First report of early blight caused by Alternaria protenta on potato in Algeria

Potato (Solanum tuberosum L.) is one of the most important vegetable crop grown in Algeria. Although less studied than late blight, early blight caused by large-spored Alternaria species including A. solani, and A. grandis is an important foliar disease on these crops under Algerian climatic conditions. Over the past few years, this disease became a major constraint on potato production in Algeria and high incidences of early blight (up to 80 %) were recorded in the north-western parts of Algeria (Bessadat et al., 2016). During years 2012-2014, surveys were carried out and sampling was performed in 12 potato growing regions from East to West and from North to South of Algeria. Two hundred and forty-seven samples with typical early blight symptoms (dark, elongated or circular lesions with concentric rings surrounded by a yellow halo) were collected. Two to three lesions perleaf were excised, surface disinfested for 2 min in 0.1% (v/v) sodium hypochlorite solution and plated on potato dextrose agar medium at 22°C. Twenty-two pure cultures were induced to sporulate by plating on V8 medium and incubating for two weeks under alternating 12 h darkness and 12 h near UV light. Cultural and morphological characteristics of the isolates [color and pigmentation of the culture, shape and size of the conidia (conidial length and width, beak length)] compared with those in the literature (Simmons, 2007) did not clearly differentiate between the large-spored Alternaria species currently reported on potato crops. For identification at the species level, partial regions of the calmodulin (cal) and RNA polymerase second largest subunit (rpb2) genes were amplified using published primer sets (Gannibal et al., 2014; Woudenberg et al., 2014) and sequenced. Maximum likelihood cluster analyses of the resulting nucleotide sequences revealed two isolates (AD82 and AD86 isolated from potato samples collected in the El Oued region) with sequences at the two loci [GenBank accession Nos. KX870505 and KX870506 (cal locus), KX870507 and KX870508 (rpb2 locus)] that shared 100% sequence homology to A. protenta isolate CBS 116696 (KJ718394, JQ646236) and were thus assigned to this species. A. protenta was previously known in Africa only on Helianthus annuus (Simmons, 1986) but isolates from S. tuberosum and S. lycopersicum collected in New Zealand and USA and formerly recognized as A. solani, were recently moved to A. protenta based on phylogeny (Woudenberg et al., 2014). To confirm pathogenicity of the two A. protenta isolates, leaves of susceptible 3-week-old potato plants were inoculated with two 10 µL drops of a 104 conidia/ml suspension. All leaves inoculated with A. protenta showed extending lesions that may reach up 50% of the leaf area at 21 dpi. To our knowledge, we report the first occurrence of A. protenta as pathogen on potato in Algeria.

Molecular cloning and biochemical characterization of a Cu,Zn-superoxide dismutase from Scedosporium apiospermum.

A Cu,Zn-superoxide dismutase has been characterized from Scedosporium apiospermum, a fungus which often colonizes the respiratory tract of patients with cystic fibrosis. Enzyme production was stimulated by iron starvation. Purification was achieved from mycelial extract from 7-day-old cultures on Amberlite XAD-16. The purified enzyme presented a relative molecular mass of 16.4 kDa under reducing conditions and was inhibited by potassium cyanide and diethyldithiocarbamate, which are two known inhibitors of Cu,Zn-SODs. Its optimum pH was 7.0 and the enzyme retained full activity after pretreatment at temperatures up to 50 degrees C. Moreover, a 450-bp fragment of the gene encoding the enzyme was amplified by PCR using degenerate primers designed from sequence alignment of four fungal Cu,Zn-SODs. Sequence data from this fragment allowed us to design primers which were used to amplify by walking-PCR the flanking regions of the known fragment. SaSODC gene (890 bp) corresponded to a 154 amino acid polypeptide with a predicted molecular mass of 15.9 kDa. A database search for sequence homology revealed for the deduced amino acid sequence 72 and 83% identity rate with Cu,Zn-SODs from Aspergillus fumigatus and Neurospora crassa, respectively. To our knowledge, this enzyme is the first putative virulence factor of S. apiospermum to be characterized

