Proteomic analysis during of spore germination of Moniliophthora perniciosa, the causal agent of witches' broom disease in cacao.

BACKGROUND: Moniliophthora perniciosa is a phytopathogenic fungus responsible for witches' broom disease of cacao trees (Theobroma cacao L.). Understanding the molecular events during germination of the pathogen may enable the development of strategies for disease control in these economically important plants. In this study, we determined a comparative proteomic profile of M. perniciosa basidiospores during germination by two-dimensional SDS-PAGE and mass spectrometry. RESULTS: A total of 316 proteins were identified. Molecular changes during the development of the germinative tube were identified by a hierarchical clustering analysis based on the differential accumulation of proteins. Proteins associated with fungal filamentation, such as septin and kinesin, were detected only 4 h after germination (hag). A transcription factor related to biosynthesis of the secondary metabolite fumagillin, which can form hybrids with polyketides, was induced 2 hag, and polyketide synthase was observed 4 hag. The accumulation of ATP synthase, binding immunoglobulin protein (BiP), and catalase was validated by western blotting. CONCLUSIONS: In this study, we showed variations in protein expression during the early germination stages of fungus M. perniciosa. Proteins associated with fungal filamentation, and consequently with virulence, were detected in basidiospores 4 hag., for example, septin and kinesin. We discuss these results and propose a model of the germination of fungus M. perniciosa. This research can help elucidate the mechanisms underlying basic processes of host invasion and to develop strategies for control of the disease.

The phytoremediation of an organic and inorganic polluted soil: A real scale experience.

A phytoremediation process with horse manure, plants (Populus alba, Cytisus scoparius, Paulownia tomentosa) and naturally growing vegetation was carried out at a real-scale in order to phytoremediate and functionally recover a soil contaminated by metals (Zn, Pb, Cd, Ni, Cu, Cr), hydrocarbons (TPH) and polychlorobiphenyls (PCB). All the plants were effective in two years in the reclamation of the polluted soil, showing an average reduction of about 35%, 40%, and 70% in metals, TPH and PCB content, respectively. As regards the plants, the poplar contributed the most to organic removal. In fact, its ability to take up and detoxify organic pollutants is well known. Paulownia tomentosa, instead, showed high metal removal. The Cytisus scoparius was the least effective plant in soil decontamination. The recovery of soil functionality was followed by enzyme activities, expressing the biochemical processes underway, and nutrient content useful for plant growth and development. Throughout the area, an enhancement of metabolic processes and soil chemical quality was observed. All the enzymatic activities showed a general increase over time (until 3-4 fold than the initial value for urease and ß-glucosidase). Moreover, Cytisus scoparius, even though it showed a lower decontamination capability, was the most effective in soil metabolic stimulation.

A real-scale soil phytoremediation.

In the present investigation, a phytoremediation process with a combination of different plant species (Populus nigra (var.italica), Paulownia tomentosa and Cytisus scoparius), and natural growing vegetation has been proposed at real-scale (10.000 m(2)) to bioremediate and functionally recover a soil historically contaminated by heavy metals and hydrocarbons. In the attempts to assess both effectiveness and evolution of the remediation system towards a natural soil ecosystem, besides the pollution parameters, also parameters describing the efficiency of the microbiological components (enzyme activities), were investigated. In 3 years, the total content of hydrocarbons and heavy metals in soil decreased with time (40 % and 20-40 %, respectively), reaching concentrations under the limit of National legislation and making the site suitable for environmental reusing. The reduction in pollutants was probably the reason of the increase in dehydrogenase (indicator of overall microbial activity), ß-glucosidase and phosphatase activities, enzymes related to C and P cycles, respectively. However, this trend was obviously due also to the increase of chemical nutrients, acting as substrate of these enzymes. Moreover, a phytotest carried out with Raphanus sativus, showed, after 3 years, a significant increase in percentage of plant growth, confirming a reduction in soil toxicity and an improvement in soil nutritional state. Therefore, this phytoremediation system seems very promising to perform both decontamination and functional recovery of a polluted soil at real-scale level.

From blooms to brooms.

Phytoplasmas enhance transmission by manipulating plant architecture. Recently, Huang et al. report that SAP05, an effector from a phytoplasma that causes witches' broom, targets host transcription factors for proteasomal degradation by binding host ubiquitin receptor RPN10. These findings provide opportunities for engineering phytoplasma-resistant plants and developing protein therapeutics.

Differential growth costs and nitrogen fixation in Cytisus multiflorus (L'Hér.) Sweet and Cytisus scoparius (L.) Link are mediated by sources of inorganic N.

