Three new species of arbuscular mycorrhizal fungi (Glomeromycota) and Acaulospora gedanensis revised.

Studies of the morphology and the 45S nuc rDNA phylogeny of three potentially undescribed arbuscular mycorrhizal fungi (phylum Glomeromycota) grown in cultures showed that one of these fungi is a new species of the genus Diversispora in the family Diversisporaceae; the other two fungi are new Scutellospora species in Scutellosporaceae. Diversispora vistulana sp. nov. came from maritime sand dunes of the Vistula Spit in northern Poland, and S. graeca sp. nov. and S. intraundulata sp. nov. originally inhabited the Mediterranean dunes of the Peloponnese Peninsula, Greece. In addition, the morphological description of spores of Acaulospora gedanensis, originally described in 1988, was emended based on newly found specimens, and the so far unknown phylogeny of this species was determined. The phylogenetic analyses of 45S sequences placed this species among Acaulospora species with atypical phenotypic and histochemical features of components of the two inner germinal walls.

Phenol and Polyaromatic Hydrocarbons Are Stronger Drivers Than Host Plant Species in Shaping the Arbuscular Mycorrhizal Fungal Component of the Mycorrhizosphere.

Changes in soil microbial communities in response to hydrocarbon pollution are critical indicators of disturbed ecosystem conditions. A core component of these communities that is functionally adjusted to the life-history traits of the host and environmental factors consists of arbuscular mycorrhizal fungi (AMF). AMF communities associated with Poa trivialis and Phragmites australis growing at a phenol and polynuclear aromatic hydrocarbon (PAH)-contaminated site and at an uncontaminated site were compared based on LSU rDNA sequencing. Dissimilarities in species composition and community structures indicated soil pollution as the main factor negatively affecting the AMF diversity. The AMF communities at the contaminated site were dominated by fungal generalists (Rhizophagus, Funneliformis, Claroideoglomus, Paraglomus) with wide ecological tolerance. At the control site, the AMF communities were characterized by higher taxonomic and functional diversity than those exposed to the contamination. The host plant identity was the main driver distinguishing the two AMF metacommunities. The AMF communities at the uncontaminated site were represented by Polonospora, Paraglomus, Oehlia, Nanoglomus, Rhizoglomus, Dominikia, and Microdominikia. Polonosporaceae and Paraglomeraceae were particularly dominant in the Ph. australis mycorrhizosphere. The high abundance of early diverging AMF could be due to the use of primers able to detect lineages such as Paraglomeracae that have not been recognized by previously used 18S rDNA primers.

Phylogenetic Review of Acaulospora (Diversisporales, Glomeromycota) and the Homoplasic Nature of Its Ornamentations.

The genus Acaulospora has undergone many updates since it was first described; however, there are some missing pieces in the phylogenetic relationships among Acaulospora species. The present review aimed to: (i) understand the evolutionary meaning of their different spore wall ornamentations; (ii) define the best molecular marker for phylogenetic inferences, (iii) address some specific issues concerning the polyphyletic nature of Acaulospora lacunosa and Acaulospora scrobiculata, and the inclusion of Kuklospora species; and (iv) update the global geographical distribution of Acaulospora species. As such, the wall ornamentation of previously described Acaulospora species was reviewed and phylogenetic analyses were carried out based on ITS and SSU-ITS-LSU (nrDNA). Moreover, the already available type material of A. sporocarpia was inspected. According to the data obtained, temperate and tropical zones are the richest in Acaulospora species. We also confirmed that A. sporocarpia does not belong to Acaulospora. Furthermore, our phylogeny supported the monophyly of Acaulospora genus, including the Kuklospora species, K. colombiana and K. kentinensis. The nrDNA phylogeny presented the best resolution and revealed the homoplasic nature of many ornamentations in Acaulospora species, pointing out their unfeasible phylogenetic signal. This review reinforces the urgency of more molecular markers, in addition to the nrDNA sequences, for the definition of a multi-locus phylogeny.

Plant association with dark septate endophytes: When the going gets tough (and stressful), the tough fungi get going.