Multi-scale simulation of the nano-metric cutting process

Molecular dynamics (MD) simulation and the finite element (FE) method are two popular numerical techniques for the simulation of machining processes. The two methods have their own strengths and limitations. MD simulation can cover the phenomena occurring at nano-metric scale but is limited by the computational cost and capacity, whilst the FE method is suitable for modelling meso- to macro-scale machining and for simulating macro-parameters, such as the temperature in a cutting zone, the stress/strain distribution and cutting forces, etc. With the successful application of multi-scale simulations in many research fields, the application of simulation to the machining processes is emerging, particularly in relation to machined surface generation and integrity formation, i.e. the machined surface roughness, residual stress, micro-hardness, microstructure and fatigue. Based on the quasi-continuum (QC) method, the multi-scale simulation of nano-metric cutting has been proposed. Cutting simulations are performed on single-crystal aluminium to investigate the chip formation, generation and propagation of the material dislocation during the cutting process. In addition, the effect of the tool rake angle on the cutting force and internal stress under the workpiece surface is investigated: The cutting force and internal stress in the workpiece material decrease with the increase of the rake angle. Finally, to ease multi-scale modelling and its simulation steps and to increase their speed, a computationally efficient MATLAB-based programme has been developed, which facilitates the geometrical modelling of cutting, the simulation conditions, the implementation of simulation and the analysis of results within a unified integrated virtual-simulation environment

Identifying Natural Products (NPs) as potential UPR inhibitors for crop protection

As far as the future of agriculture is concerned, one major challenge will be to face an expected increase in health risks due to pesticides together with a lower efficiency of crop treatments. Therefore it is today necessary to develop new strategies to enhance the effectiveness and sustainability of current control methods. The so-called “Alternaria Leaf Spot“ is a common disease of crucifers caused by the fungal pathogen Alternaria brassicicola which affects different crops including cabbage, kale, Brussels sprout, cauliflower and broccoli. Indole phytoalexins camalexin and brassinin play in planta a key role in crop protection against this necrotrophic agent. However it has been shown that mutants become phytoalexin-resistant by activating at least three signaling pathways named as Cell Wall Integrity (CWI), High Osmolarity Glycerol (HOG) and Unfolded Protein Response (UPR) [1,2]. The latter is particularly involved in the fungus protection against phytoalexins since UPR deficient avirulent mutants of A. brassicicola appear as hypersensitive to camalexin and brassinin [3]. Since very few UPR inhibitors such as the synthetic STF-083010 [4] are known we decided to develop an original screening assay, detecting the production of a HAC1 fluorescence-induced protein, i.e. a transcriptional activator involved in the UPR pathway, in Saccharomyces cerevisiae cultures (Figure 1). The preliminary screening of an in-house NPs library [c.a. 70 compounds (polyphenols, terpenoids and alkaloids)] clearly revealed aescin (Aesculus hippocastanum)] as a potential UPR inhibitor

Pan-European early switch/early discharge opportunities exist for hospitalised patients with methicillin-resistant <em>Staphylococcus</em> <em>aureus</em> complicated skin and soft-tissue infections

AbstractThe objective of this study was to document pan-European real-world treatment patterns and healthcare resource use and estimate opportunities for early switch (ES) from intravenous (IV) to oral antibiotics and early discharge (ED) in hospitalized patients with methicillin-resistant Staphylococcus aureus (MRSA) complicated skin and soft tissue infections (cSSTIs). This retrospective observational medical chart review study enrolled 342 physicians across 12 European countries who collected data from 1542 patients with documented MRSA cSSTI who were hospitalized (July 2010 to June 2011) and discharged alive (by July 2011). Data included clinical characteristics and outcomes, hospital length of stay (LOS), MRSA-targeted IV and oral antibiotic use, and ES and ED eligibility according to literature-based and expert-validated criteria. The most frequent initial MRSA-active antibiotics were vancomycin (50.2%), linezolid (15.1%), clindamycin (10.8%), and teicoplanin (10.4%). Patients discharged with MRSA-active antibiotics (n = 480) were most frequently prescribed linezolid (42.1%) and clindamycin (19.8%). IV treatment duration (9.3 ± 6.5 vs. 14.6 ± 9.9 days; p <0.001) and hospital LOS (19.1 ± 12.9 vs. 21.0 ± 18.2 days; p 0.162) tended to be shorter for patients switched from IV to oral treatment than for patients who received IV treatment only. Of the patients, 33.6% met ES criteria and could have discontinued IV treatment 6.0 ± 5.5 days earlier, and 37.9% met ED criteria and could have been discharged 6.2 ± 8.2 days earlier. More than one-third of European patients hospitalized for MRSA cSSTI could be eligible for ES and ED, resulting in substantial reductions in IV days and bed-days, with potential savings of €2000 per ED-eligible patient

