Taxonomic status of Monotropastrum humile, with special reference to M. humile var. glaberrimum (Ericaceae, Monotropoideae)

Taxonomic treatment of the achlorophyllous monotropoid plant Monotropastrum humile is still unclear and confusing because of the lack of detailed morphological analyses and molecular phylogeny. In particular, the taxonomic status of a glabrous variety, M. humile var. glaberrimum, is under debate. Our detailed examination of the morphological characteristics of living plants revealed that M. humile var. glaberrimum can be easily distinguished from the putative conspecific taxon M. humile var. humile by characteristics not previously recognized, namely the shape and color of the floral disc. Most morphological features characterizing Cheilotheca were also found in M. humile var. glaberrimum. Moreover, there was considerable nucleotide differentiation in the internal transcribed spacer (ITS)2 sequences of M. humile var. humile and var. glaberrimum. Molecular analysis of the phylogenetic relationship of M. humile var. humile, var. glaberrimum, and other monotropoids using ITS2 sequences showed that two varieties of M. humile formed a monophyletic clade with a member of a different genus, Monotropa L., but obvious phylogenetic relationships among these three taxa were not obtained. Thus we conclude that Monotropastrum humile var. glaberrimum should be treated as a distinct species. However, the generic affiliation of M. humile var. glaberrimum could not be determined because of its intermediate character state combination and the insufficient characterization of related species. We strongly suggest that Monotropastrum as a whole needs re-evaluation.     

Introduction of asymbiotically propagated seedlings of <Emphasis Type="Italic">Cephalanthera falcata</Emphasis> (Orchidaceae) into natural habitat and investigation of colonized mycorrhizal fungi

Asymbiotic seedling propagation and introduction of seedlings into a natural habitat were achieved for Cephalanthera falcata. For immature seeds collected 65 days after pollination, high germination rate (av. 50%) was achieved on Hyponex agar medium plates. Root development occurred in about 10% of the protocorms 5 months after seed sowing. Rooted protocorms were transferred to a culture bottle containing 100 ml of the Hyponex agar medium and incubated continually. In about 30% of the transferred individuals, shoot height reached 1.5–2 cm 8 months after the transfer. After acclimatization in wet vermiculite at 4°C for 6 months, 135 individuals were planted in a natural stand of C. falcata in mid February 2001. Shoot appearance rate was 44.4% at the first year and flowering was noted in some plants. At the fifth year, shoots with an average height of 21.6 cm still appeared in four plants, and flowering was noted in three of them. Colonization of mycorrhizal fungi was examined in two of them as well as one wild plant, in which the mycorrhizal fungi were identified to be in Thelephoraceae or Russulaceae. These fungi are known to form ectomycorrhiza with trees, and thus a tripartnership symbiosis consisting of C. falcata, mycorrhizal fungi and trees was suggested. The involvement of ectomycorrhizal fungi might be the reason for the low survival rate in the field experiment, because the distribution of ectomycorrhizal fungi relevant to this orchid is assumed to be uneven. The possibility of introducing artificially propagated orchids into natural habitats was discussed.     

拮抗植物病原真菌的纤维素/木质素降解菌株的筛选及鉴定

【背景】秸秆还田在改善土壤肥力和丰富营养等方面有重要作用,但是也存在秸秆难以快速降解利用和病原真菌病害威胁的问题。【目的】为解决还田秸秆的降解和病原真菌病害问题,从长期秸秆还田地区采集样品,从中筛选出具有降解秸秆和抑菌功能的菌株。【方法】采用稀释分离法、苯胺蓝染色法和刚果红染色法等对秸秆高效降解菌株进行筛选,通过16S rRNA基因测序及构建系统发育树进行菌株鉴定。采用对峙培养法测定筛选到的秸秆降解菌株对玉米大斑病菌(Setostphaeria turcica)、梨黑斑病菌(Alternaria kikuchiana)、马铃薯早疫病菌(Alternaria solani)、链格孢属真菌(Alternaria alternata) ACCC38230和ACCC38231等5种供试植物病原真菌的抑制作用。以玉米大斑病菌(Setostphaeria turcica)为后期供试植物病原真菌,测定拮抗菌株代谢产物的抑菌能力;通过观察拮抗菌株粗提液对玉米大斑病菌分生孢子萌发和菌丝生长的影响,从而测定菌株对病原真菌的抑制作用。【结果】从秸秆还田土壤中分离筛选获得3株高效降解纤维素及木质素菌株并命名为JY122、ZY133和JY215。通过形态学观察和16S rRNA基因测序鉴定上述菌株全部为芽孢杆菌属(Bacillus)。通过系统发育树分析结果表明,JY122与蜡样芽孢杆菌(Bacillus cereus)相似性为99.4%,ZY133与枯草芽孢杆菌(Bacillus subtilis)相似性为100%,JY215与贝莱斯芽孢杆菌(Bacillus velezensis)相似性为99.1%。平板对峙实验结果显示,JY122、ZY133和JY215菌株对不同种属的植物病原真菌均有较强的抑制作用,抑制率高达43.74%-67.54%。此外,上述芽孢杆菌代谢产物具有抑菌活性及很强的热稳定性,经95℃处理后依然具有良好的抑菌效果。【结论】筛选获得的JY122、JY133和JY215菌株具有高效降解纤维素/木质素能力,抑制多种植物病原真菌生长,代谢产物抑菌能力强且热稳定性高。这为玉米秸秆还田提供菌株资源,也为进一步解决秸秆还田难点提供了新方法和思路。     

