The Distribution of Cytoplasm and Nuclei within the Extra-radical Mycelia in Glomus intraradices, a Species of Arbuscular Mycorrhizal Fungi.

Nuclear distribution within the extra-radical fungal structures and during spore production in the arbuscular mycorrhizae fungus Glomus intraradices was examined using an in vitro monoxenic culture system. A di-compartmental monoxenic culture system was modified using a nitrocellulose membrane and a coverglass slip for detailed observations. Nuclear distribution was observed using the fluorescent DNA binding probes SYBR Green I and DAPI. Both septate and non-septate mycelial regions were observed, but cytoplasmic contents were only found within non-septate mycelia. Nuclear fluorescent staining revealed that the non-septate hyphal region contained nuclei only with cytoplasm, and that nuclear distribution was limited by septa. Swollen hyphal bodies were often associated with septate and empty-looking hyphae. Cytoplasmic contents filled the swollen hyphal body from the non-septate hyphal region following removal of the septa. As a consequence, the swollen body developed into a new spore. These observations provide understanding about the distribution of AM fungal nuclei within extra-radical mycelia and during spore formation. The results suggest a mechanism by which the development of a cytoplasm-containing mycelium is controlled by the formation or removal of septa to efficiently maintain and proliferate essential contents. This mechanism may provide a survival strategy to the fungus.

The mitochondrial genome sequence of the arbuscular mycorrhizal fungus Glomus intraradices isolate 494 and implications for the phylogenetic placement of Glomus.

* We have determined the complete mitochondrial genome sequence of an isolate of Glomus intraradices, a widespread and well-studied species of arbuscular mycorrhizal fungus. * The total genomic DNA of 24 spores from an in vitro root organ culture of the Swiss isolate G. intraradices 494 was amplified by multiple displacement and sequenced using the Roche 454 FLX platform. Contigs were joined by PCR and Sanger sequencing. * The circular genome map of 70 606 bp has a G + C content of 37.2%. All the standard fungal mitochondrial genes are present and encoded on the same strand. There are 26 introns and five complete LAGLIDADG homing endonuclease genes. There is no evidence of substantial sequence variation. * A well-supported phylogeny based on 14 mitochondrially encoded proteins indicates that the Glomeromycota are not the sister group of the Dikarya.

Improved PCR primers for the detection and identification of arbuscular mycorrhizal fungi.

A set of PCR primers that should amplify all subgroups of arbuscular mycorrhizal fungi (AMF, Glomeromycota), but exclude sequences from other organisms, was designed to facilitate rapid detection and identification directly from field-grown plant roots. The small subunit rRNA gene was targeted for the new primers (AML1 and AML2) because phylogenetic relationships among the Glomeromycota are well understood for this gene. Sequence comparisons indicate that the new primers should amplify all published AMF sequences except those from Archaeospora trappei. The specificity of the new primers was tested using 23 different AMF spore morphotypes from trap cultures and Miscanthus sinensis, Glycine max and Panax ginseng roots sampled from the field. Non-AMF DNA of 14 plants, 14 Basidiomycota and 18 Ascomycota was also tested as negative controls. Sequences amplified from roots using the new primers were compared with those obtained using the established NS31 and AM1 primer combination. The new primers have much better specificity and coverage of all known AMF groups.

Molecular identification of arbuscular mycorrhizal fungal spores collected in Korea.

Arbuscular mycorrhizas (AM) have mutualistic symbiosis with plants and thus efforts have been placed on application of these symbiotic relationships to agricultural and environmental fields. In this study, AM fungi were collected from 25 sites growing with 16 host plant species in Korea and cultured with Sorghum bicolor in greenhouse condition. AM fungal spores were extracted and identified using both morphological and molecular methods. Using morphological characters, total 15 morpho-speices were identified. DNA was extracted from single spore of AM fungi and a partial region on 18S rDNA was amplified using nested PCR with AM fungal specific primers AML1/AML2. A total of 36 18S rDNA sequences were analyzed for phylogenetic analysis and 15 groups of AM fungi were identified using both morphological and molecular data of spores. Among the species, 4 species, Archaeospora leptoticha, Scutellospora castanea, S. cerradensis, S. weresubiae were described for the first time in Korea and two species in Glomus and a species in Gigaspora were not identified. Morphological and molecular identification of AM fungal spores in this study would help identify AM fungal community colonizing roots.

Effects of interspecific interactions of arbuscular mycorrhizal fungi on growth of soybean and corn.

Growth responses of Zea mays and Glycine max to colonization by mixture of combination of three species of arbuscular mycorrhizal (AM) fungi, two species of Glomus and a species of Scutellospora were compared. In Zea mays, plants inoculated with single species of AM fungi showed significantly higher in dry weight than non-mycorrhizal plant for all three AM fungal species. Also, growth of plants inoculated with spores of two species of AM fungi was significantly higher than nonmycorrhizal control except for plants inoculated with two Glomus species. When three species of AM fungi were inoculated, the plants showed the highest growth. In Glycine max, plants with single AM fungal species inoculation were not significantly different in plant growth from nonmycorrhizal plants. When the plants were inoculated with combination of two or more AM fungal species, their growth significantly increased compared to nonmycorrhizal plants. In both plant species, mycorrhizal root colonization by Scutellospora species was significantly lower than by Glomus species.