Aflatoxins are natural scavengers of reactive oxygen species

The role of aflatoxins (AFs) in the biology of producing strains, Aspergillus sect. Flavi, is still a matter of debate. Over recent years, research has pointed to how environmental factors altering the redox balance in the fungal cell can switch on the synthesis of AF. Notably, it has been known for decades that oxidants promote AF synthesis. More recent evidence has indicated that AF synthesis is controlled at the transcriptional level: reactive species that accumulate in fungal cells in the stationary growth phase modulate the expression of aflR, the main regulator of AF synthesis—through the oxidative stress related transcription factor AP-1. Thus, AFs are largely synthesized and secreted when (i) the fungus has exploited most nutritional resources; (ii) the hyphal density is high; and (iii) reactive species are abundant in the environment. In this study, we show that AFs efficiently scavenge peroxides and extend the lifespan of E. coli grown under oxidative stress conditions. We hypothesize a novel role for AF as an antioxidant and suggest its biological purpose is to extend the lifespan of AFs-producing strains of Aspergillus sect. Flavi under highly oxidizing conditions such as when substrate resources are depleted, or within a host.

Response of arbuscular mycorrhizal fungal community in soil and roots to grazing differs in a wetland on the Qinghai-Tibet plateau

Grazing as one of the most important disturbances affects the abundance, diversity and community composition of arbuscular mycorrhizal (AM) fungi in ecosystems, but the AM fungi in response to grazing in wetland ecosystems remain poorly documented. Here, we examined AM fungi in roots and soil in grazing and non-grazing plots in Zoige wetland on the Qinghai-Tibet plateau. Grazing significantly increased AM fungal spore density and glomalin-related soil proteins, but had no significant effect on the extra radical hyphal density of AM fungi. While AM fungal richness and community composition differed between roots and soil, grazing was found to influence only the community composition in soil. This study shows that moderate grazing can increase the biomass of AM fungi and soil carbon sequestration, and maintain the AM fungal diversity in the wetland ecosystem. This finding may enhance our understanding of the AM fungi in response to grazing in the wetland on the Qinghai-Tibet plateau.

Simulated folivory increases vertical transmission of fungal endophytes that deter herbivores and alter tolerance to herbivory in Poa autumnalis

Background and Aims The processes that maintain variation in the prevalence of symbioses within host populations are not well understood. While the fitness benefits of symbiosis have clearly been shown to drive changes in symbiont prevalence, the rate of transmission has been less well studied. Many grasses host symbiotic fungi (Epichloë spp.), which can be transmitted vertically to seeds or horizontally via spores. These symbionts may protect plants against herbivores by producing alkaloids or by increasing tolerance to damage. Therefore, herbivory may be a key ecological factor that alters symbiont prevalence within host populations by affecting either symbiont benefits to host fitness or the symbiont transmission rate. Here, we addressed the following questions: Does symbiont presence modulate plant tolerance to herbivory? Does folivory increase symbiont vertical transmission to seeds or hyphal density in seedlings? Do plants with symbiont horizontal transmission have lower rates of vertical transmission than plants lacking horizontal transmission? Methods We studied the grass Poa autumnalis and its symbiotic fungi in the genus Epichloë. We measured plant fitness (survival, growth, reproduction) and symbiont transmission to seeds following simulated folivory in a 3-year common garden experiment and surveyed natural populations that varied in mode of symbiont transmission. Key Results Poa autumnalis hosted two Epichloë taxa, an undescribed vertically transmitted Epichloë sp. PauTG-1 and E. typhina subsp. poae with both vertical and horizontal transmission. Simulated folivory reduced plant survival, but endophyte presence increased tolerance to damage and boosted fitness. Folivory increased vertical transmission and hyphal density within seedlings, suggesting induced protection for progeny of damaged plants. Across natural populations, the prevalence of vertical transmission did not correlate with symbiont prevalence or differ with mode of transmission. Conclusions Herbivory not only mediated the reproductive fitness benefits of symbiosis, but also promoted symbiosis prevalence by increasing vertical transmission of the fungus to the next generation. Our results reveal a new mechanism by which herbivores could influence the prevalence of microbial symbionts in host populations.

Detection of white head symptoms of panicle blast caused by Pyricularia oryzae using cut-flower dye

Background Breeding of rice with panicle resistance to rice blast disease caused by Pyricularia oryzae is a challenge towards sustainable rice production. Methods for accurate estimation of disease severity can support breeding. White head symptoms are a commonly used index of panicle blast in the field. As the development mechanism of this symptom remains unclear, we used cut-flower dye (CFD) solution to visualize the infected panicle tissues. Results CFD delineated the edge of white head symptoms in rice panicles artificially infected with P. oryzae. Hyphae within the tissues were confirmed through staining with a fluorescent wheat germ agglutinin conjugate. Hyphal density was obviously diminished at the dye edge. Growing hyphae preferred to move along the vascular bundles; infected tissues lost the ability to transport water, leading to white head formation. By marking the edge of the white heads, this simple dyeing technique precisely reveals the extent of infection. Further, digital imaging allowed dried samples to be stored and reassessed later. Conclusions The CFD detection technique served as a powerful tool for estimating disease severity by color, as it clearly revealed lesions in both the panicles and leaves. Combined with reliable methods for artificial inoculation and observation of infecting hyphae, this technique will advance the research and breeding of panicle blast-resistant rice.

