The oxygen concentration in cultures modulates protein expression and enzymatic antioxidant responses in Metarhizium lepidiotae conidia

Conidia from Metarhizium spp. are used for integrated pest control; however, environmental factors diminish the effectivity of these programs. Several approaches tried to improve conidia resistance to overcome this limitation, although little is known about the mechanisms involved in this effect. Here we measured the activity of antioxidant enzymes and conidia virulence, comparing the proteomic profiles of Metarhiziumlepidiotae CP-OAX conidia produced under normal (21% O₂) and high oxygen atmospheres (pulses with 30% O₂). We detected a higher virulence against Tenebrio molitor larvae, in addition to an increase in ultraviolet light tolerance in conidia produced under 30% O₂, which correlates with increased glutathione reductase activity. Two-dimensional gel electrophoresis (2D SDS–PAGE) of proteins extracted in conidia harvested from both experimental conditions revealed a group of proteins that was observed only in conidia from oxidant atmospheres. Some of those proteins were directly involved in oxidative stress responses, whereas others were involved in conidial virulence, thermo-tolerance, and the central metabolism. Thus, a high atmospheric oxygen concentration (30%) activates antioxidant defence and general stress response mechanisms involved in conidia resistance to adverse environmental factors, which can ultimately translate into higher effectivity for the use of entomopathogenic fungi conidia in pest control.     

Cold-adapted enzymes produced by fungi from terrestrial and marine Antarctic environments

Antarctica is the coldest, windiest, and driest continent on Earth. In this sense, microorganisms that inhabit Antarctica environments have to be adapted to harsh conditions. Fungal strains affiliated with Ascomycota and Basidiomycota phyla have been recovered from terrestrial and marine Antarctic samples. They have been used for the bioprospecting of molecules, such as enzymes. Many reports have shown that these microorganisms produce cold-adapted enzymes at low or mild temperatures, including hydrolases (e.g. α-amylase, cellulase, chitinase, glucosidase, invertase, lipase, pectinase, phytase, protease, subtilase, tannase, and xylanase) and oxidoreductases (laccase and superoxide dismutase). Most of these enzymes are extracellular and their production in the laboratory has been carried out mainly under submerged culture conditions. Several studies showed that the cold-adapted enzymes exhibit a wide range in optimal pH (1.0–9.0) and temperature (10.0–70.0?°C). A myriad of methods have been applied for cold-adapted enzyme purification, resulting in purification factors and yields ranging from 1.70 to 1568.00-fold and 0.60 to 86.20%, respectively. Additionally, some fungal cold-adapted enzymes have been cloned and expressed in host organisms. Considering the enzyme-producing ability of microorganisms and the properties of cold-adapted enzymes, fungi recovered from Antarctic environments could be a prolific genetic resource for biotechnological processes (industrial and environmental) carried out at low or mild temperatures.     

Fungal Communities on Rock Surfaces in Demänovská Ice Cave and Demänovská Cave of Liberty (Slovakia)

This paper is the first geomycological report regarding the fungal communities on rock surfaces in the Demänovská Ice Cave and the Demänovská Cave of Liberty, Slovakia. The samples were collected in June 2014 from five locations from inside both the caves by using sterile swabs wetted with physiological saline (0.85% NaCl). The density of epilithic fungi isolated from the Demänovská Ice Cave ranged from 238.7 to 575.1 CFU (colony-forming units) per m² of the rock surface, and from the Demänovská Cave of Liberty ranged from 88.6 to 347 CFU. Seventeen different free-living culturable fungi (15 filamentous fungi, one yeast, and one yeast-like fungus) were isolated from the rock surfaces of both caves. Generally, Cladosporium cladosporioides and Aspergillus flavus were the most frequently cultured species from the Demänovská Ice Cave and the Demänovská Cave of Liberty, respectively. Free-living fungi found on the rock surfaces of both caves can lead to their slow biodegradation.     

Infestation of maize by Prostephanus truncatus initiated by male-produced pheromone

Delta traps baited with maize cobs, which were infested each with one male Prostephanus truncatus (Horn) (Col.: Bostrichidae), were distributed in southern Benin and collected after one, two, three and four weeks. The numbers of P. truncatus caught during the different trapping periods were not significantly different. Sixty-four percent of the trapped P. truncatus were females. Females attracted during the one-week trapping period produced a mean of 6.9 progeny during the seven days. The sex ratio of the progeny was 1:1. Trap catches with the infested cobs were on average 13 times lower than catches with 2 mg of the artificial pheromone. Estimation of P. truncatus densities in a maize store at the beginning of the storage period (based on laboratory data) revealed that small initial numbers of P. truncatus, possibly attracted by a single male, sufficed to initiate high infestation rates later in the storage season.     