Dark septate endophytes (DSEs) comprise a diverse and ubiquitous group of fungal generalists with broad habitat niches that robustly colonize the roots of plants in stressful environments. DSEs possess adaptation strategies that determine their high tolerance to heavy metal (HM) contamination, drought, and salinity. Most DSEs developed efficient melanin-dependent and melanin-independent mechanisms of HM detoxification and osmoprotection, including intracellular immobilization and extracellular efflux of HMs and excess ions, and the scavenging of reactive oxygen species. DSEs form mutualistic relationship with plants according to the hypothesis of "habitat-adapted associations", supporting the survival of their hosts under stressful conditions. As saprophytes, DSEs mineralize a complex soil substrate improving plants' nutrition and physiological parameters. They can protect the host plant from HMs by limiting HM accumulation in plant tissues and causing their sequestration in root cell walls as insoluble compounds, preventing further HM translocation to shoots. The presence of DSE in drought-affected plants can substantially ameliorate the physiology and architecture of root systems, improving their hydraulic properties. Plant growth-promoting features, supported by the versatility and easy culturing of DSEs, determine their high potential to enhance phytoremediation and revegetation projects for HM-contaminated, saline, and desertic lands reclamation.

Three new species of arbuscular mycorrhizal fungi of the genus Diversispora from maritime dunes of Poland.

Three new species of arbuscular mycorrhizal fungi of the genus Diversispora (phylum Glomeromycota) were described based on their morphology and molecular phylogeny. The phylogeny was inferred from the analyses of the partial 45S rDNA sequences (18S-ITS-28S) and the largest subunit of RNA polymerase II (rpb1) gene. These species were associated in the field with plants colonizing maritime sand dunes of the Baltic Sea in Poland and formed mycorrhiza in single-species cultures.

A new order, Entrophosporales, and three new Entrophospora species in Glomeromycota.

As a result of phylogenomic, phylogenetic, and morphological analyses of members of the genus Claroideoglomus, four potential new glomoid spore-producing species and Entrophospora infrequens, a new order, Entrophosporales, with one family, Entrophosporaceae (=Claroideoglomeraceae), was erected in the phylum Glomeromycota. The phylogenomic analyses recovered the Entrophosporales as sister to a clade formed by Diversisporales and Glomeraceae. The strongly conserved entrophosporoid morph of E. infrequens, provided with a newly designated epitype, was shown to represent a group of cryptic species with the potential to produce different glomoid morphs. Of the four potential new species, three enriched the Entrophosporales as new Entrophospora species, E. argentinensis, E. glacialis, and E. furrazolae, which originated from Argentina, Sweden, Oman, and Poland. The fourth fungus appeared to be a glomoid morph of the E. infrequens epitype. The physical association of the E. infrequens entrophosporoid and glomoid morphs was reported and illustrated here for the first time. The phylogenetic analyses, using nuc rDNA and rpb1 concatenated sequences, confirmed the previous conclusion that the genus Albahypha in the family Entrophosporaceae sensu Oehl et al. is an unsupported taxon. Finally, the descriptions of the Glomerales, Entrophosporaceae, and Entrophospora were emended and new nomenclatural combinations were introduced.

New Glomeromycotan Taxa, Dominikia glomerocarpica sp. nov. and Epigeocarpum crypticum gen. nov. et sp. nov. From Brazil, and Silvaspora gen. nov. From New Caledonia.

Examination of fungal specimens collected in the Atlantic rain forest ecosystems of Northeast Brazil revealed many potentially new epigeous and semihypogeous glomerocarp-producing species of the phylum Glomeromycota. Among them were two fungi that formed unorganized epigeous glomerocarps with glomoid spores of almost identical morphology. The sole structure that distinguished the two fungi was the laminate layer 2 of their three-layered spore wall, which in spores of the second fungus crushed in PVLG-based mountants contracted and, consequently, transferred into a crown-like structure. Surprisingly, phylogenetic analyses of sequences of the 18S-ITS-28S nuc rDNA and the rpb1 gene indicated that these glomerocarps represent two strongly divergent undescribed species in the family Glomeraceae. The analyses placed the first in the genus Dominikia, and the second in a sister clade to the monospecific generic clade Kamienskia with Kamienskia bistrata. The first species was described here as Dominikia glomerocarpica sp. nov. Because D. glomerocarpica is the first glomerocarp-forming species in Dominikia, the generic description of this genus was emended. The very large phylogenetic distance and the fundamental morphological differences between the second species and K. bistrata suggested us to introduce a new genus, here named as Epigeocarpum gen. nov., and name the new species Epigeocarpum crypticum sp. nov. In addition, our analyses also focused on an arbuscular mycorrhizal fungus originally described as Rhizophagus neocaledonicus, later transferred to the genus Rhizoglomus. The analyses indicated that this species does not belong to any of these two genera but represents a new clade at the rank of genus in the Glomeraceae, here described as Silvaspora gen. nov.