Increasing endophyte alkaloid expression in tall fescue by selecting for increased endophyte hyphal density

Two AR542-endophyte infected tall fescue breeding pools were subjected to one cycle of selection for increased endophyte hyphal density. Hyphal density was assessed subjectively by microscopic examination of the leaf sheath. Levels of alkaloids produced by both the original and selected populations were subsequently measured during summer. Levels of lolines and peramine alkaloids increased significantly in one breeding pool (by 40% and 29%, respectively), with non-significant increases (by 24% and 2%) in the other breeding pool. The apparent correlation between observed hyphal density and levels of alkaloids supports the hypothesis that levels of lolines and peramine produced by fungal endophytes in their host grass are at least partly due to endophyte hyphal mass in the host sheath tissue. Measured lolines in one breeding pool were 21 times greater than the other, while sheath hyphal density was less. Possible explanations include contrasting patterns of hyphal colonisation beyond the sheath, and/or variation in the quantity of lolines produced per unit hyphal mass. Keywords: Neotyphodium coenophalium, endophyte, Festuca arundinacea, tall fescue, peramine, lolines, hyphal mass

Topical Caspofungin for Treatment of Keratitis Caused by Candida albicans in a Rabbit Model

ABSTRACT Candida albicans is the most frequent cause of fungal keratitis in temperate regions. Caspofungin has potent activity against Candida spp. in a variety of clinical settings. Little is known, however, about its activity against fungal keratitis. We compared the efficacy of topical caspofungin with that of topical amphotericin B (AMB) in a rabbit model of experimental keratomycosis. Keratitis was induced with a standardized inoculum of Candida albicans (SC 5314) placed on the debrided cornea. Twenty-four hours after infection, animals were randomly assigned to treatment with 0.15% caspofungin, 0.5% caspofungin, 0.15% AMB, and a saline control (n = 12 rabbits in each group). For the first 12 h, treatment was repeated every 30 min and, after a 12-h pause, was resumed at hourly intervals for another 12 h. The animals were examined and killed 12 h after administration of the last dose. Treatment effects were evaluated by clinical assessment, fungal culture, and histopathology. Drug treatment significantly reduced corneal fungal recovery from 3.78 log10 CFU in saline-treated animals to 2.97, 1.76, and 1.18 log10 CFU in animals treated with 0.15% caspofungin, 0.5% caspofungin, and 0.15% AMB, respectively. By histopathology, the mean hyphal density was significantly lower in the corneas of treated animals than in those of the controls; there was no difference in hyphal densities between the different treatment groups. The depth of corneal invasion was not significantly reduced by the antifungal treatments. By clinical assessment, keratitis progressed in animals treated with saline, whereas disease progression was inhibited by all drug treatment regimens. In our rabbit model, 0.5% caspofungin was as effective as 0.15% AMB for the topical treatment of Candida keratitis. The potential clinical efficacy of caspofungin awaits further investigation.

Ethylene is synthesized by vegetative mycelium in surface cultures of Penicillium cyclopium Westling

Surface cultures of Penicillium cyclopium, grown in a closed aeration system on a semisolid minimal medium with 2-keto-glutarate, glutamate or glucose as carbon source, produced ethylene in two phases. The first was associated with aerial hyphae formation; the second, with conidiation. The maximum rate of ethylene synthesis coincided with intensive oxygen consumption whereas the cessation and reutilization of ethylene occurred together with a low respiration rate. After the removal of the aerial mycelium, vegetative hyphae continued ethylene production up to 50% of the value reached in intact cultures. Neither detached aerial mycelium with penicilli nor pure conidia produced ethylene. The content of ethylene in the medium of a producing culture was four-fold higher than would correspond to its concentration in the atmosphere above the culture. In the upper agar layer (1.25 mm), a high pH (8.8) and 30 mM [Formula: see text]was found, presumably because of mycelium autolysis. The pH in the layer 3 mm below the surface was 2.4. In lower layers (up to 2 mm beneath the last vegetative hyphae), ammonium ions were completely depleted. The phosphate content was inversely proportional to the hyphal density. We suggest that the low phosphate concentration (0.06 mM) in the upper layers permitted ethylene formation.Key words: conidiation, vegetative hyphae, ethylene, ion gradients, surface culture.

Cadmium alters the growth of the ectomycorrhizal fungus Paxillus involutes: a new growth model accounts for changes in branching

Cadmium reduced the growth rate of Paxillus involutus in pure cultures either on agar or liquid medium. On Cd-containing agar most of the mycelium grew submerged rather than on the surface as occurs on Cd-free agar. Cadmium increased hyphal density by both increasing the number of laterals at a branch point and decreasing the distance between branch points. These variables were included in a new model to determine the specific growth rate on the basis of mycelial length for the fungus growing on agar. The degree to which Cd reduced the specific growth rate was the same whether based on mycelial length from agar plates according to the new model or on mycelial mass from a liquid medium. The new model for specific growth rate (length) derived from agar cultures is particularly suited to those situations where a growth-modifying agent alters the branching frequency and the distance between branch points.

Growth of Verticillium dahliae Kleb. hyphae and of bacteria along the roots of rape (Brassica napus L.) seedlings

Bacterial density and the hyphal density of Verticillium dahliae were determined along the roots of sand-grown rape seedlings. Verticillium dahliae microsclerotia were trapped in nylon nets which were placed in front of the growing root tip. The growing hyphae on the nets were stained with fluorescein diacetate and rhodamine B isothiocyanate. Bacteria on the petri dish lids adjacent to the nets were stained with acridine orange. Both fungal and bacterial densities increased exponentially over a distance of 15–20 mm from the root tip. Further back along the root, the densities of microorganisms stabilized, but in some cases the fluorescein diacetate active hyphal density increased. The hyphal densities stopped increasing before bacterial densities, suggesting an increasing bacterial competition for root exudates as their population grew. The hyphal activity, measured as the proportion of flurescein diacetate to rhodamine B isothioicyanate stained hyphae, increased during the exponential growth phase of the bacteria, then decreased, and finally increased again as the lateral roots appeared.