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。     

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。     

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。     

植物与菌根菌

菌根是土壤中的真菌与高等植物根系所形成的共生体。共生真菌从植物体内获得碳水化合物等营养物质,而植物根也从真菌那里得到所需的营养元素和水,从而互利共生,共同进化。有趣的是,有些真菌对于一种植物来说是菌根菌,而对另一种植物来说可能是寄生菌或病原菌。据统计,种子植物中９５％以上的种能与接合菌、子囊菌、担子菌等亚门的真菌共生,形成菌根。菌根学者习惯上根据其形态解剖学特征,将菌根分为三大类：外生菌根、内生菌根、内外生菌根。外生菌根是菌根真菌菌丝体包围植物尚未木栓化的营养根形成的,其菌丝体不侵染到皮层细胞内…     

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。     

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。     

兰科植物菌根真菌的研究进展

兰科（Orehidaceae）是仅次于菊科（Composltae）的一个兴盛、复杂的植物类群,广布于全球,多数种是著名的药用植物和珍贵花卉。兰科植物具有三大特点：第一、其花形状奇特,色彩艳丽,芳香宜人,授粉机制独特而复杂;第二、种子细小,仅具未分化的原胚;第三、在生活史中,与真菌共生形成内生苗根。因而,长期以来受到人们的普遍喜爱,带来了较高的商业利益,科学家,园艺工作者也从各个角度对兰科植物作了大量的研究。本文从兰科菌根的共生物之———菌根真菌的角度对有关其分类及与植物之间的专一性的研究状况进行了分析讨论,旨在为从…     

真菌在兰科植物种子萌发生长中的作用及相互关系

本文从真菌在兰科种子萌发中的应用、真菌侵染的过程、种子和根系消化真菌的途径及共生真菌的分布等方面,综述了真菌与兰科植物种子萌发及菌根关系的研究进展,讨论了该研究的有关问题及真菌在兰科种子萌发中应用的前景。     

真菌在兰科植物种子萌发生长中的作用及相互关系

本文从真菌在兰科种子萌发中的应用、真菌侵染的过程、种子和根系消化真菌的途径及共生真菌的分布等方面,综述了真菌与兰科植物种子萌发及菌根关系的研究进展,讨论了该研究的有关问题及真菌在兰科种子萌发中应用的前景。     

美花石斛菌根真菌接菌方式与接种效应初步研究

自然条件下,兰科菌根真菌的共生对于兰科植物种子萌芽和植株生长是必不可少的。为探讨有益共生真菌对兰科植物生长的促进作用,特别是在实验室环境下接菌方式的改变与接种效应直接的联系,本研究从野生美花石斛(Dendrobium loddigesii Rolfe.)新鲜营养根中分离、筛选出3种菌根真菌（M1、M2和M3）,采用单一接菌和混合接菌的接种方式,分析3个菌株及其不同接种方式对美花石斛生长的影响。研究得到优势菌株M1和M3,并证实混合接菌对美花石斛的生物量增长具有较好的正效应,两两混合接种方式M1-M2、M2-M3及3个混合接种方式M1-M2-M3均能较好的促进美花石斛生物量的积累。充分发挥混合接菌对兰科植物生长发育所产生的效能,提高生产效率,具有较强的现实意义。     

兰科石斛属植物菌根真菌研究进展

石斛属(Dendrobium)隶属于兰科(Orchidaceae)树兰亚科(Epidendroideae)石斛兰族(Dendrobiinae),是兰科最大的属之一,终生附生于树上或岩石上。石斛属很多种类具有很高的药用价值与观赏价值。由于人为过度采挖和野生生境的破坏,使得野生石斛资源濒临灭绝。石斛属植物为典型的兰科菌根植物,在自然条件下需要与真菌共生,才能完成生活史。菌根真菌对于石斛属植物的种子萌发和植株生长具有重要的作用。对石斛属植物菌根的形成、菌根真菌的作用、菌根真菌多样性及菌根技术在石斛属植物中的应用做了评述,并对今后的研究内容和重点提出了一些思路。     

