We present a review of some of these neglected publications. The most important discovery comes from the mycorrhizal studies on G. elata, in which the researchers concluded that those fungi species required to stimulate seed germination are different from those that facilitate the growth of G. elata beyond seedling stages. In addition, presence of the mycorrhizal fungi associated with vegetative growth of post-seedling G. elata hindered the germination of seeds. These phenomena were unreported prior to these studies. Furthermore, orchid mycorrhizal studies in China differ from the mainstream orchid studies in that many epiphytic species (in the genus of Dendrobium, as medicinal herbs) were investigated as well as terrestrial orchids (mostly in the genus Cymbidium, as traditional horticultural species). The different responses between epiphytic and terrestrial orchid seeds to fungi derived from roots suggest that epiphytic orchids may have a more general mycorrhizal relationship with fungi than do terrestrial orchid species during the seed germination stage. To date, orchid mycorrhizal research in China has had a strongly commercial purpose. We suggest that this continuing research on orchid mycorrhizal relationships are a solid foundation for further research that includes more rare and endangered taxa, and more in-situ studies to assist conservation and restoration of the endangered orchids. Knowledge on the identities and roles of mycorrhizal fungi of orchids holds one of the keys to successful restoration and sustainable use of Chinese orchids.     

Partial mycoheterotrophy in rhizoctonia-associated orchid Cheirostylis liukiuensis

Partially mycoheterotrophic plant species obtain organic carbon, via both photosynthesis and mycorrhizal symbiosis. In this study, we investigated the mycorrhizal fungi association and nutritional mode of Cheirostylis liukiuensis, which is suspected to be a partial mycoheterotrophic plant, due to its characteristic reduced underground organs, low-light growth environment, and some fully mycoheterotrophic species in the phylogenetically related genera. Molecular analysis of the dominant mycobiont and stable isotope analysis suggested that C. liukiuensis is a partial mycoheterotrophic plant predominantly associate with non-ectomycorrhizal Ceratobasidiaceae fungi. As examples of partial mycoheterotrophic orchids exploiting non-ectomycorrhizal rhizoctonia are still limited, this study provides valuable information on the nutritional modes of green orchids.     

Untangling above‐ and belowground mycorrhizal fungal networks in tropical orchids

Orchids typically depend on fungi for establishment from seeds, forming mycorrhizal associations with basidiomycete fungal partners in the polyphyletic group rhizoctonia from early stages of germination, sometimes with very high specificity. This has raised important questions about the roles of plant and fungal phylogenetics, and their habitat preferences, in controlling which fungi associate with which plants. In this issue of Molecular Ecology, Martos et al. (2012) report the largest network analysis to date for orchids and their mycorrhizal fungi, sampling a total of over 450 plants from nearly half the 150 tropical orchid species on Reunion Island, encompassing its main terrestrial and epiphytic orchid genera. The authors found a total of 95 operational taxonomic units of mycorrhizal fungi and investigated the architecture and nestedness of their bipartite networks with 73 orchid species. The most striking finding was a major ecological barrier between above‐ and belowground mycorrhizal fungal networks, despite both epiphytic and terrestrial orchids often associating with closely related taxa across all three major lineages of rhizoctonia fungi. The fungal partnerships of the epiphytes and terrestrial species involved a diversity of fungal taxa in a modular network architecture, with only about one in ten mycorrhizal fungi partnering orchids in both groups. In contrast, plant and fungal phylogenetics had weak or no effects on the network. This highlights the power of recently developed ecological network analyses to give new insights into controls on plant–fungal symbioses and raises exciting new hypotheses about the differences in properties and functioning of mycorrhiza in epiphytic and terrestrial orchids.     

Strong phylogenetic congruence between Tulasnella fungi and their associated Drakaeinae orchids