Shrubby legumes in Mediterranean-type ecosystems face strong nutrient limitations that worsen in summer, when water is absent. Nitrogen-fixing legumes are likely to be able to switch between soil N and atmospheric N (N2 ) sources to adjust the C costs of N acquisition in different seasons. We investigated the utilisation of different inorganic N sources by two indigenous shrubby legumes (Cytisus multiflorus and Cytisus scoparius). Plant performance in terms of photosynthesis and biomass production was also analysed. Plants were cultivated in sterile river sand supplied with Hoagland nutrient solution, grown in N-free nutrient solution and inoculated with effective rhizobial strains from nodules of adult plants of the same species. A second treatment consisted of plants given 500 µm NH4 NO3 added into the nutrient solution. In a third treatment, plants were watered with another source of N (500 µm NH4 NO3 ) as well as being inoculated with effective rhizobial strains. The application of NH4 NO3 to the legumes resulted in a larger increase in plant dry matter. Carbon construction costs were higher in plants supplied with mineral and symbiotic N sources and always higher in the endemic C. multiflorus. Differences in photosynthesis rates were only observed between species, regardless of the N source. Non-fertilised inoculated plants had more effective root nodules and a clear dependence on N2 fixation. We propose that the ability of C. scoparius to change N source makes it a plastic species, which would account for its broader distribution in nature.

(+)-2,3-dehydro-10-oxo-alpha-isosparteine in Uresiphita reversalis larvae fed on Cytisus monspessulanus leaves.

Quinolizidine alkaloids, found in the leaves of Cytisus monspessulanus L. (Leguminosae), were characterized in the cuticle of larvae of the pyralid moth Uresiphita reversalis (Lepidoptera: Pyralidae) when the latter were fed on this weed. By GC-MS analysis of the methanolic extracts of the cuticle, four quinolizidine alkaloids, N-methylcytisine, cytisine, aphylline and anagyrine, were identified as possible defense substances. In addition, the quinolizidine alkaloid, (+)-2,3-dehydro-10-oxo-alpha-isosparteine was characterized in both the insect and host plant.

Influence of plant root exudates on the mobility of fuel volatile compounds in contaminated soils.

Vegetation and its associated microorganisms play an important role in the behaviour of soil contaminants. One of the most important elements is root exudation, since it can affect the mobility, and therefore, the bioavailability of soil contaminants. In this study, we evaluated the influence of root exudates on the mobility of fuel derived compounds in contaminated soils. Samples of humic acid, montmorillonite, and an A horizon from an alumi-umbric Cambisol were contaminated with volatile contaminants present in fuel: oxygenates (MTBE and ETBE) and monoaromatic compounds (benzene, toluene, ethylbenzene and xylene). Natural root exudates obtained from Holcus lanatus and Cytisus striatus and ten artificial exudates (components frequently found in natural exudates) were added to the samples, individually and as a mixture, to evaluate their effects on contaminant mobility. Fuel compounds were analyzed by headspace-gas chromatography-mass spectrometry. In general, the addition of natural and artificial exudates increased the mobility of all contaminants in humic acid. In A horizon and montmorillonite, natural or artificial exudates (as a mixture) decreased the contaminant mobility. However, artificial exudates individually had different effects: carboxylic components increased and phenolic components decreased the contaminant mobility. These results established a base for developing and improving phytoremediation processes of fuel-contaminated soils.

Metal uptake by native plants and revegetation potential of mining sulfide-rich waste-dumps.

Waste dumps resulting from metal exploitation create serious environmental damage, providing soil and water degradation over long distances. Phytostabilization can be used to remediate these mining sites. The present study aims to evaluate the behavior of selected plant species (Erica arborea, Ulex europaeus, Agrostis delicatula, and Cytisus multiflorus) that grow spontaneously in three sulfide-rich waste-dumps (Lapa Grande, Cerdeirinha, and Penedono, Portugal). These sites represent different geological, climatic and floristic settings. The results indicate distinctive levels and types of metal contamination: Penedono presents highest sulfate and metal contents, especially As, with low levels of Fe. In contrast, at Lapa Grande and Cerdeirinha Fe, Mn, and Zn are the dominant metals. In accordance, each waste dump develops a typical plant community, providing a specific vegetation inventory. At Penedono, Agrostis delicatula accumulates As, Pb, Cu, Mn, and Zn, showing higher bioaccumulation factors (BF) for Mn (32.1) and As (24.4). At Cerdeirinha, Ulex europaeus has the highest BF for Pb (984), while at Lapa Grande, Erica arborea presents high BF for Mn (9.8) and Pb (8.1). Regarding TF, low values were obtained for most of the metals, especially As (TF < 1). Therefore, the results obtained from representative plant species suggest appropriate behavior for phytostabilization measures.