Mycorrhizal diversity in Apostasia (Orchidaceae) indicates the origin and evolution of orchid mycorrhiza

We demonstrated that "orchid mycorrhiza," a specialized mycorrhizal type, appeared in the common ancestor of the largest plant family Orchidaceae and that the fungal partner shifted from Glomeromycota to a particular clade of Basidiomycota in association with this character evolution. Several unique mycorrhizal characteristics may have contributed to the diversification of the family. However, the origin of orchid mycorrhiza and the diversity of mycobionts across orchid lineages still remain obscure. In this study, we investigated the mycorrhizae of five Apostasia taxa, members of the earliest-diverging clade of Orchidaceae. The results of molecular identification using nrDNA ITS and LSU regions showed that Apostasia mycorrhizal fungi belong to families Botryobasidiaceae and Ceratobasidiaceae, which fall within the order Cantharellales of Basidiomycota. Most major clades in Orchidaceae also form mycorrhizae with members of Cantharellales, while the sister group and other closely related groups to Orchidaceae (i.e., Asparagales except for orchids and the "commelinid" families) ubiquitously form symbioses with Glomeromycota to form arbuscular mycorrhizae. This pattern of symbiosis indicates that a major shift in fungal partner occurred in the common ancestor of the Orchidaceae.     

兰科植物种子共生萌发真菌多样性及共生萌发机制研究进展

自然环境下,兰科植物种子细小无胚乳,需要和适宜的真菌共生才能萌发,因而与真菌有天然的共生关系。自身繁殖率低加之近年来栖息地环境破坏导致兰科植物资源更加濒危,而通过筛选适合的真菌进行种子的共生萌发可以有效地实现兰科植物的种质保育及濒危种类野生居群的生态恢复。本文对地生型、附生型以及腐生型等兰科植物已发现的萌发真菌的多样性进行了系统地梳理,发现担子菌门的胶膜菌科、角担菌科以及蜡壳耳目真菌为已报道共生萌发真菌的主要类群;同时对兰科植物种子的共生萌发机制,包括形态学机制、营养机制和分子机制等方面的相关研究进行了归纳论述,但是当前关于兰科植物和真菌互作机制方面的研究还相对较少,许多问题需要进一步明确。本文对共生萌发真菌在兰科植物保育和繁育中的应用以及共生萌发机制的研究等方面具有一定的参考价值。     

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.     

Chlorophyllous and Achlorophyllous Specimens of <Emphasis Type="Italic">Epipactis microphylla</Emphasis> (Neottieae,Orchidaceae) Are Associated with Ectomycorrhizal Septomycetes,including Truffles

Mycoheterotrophic species (i.e., achlorophyllous plants obtaining carbon from their mycorrhizal fungi) arose many times in evolution of the Neottieae, an orchid tribe growing in forests. Moreover, chlorophyllous Neottieae species show naturally occurring achlorophyllous individuals. We investigated the fungal associates of such a member of the Neottieae, Epipactis microphylla, to understand whether their mycorrhizal fungi predispose the Neottieae to mycoheterotrophy. Root symbionts were identified by sequencing the fungal ITS of 18 individuals from three orchid populations, including achlorophyllous and young, subterranean individuals. No rhizoctonias (the usual orchid symbionts) were recovered, but 78% of investigated root pieces were colonized by Tuber spp. Other Pezizales and some Basidiomycetes were also found. Using electron microscopy, we demonstrated for the first time that ascomycetes, especially truffles, form typical orchid mycorrhizae. All identified fungi (but one) belonged to taxa forming ectomycorrhizae on tree roots, and four of them were even shown to colonize surrounding trees. This is reminiscent of mycoheterotrophic orchid species that also associate with ectomycorrhizal fungi, although with higher specificity. Subterranean and achlorophyllous E. microphylla individuals thus likely rely on tree photosynthates, and a partial mycoheterotrophy in individuals plants can be predicted. We hypothesize that replacement of rhizoctonias by ectomycorrhizal symbionts in Neottieae entails a predisposition to achlorophylly.     

