Blast disease in ryegrass is similar to rice blast in regard to infection cell biology

Magnaporthe oryzae has been devastating to various grasses. This is especially true in food crops around the globe. Resistance has been implemented but has not been completely successful due to the variability of the fungus. It is hypothesized that this is due in part to the presence of disposable mini-chromosomes. Ryegrass blast pathogen is closely related to the dangerous, emerging wheat blast pathogen, and understanding it can help us understand wheat blast as well as pathogen variability. The ryegrass pathogen on its own recently emerged as a serious threat to golf courses and sports fields in the U.S. Hypothesis: The ryegrass pathogen uses the same biotrophic (live-cell) invasion strategy as the extensively-studied rice blast pathogen

Improved Vectors for Selecting Resistance to Hygromycin

Resistance to hygromycin B is an important dominant selectable marker in fungal transformation. Our goal was to improve vectors for hygromycin selection by making the gene more compact, by eliminating sites for commonly used restriction enzymes, and by subcloning the modified gene into convenient vectors. These improvements were made by modifying pCSN43 (Staben et al. 1989 Fungal Genetics Newsl. 36:79-81) through three rounds of megaprimer mutagenesis (Aiyar and Leis, 1993 Biotechniques 14:366-368 ), a technique based on polymerase chain reaction amplification. Plasmid pCSN43 has a 2.4 kb SalI fragment containing the bacterial hph gene (Gritz and Davies, 1983 Gene 25:179-188), encoding hygromycin B phosphotransferase, under control of the Aspergillus nidulans trpC promoter and terminator (Mullaney et al. 1985 MGG 199:37-45

Conidial Morphogenesis and Septin-Mediated Plant Infection Require Smo1, a Ras GTPase-Activating Protein in Magnaporthe oryzae

The pathogenic life cycle of the rice blast fungus Magnaporthe oryzae involves a series of morphogenetic changes, essential for its ability to cause disease. The smo mutation was identified > 25 years ago, and affects the shape and development of diverse cell types in M. oryzae, including conidia, appressoria, and asci. All attempts to clone the SMO1 gene by map-based cloning or complementation have failed over many years. Here, we report the identification of SMO1 by a combination of bulk segregant analysis and comparative genome analysis. SMO1 encodes a GTPase-activating protein, which regulates Ras signaling during infection-related development. Targeted deletion of SMO1 results in abnormal, nonadherent conidia, impaired in their production of spore tip mucilage. Smo1 mutants also develop smaller appressoria, with a severely reduced capacity to infect rice plants. SMO1 is necessary for the organization of microtubules and for septin-dependent remodeling of the F-actin cytoskeleton at the appressorium pore. Smol physically interacts with components of the Ras2 signaling complex, and a range of other signaling and cytoskeletal components, including the four core septins. SMO1 is therefore necessary for the regulation of RAS activation required for conidial morphogenesis and septin-mediated plant infection

Geophysical research of archaeological sites with presumed ironworking activity on the example of the sites of Bakovčice, Nađbarice, and Ždala

