Interkingdom Gene Transfer of a Hybrid NPS/PKS from Bacteria to Filamentous Ascomycota

Nonribosomal peptides (NRPs) and polyketides (PKs) are ecologically important secondary metabolites produced by bacteria and fungi using multidomain enzymes called nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs), respectively. Previous phylogenetic analyses of fungal NRPSs and PKSs have suggested that a few of these genes were acquired by fungi via horizontal gene transfer (HGT) from bacteria, including a hybrid NPS/PKS found in Cochliobolus heterostrophus (Dothideomycetes, Ascomycota). Here, we identify this hybrid gene in fungi representing two additional classes of Ascomycota (Aspergillus spp., Microsporum canis, Arthroderma spp., and Trichophyton spp., Eurotiomycetes; Chaetomium spp. and Metarhizium spp., Sordariomycetes) and use phylogenetic analyses of the most highly conserved domains from NRPSs (adenylation (A) domain) and PKSs (ketoacyl synthase (KS) domain) to examine the hypothesis that the hybrid NPS7/PKS24 was acquired by fungi from bacteria via HGT relatively early in the evolution of the Pezizomycotina. Our results reveal a unique ancestry of the A domain and KS domain in the hybrid gene relative to known fungal NRPSs and PKSs, provide strong evidence for HGT of the hybrid gene from a putative bacterial donor in the Burkholderiales, and suggest the HGT event occurred early in the evolution of the filamentous Ascomycota

Reducing Legionella Colonization of Water Systems with Monochloramine

Monochloramine reduced colonization in building hot water systems

A window into fungal endophytism in Salicornia europaea: deciphering fungal characteristics as plant growth promoting agents

Aim Plant-endophytic associations exist only when equilibrium is maintained between both partners. This study analyses the properties of endophytic fungi inhabiting a halophyte growing in high soil salinity and tests whether these fungi are beneficial or detrimental when non-host plants are inoculated. Method Fungi were isolated from Salicornia europaea collected from two sites differing in salinization history (anthropogenic and naturally saline) and analyzed for plant growth promoting abilities and non-host plant interactions. Results Most isolated fungi belonged to Ascomycota (96%) including dematiaceous fungi and commonly known plant pathogens and saprobes. The strains were metabolically active for siderophores, polyamines and indole-3-acetic acid (mainly Aureobasidium sp.) with very low activity for phosphatases. Many showed proteolytic, lipolytic, chitinolytic, cellulolytic and amylolytic activities but low pectolytic activity. Different activities between similar fungal species found in both sites were particularly seen for Epiccocum sp., Arthrinium sp. and Trichoderma sp. Inoculating the non-host Lolium perenne with selected fungi increased plant growth, mainly in the symbiont (Epichloë)-free variety. Arthrinium gamsii CR1-9 and Stereum gausapatum ISK3-11 were most effective for plant growth promotion. Conclusions This research suggests that host lifestyle and soil characteristics have a strong effect on endophytic fungi, and environmental stress could disturb the plant-fungi relations. In favourable conditions, these fungi may be effective in facilitating crop production in non-cultivable saline lands

Effects of fluoxetine on functional outcomes after acute stroke (FOCUS): a pragmatic, double-blind, randomised, controlled trial

Background Results of small trials indicate that fluoxetine might improve functional outcomes after stroke. The FOCUS trial aimed to provide a precise estimate of these effects. Methods FOCUS was a pragmatic, multicentre, parallel group, double-blind, randomised, placebo-controlled trial done at 103 hospitals in the UK. Patients were eligible if they were aged 18 years or older, had a clinical stroke diagnosis, were enrolled and randomly assigned between 2 days and 15 days after onset, and had focal neurological deficits. Patients were randomly allocated fluoxetine 20 mg or matching placebo orally once daily for 6 months via a web-based system by use of a minimisation algorithm. The primary outcome was functional status, measured with the modified Rankin Scale (mRS), at 6 months. Patients, carers, health-care staff, and the trial team were masked to treatment allocation. Functional status was assessed at 6 months and 12 months after randomisation. Patients were analysed according to their treatment allocation. This trial is registered with the ISRCTN registry, number ISRCTN83290762. Findings Between Sept 10, 2012, and March 31, 2017, 3127 patients were recruited. 1564 patients were allocated fluoxetine and 1563 allocated placebo. mRS data at 6 months were available for 1553 (99·3%) patients in each treatment group. The distribution across mRS categories at 6 months was similar in the fluoxetine and placebo groups (common odds ratio adjusted for minimisation variables 0·951 [95% CI 0·839–1·079]; p=0·439). Patients allocated fluoxetine were less likely than those allocated placebo to develop new depression by 6 months (210 [13·43%] patients vs 269 [17·21%]; difference 3·78% [95% CI 1·26–6·30]; p=0·0033), but they had more bone fractures (45 [2·88%] vs 23 [1·47%]; difference 1·41% [95% CI 0·38–2·43]; p=0·0070). There were no significant differences in any other event at 6 or 12 months. Interpretation Fluoxetine 20 mg given daily for 6 months after acute stroke does not seem to improve functional outcomes. Although the treatment reduced the occurrence of depression, it increased the frequency of bone fractures. These results do not support the routine use of fluoxetine either for the prevention of post-stroke depression or to promote recovery of function. Funding UK Stroke Association and NIHR Health Technology Assessment Programme

Comparison of Alternaria spp. collected in Italy from apple with A. mali and other AM-toxin producing strains.