The rare terrestrial orchid Nervilia nipponica consistently associates with a single group of novel mycobionts

Nervilia nipponica is a tuberous terrestrial orchid that has a highly restricted distribution within common secondary evergreen forest communities in central and western Japan. Such a limited occurrence could be attributable to a requirement for a specific mycorrhizal fungus. As part of a broader examination of this hypothesis, we sought to elucidate the mycorrhizal associations of N. nipponica. Seventy-five samples of mycorrhizae from forty individuals were collected at ten populations throughout the orchid’s range in Japan. The identity of mycorrhizal fungi was investigated by sequencing two genetic markers (nrDNA ITS and nrDNA 28S LSU) and their relationships were assessed via phylogenetic analyses. The most frequently encountered mycorrhizal fungi consisted of four closely related Agaricomycetes that infected an average of 78.7 % of individuals per population. All four formed a discrete, monophyletic clade with low sequence homology to other fungi registered in GenBank, indicating that they belong to a novel, unnamed family. Two additional fungal groups, belonging to Ceratobasidiaceae and “Group B” Sebacinales, were found in 22.0 and 21.5 % of individuals per population, respectively. The orchid probably uses these two groups opportunistically, because they were found at lower densities and always in combination with the unidentified Agaricomycete. These findings suggest that a group of novel Agaricomycete fungi constitutes the dominant mycobiont of N. nipponica.     

Mycorrhizal symbioses found in roots of fern and its relatives in Korea

Mycorrhizal symbioses were found in the roots of 45 out of 59 species of pteridophytes collected in Korea. The mycorrhizal fungi were colonized in the root cortical cells, primarily in terrestrial species, but rarely in epiphytic or aquatic pteridophytes. Mycorrhizae that are typically found in orchid colonized the roots of the epiphytic pteridophytes, but not in other species. These were the first observations of orchid mycorrhizae in pteridophytes. Arbuscular mycorrhizal fungi were examined after staining, then confirmed with PCR, using a specific primer. This is the first report of arbuscular mycorrhizal colonization in the roots of pteridophyte species in Asia.     

Variation in mycorrhizal performance in the epiphytic orchid <Emphasis Type="Italic">Tolumnia variegata in vitro</Emphasis>: the potential for natural selection

Symbiotic seed germination is a critical stage in orchid life histories. Natural selection may act to favor plants that efficiently use mycorrhizal fungi. However, the necessary conditions for natural selection – variation, heritability, and differences in fitness – have not been demonstrated for either orchid or fungus. With the epiphytic orchid Tolumnia variegata as a model system, we ask the following questions: (1) Do seeds from different individuals in a population differ in germination and seedling development in the presence of the same fungi? (2) Do different mycorrhizal fungi (Ceratobasidium spp.) differ in ability to stimulate seed germination and growth in T. variegata? And (3) are the Ceratobasidium isolates that best induce seed germination and seedling development more closely related to each other than to isolates that are less effective? We performed symbiotic seed germination experiments in vitro. The experiments were done using mycorrhizal fungi isolated from T. variegata; relationships among the fungi were inferred from nuclear ribosomal ITS sequences. We found significant variation for both symbiotic germination and seedling growth among biparental seed crops obtained from a population of T. variegata plants. Differences among Ceratobasidium fungi in seed germination were significant. The fungi that induced highest seed germination and seedling development belonged to two of four clades of Ceratobasidium. The two experiments show that there is potential for natural selection to act on orchid–fungus relationships. Given that orchids vary in performance, and that mycorrhizal fungi are not geographically distributed homogeneously, mycorrhizae may affect population size, distribution and evolution of orchids.