Septoglomus jasnowskae and Septoglomus turnauae,two new species of arbuscular mycorrhizal fungi (Glomeromycota)

Phylogenetic analyses of SSU-ITS-LSU nrDNA sequences and morphological studies of spores and mycorrhizae confirmed our supposition of finding two new species of arbuscular mycorrhizal fungi of the genus Septoglomus in the phylum Glomeromycota. Morphologically, the first species, named S. jasnowskae, is distinguished by its pale yellow to brownish yellow, small spores with a 2-layered spore wall, of which the colourless outer layer 1 stains dark in Melzer’s reagent and layer 2 is laminate. The spores usually arise in loose clusters. The structures most distinguishing S. turnauae are its two coloured laminate layers in the 4-layered spore wall. In the field S. jasnowskae was associated with roots of Ammophila arenaria and an unrecognized plant species colonizing maritime dunes of the Mediterranean Sea near Thessalonica (Greece) and Calella (Spain), respectively, and S. turnauae formed mycorrhiza with a Cistus sp. (Cistaceae) growing in the soil of a mine located in Sulcis-Iglesiente, SW-Sardinia, Italy. In single-species cultures with Plantago lanceolata as host plant, the mycorrhiza of S. jasnowskae consisted of arbuscules, hyphae and vesicles, and that of S. turnauae comprised arbuscules and hyphae only.     

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。     

Glomus drummondii and G. walkeri,two new species of arbuscular mycorrhizal fungi (Glomeromycota)

Two new ectocarpic arbuscular mycorrhizal fungal species, Glomus drummondii and G. walkeri (Glomeromycota), found in maritime sand dunes of northern Poland and those adjacent to the Mediterranean Sea are described and illustrated. Mature spores of G. drummondii are pastel yellow to maize yellow, globose to subglobose, (58–)71(–85) μm diam, or ovoid, 50–80 × 63–98 μm. Their wall consists of three layers: an evanescent, hyaline, short-lived outermost layer, a laminate, smooth, pastel yellow to maize yellow middle layer, and a flexible, smooth, hyaline innermost layer. Spores of G. walkeri are white to pale yellow, globose to subglobose, (55–)81(–95) μm diam, or ovoid, 60–90 × 75–115 μm, and have a spore wall composed of three layers: a semi-permanent, hyaline outermost layer, a laminate, smooth, white to pale yellow middle layer, and a flexible, smooth, hyaline innermost layer. In Melzer's reagent, only the inner- and outermost layers stain reddish white to greyish rose in G. drummondii and G. walkeri, respectively. Both species form vesicular–arbuscular mycorrhizae in one-species cultures with Plantago lanceolata as the host plant. Phylogenetic analyses of the ITS and parts of the LSU of the nrDNA of spores placed both species in Glomus Group B sensu Schüßler et al. [Schüßler A, Schwarzott D, Walker C, 2001. A new fungal phylum, the Glomeromycota: phylogeny and evolution. Mycolological Research 105: 1413-1421.]     

Glomus africanum and G. iranicum, two new species of arbuscular mycorrhizal fungi (Glomeromycota)

Two new arbuscular mycorrhizal fungal species (Glomeromycota) of genus Glomus, G. africanum and G. iranicum, are described and illustrated. Both species formed spores in loose clusters and singly in soil and G. iranicum sometimes inside roots. G. africanum spores are pale yellow to brownish yellow, globose to subglobose, (60-)87(-125) μm diam, sometimes ovoid to irregular, 80-110 x 90-140 μm. The spore wall consists of a semipermanent, hyaline, outer layer and a laminate, smooth, pale yellow to brownish yellow, inner layer, which always is markedly thinner than the outer layer. G. iranicum spores are hyaline to pastel yellow, globose to subglobose, (13-)40(-56) μm diam, rarely egg-shaped, prolate to irregular, 39-54 x 48-65 μm. The spore wall consists of three smooth layers: one mucilaginous, short-lived, hyaline, outermost; one permanent, semirigid, hyaline, middle; and one laminate, hyaline to pastel yellow, innermost. Only the outermost spore wall layer of G. iranicum stains red in Melzer's reagent. In the field G. africanum was associated with roots of five plant species and an unrecognized shrub colonizing maritime sand dunes of two countries in Europe and two in Africa, and G. iranicum was associated with Triticum aestivum cultivated in southwestern Iran. In one-species cultures with Plantago lanceolata as the host plant G. africanum and G. iranicum formed arbuscular mycorrhizae. Phylogenetic analyses of partial SSU sequences of nrDNA placed the two new species in Glomus group A. Both species were distinctly separated from sequences of described Glomus species.     