Differences in the effects of single and mixed species of AMF on the growth and oxidative stress defense in Lolium perenne exposed to hydrocarbons.

Arbuscular mycorrhizal fungi (AMF) are ubiquitous mutualistic plant symbionts that promote plant growth and protect them from abiotic stresses. Studies on AMF-assisted phytoremediation have shown that AMF can increase plant tolerance to the presence of hydrocarbon contaminants by improving plant nutrition status and mitigating oxidative stress. This work aimed to evaluate the impact of single and mixed-species AMF inocula (Funneliformis caledonium, Diversispora varaderana, Claroideoglomus walkeri), obtained from a contaminated environment, on the growth, oxidative stress (DNA oxidation and lipid peroxidation), and activity of antioxidative enzymes (superoxide dismutase, catalase, peroxidase) in Lolium perenne growing on a substrate contaminated with 0/0-30/120 mg phenol/polynuclear aromatic hydrocarbons (PAHs) kg-1. The assessment of AMF tolerance to the presence of contaminants was based on mycorrhizal root colonization, spore production, the level of oxidative stress, and antioxidative activity in AMF spores. In contrast to the mixed-species AMF inoculum, single AMF species significantly enhanced the growth of host plants cultured on the contaminated substrate. The effect of inoculation on the level of oxidative stress and the activity of antioxidative enzymes in plant tissues differed between the AMF species. Changes in the level of oxidative stress and the activity of antioxidative enzymes in AMF spores in response to contamination also depended on AMF species. Although the concentration of phenol and PAHs had a negative effect on the production of AMF spores, low (5/20 mg phenol/PAHs kg-1) and medium (15/60 mg phenol/PAHs kg-1) substrate contamination stimulated the mycorrhizal colonization of roots. Among the studied AMF species, F. caledonium was the most tolerant to phenol and PAHs and showed the highest potential in plant growth promotion. The results presented in this study might contribute to the development of functionally customized AMF-assisted phytoremediation strategies with indigenous AMF, more effective than commercial AMF inocula, as a result of their selection by the presence of contaminants.

Native Arbuscular Mycorrhizal Fungi Characterization from Saline Lands in Arid Oases, Northwest China.

Arbuscular mycorrhizal fungi (AMF) colonize land plants in almost every ecosystem, even in extreme conditions, such as saline soils. In the present work, we report the mycorrhizal capacity of rhizosphere soils collected in the dry desert region of the Minqin Oasis, located in the northwest of China (Gansu province), which is characterized by several halophytes. Lycium spp. and Peganum nigellastrum were used as trap plants in a greenhouse experiment to identify autochthonous AMF associated with the halophytes' rhizospheres. Morphological observations showed the typical AMF structures inside roots. Twenty-six molecularly distinct AMF taxa were recovered from soil and root DNA. The taxonomical diversity mirrors the several AMF adapted to extreme environmental conditions, such as the saline soil of central China. Knowledge of the AMF associated with halophytes may contribute to select specific fungal isolates to set up agriculture strategies for protecting non-halophyte crop plants in saline soils.

Arbuscular mycorrhizal and microbial profiles of an aged phenol-polynuclear aromatic hydrocarbon-contaminated soil.

Arbuscular mycorrhizal fungi (AMF) are ubiquitous, obligatory plant symbionts that have a beneficial influence on plants in contaminated environments. This study focused on evaluating the biomass and biodiversity of the AMF and microbial communities associated with Poa trivialis and Phragmites australis plants sampled at an aged site contaminated with phenol and polynuclear aromatic hydrocarbons (PAHs) and an uncontaminated control site. We analyzed the soil phospholipid fatty acid profile to describe the general structure of microbial communities. PCR-denaturing gradient gel electrophoresis with primers targeting the 18S ribosomal RNA gene was used to characterize the biodiversity of the AMF communities and identify dominant AMF species associated with the host plants in the polluted and control environments. The root mycorrhizal colonization and AMF biomass in the soil were negatively affected by the presence of PAHs and phenol, with no significant differences between the studied plant species, whereas the biodiversity of the AMF communities were influenced by the soil contamination and plant species. Soil contamination was more detrimental to the biodiversity of AMF communities associated with Ph. australis, compared to P. trivialis. Both species favored the development of different AMF species, which might be related to the specific features of their different root systems and soil microbial communities. The contaminated site was dominated by AMF generalists like Funneliformis and Rhizophagus, whereas in the control site Dominikia, Archaeospora, Claroideoglomus, Glomus, and Diversispora were also detected.