春兰菌根真菌CL-3菌株的鉴定及共生体系的构建

采用分离自野生春兰(Cymbidium goeringii)根部的真菌CL-3菌株, 进行了春兰内生菌根真菌的人工接种、再分离及其共生培养研究。通过对CL-3菌株形态学观察和ITS序列同源性分析, 发现该菌株ITS序列与Acremonium strictum的亲缘关系最为接近, 序列同源性为100%。用CL-3菌株接种春兰组培苗, 接种后2个月可从组培苗中再分离获得该菌株, 且CL-3菌株处理苗的鲜重增长率达80.5%, 经方差分析, 与对照相比有显著差异。通过石蜡切片和染色, 在已接种的组培苗的根部组织中可观察到CL-3菌株存在。表明CL-3菌株能与组培幼苗成功建立共生培养体系。     

Isolation and identification of mycorrhizal fungi associating with an achlorophyllous plant, Epipogium roseum (Orchidaceae)

The identity of mycorrhizal fungi associated with the achlorophyllous orchid Epipogium roseum was investigated by DNA analysis. The fungi were isolated from each coiled hypha (peloton), and the ITS region of nuclear rDNA was sequenced. Phylogenetic analysis based on the neighbor-joining method showed that all the isolates clustered with fungi belonging to Psathyrella or Coprinus in Coprinaceae. Those fungi are known as saprobes, using dead organic materials for a nutritive source. Large colonies of this orchid were frequently found around tree stumps or fallen logs. In such colonies, these decaying wood materials would be used as a large and persistent carbon source for the growth of this orchid. This is the first report of Coprinaceae as an orchid mycorrhizal fungi.     

Innate attractant for long‐tongued bumblebee,Bombus diversus diversus in floral scent of bumblebee‐pollinated epiparasitic plant,Monotropastrum humile (Ericaceae)

The epiparasitic plant Monotropastrum humile (D. Don) Hara is pollinated by workers of the long‐tongued bumblebee, Bombus diversus diversus (Smith). The interaction between the floral scents of this plant and bumblebee was investigated using electrophysiological and behavioral techniques. Three components (linalool, α‐terpineol and geraniol) elicited strong responses in antennae from B. d. diversus workers and are innate attractants, suggesting floral scents play an important role in attracting these pollinators and successful pollination.     

独蒜兰提取物对植物病原真菌的抑菌活性

【目的】研究独蒜兰假鳞茎乙醇提取物对植物病原真菌的抑菌作用,为植物源杀菌剂的开发提供依据。【方法】采用菌丝生长速率法,研究独蒜兰假鳞茎乙醇提取物对15种植物病原菌的抑制活性;以西瓜尖孢镰刀菌作为供试菌,进一步研究该提取物对病原真菌的菌丝干重、细胞膜、过氧化氢酶（CAT）、过氧化物酶（POD）和超氧化物歧化酶（SOD）等的影响。【结果】独蒜兰假鳞茎提取物对辣椒疫霉病菌、西瓜尖孢镰刀菌、番茄灰霉病菌和非洲隐地疫霉菌的抑菌效果明显,其EC₅₀值分别为0.849、0.782、0.813和1.161 mg·mL^-1;经独蒜兰提取物处理后的西瓜尖孢镰刀菌菌丝干重随着药剂浓度的增加而减少;细胞膜丙二醛含量和相对电导率增加;菌丝体细胞内CAT、POD和SOD 3种保护酶活性增加。【结论】独蒜兰假鳞茎提取物对植物病原真菌具有较好的抑菌活性,其抑菌作用可能与其干扰菌丝生长、使菌丝细胞膜正常功能受损等有关。     