Since 1999, a disease of apple caused by an Alternaria sp. has been affecting orchards in northern Italy resulting in necrotic spots on leaves and on fruit. Forty-four single-spored isolates were obtained from diseased plant materials to investigate the diversity of this fungus in Italy and to compare these isolates to isolates of Alternaria associated with apple disease in previous studies, including A. mali, causal agent of apple blotch. All isolates, including the reference strains, were tested for pathogenicity utilizing in vitro bioassays on detached leaf or on fruit ('Golden Delicious'). In addition, morphological characterizations were conducted describing both the three-dimensional sporulation pattern and the colony morphology of each isolate. In order to assess the genetic diversity within the Italian Alternaria population, sequence characterization of specific loci and anonymous regions (endoPG, OPA1-3, OPA2-1, and OPA10-2) and genetic fingerprinting based on amplified fragment length polymorphism and inter simple sequence repeat markers were performed. The single spore isolates exhibited differential pathogenicity, which did not correlate with the morphological groupings or to groupings defined by molecular approaches. Moreover, 10 pathogenic isolates out of the 44 single-spored tested were positive for the host-specific AM-toxin gene based upon polymerase chain reaction amplification using specific primers for the AM-toxin gene. This suggests that the production of the AM-toxin may be involved in pathogenesis by some of the Italian isolates of A. alternata from apple. However, this research also suggests that a number of different Alternaria genotypes and morphotypes may be responsible for the apple disease in Italy and that a single taxon cannot be defined as the sole causal agent

Degradation of aflatoxin B1 from naturally contaminated maize using the edible fungus Pleurotus ostreatus

Aflatoxins are highly carcinogenic secondary metabolites that can contaminate approximately 25% of crops and that cause or exacerbate multiple adverse health conditions, especially in Sub-Saharan Africa and South and Southeast Asia. Regulation and decontamination of aflatoxins in high exposure areas is lacking. Biological detoxification methods are promising because they are assumed to be cheaper and more environmentally friendly compared to chemical alternatives. White-rot fungi produce non-specific enzymes that are known to degrade aflatoxin in in situ and ex situ experiments. The aims of this study were to (1) decontaminate aflatoxin-B-1-(AFB(1)) in naturally contaminated maize with the edible, white-rot fungus Pleurotus ostreatus (oyster mushroom) using a solid-state fermentation system that followed standard cultivation techniques, and to (2) and to assess the risk of mutagenicity in the resulting breakdown products and mushrooms. Vegetative growth and yield characteristics of P. ostreatus were not inhibited by the presence of-AFB(1).-AFB(1) was degraded by up to 94% by the Blue strain. No aflatoxin could be detected in P. ostreatus mushrooms produced from-AFB(1)-contaminated maize. Moreover, the mutagenicity of breakdown products from the maize substrate, and reversion of breakdown products to the parent compound, were minimal. These results suggest that P. ostreatus significantly degrades-AFB(1) in naturally contaminated maize under standard cultivation techniques to levels that are acceptable for some livestock fodder, and that using P. ostreatus to bioconvert crops into mushrooms can reduce-AFB(1)-related losses.University of Arizona Green Fund [GF 15.31]Open Access Journal.This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Fungal communities on speleothem surfaces in Kartchner Caverns, Arizona, USA.

Kartchner Caverns, located near Benson, Arizona, USA, is an active carbonate cave that serves as the major attraction for Kartchner Caverns State Park. Low-impact development and maintenance have preserved prediscovery macroscopic cavern features and minimized disturbances to biological communities within the cave.. The goal of this study was to examine fungal diversity in Kartchner Caverns on actively-forming speleothem surfaces. Fifteen formations were sampled from five sites across the cave. Richness was assessed using standard culture-based fungal isolation techniques. A culture-independent analysis using denaturing gradient gel electrophoresis (DGGE) was used to assay evidence of community homogeneity across the cave through the separation of 18S rDNA amplicons from speleothem community DNA. The culturing effort recovered 53 distinct morphological taxonomic units (MTUs), corresponding to 43 genetic taxonomic units (GTUs) that represented 21 genera. From the observed MTU accumulation curve and the projected total MTU richness curve, it is estimated that 51 percent of the actual MTU richness was recovered. The most commonly isolated fungi belonged to the genera Penicillium, Paecilomyces, Phialophora, and Aspergillus. This culturebased analysis did not reveal significant differences in fungal richness or number of fungi recovered across sites. Cluster analysis using DGGE band profiles did not reveal distinctive groupings of speleothems by sample site. However, canonical correspondence analysis (CCA) analysis of culture-independent DGGE profiles showed a significant effect of sampling site and formation type on fungal community structure. Taken together, these results reveal that diverse fungal communities exist on speleothem surfaces in Kartchner Caverns, and that these communities are not uniformly distributed spatially. Analysis of sample saturation indicated that more sampling depth is required to uncover the full scale of mycological richness across spelothem surfaces