U okviru opsežnoga terenskog pregleda obavljenoga za potrebe projekta TransFER prikupljeni površinski nalazi ukazivali su kako su se na položajima Bakovčice – Velike livade 1, Nađbarice 1 i Ždala – Telek odvijale aktivnosti povezane s proizvodnjom željeza. Uz fragmente talioničke zgure nesumnjivo povezane s proizvodnjom željeza, među površinskim nalazima pronađeni su brojni ulomci keramike iz različitih povijesnih razdoblja. S ciljem jasnijega definiranja karaktera nalazišta odabranih na osnovi analiza površinskih nalaza, pristupilo se neinvazivnim, geofizičkim istraživanjima primjenom magnetske metode. U svim se slučajevima radi o površinama s intenzivnom poljoprivrednom obradom zemljišta, pa se mogla očekivati slabija očuvanost arheoloških ostataka i preme tome i slabije magnetske anomalije. To je bio dosta ozbiljan izazov za arheološki rječitu obradu magnetograma, ali je na više mjesta potvrđeno postojanje ostataka željezarskih aktivnosti in situ ispod razine oranja, dok se ostaci željezarskih djelatnosti u velikoj mjeri nalaze u sloju oranice. Na osnovi svih karakterističnih magnetskih anomalija, uspjelo se odrediti približne granice područja aktivnosti za proizvodnju željeza. Ovo je geofizičko istraživanje značajno za teoretska razmatranja korelacija rezultata terenskoga pregleda i geofizičkih istraživanja za usavršavanje metodologije arheološke prospekcije kao i za određivanje mjera zaštite arheološke baštine, budući da potvrđuje već dobro poznatu činjenicu da arheološki ostaci na primarnome mjestu na poljoprivrednim površinama sa sve intenzivnijom obradom zemljišta s vremenom postepeno nestaju.The surface finds collected in an extensive field review carried out for the needs of the TransFER project pointed to iron production activities in the locations of Bakovčice – Velike Livade 1, Nađbarice 1, and Ždala – Telek. The surface finds included fragments of smelting slag, undoubtedly associated with iron production, but also numerous potsherds from different historical periods. In order to define more clearly the character of the sites chosen after the analyses of surface finds, it was decided to undertake non-invasive geophysical research using the magnetic method. These are all areas with intensive agricultural activity, so it was expected that there would be poor preservation of archaeological remains and, therefore, weaker magnetic anomalies. This was a rather serious challenge for the archaeologically eloquent processing of magnetograms, but the existence of the remains of ironworks activities in situ below the plough zone was confirmed in several places, while the remains of ironworks activities are largely found in the arable layer. On the basis of all the characteristic magnetic anomalies, they managed to determine the approximate boundaries of the area of iron production activity. This geophysical survey is significant for the theoretical considerations of the correlation between the results of field surveys and geophysical research for improving the methodology of archaeological prospecting and for determining the measures for the protection of archaeological heritage, since it confirms the well-known fact that the archaeological remains on the primary location in agricultural areas with ever more intensive cultivation gradually disappear over time

Irritable bowel syndrome - An inflammatory disease involving mast cells

Irritable bowel syndrome (IBS) is traditionally defined as a functional disorder - that is the presence of symptoms in the absence of demonstrable pathological abnormalities. In recent times, low grade inflammatory infiltrates in both the small and large bowel of some patients with IBS - often rich in mast cells, along with serological markers of low grade inflammation have focussed attention on IBS as an inflammatory disease. The observation that mast cells often lie in close association to enteric neurons, and in-vitro and in-vivo animal studies demonstrating that mast cell mediators may influence enteric motility provides a biologically plausible causal mechanism in IBS. Pilot studies on patients with IBS using the mast cell stabiliser sodium cromoglycate ('proof of concept') have been encouraging. The essential question remains why mast cells infiltrate the bowel of IBS patients. A disturbance of the 'brain-gut axis' is the current favoured hypothesis, whereby childhood stress or psychiatric comorbidity act via neuro-immune mechanisms to modulate low grade inflammation. An alternative hypothesis is that food allergy may be responsible. Serum specific IgE, and skin prick tests are not elevated in IBS patients, suggesting type 1 IgE mediated food allergy is not the cause. However questionnaire based studies indicate IBS patients have higher rates of atopic disease, and increased bronchial reactivity to methacholine has been demonstrated. In this review, we highlight the potential role of mast cells in IBS, and current and future research directions into this intriguing condition

Multiple Translocation of the AVR-Pita Effector Gene among Chromosomes of the Rice Blast Fungus Magnaporthe oryzae and Related Species

Magnaporthe oryzae is the causal agent of rice blast disease, a devastating problem worldwide. This fungus has caused breakdown of resistance conferred by newly developed commercial cultivars. To address how the rice blast fungus adapts itself to new resistance genes so quickly, we examined chromosomal locations of AVR-Pita, a subtelomeric gene family corresponding to the Pita resistance gene, in various isolates of M. oryzae (including wheat and millet pathogens) and its related species. We found that AVR-Pita (AVR-Pita1 and AVR-Pita2) is highly variable in its genome location, occurring in chromosomes 1, 3, 4, 5, 6, 7, and supernumerary chromosomes, particularly in rice-infecting isolates. When expressed in M. oryzae, most of the AVR-Pita homologs could elicit Pita-mediated resistance, even those from non-rice isolates. AVR-Pita was flanked by a retrotransposon, which presumably contributed to its multiple translocation across the genome. On the other hand, family member AVR-Pita3, which lacks avirulence activity, was stably located on chromosome 7 in a vast majority of isolates. These results suggest that the diversification in genome location of AVR-Pita in the rice isolates is a consequence of recognition by Pita in rice. We propose a model that the multiple translocation of AVR-Pita may be associated with its frequent loss and recovery mediated by its transfer among individuals in asexual populations. This model implies that the high mobility of AVR-Pita is a key mechanism accounting for the rapid adaptation toward Pita. Dynamic adaptation of some fungal plant pathogens may be achieved by deletion and recovery of avirulence genes using a population as a unit of adaptation