The study of congruency between phylogenies of interacting species can provide a powerful approach for understanding the evolutionary history of symbiotic associations. Orchid mycorrhizal fungi can survive independently of orchids making cospeciation unlikely, leading us to predict that any congruence would arise from host-switches to closely related fungal species. The Australasian orchid subtribe Drakaeinae is an iconic group of sexually deceptive orchids that consists of approximately 66 species. In this study, we investigated the evolutionary relationships between representatives of all six Drakaeinae orchid genera (39 species) and their mycorrhizal fungi. We used an exome capture dataset to generate the first well-resolved phylogeny of the Drakaeinae genera. A total of 10 closely related Tulasnella Operational Taxonomic Units (OTUs) and previously described species were associated with the Drakaeinae orchids. Three of them were shared among orchid genera, with each genus associating with 1–6 Tulasnella lineages. Cophylogenetic analyses show Drakaeinae orchids and their Tulasnella associates exhibit significant congruence (p < 0.001) in the topology of their phylogenetic trees. An event-based method also revealed significant congruence in Drakaeinae–Tulasnella relationships, with duplications (35), losses (25), and failure to diverge (9) the most frequent events, with minimal evidence for cospeciation (1) and host-switches (2). The high number of duplications suggests that the orchids speciate independently from the fungi, and the fungal species association of the ancestral orchid species is typically maintained in the daughter species. For the Drakaeinae–Tulasnella interaction, a pattern of phylogenetic niche conservatism rather than coevolution likely explains the observed phylogenetic congruency in orchid and fungal phylogenies. Given that many orchid genera are characterized by sharing of fungal species between closely related orchid species, we predict that these findings may apply to a wide range of orchid lineages.     

The importance of associations with saprotrophic non-Rhizoctonia fungi among fully mycoheterotrophic orchids is currently under-estimated: novel evidence from sub-tropical Asia

Background and Aims Most fully mycoheterotrophic (MH) orchids investigated to date are mycorrhizal with fungi that simultaneously form ectomycorrhizas with forest trees. Only a few MH orchids are currently known to be mycorrhizal with saprotrophic, mostly wood-decomposing, fungi instead of ectomycorrhizal fungi. This study provides evidence that the importance of associations between MH orchids and saprotrophic non-Rhizoctonia fungi is currently under-estimated.Methods Using microscopic techniques and molecular approaches, mycorrhizal fungi were localized and identified for seven MH orchid species from four genera and two subfamilies, Vanilloideae and Epidendroideae, growing in four humid and warm sub-tropical forests in Taiwan. Carbon and nitrogen stable isotope natural abundances of MH orchids and autotrophic reference plants were used in order to elucidate the nutritional resources utilized by the orchids.Key Results Six out of the seven MH orchid species were mycorrhizal with either wood- or litter-decaying saprotrophic fungi. Only one orchid species was associated with ectomycorrhizal fungi. Stable isotope abundance patterns showed significant distinctions between orchids mycorrhizal with the three groups of fungal hosts.Conclusions Mycoheterotrophic orchids utilizing saprotrophic non-Rhizoctonia fungi as a carbon and nutrient source are clearly more frequent than hitherto assumed. On the basis of this kind of nutrition, orchids can thrive in deeply shaded, light-limiting forest understoreys even without support from ectomycorrhizal fungi. Sub-tropical East Asia appears to be a hotspot for orchids mycorrhizal with saprotrophic non-Rhizoctonia fungi.     

Diversity of mycorrhizal fungi of terrestrial orchids: compatibility webs, brief encounters, lasting relationships and alien invasions

The diversity of mycorrhizal fungi associated with an introduced weed-like South African orchid (Disa bracteata) and a disturbance-intolerant, widespread, native West Australian orchid (Pyrorchis nigricans) were compared by molecular identification of the fungi isolated from single pelotons. Molecular identification revealed both orchids were associated with fungi from diverse groups in the Rhizoctonia complex with worldwide distribution. Symbiotic germination assays confirmed the majority of fungi isolated from pelotons were mycorrhizal and a factorial experiment uncovered complex webs of compatibility between six terrestrial orchids and 12 fungi from Australia and South Africa. Two weed-like (disturbance-tolerant rapidly spreading) orchids — D. bracteata and the indigenous Australian Microtis media, had the broadest webs of mycorrhizal fungi. In contrast, other native orchids had relatively small webs of fungi (Diuris magnifica and Thelymitra crinita), or germinated exclusively with their own fungus (Caladenia falcata and Pterostylis sanguinea). Orchids, such as D. bracteata and M. media, which form relationships with diverse webs of fungi, had apparent specificity that decreased with time, as some fungi had brief encounters with orchids that supported protocorm formation but not subsequent seedling growth. The interactions between orchid mycorrhizal fungi and their hosts are discussed.     