Molecular diversity of arbuscular mycorrhizal fungi in onion roots from organic and conventional farming systems in the Netherlands

Diversity and colonization levels of naturally occurring arbuscular mycorrhizal fungi (AMF) in onion roots were studied to compare organic and conventional farming systems in the Netherlands. In 2004, 20 onion fields were sampled in a balanced survey between farming systems and between two regions, namely, Zeeland and Flevoland. In 2005, nine conventional and ten organic fields were additionally surveyed in Flevoland. AMF phylotypes were identified by rDNA sequencing. All plants were colonized, with 60% for arbuscular colonization and 84% for hyphal colonization as grand means. In Zeeland, onion roots from organic fields had higher fractional colonization levels than those from conventional fields. Onion yields in conventional farming were positively correlated with colonization level. Overall, 14 AMF phylotypes were identified. The number of phylotypes per field ranged from one to six. Two phylotypes associated with the Glomus mosseae–coronatum and the G. caledonium–geosporum species complexes were the most abundant, whereas other phylotypes were infrequently found. Organic and conventional farming systems had similar number of phylotypes per field and Shannon diversity indices. A few organic and conventional fields had larger number of phylotypes, including phylotypes associated with the genera Glomus-B, Archaeospora, and Paraglomus. This suggests that farming systems as such did not influence AMF diversity, but rather specific environmental conditions or agricultural practices. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Authors Guillermo A. Galván and István Parádi contributed equally to this research and share first co-authorship.     

Sequences and interactions of mycorrhizal fungi on birch

Summary Soil cores collected under a birch tree (Betula pubescens) on an experimental plot showed a progressive change in types of sheathing mycorrhiza with distance from the tree base. Seedlings grown in cores in a glasshouse also developed different mycorrhizal types depending on distance from the tree at which the cores were taken, but the types on seedlings were often different from those in the parent cores. When cores were taken directly beneath fruitbodies and sown to birch in a glasshouse, seedlings developed mycorrhizas of Laccaria, Inocybe and Hebeloma in cores from beneath these fruitbodies, but they seldom developed Lactarius mycorrhizas and never developed Leccinum mycorrhizas in cores taken beneath these fruitbodies. Similarly, when seedlings were grown in soils supplemented with vermiculite-peat inocula in a glasshouse, Laccaria and Hebeloma readily formed mycorrhizas, butLactarius pubescens seldom did so and Leccinum andAmanita muscaria never dit so. Yet all these fungi form mycorrhizas on birch seedlings in aseptic conditions.The results suggest a distinction between early stage and late stage mycorrhizal fungi of birch. Early stage fungi readily infect seedlings from resident or introduced inoculum in normal, unsterile soil, whereas late stage fungi do not readily form mycorrhizas in these conditions.     

Production of B-group vitamins by mycorrhizal fungi and actinomycetes isolated from the root zone of pine (Pinus sylvestris L.)

Summary Studies were carried out on synthesis of B-group vitamins by mycorrhizal fungi and actinomycetes (Streptomyces sp.) derived from soil, rhizosphere and mycorrhizosphere of pine.None of the fungal isolates produced biotin. The vitamin produced in largest amounts by the mycorrhizal fungi was thiamin.In general more actinomycetes isolated from the rhizosphere than from the root free soil produced B-group vitamins. This was particularly true for thiamin.The amount of vitamins produced was higher in actinomycetes than the amounts produced by the mycorrhizal fungi.This research was carried out under problem MR.II. 16 coordinated by the Institute of Dendrology, Polish Academy of Sciences.     

