Eleonora Egidi

One fungus, which genes? Development and assessment of universal primers for potential secondary fungal DNA barcodes

The aim of this study was to assess potential candidate gene regions and corresponding universal primer pairs as secondary DNA barcodes for the fungal kingdom, additional to ITS rDNA as primary barcode. Amplification efficiencies of 14 (partially) universal primer pairs targeting eight genetic markers were tested across > 1 500 species (1 931 strains or specimens) and the outcomes of almost twenty thousand (19 577) polymerase chain reactions were evaluated. We tested several well-known primer pairs that amplify: i) sections of the nuclear ribosomal RNA gene large subunit (D1–D2 domains of 26/28S); ii) the complete internal transcribed spacer region (ITS1/2); iii) partial β -tubulin II (TUB2); iv) γ-actin (ACT); v) translation elongation factor 1-α (TEF1α); and vi) the second largest subunit of RNA-polymerase II (partial RPB2, section 5–6). Their PCR efficiencies were compared with novel candidate primers corresponding to: i) the fungal-specific translation elongation factor 3 (TEF3); ii) a small ribosomal protein necessary for t-RNA docking; iii) the 60S L10 (L1) RP; iv) DNA topoisomerase I (TOPI); v) phosphoglycerate kinase (PGK); vi) hypothetical protein LNS2; and vii) alternative sections of TEF1α. Results showed that several gene sections are accessible to universal primers (or primers universal for phyla) yielding a single PCR-product. Barcode gap and multi-dimensional scaling analyses revealed that some of the tested candidate markers have universal properties providing adequate infra- and inter-specific variation that make them attractive barcodes for species identification. Among these gene sections, a novel high fidelity primer pair for TEF1α, already widely used as a phylogenetic marker in mycology, has potential as a supplementary DNA barcode with superior resolution to ITS. Both TOPI and PGK show promise for the Ascomycota, while TOPI and LNS2 are attractive for the Pucciniomycotina, for which universal primers for ribosomal subunits often fail.

Phylogeny and taxonomy of meristematic rock-inhabiting black fungi in the Dothideomycetes based on multi-locus phylogenies

The last decade has revealed an unexpected fungal diversity associated with natural rocks, often collected in environments influenced by harsh climatic conditions. Yet the phylogenetic affiliations and the taxonomy of many of these extreme fungi, mainly within Dothideomycetes, the largest class of Ascomycota, have only partially been described. In the present study we confirm that most rock inhabiting-fungi (RIF) are highly polyphyletic among Dothideomycetidae, mainly within the order Capnodiales, an order otherwise incorporating several families of major plant pathological importance. Novel taxa were identified within the two major and distinct clades of Teratosphaeriaceae, both comprising meristematic black fungi. Thirty one novel species and 13 new genera are proposed, based on ITS and partial nucLSU, RPB2 and BT2 sequences.

Biodiversity, evolution and adaptation of fungi in extreme environments

Fungi play irreplaceable roles for ecosystem functioning. They may adopt different lifestyles, for example saprotrophs, symbionts or parasites: some species are cosmopolitan with a wide distribution and others, thanks to their ecological plasticity, may adapt to harsh environments precluded to most of life forms. In stressing conditions, their role is even more crucial for the recycling of organic matter or favoring nutrients uptake. When the conditions become really extreme and competition is low, fungi focus on extremotolerance and evolve peculiar competences to exploit natural or xenobiotic resources in the particular constrains imposed by the environment. This paper focuses on three different cases of fungal life in the extremes: hydrocarbon-polluted sites, extremely acidic substrates, and littoral dunes, aiming to give few but significant examples of the role of these fascinating organisms in peculiar habitats and the valuable biotechnological potentialities of the abilities they have evolved in response to such constrains.