寄主植物和接种物对丛枝菌根菌接种势的影响(英文)

在盆栽条件下研究了丛枝菌根菌(arbuscular mycorrhizal fungi,AMF)单孢、多孢和菌根根段接种物及其寄主植物烟草(Nicotiana tabacum L.)、苏丹草(Sorghum sudanense(Piper)Stapf)和三叶草(Trifoliumrepens L.)对AMF Glomus macrocarpum Tul ＆ Tul、Glomus mosseae(Nicol ＆ Gerd.)Gerdemann ＆ Trappe、Glomusversiforme(Karsten)Berch和Sclerocystis sinuosa Gerdemann ＆ Bakhi发育和接种势值(value of inoculum poten-tial,VIP)的影响。用50个孢子或0.5g菌根根段接种物处理的感染迟缓期为28d,大大短于单孢接种处理(64d);前者的VIP和根系感染百分率(percentage of root colonization,PRC)均高于后者。50个孢子和0.5g菌根根段接种物两处理之间仅在感染的早期阶段存在差异。0.5g菌根根段接种物(烟草上G.versiforme菌除外)接种处理的VIP大于其他两种接种物的处理;而50个孢子或0.5g菌根根段接种物处理的PRC直至接种后70d仍高于单孢接种处理。三叶草上AMF的VIP显著大于其他两种寄主植物上的VIP。G.mosseae、G.versiforme和S.sinuosa的VIP则大于G.macrocarpum的处理。这表明不同AMF菌种能产生不同接种势值的接种物。烟草则是用于单孢接种和接种物生产的良好寄主植物。     

Hormonema schizolunatum, a new species of dothideaceous black yeasts from phyllosphere

Two black yeast isolates from plants from the Canary Islands (Spain) are described and illustrated. Absence of Woronin bodies at simple septal pores, local coralloid terminal hyphal cells, indeterminate thallus maturation, the presence of budding cells and local conversion to meristematic growth all indicate a relationship to the Dothideaceae (Dothideales, Ascomycota). Morphological properties were consistent with the genus Hormonema Lagerberg & Melin, as defined by presence of percurrent conidiogenous loci alongside undifferentiated hyphae, and results of PCR-ribotyping supported this classification. The isolates were judged to belong to a hitherto undescribed species, characterized in particular by curved conidia soon developing transverse septa. The physiological profile of this species is also described.     

The incorporation of mannoproteins in the cell wall of S. cerevisiae and filamentous Ascomycetes

In yeast, glucanase extractable cell wall proteins are anchored to the plasma membrane at an intermediate stage in their biogenesis via a glycosylphosphatidylinositol (GPI) moiety before they become anchored to the wall glucan via a 1,6-glucan linkage. The mechanism of the membrane processing step of cell wall proteins is not known. Here, we report that Ascomycete filamentous fungi involved in food spoilage such as Aspergillus, Paecilomyces and Penicillium, also contain GPI membrane-anchored proteins some of which are processed by an endogenous phospholipase C activity. Furthermore, similar to the situation in yeast, their cell walls contain mannoproteins which are linked to the glucan backbone through a 1,6-glucan linkage. Interestingly, one mould which contains a significant amount of non covalently linked 1,6-glucosylated cell wall proteins, is much more sensitive towards 1,3-glucanases and membrane perturbing peptides than the others.     

Allelic variation at a hypervariable compound microsatellite locus in the ascomycete Ascochyta rabiei

The genome of the fungal chickpea pathogen Ascochyta rabiei was screened for polymorphisms by microsatellite-primed PCR. While ethidium-bromide staining of electrophoretically separated amplification products showed only limited polymorphism among 24 Tunisian A. rabiei isolates, Southern hybridization of purified PCR fragments to restriction digests of fungal DNA revealed polymorphic DNA fingerprints. One particular probe that gave rise to a hypervariable single-locus hybridization signal was cloned from the Syrian isolate AA6 and sequenced. It contained a large compound microsatellite harbouring the penta- and decameric repeat units (CATTT)_n, (CATTA)_n, (CATATCATTT)_n and (TATTT)_n. We call this locus ArMS1 (Ascochyta rabiei microsatellite 1). Unique flanking sequences were used to design primer pairs for locus- specific microsatellite amplification and direct sequencing of additional ArMS1 alleles from Tunisian and Pakistani isolates. A high level of sequence variation was observed, suggesting that multiple mutational mechanisms have contributed to polymorphism. Hybridization and PCR analyses were performed on the parents and 62 monoascosporic F₁ progeny derived from a cross between two different mating types of the fungus. Progeny alleles could be traced back to the parents, with one notable exception, where a longer than expected fragment was observed. Direct sequencing of this new length allele revealed an alteration in the copy number of the TATTT repeat [(TATTT)₅₃ to (TATTT)₆₅], while the remainder of the sequence was unchanged. Received: 11 March 1997 / Accepted: 21 June 1997