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。     

Utilisation of carbon substrates by multiple genotypes of ericoid mycorrhizal fungal endophytes from eastern Australian Ericaceae

The abilities of six genotypes of two putative Helotiales ascomycete ericoid mycorrhizal fungal taxa from Woollsia pungens and Leucopogon parviflorus (Ericaceae) to utilise glucose, galactose, mannose, cellobiose, carboxymethylcellulose, crystalline cellulose, starch and xylan as sole carbon sources were tested in axenic liquid culture. With the exception of all taxon II isolates on carboxymethylcellulose, all genotypes of both taxa produced measurable biomass on all substrates. Significant intraspecific variation was observed in biomass production on all substrates. While pooled data for all genotypes of each taxon revealed significant interspecific differences in biomass production on carboxymethylcellulose, glucose, cellobiose, and starch, mean biomass production for each taxon on the latter three substrates differed less than threefold, suggesting that the saprotrophic abilities of the two taxa are broadly similar.     

The contributions of mycorrhizal fungi to the determination of plant community structure

While it is now widely accepted, even by ecologists, that most plants in the majority of ecosystems are infected by mycorrhizal fungi, few experiments have been designed to investigate the function of the mutualism at the community level. Those involved with mycorrhizal research have been largely preoccupied with questions of the mineral, particularly phosphorus, nutrition of individual plants, while plant community ecologists have too often found it convenient, even when acknowledging the presence of infection, to ignore its possible function in the ecosystem. This presentation examines a selected number of seminal papers written by plant community ecologists and highlights some of ‘the most striking mysteries’ which they reveal. It describes experiments designed to determine whether knowledge of the presence and activity of the mycorrhizal mycelium can help us to unravel the ‘mysteries’ which they define. It is revealed that by having direct adverse effects upon seedlings of many ‘r’ selected species, while at the same time being beneficial, if not essential, to those that are ‘K’ selected, the activities of the mycelium of VA fungi have a direct bearing upon community composition. The extent to which ‘turf compatibility’ is actually a reflection of the compatibility of plant species with the VA mycorrhizal mycelium is discussed and the possible role of the mycelium in consigning some species to the ruderal habit is considered. It is concluded that those attempting scientifically to understand, or managerially to manipulate, plant communities, without recognizing the role of the mycorrhizal mycelium, do so at their peril, and it is recommended that scientists involved in research on mycorrhiza extend their vision beyond the limited horizons which are currently so often defined by considerations of the phosphorus nutrition of individual host plants.     

Distribution of dominant arbuscular mycorrhizal fungi among five plant species in undisturbed vegetation of a coastal grassland

Most plant species in mixed grassland vegetation are colonized by arbuscular mycorrhizal (AM) fungi. Previous studies have reported differences in host preferences among AM fungi, although the fungi are known to lack host specificity. In the present study, the distribution of phylogenetic groups of AM fungi belonging to a clade of Glomus species was studied in five plant species from a coastal grassland in Denmark. The occurrence of the fungi was determined by PCR analyses of fungal large subunit ribosomal DNA sequences amplified from root fragments using a specific primer set. The results showed that the dominant Glomus species were able to colonize all the studied plant species, supporting the view that the AM fungi represent a large underground interconnecting mycelial network.     

Production of auxins and gibberellin-like substances by mycorrhizal fungi,bacteria and actinomycetes isolated from soil and the mycorrhizosphere of pine (Pinus silvestris L.)

Summary Production of auxins and gibberellin-like substances by mycorrhizal fungi, bacteria and actinomycetes isolated from the mycorrhizosphere of Scots pine was studied. Chromatography and biossays were used.Most of the organisms required tryptophan for auxins production. The highest biological activity exhibited substances located at R_f 0.2–0.4.The organisms produced minute amounts of gibberellin-like substances which appeared at different R_f values. It was stated that auxins production is much more common among the root zone organisms of pine than the production of gibberellin-like substances.This research was carried out under problem MR.II.16 coordinated by the Institute of Dendrology, Polish Academy of Sciences.     

Effects of arbuscular mycorrhizal fungi and P-solubilizing Pseudomonas fluorescence (ATCC-17400) on morphological traits and mineral content of sesame

Sesame (Sesamum indicum L.) is an important staple crop of the family Pedaliaceae. The commercial production of sesame is still dependent on the applications of chemical fertilizers. Mycorrhiza inoculum resulted in better morphological and biochemical traits in vegetables. Thus, here the outcome of arbuscular mycorrhizal fungi (AMF) and Pseudomonas fluorescence (ATCC-17400) inoculation was studied in the pot culture experiment. Primarily, there seems to be a promising opportunity of AMF in sesame under pot and field trials because of enhanced morphological parameters, especially root weight, and disparities in nutrients and metabolites. The AMF appears to be an option to boost plant growth, mineral content, and sesame yield. The AMF treatment with Pseudomonas fluorescence strain (ATCC-17400) determined the maximum values for the morphological traits and mineral content. Overall, our study highlights mycorrhizal fungi and other microbes efficacy in achieving a successful sesame production.     