Distribution of large-spored Alternaria species associated with potato and tomato early blight according to hosts and bioclimatic regions of Algeria

Potato and tomato are important crops in Algerian agriculture, and both are threatened by abiotic and biotic stresses, and early blight is a major disease affecting both crops. Surveys carried out from 2012 to 2015 in 12 major growing regions for these crops yielded a total of 247 Alternaria isolates having morphological and cultural characteristics of sections Alternaria and Porri. Since early blight symptoms and morphological characteristics of the isolates did not allow sharp distinction between the different large-spored species of Alternaria, the isolates in section Porri, often considered primary causes of the diseases, were selected for molecular characterization by diagnostic PCR using specific primers. This allowed species identification of 147 Alternaria isolates as A. solani, A. protenta, A. grandis or A. linariae. These species were present on potato and tomato crops at varying frequencies, depending on the hosts and on bioclimatic locations. Pathogenicity tests for the four species, on detached leaflets and whole seedlings, showed that all were pathogenic to potato and tomato, with varying virulence. These results suggest that parasitic specialization of these Alternaria species on solanaceous plants should be reconsidered

First report of early blight caused by Alternaria linariae on potato in Algeria

Potato (Solanum tuberosum L.) occupies a leading place in Algerian agriculture where it is grown over an area of approximatively 160,000 ha and can be planted and harvested in virtually any month of the year. However, this production remains threatened by early blight disease caused by Alternaria species including A. solani, A. grandis and A. protenta (Bessadat et al., 2017; Ayad et al., 2017). High incidences of early blight (up to 80 %) have previously been observed in north-western parts of Algeria on potato as well as tomato (Bessadat et al., 2017). Potato disease surveys were carried out between 2012 and 2015 with sampling performed in twelve potato growing regions of Algeria. One hundred and sixty-four leaf samples exhibiting typical early blight symptoms (dark, elongated or circular lesions with concentric rings surrounded by a yellow halo) were collected. Small pieces (3-4 mm2) were cut off from the lesion edges, surface disinfested and plated on potato dextrose agar medium at 22°C. From the isolates obtained, eighty-two exhibited morphological characteristics associated with species of Alternaria belonging to the section Porri. Twenty-two pure cultures were induced to sporulate by plating on V8 medium and incubating for two weeks under alternating 12 h darkness and 12 h near-UV light. Although most of the isolates had typical morphological and sporulation characteristics of A. solani and A. grandis, five isolates, originating from north Algeria, produced conidia whose body and beak length (up to 108 µm and 225 µm, respectively) were in the range of those described for A. linariae (Gannibal et al., 2014). For confirmation of the identity of these isolates at the species level, partial region of the calmodulin (cal) gene was amplified using published primer set (Gannibal et al., 2014) and sequenced. Maximum-likelihood cluster analysis of the resulting nucleotide sequences (GenBank accession Nos. MH243795, MH243769, MH243793, MH243789, MH243794) and additional reference sequences of species within the section Porri confirmed that the five isolates (DA01, DA02, DA03 from the wilaya Tipaza and DA06, DA07 from the wilaya Alger) could be assigned to A. linariae with 100% sequence similarity to a reference strain (CBS 109161; GenBank accession No. JQ646254) (Woudenberg et al., 2014). To confirm pathogenicity of the five A. linariae isolates, 3-week-old leaves of susceptible varieties of tomato (Marmande and St Pierre) and potato (Spunta and Sarpomira), cultivated in pot under greenhouse at 28°C with 16h/day light, were inoculated by depositing 20 µl drops of a 104 conidia/ml suspension. Irrespective of the tested plant species and variety, all inoculated leaves showed extending lesions that may reach up 60% of the leaf area at 21 days post-inoculation. No symptom was observed on control plants treated with distilled water. Together with A. solani, A. linariae is considered as the main causal agent of tomato early blight (Gannibal et al., 2014) but it has never been described on potato. To our knowledge, this is the first report for the occurrence of A. linariae on potato. The fact that potato and tomato fields often co-exist in close proximity in northwestern Algeria with farmers even using tomato in rotation with potato may favor the development of A. linariae on the latter plant specie