Continental-scale distribution and diversity of Ceratobasidium orchid mycorrhizal fungi in Australia

Background and AimsMycorrhizal fungi are a critical component of the ecological niche of most plants and can potentially constrain their geographical range. Unlike other types of mycorrhizal fungi, the distributions of orchid mycorrhizal fungi (OMF) at large spatial scales are not well understood. Here, we investigate the distribution and diversity of Ceratobasidium OMF in orchids and soils across the Australian continent.MethodsWe sampled 217 Ceratobasidium isolates from 111 orchid species across southern Australia and combined these with 311 Ceratobasidium sequences from GenBank. To estimate the taxonomic diversity of Ceratobasidium associating with orchids, phylogenetic analysis of the ITS sequence locus was undertaken. Sequence data from the continent-wide Australian Microbiome Initiative were used to determine the geographical range of operational taxonomic units (OTUs) detected in orchids, with the distribution and climatic correlates of the two most frequently detected OTUs modelled using MaxEnt.Key ResultsWe identified 23 Ceratobasidium OTUs associating with Australian orchids, primarily from the orchid genera Pterostylis, Prasophyllum, Rhizanthella and Sarcochilus. OTUs isolated from orchids were closely related to, but distinct from, known pathogenic fungi. Data from soils and orchids revealed that ten of these OTUs occur on both east and west sides of the continent, while 13 OTUs were recorded at three locations or fewer. MaxEnt models suggested that the distributions of two widespread OTUs are correlated with temperature and soil moisture of the wettest quarter and far exceeded the distributions of their host orchid species.ConclusionsCeratobasidium OMF with cross-continental distributions are common in Australian soils and frequently have geographical ranges that exceed that of their host orchid species, suggesting these fungi are not limiting the distributions of their host orchids at large spatial scales. Most OTUs were distributed within southern Australia, although several OTUs had distributions extending into central and northern parts of the continent, illustrating their tolerance of an extraordinarily wide range of environmental conditions.     

Saprotrophic fungal mycorrhizal symbionts in achlorophyllous orchids: Finding treasures among the ‘molecular scraps’?

Mycoheterotrophic plants are achlorophyllous plants that obtain carbon from their mycorrhizal fungi. They are usually considered to associate with fungi that are (1) specific of each mycoheterotrophic species and (2) mycorrhizal on surrounding green plants, which are the ultimate carbon source of the entire system. Here we review recent works revealing that some mycoheterotrophic plants are not fungal-specific, and that some mycoheterotrophic orchids associate with saprophytic fungi. A re-examination of earlier data suggests that lower specificity may be less rare than supposed in mycoheterotrophic plants. Association between mycoheterotrophic orchids and saprophytic fungi arose several times in the evolution of the two partners. We speculate that this indirectly illustrates why transition from saprotrophy to mycorrhizal status is common in fungal evolution. Moreover, some unexpected fungi occasionally encountered in plant roots should not be discounted as ‘molecular scraps’, since these facultatively biotrophic encounters may evolve into mycorrhizal symbionts in some other plants.Key words: endophytic fungi, evolution of mycorrhizae, mycoheterophy, mycorrhizae, saprophytic fungi, specificityConsiderable advances were recently made in the ecology of achlorophyllous, heterotrophic plants that obtain carbon from their mycorrhizal fungi (Fig. 1). Most plants have contact with soil through mycorrhizal symbioses, in which roots associate with a suitable fungal partner. Fungi utilize soil mineral nutrients, and while sharing them with host plants, they generally receive carbon as a reward. In contrast, some achlorophyllous plants living in the shaded forest understorey have reversed the process. They receive carbon from their mycorrhizal fungi exclusively, hence the designation ‘mycoheterotrophic’ (MH) plants.¹ Mycoheterotrophy has appeared several times during the evolution of land plants, and more than 20 times among orchids that encompass half of all MH species.² In the last decade, the development of molecular tools has enabled researchers to identify many fungal symbionts, which are often uncultivable. The fungi occurring in the densely colonized roots of MH species often produce a stronger PCR signal than any fungal contaminant, making molecular tools very effective for this field of study.Open in a separate windowFigure 1Wullschlaegelia aphylla, a mycoheterotrophic orchid unspecifically associated with saprotrophic Mycena and Gymnopus species. (A) Whole plant at flowering time, with reduced, tuberoid root system at that period. (B) Section of mycorrhizal root showing intracellular hyphal pelotons at early stage (p), or late stage (undergoing lysis, lp); among orchids, the colonization of dead cortical cell (cc) is a unique feature to some saprotrophic fungi (picture by A. Faccio, University of Torino).