Arabidopsis heterotrimeric G-protein regulates cell wall defense and resistance to necrotrophic fungi

The Arabidopsis heterotrimeric G-protein controls defense responses to necrotrophic and vascular fungi. The agb1 mutant impaired in the Gβ subunit displays enhanced susceptibility to these pathogens. Gβ/AGB1 forms an obligate dimer with either one of the Arabidopsis Gγ subunits (γ1/AGG1 and γ2/AGG2). Accordingly, we now demonstrate that the agg1 agg2 double mutant is as susceptible as agb1 plants to the necrotrophic fungus Plectosphaerella cucumerina. To elucidate the molecular basis of heterotrimeric G-protein-mediated resistance, we performed a comparative transcriptomic analysis of agb1-1 mutant and wild-type plants upon inoculation with P. cucumerina. This analysis, together with metabolomic studies, demonstrated that G-protein-mediated resistance was independent of defensive pathways required for resistance to necrotrophic fungi, such as the salicylic acid, jasmonic acid, ethylene, abscisic acid, and tryptophan-derived metabolites signaling, as these pathways were not impaired in agb1 and agg1 agg2 mutants. Notably, many mis-regulated genes in agb1 plants were related with cell wall functions, which was also the case in agg1 agg2 mutant. Biochemical analyses and Fourier Transform InfraRed (FTIR) spectroscopy of cell walls from G-protein mutants revealed that the xylose content was lower in agb1 and agg1 agg2 mutants than in wild-type plants, and that mutant walls had similar FTIR spectratypes, which differed from that of wild-type plants. The data presented here suggest a canonical functionality of the Gβ and Gγ1/γ2 subunits in the control of Arabidopsis immune responses and the regulation of cell wall composition.

Multiple avirulence paralogues in cereal powdery mildew fungi may contribute to parasite fitness and defeat of plant resistance

Powdery mildews, obligate biotrophic fungal parasites on a wide range of important crops, can be controlled by plant resistance (R) genes, but these are rapidly overcome by parasite mutants evading recognition. It is unknown how this rapid evolution occurs without apparent loss of parasite fitness. R proteins recognize avirulence (AVR) molecules from parasites in a gene-for-gene manner and trigger defense responses. We identify AVRa10 and AVRk1 of barley powdery mildew fungus, Blumeria graminis f sp hordei (Bgh), and show that they induce both cell death and naccessibility when transiently expressed in Mla10 and Mlk1 barley (Hordeum vulgare) varieties, respectively. In contrast with other reported fungal AVR genes, AVRa10 and AVRk1 encode proteins that lack secretion signal peptides and enhance infection success on susceptible host plant cells. AVRa10 and AVRk1 belong to a large family with mayor que30 paralogues in the genome of Bgh, and homologous sequences are present in other formae speciales of the fungus infecting other grasses. Our findings imply that the mildew fungus has a repertoire of AVR genes, which may function as effectors and contribute to parasite virulence. Multiple copies of related but distinct AVR effector paralogues might enable populations of Bgh to rapidly overcome host R genes while maintaining virulence.

The Interactive Effects of Temperature and Osmotic Potential on the Growth of Aquatic Isolates of Fusarium culmorum.

The mycelial growth of 10 Fusarium culmorum strains isolated from water of the Andarax riverbed in the provinces of Granada and Almeria in southeastern Spain was tested on potato-dextroseagar adjusted to different osmotic potentials with either KCl or NaCl (−1.50 to−144.54 bars) at 10◦C intervals ranging from15◦ to 35◦C. Fungal growth was determined by measuring colony diameter after 4 d of incubation. Mycelial growth was maximal at 25◦C. The quantity and capacity of mycelial growth of F. culmorum were similar at 15 and 25◦C, with maximal growth occurring at −13.79 bars water potential and a lack of growth at 35◦C. The effect of water potential was independent of salt composition. The general growth pattern of Fusarium culmorum growth declined at potentials below −13.79 bars. Fungal growth at 25◦C was always greater than growth at 15◦C, at all of the water potentials tested. Significant differences were observed in the response ofmycelia to water potential and temperature as main and interactive effects. The number of isolates that showed growth was increasingly inhibited as the water potential dropped, but some growth was still observable at −99.56 bars. These findings could indicate that F. culmorum strains isolated from water have a physiological mechanism that permits survival in environments with low water potential. Propagules of Fusarium culmorum are transported long distances by river water, which could explain the severity of diseases caused by F.culmorum on cereal plants irrigated with river water and its interaction under hydric stress ormoderate soil salinity. The observed differences in growth magnitude and capacity could indicate that the biological factors governing potential and actual growth are affected by osmotic potential in different ways.

Toxicity of ZnO nanoparticles, ZnO bulk and ZnCl2 on earthworms in a spiked natural soil and toxicological effects of leachates on aquatic organisms

The present study assessed the uptake and toxicity of ZnO nanoparticles (NPs), ZnO bulk, and ZnCl2 salt in earthworms in spiked agricultural soils. In addition, the toxicity of aqueous extracts to Daphnia magna and Chlorella vulgaris was analyzed to determine the risk of these soils to the aquatic compartment. We then investigated the distribution of Zn in soil fractions to interpret the nature of toxicity. Neither mortality nor differences in earthworm body weight were observed compared with the control. The most sensitive end point was reproduction. ZnCl2 was notably toxic in eliminating the production of cocoons. The effects induced by ZnO-NPs and bulk ZnO on fecundity were similar and lower than those of the salt. In contrast to ZnO bulk, ZnO-NPs adversely affected fertility. The internal concentrations of Zn in earthworms in the NP group were greater than those in the salt and bulk groups, although bioconcentration factors were consistently <1. No relationship was found between toxicity and internal Zn amounts in earthworms. The results from the sequential extraction of soil showed that ZnCl2 displayed the highest availability compared with both ZnO. Zn distribution was consistent with the greatest toxicity showed by the salt but not with Zn body concentrations. The soil extracts from both ZnO-NPs and bulk ZnO did not show effects on aquatic organisms (Daphnia and algae) after short-term exposure. However, ZnCl2 extracts (total and 0.45-μm filtered) were toxic to Daphnia.

Simultaneous determination of multiclass emerging contaminants in aquatic plants by ultrasound assisted matrix solid phase dispersion and GC-MS

A multiresidue method was developed for the simultaneous determination of 31 emerging contaminants (pharmaceutical compounds, hormones, personal care products, biocides and flame retardants) in aquatic plants. Analytes were extracted by ultrasound assisted-matrix solid phase dispersion (UA-MSPD) and determined by gas chromatography-mass spectrometry after sylilation. The method was validated for different aquatic plants (Typha angustifolia, Arundo donax and Lemna minor) and a semiaquatic cultivated plant (Oryza sativa) with good recoveries at concentrations of 100 and 25 ng g-1 wet weight, ranging from 70 to 120 %, and low method detection limits (0.3 to 2.2 ng g-1 wet weight). A significant difference of the chromatographic response was observed for some compounds in neat solvent versus matrix extracts and therefore quantification was carried out using matrix-matched standards in order to overcome this matrix effect. Aquatic plants taken from rivers located at three Spanish regions were analyzed and the compounds detected were parabens, bisphenol A, benzophenone-3, cyfluthrin and cypermethrin. The levels found ranged from 6 to 25 ng g-1 wet weight except for cypermethrin that was detected at 235 ng g-1 wet weight in Oryza sativa samples.