Rhizoglomus dalpeae, R. maiae, and R. silesianum, new species.

We examined three arbuscular mycorrhizal fungi (AMF; phylum Glomeromycota) producing glomoid spores. The mode of formation and morphology of these spores suggested that they represent undescribed species in the genus Rhizoglomus of the family Glomeraceae. Subsequent morphological studies of the spores and molecular phylogenetic analyses of sequences of the nuc rDNA small subunit (18S), internal transcribed spacer (ITS1-5.8S-ITS2 = ITS), and large subunit (28S) region (= 18S-ITS-28S) confirmed the suggestion and indicated that the fungi strongly differ from all previously described Rhizoglomus species with known DNA barcodes. Consequently, the fungi were described here as new species: R. dalpeae, R. maiae, and R. silesianum. Two of these species lived hypogeously in the field in habitats subjected to strong environmental stresses. Rhizoglomus dalpeae originated from an inselberg located within Guineo-Sudanian transition savanna zone in Benin, West Africa, where the temperature of the inselberg rock during a 5-mo drought ranges from 40 to 60 C. Rhizoglomus silesianum originated from a coal mine spoil heap in Poland, whose substrate is extremely poor in nutrients, has unfavorable texture, and may heat up to 50 C. By contrast, R. maiae was found in more favorable habitat conditions. It produced an epigeous cluster of spores among shrubs growing in a tropical humid reserve in Brazil. Moreover, the compatibility of phylogenies of species of the family Glomeraceae reconstructed from analyses of sequences of 18S-ITS-28S and the largest subunit of RNA polymerase II (RPB1) gene was discussed.

Cohorts of arbuscular mycorrhizal fungi (AMF) in Vitis vinifera, a typical Mediterranean fruit crop.

In field conditions, grapevine roots normally are colonized by arbuscular mycorrhizal fungi (AMF). However, little is published, from either morphological or molecular studies, on the species composition of these symbionts in production vineyards. The AMF biodiversity of two Piedmont vineyards (at Neive and Lessona), characterized by different soil features, was investigated by morphological and molecular analyses. Several morphotypes were identified from the two vineyard soils. Community composition of AMF, both in soil and root samples, was then analysed with molecular approach to amplify a portion (550 bp) of AM fungal SSU rDNA. Phylogenetic analyses show a different distribution of sequences from the two sites in the main glomeromycotan groups. In the Neive site, the Glomeraceae group A is the only one well represented whereas more groups were found at Lessona. Among the more representative operational taxonomic units (OTUs), only one related to the Glomus irregulare phylotype was shared between the two vineyard soils. The data obtained in this work together with similar results in literature on this important fruit crop reinforce the concept that the general AMF assemblage structure and composition in vineyards might be influenced more by soil type than by host plant features (age, vegetative stages) or management practices.

Retrotransposon-based S-SAP as a platform for the analysis of genetic variation and linkage in globe artichoke.

A high copy number of retrotransposon sequences are present and widely dispersed in plant genomes. Their activity generates a considerable degree of sequence polymorphism. Here, we report the cloning of CYRE-5, a long-terminal repeat carrying retrotransposon-like sequence in Cynara cardunculus L., and its exploitation to develop a DNA fingerprinting assay across 22 accessions, including both cultivated (globe artichoke and cultivated cardoon) and wild (wild cardoon) types. The effectiveness of the sequence-specific amplified polymorphism (S-SAP) platform is compared with that of amplified fragment length polymorphism (AFLP). A genetic linkage analysis, based on a hybrid population between 2 globe artichoke varietal types, resulted in the inclusion of 29 S-SAP loci in the core genetic map, confirming their dispersed distribution across the globe artichoke genome.