Molecular detection,community structure and phylogeny of ericoid mycorrhizal fungi

Through traditional culturing and molecular characterization, we have determined that five putative species and 2 polyphyletic assemblages of fungi produce ericoid mycorrhizae in Gaultheria shallon, other Ericaceae and Epacridaceae. Using phylogenetic analysis of ITS2 sequences in GenBank, we have confirmed that most of these fungi occur in North America, Europe, and Australia. The low recovery rate of culturable ericoid mycorrhizal fungi from Gaultheria shallon may partly be explained by the fact that most mycorrhizal root segments contain an unculturable basidiomycete, revealed by direct amplification, cloning, and sequencing of LSU fungal DNA from root. Molecular characterization and phylogenetic analysis are powerful tools in revealing the geographic distribution and identity of ericoid mycorrhizal fungi.     

Vesicular-arbuscular mycorrhizal fungi in coastal dune plant communities I. Spore formation ofGlomus spp. predominates under a patch ofElymus mollis

Vesicular-arbuscular (VA) mycorrhizal fungi in pure patches of coastal dune plantsElymus mollis, Wedelia prostrata andZoysia macrostachya were examined for frequency of occurrence and number of spores of VA mycorrhizal fungi over one year. Six species in three genera of VA mycorrhizal fungi were recovered. Under a patch ofE. mollis, spores ofAcaulospora sp. 1,Glomus tortuosum, Glomus sp. 1,Glomus sp. 2 andScutellospora gregaria were recovered. Spores ofGlomus spp. were most common. In patches ofW. prostrata andZ. macrostachya spores ofAcaulospora sp. 1,G. tortuosum, Glomus sp. 1,Glomus sp. 2,S. gregaria andScutellospora sp. 1 were found.Contribution No. 112, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba.     

丛枝菌根真菌对紫茎泽兰生长与繁殖的影响

【目的】紫茎泽兰是一种世界性的恶性入侵杂草,侵入我国后迅速扩散至多个省市,严重阻碍了当地农、林、牧业的发展。研究丛枝菌根真菌对紫茎泽兰生长和繁殖的影响,解析土壤中丛枝菌根真菌在紫茎泽兰入侵、传播和扩散过程中的作用,能为开展紫茎泽兰防控工作提供参考。【方法】通过盆栽实验,测定接种丛枝菌根真菌的紫茎泽兰株高、叶片数、总叶面积、节间距、地径,并统计单株花序数、单花序种子量、种子量、种子千粒重、含水率、发芽率、发芽势、发芽指数等。【结果】丛枝菌根真菌对紫茎泽兰的株高、地径有极显著影响(P0.01),接种丛枝菌根真菌后,紫茎泽兰的单株花序数、单花序种子数、总种子量分别增加了5.03、1.51、7.64倍;种子的长度、宽度、含水率、发芽率、发芽势、发芽指数分别增加了15.24%、23.08%、19.46%、29.22%、391.01%、183.56%(P0.05)。【结论】丛枝菌根真菌可显著促进紫茎泽兰生长,控制丛枝菌根真菌的数量可有效抑制紫茎泽兰生长,并抑制紫茎泽兰花序数、种子量及种子的生命力,降低其扩散蔓延速度。     

Isolation and identification of mycorrhizal fungi associated with <Emphasis Type="Italic">Stigmatodactylus sikokianus</Emphasis> (Maxim. ex Makino) Rauschert (Orchidaceae)

The mycorrhizal fungi of Stigmatodactylus sikokianus (Orchidaceae) were isolated and identified to be nearly related to Sebacina spp. in Sebacinaceae (Basidiomycota) by a neighbor-joining phylogenetic analysis based on the sequences of the ITS region of nuclear rDNA. In spite of the geographically separated samplings, high sequence similarity was found among the obtained DNA sequences, which suggested that S. sikokianus might be highly specialized to the group of fungi. It is known that Sebacina spp. are saprobes or ectomycorrhiza-forming fungi. The mycorrhizal fungi of S. sikokianus were regarded to be saprobic from the environment of their habitats.