Multiple Plant Surface Signals are Sensed by Different Mechanisms in the Rice Blast Fungus for Appressorium Formation

Surface recognition and penetration are among the most critical plant infection processes in foliar pathogens. In Magnaporthe oryzae, the Pmk1 MAP kinase regulates appressorium formation and penetration. Its orthologs also are known to be required for various plant infection processes in other phytopathogenic fungi. Although a number of upstream components of this important pathway have been characterized, the upstream sensors for surface signals have not been well characterized. Pmk1 is orthologous to Kss1 in yeast that functions downstream from Msb2 and Sho1 for filamentous growth. Because of the conserved nature of the Pmk1 and Kss1 pathways and reduced expression of MoMSB2 in the pmk1 mutant, in this study we functionally characterized the MoMSB2 and MoSHO1 genes. Whereas the Momsb2 mutant was significantly reduced in appressorium formation and virulence, the Mosho1 mutant was only slightly reduced. The Mosho1 Momsb2 double mutant rarely formed appressoria on artificial hydrophobic surfaces, had a reduced Pmk1 phosphorylation level, and was nonresponsive to cutin monomers. However, it still formed appressoria and caused rare, restricted lesions on rice leaves. On artificial hydrophilic surfaces, leaf surface waxes and primary alcohols-but not paraffin waxes and alkanes- stimulated appressorium formation in the Mosho1 Momsb2 mutant, but more efficiently in the Momsb2 mutant. Furthermore, expression of a dominant active MST7 allele partially suppressed the defects of the Momsb2 mutant. These results indicate that, besides surface hydrophobicity and cutin monomers, primary alcohols, a major component of epicuticular leaf waxes in grasses, are recognized by M. oryzae as signals for appressorium formation. Our data also suggest that MoMsb2 and MoSho1 may have overlapping functions in recognizing various surface signals for Pmk1 activation and appressorium formation. While MoMsb2 is critical for sensing surface hydrophobicity and cutin monomers, MoSho1 may play a more important role in recognizing rice leaf waxes

A Novel Pathogenicity Gene Is Required in the Rice Blast Fungus to Suppress the Basal Defenses of the Host

For successful colonization and further reproduction in host plants, pathogens need to overcome the innate defenses of the plant. We demonstrate that a novel pathogenicity gene, DES1, in Magnaporthe oryzae regulates counter-defenses against host basal resistance. The DES1 gene was identified by screening for pathogenicity-defective mutants in a T-DNA insertional mutant library. Bioinformatic analysis revealed that this gene encodes a serine-rich protein that has unknown biochemical properties, and its homologs are strictly conserved in filamentous Ascomycetes. Targeted gene deletion of DES1 had no apparent effect on developmental morphogenesis, including vegetative growth, conidial germination, appressorium formation, and appressorium-mediated penetration. Conidial size of the mutant became smaller than that of the wild type, but the mutant displayed no defects on cell wall integrity. The Δdes1 mutant was hypersensitive to exogenous oxidative stress and the activity and transcription level of extracellular enzymes including peroxidases and laccases were severely decreased in the mutant. In addition, ferrous ion leakage was observed in the Δdes1 mutant. In the interaction with a susceptible rice cultivar, rice cells inoculated with the Δdes1 mutant exhibited strong defense responses accompanied by brown granules in primary infected cells, the accumulation of reactive oxygen species (ROS), the generation of autofluorescent materials, and PR gene induction in neighboring tissues. The Δdes1 mutant displayed a significant reduction in infectious hyphal extension, which caused a decrease in pathogenicity. Notably, the suppression of ROS generation by treatment with diphenyleneiodonium (DPI), an inhibitor of NADPH oxidases, resulted in a significant reduction in the defense responses in plant tissues challenged with the Δdes1 mutant. Furthermore, the Δdes1 mutant recovered its normal infectious growth in DPI-treated plant tissues. These results suggest that DES1 functions as a novel pathogenicity gene that regulates the activity of fungal proteins, compromising ROS-mediated plant defense