Four new species of arbuscular mycorrhizal fungi (Glomeromycota) associated with endemic plants from ultramafic soils of New Caledonia

Four new species of arbuscular mycorrhizal (AM) fungi (Glomeromycota) were isolated from the rhizosphere of endemic metallophytic plants in ultramafic soils in New Caledonia (South Pacific) and propagated on Sorghum vulgare. Acaulospora saccata and A. fragilissima are placed in the Acaulosporaceae, Scutellospora ovalis in the Gigasporaceae, and Rhizophagus neocaledonicus in the Glomeraceae. The novelty of these species is supported by morphological characters of spores and phylogenetic analyses of sequences of the rDNA region, comprising partial small subunit rRNA gene, the internal transcribed spacers, 5.8S rRNA gene, and the partial large subunit rRNA gene. New Caledonia is known for its high degree of endemism in plants, which is due to its geographic position and geological history. This is the first taxonomic study exploring local Glomeromycota of this island, which may help to address the question of possible AMF endemism in future studies.     

丛枝菌根真菌最新分类系统与物种多样性研究概况

丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)是自然界分布最广泛的一类植物共生真菌,能够与大部分高等植物的根系形成共生关系.由于它们在农林、环境等领域的巨大应用潜力,国内外关于AMF物种多样性的研究一直受到较高的关注.然而,AMF专性共生的特征以及研究方法不够理想等因素长期阻碍了AMF物种多样性的研究进展.近年来,研究方法的改进与新技术的应用为AMF物种多样性的研究提供了极好的机遇.简述了AMF的最新分类系统及全球物种数量、AMF物种多样性影响因素以及AMF物种多样性研究方法三个方面的研究进展,并分析了今后在AMF物种多样性相关领域值得关注的研究方向.     

Vegetative compatibility and anastomosis formation within and among individual germlings of tropical isolates of arbuscular mycorrhizal fungi (Glomeromycota)

Hyphal anastomoses which play a key role in the formation of interconnected mycorrhizal networks and in genetic exchange among compatible individuals have been studied in a limited number of species and isolates of arbuscular mycorrhizal fungi (AMF), mainly in symbiotic mycelium. In this work, the occurrence and frequency of anastomosis between hyphae of the same and different germlings were assessed in tropical isolates belonging to Acaulospora, Claroideoglomus, Gigaspora, Glomus, Rhizophagus and Scutellospora. Germlings belonging to Acaulospora, Claroideoglomus, Glomus and Rhizophagus formed perfect hyphal fusions, with frequencies ranging from 9.29?±?3.01 to 79.84?±?4.39 % within the same germling and from 14.02?±?7.36 to 91.41?±?3.92 % between different germlings. Rare fusions, occurring within the same hypha, were detected in Gigaspora species, and no anastomoses were observed in Scutellospora species. The consistent detection of nuclei in perfect fusions suggests that nuclear migration is active both within and between germlings. Present data on anastomosis formation, nuclear migration and germling viability in tropical isolates of AMF widen our knowledge on the extensive and consistent occurrence of successful hyphal fusions in this group of beneficial symbionts. The ability to anastomose and establish protoplasm flow, fundamental for the maintenance of physiological and genetic continuity, may produce important fitness consequences for the obligately biotrophic AMF.     

Molecular characterization of chromosome termini of the arbuscular mycorrhizal fungus Glomus intraradices (Glomeromycota)

The minimum chromosome number of Glomus intraradices was assessed through cloning and sequencing of the highly divergent telomere-associated sequences (TAS) and by pulsed field gel electrophoresis (PFGE). The telomere of G. intraradices, as in other filamentous fungi, consists of TTAGGG repeats, this was confirmed using Bal31 nuclease time course reactions. Telomere length was estimated to be roughly 0.9 kb by Southern blots on genomic DNA and a telomere probe. We have identified six classes of cloned chromosomal termini based on the TAS. An unusually high genetic variation was observed within two of the six TAS classes. To further assess the total number of chromosome termini, we used telomere fingerprinting. Surprisingly, all hybridization patterns showed smears, which demonstrate that TAS are remarkably variable in the G. intraradices genome. These analyses predict the presence of at least three chromosomes in G. intraradices while PFGE showed a pattern of four bands ranging from 1.2 to 1.5 Mb. Taken together, our results indicate that there are at least four chromosomes in G. intraradices but there are probably more. The information on TAS and telomeres in the G. intradicies will be essential for making a physical map of the G. intraradices genome and could provide molecular markers for future studies of genetic variation among nuclei in these multigenomic fungi.     