Mountain tips as reservoirs for new rock-fungal entities: Saxomyces gen. nov. and four new species from the Alps

As part of a worldwide sampling nine black fungi were isolated from rocks collected in four distinct sites of the Alps at high altitudes. Based on a nucSSU, nucLSU and mtSSU multi-locus phylogeny, seven of them were found to cluster into a distinct and well-supported clade in a basal position within the Class Dothideomycetes. As in other rock fungi these new groups of isolates were characterized by a meristematic growth and a scarcely differentiated morphology with highly melanized and thick-walled toruloid hyphae. Nonetheless, few peculiar characters were also observed as convoluted hyphal tips and the production of spherical propagules. The new genus and species Saxomyces alpinus and S. penninicus, are here described based on morphological and molecular data, in a yet to be defined order of the Dothideomycetes. The remaining two black fungi clustered in Cryomyces, a genus previously exclusively found in rocks from the McMurdo Dry Valleys in Antarctica. These two isolates were genetically distant from other Antarctic Cryomyces species based on ITS sequences, and they showed a peculiar morphology; they are here described as the new species C. montanus and C. funiculosus. Implications of our results on the evolution, adaptation and dispersal of rock-inhabiting fungi under extreme conditions are discussed.

Fire regime, not time-since-fire, affects soil fungal community diversity and composition in temperate grasslands

Frequent burning is commonly undertaken to maintain diversity in temperate grasslands of southern Australia. How burning affects below-ground fungal community diversity remains unknown. We show, using a fungal rDNA metabarcoding approach (Illumina MiSeq), that the fungal community composition was influenced by fire regime (frequency) but not time-since-fire. Fungal community composition was resilient to direct fire effects, most likely because grassland fires transfer little heat to the soil. Differences in the fungal community composition due to fire regime was likely due to associated changes that occur in vegetation with recurrent fire, via the break up of obligate symbiotic relationships. However, fire history only partially explains the observed dissimilarity in composition among the soil samples, suggesting a distinctiveness in composition in each grassland site. The importance of considering changes in soil microbe communities when managing vegetation with fire is highlighted.

Draft Genome Sequence of Enterobacter ludwigii NCR3, a Heavy Metal–Resistant Rhizobacterium

ABSTRACT We report here the draft genome of Enterobacter ludwigii NCR3, a Gram-negative bacterium isolated from the Carpobrotus rossii (Haw.) Schwantes rhizosphere. The analysis of the ~4.8-Mb draft genome shows that this strain harbors several genes associated with heavy metal resistance and plant growth–promoting activity, suggesting its potential application in microbe-assisted phytoremediation.

Draft Genome Sequence of Bacillus cereus LCR12, a Plant Growth–Promoting Rhizobacterium Isolated from a Heavy Metal–Contaminated Environment

ABSTRACT Bacillus cereus LCR12 is a plant growth–promoting rhizobacterium, isolated from a heavy metal–contaminated environment. The 6.01-Mb annotated genome sequence provides the genetic basis for revealing its potential application to remediate contaminated soils in association with plants.

Antarctic cryptoendolithic fungal communities are highly adapted and dominated by Lecanoromycetes and Dothideomycetes

Endolithic growth is one of the most spectacular microbial adaptations to extreme environmental constraints and the predominant life-form in the ice-free areas of Continental Antarctica. Although Antarctic endolithic microbial communities are known to host among the most resistant and extreme-adapted organisms, our knowledge on microbial diversity and composition in this peculiar niche is still limited. In this study, we investigated the diversity and structure of the fungal assemblage in the cryptoendolithic communities inhabiting sandstone using a meta-barcoding approach targeting the fungal Internal Transcribed Sequence region 1 (ITS1). Samples were collected from 14 sites in the Victoria Land, along an altitudinal gradient ranging from 1,000 to 3,300 m a.s.l. and from 29 to 96 km distance to coast. Our study revealed a clear dominance of a ‘core’ group of fungal taxa consistently present across all the samples, mainly composed of lichen-forming and Dothideomycetous fungi. Pareto-Lorenz curves indicated a very high degree of specialization (F0 approximately 95%), suggesting these communities are highly adapted but have limited ability to recover after perturbations. Overall, both fungal community biodiversity and composition did not show any correlation with the considered abiotic parameters, potentially due to strong fluctuations of environmental conditions at local scales.

Draft Genome Sequence of Rhodococcus erythropolis NSX2, an Actinobacterium Isolated from a Cadmium-Contaminated Environment

ABSTRACT Rhodococcus erythropolis NSX2 is a rhizobacterium isolated from a heavy metal–contaminated environment. The 6.2-Mb annotated genome sequence shows that this strain harbors genes associated with heavy-metal resistance and xenobiotics degradation.