Ancestral lineages of arbuscular mycorrhizal fungi (Glomales)

Using new and existing 18S rRNA sequence data, we show that at least five species of glomalean fungi lie outside the previously defined families and diverged very early in the evolution of that group. These five fungi would have been missed by many previous ecological studies because their sequences are not well matched to available taxon-specific primers and they do not stain well with the standard reagents used for morphological analysis. Based upon spore morphology, these species are currently assigned to Glomus and Acaulospora, and two of the species are dimorphic, exhibiting spore stages of both genera. This suggests that dimorphic spores are the ancestral state for the order and that one or the other morphology was lost in various lineages. Our analyses also show that Geosiphon pyriforme, a symbiont with cyanobacteria, is not necessarily a sister group of the Glomales; instead, it may be derived from mycorrhizal ancestors.     

Arbuscular mycorrhizal fungi (Glomeromycota) communities in tropical savannas of Roraima,Brazil

Savanna vegetation in the northern region of Brazil is jeopardized by several anthropogenic activities including cattle ranching and extensive agriculture, and soil biota of these ecosystems is virtually unknown. The soils in savannas are poor in nutrients, very acidic, and subject to drought, and under these conditions, arbuscular mycorrhizal fungi (AMF) are likely to play a key role on plant nutrition and improving soil structure. In this study, we surveyed AMF communities in five savanna locations in Roraima state, Northern Brazil. AMF species were identified using two approaches: field collected spores and trap cultures. Twenty-three AMF species were identified, including 21 species in field samples, 8 species in trap cultures, of which 15 and 2 were unique to field and trap culture samples, respectively. Gigaspora margarita, Dentiscutata heterogama, and Glomus sp1 were the most frequent species recovered from all locations. AMF communities were dominated by members of Gigasporaceae that accounted for 50 to 87% of the total species richness within each location. Spore numbers differed across locations and ranged from 5 to 25 spores 100 cm^?3 soil. Redundancy analysis indicated that soil organic matter was the only selected predictor among soil parameters and correlated positively with occurrence of Glomus heterosporum. We conclude that savannas in Roraima harbor a high sporulating AMF species richness with communities dominated by members of Gigasporaceae and that organic carbon is an important edaphic factor influencing AMF community composition in this ecosystem.     

Cd-tolerant arbuscular mycorrhizal (AM) fungi from heavy-metal polluted soils

Spores of arbuscular mycorrhizal (AM) fungi were isolated from two heavy-metal polluted soils in France via trap culture with leek (Allium porrum L.). Preliminary identification showed that the predominant spore type of both cultures (P2 and Cd40) belongs to the Glomus mosseae group. Their sensitivity to cadmium was compared to a laboratory reference strain (G. mosseae) by in vitro germination tests with cadmium nitrate solutions at a range of concentrations (0 to 100 mg L^–1) as well as extracts from a metal-polluted and unpolluted soils. Both cultures of AM fungi from heavy-metal polluted soils were more tolerant to cadmium than the G. mosseae reference strain. The graphically estimated EC₅₀ was 0.8 mg L^–1 Cd (concentration added to the test device) for G. mosseae and 7 mg L^–1 for P2 culture, corresponding to effective Cd concentrations of approximately 50–70 g L^–1 and 200–500 g L^–1, respectively. The extract of the metal-polluted soil P2 decreased germination of spores from the reference G. mosseae but not from P2 culture. However, the extracts of two unpolluted soils with different physico-chemical characteristics did not affect G. mosseae, whereas germination of P2 spores was markedly decreased in the presence of one of the extracts. These results indicate a potential adaptation of AM fungi to elevated metal concentrations in soil. The tested spores may be considered as metal-tolerant ecotypes. Spore germination results in presence of soil extracts show the difficulty of assessing the ecotoxic effect of metals on AM fungi without considering other soil factors that may interfere in spore germination and hyphal extension.     

Biolistic transformation of arbuscular mycorrhizal fungi

Gene transfer systems have proved effective for the transformation of a range of organisms for both fundamental and applied studies. Biolistic transformation is a powerful method for the gene transfer into various organisms and tissues that have proved recalcitrant to more conventional means. For fungi, the biolistic approach is particularly effective where protoplasts are difficult to obtain and/or the organisms are difficult to culture. This is particularly applicable to arbuscular mycorrhizal (AM) fungi, being as they are obligate symbionts that can only be propagated in association with intact plants or root explants. Furthermore, these fungi are aseptate and protoplasts cannot be released. Recent advancements in gene transformation systems have enabled the use of biolistic technology to introduce foreign DNA linked to molecular markers into these fungi. In this review we discuss the development of transformation strategies for AM fungi by biolistics and highlight the areas of this technology which require further development for the stable transformation of these elusive organisms.     

Arbuscular mycorrhizal fungi (Glomeromycota) harbour ancient fungal tubulin genes that resemble those of the chytrids (Chytridiomycota)

The genes encoding alpha- and beta-tubulins have been widely sampled in most major fungal phyla and they are useful tools for fungal phylogeny. Here, we report the first isolation of alpha-tubulin sequences from arbuscular mycorrhizal fungi (AMF). In parallel, AMF beta-tubulins were sampled and analysed to identify the presence of paralogs of this gene. The AMF alpha-tubulin amino acid phylogeny was congruent with the results previously reported for AMF beta-tubulins and showed that AMF tubulins group together at a basal position in the fungal clade and showed high sequence similarities with members of the Chytridiomycota. This is in contrast with phylogenies for other regions of the AMF genome. The amount and nature of substitutions are consistent with an ancient divergence of both orthologs and paralogs of AMF tubulins. At the amino acid level, however, AMF tubulins have hardly evolved from those of the chytrids. This is remarkable given that these two groups are ancient and the monophyletic Glomeromycota probably diverged from basal fungal ancestors at least 500 million years ago. The specific primers we designed for the AMF tubulins, together with the high molecular variation we found among the AMF species we analysed, make AMF tubulin sequences potentially useful for AMF identification purposes.     

Genetic processes in arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi (Glomeromycota) colonize roots of the majority of land plants and facilitate their mineral nutrient uptake. Consequently, AM fungi play an important role in terrestrial ecosystems and are becoming a component of sustainable land management practices. The absence of sexual reproductive structures in modern Glomeromycota combined with their long evolutionary history suggest that these fungi may represent an ancient asexual lineage of great potential interest to evolutionary biology. However, many aspects of basic AM fungal biology, including genome structure, within-individual genetic variation, and reproductive mode are poorly understood. These knowledge gaps hinder research on the mechanisms of AM fungal interactions with individual plants and plant communities, and utilization of AM fungi in agricultural practices. I present here the current state of research on the reproduction in AM fungi and indicate what new findings can be expected in the future.     

丛枝菌根真菌系统分类及群落研究技术进展

丛枝菌根真菌（AMF）是自然生态系统重要的组成部分,能与植物根系形成互惠共生体.传统的AMF分类主要依赖于对土壤无性孢子的形态鉴定,具有一定的局限性.近年来基于核酸分析的分子鉴定技术使AMF的分类更具科学性和准确性,补充和完善了基于孢子形态鉴定所建立的分类系统.AMF群落研究依赖于AMF的分类鉴定,主要包括孢子形态鉴定和分子生物学分析两类研究法.本文综述了AMF的分类系统和群落研究方法,着重介绍了近年来应用较多的AMF群落研究的分子生物学技术.作者认为,采取形态与分子相结合的办法将有助于推动AMF群落研究和AMF自然分类系统的建立和完善.     

Enkianthus campanulatus (Ericaceae) is commonly associated with arbuscular mycorrhizal fungi

Enkianthus is the most basal extant genus in the phylogeny of ericaceous plants. Its members harbor arbuscular mycorrhiza (AM)-like hyphal structures in their roots but, as yet, no study has surveyed the AM fungal species component. Roots from six species of Enkianthus were collected from five distantly located sites in Japan. Intracellular hyphal coils were observed in the root cortical cells of all species. Fungal DNA sequences of the small subunit ribosomal RNA gene were obtained from 73 of 75 segments of Enkianthus campanulatus roots by PCR using either AML2 or NS31/AM1primer pairs. Results indicated that all E. campanulatus trees were extensively associated with Glomus spp. A phylogenetic analysis showed that 71 root segments harbored fungi belonging to Glomus group A. Among eight delineated clades, seven did not nest with any known AM fungal species. One clade was detected in all roots at all sites at relatively high frequencies, but the rest were detected sporadically at each site. The placement of sequences from distantly located sites into a single clade without known AM fungal species suggests the common association of E. campanulatus with particular AM fungal taxa.