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Chemical warfare between the host and the pathogen plays a crucial role in plant-necrotrophic pathogen interactions, but examples of its involvement in quantitative disease resistance in plants are poorly documented. In the Daucus carota-Alternaria dauci pathosystem, the novel toxin aldaulactone has been identified as a key factor in both fungal pathogenicity and the carrot’s partial resistance to the pathogen. Bioinformatic analyses have pinpointed a secondary metabolism gene cluster that harbors two polyketide synthase genes, AdPKS7 and AdPKS8 , that are likely responsible for the biosynthesis of aldaulactone. Here, we present the functional validation of AdPKS7 and AdPKS8 as genes responsible for aldaulactone production in A. dauci . We generated A. dauci knock-out mutants for AdPKS7 and AdPKS8 by replacing essential domains with a hygromycin resistance gene, marking the first reported case of genetic manipulation in A. dauci . Following transformation, the mutants were analyzed for toxin production via HPLC-UV and assessed for pathogenicity in planta . Aldaulactone production was abolished in all PKS mutants, which also exhibited significantly reduced pathogenicity on H1-susceptible carrot leaves. These findings confirm the roles of AdPKS7 and AdPKS8 in aldaulactone biosynthesis and their contribution to fungal pathogenicity.

Alternaria species have often been reported as plant pathogens and are commonly isolated from diseased plant hosts. During an investigation of this genus in Algeria, seven Embellisia -like isolates were collected from Apiaceae. Phylogenetic analysis using sequences at four loci, the internal transcribed spacer of the rDNA region (ITS), glyceraldehyde-3-phosphate dehydrogenase ( gpd ), translation elongation factor 1-alpha (tef1), and RNA polymerase second largest subunit (rpb2), revealed that these isolates grouped into three sections, namely Embellisia , Embellisioides , and Eureka . Four isolates had significant differences with their closest species and were determined to be new species, namely Alternaria longiformis and A. radicicola spp. nov. The three other isolates of section Eureka were identified as A. eureka and A. hungarica , the latter species being described as a new record in Algeria. Detailed descriptions of new species are provided based on colony color, aspect, diameter, conidial size, septa, sporulation patterns and compared with other relevant Alternaria species within same sections. All these species were weakly pathogenic on carrot, coriander, and fennel under greenhouse experiments. Apiaceae may constitute a reservoir of Alternaria species that could represent potential pathogens for other plant families.

Leaf blight and spot caused by Stemphylium spp. and Alternaria spp. are the most common destructive tomato diseases in north-western Algeria. During 2018 growing seasons, more than 30% of samples collected from plants grown in greenhouses or open fields were infected with Stemphylium. Initial symptoms were small, multiple, irregular to oval, yellow leaf spots, which enlarged to brown lesions later. In these lesions, Stemphylium mostly co-occurred with Alternaria spp. Twenty nine Stemphylium isolates were characterized based on morphological features, and multi-locus phylogenies using ITS, gpd, and cmdA genomic loci. Five Stemphylium species (S. lycopersici, S. gracilariae, S. eturmiunum, S. vesicarium, S. lycii) were associated with tomato leaf spot, of which S. lycii is a new report for tomato. Pathogenicity tests on healthy 2-months-old tomato seedlings reproduced symptoms similar to those observed in tomato crops. The tested fungus isolates differed in pathogenicity. Two isolates of S. lycopersici were more aggressive than those of the other species, causing major lesions on tomato plants. The five identified Stemphylium species are reported for the first time as new pathogens for tomato in Algeria, and S. lycopersici, S. gracilariae, S. eturmiunum, and S. lycii as new species of Algerian mycoflora.

Alternaria brassicicola causes black spot disease in Brassicaceae . During host infection, this necrotrophic fungus is exposed to various antimicrobial compounds, such as the phytoalexin brassinin which is produced by many cultivated Brassica species. To investigate the cellular mechanisms by which this compound causes toxicity and the corresponding fungal adaptive strategies, we first analyzed fungal transcriptional responses to short-term exposure to brassinin and then used additional functional approaches. This study supports the hypothesis that indolic phytoalexin primarily targets mitochondrial functions in fungal cells. Indeed, we notably observed that phytoalexin treatment of A. brassicicola disrupted the mitochondrial membrane potential and resulted in a significant and rapid decrease in the oxygen consumption rates. Secondary effects, such as Reactive oxygen species production, changes in lipid and endoplasmic reticulum homeostasis were then found to be induced. Consequently, the fungus has to adapt its metabolism to protect itself against the toxic effects of these molecules, especially via the activation of high osmolarity glycerol and cell wall integrity signaling pathways and by induction of the unfolded protein response.

Background MCC/eisosomes are membrane microdomains that have been proposed to participate in the plasma membrane function in particular by regulating the homeostasis of lipids, promoting the recruitment of specific proteins and acting as provider of membrane reservoirs. Results Here we showed that several potential MCC/eisosomal protein encoding genes in the necrotrophic fungus A. brassicicola were overexpressed when germinated spores were exposed to antimicrobial defence compounds, osmotic and hydric stresses, which are major constraints encountered by the fungus during the plant colonization process. Mutants deficient for key MCC/eisosome components did not exhibit any enhanced susceptibility to phytoalexins and to applied stress conditions compared to the reference strain, except for a slight hypersensitivity of the ∆∆abpil1a-abpil1b strain to 2 M sorbitol. Depending on the considered mutants, we showed that the leaf and silique colonization processes were impaired by comparison to the wild-type, and assumed that these defects in aggressiveness were probably caused by a reduced appressorium formation rate. Conclusions This is the first study on the role of MCC/eisosomes in the pathogenic process of a plant pathogenic fungus. A link between these membrane domains and the fungus ability to form functional penetration structures was shown, providing new potential directions for plant disease control strategies.

MCC/eisosomes are membrane microdomains that have been proposed to participate in the plasma membrane function in particular by regulating the homeostasis of lipids, promoting the recruitment of specific proteins and acting as provider of membrane reservoirs. Here we showed that several potential MCC/eisosomal protein encoding genes in the necrotrophic fungus A. brassicicola were overexpressed when germinated spores were exposed to antimicrobial defence compounds, osmotic and hydric stresses, which are major constraints encountered by the fungus during the plant colonization process. Mutants deficient for key MCC/eisosome components did not exhibit any enhanced susceptibility to phytoalexins and to applied stress conditions compared to the reference strain, except for a slight hypersensitivity of the [DELA][DELA]abpil1a-abpil1b strain to 2 M sorbitol. Depending on the considered mutants, we showed that the leaf and silique colonization processes were impaired by comparison to the wild-type, and assumed that these defects in aggressiveness were probably caused by a reduced appressorium formation rate. This is the first study on the role of MCC/eisosomes in the pathogenic process of a plant pathogenic fungus. A link between these membrane domains and the fungus ability to form functional penetration structures was shown, providing new potential directions for plant disease control strategies.

ABSTRACTAlternaria brassicicola causes dark spot (or black spot) disease, which is one of the most common and destructive fungal diseases of Brassicaceae spp. worldwide. Here, we report the draft genome sequence of strain Abra43. The assembly comprises 29 scaffolds, with an N50 value of 2.1 Mb. The assembled genome was 31,036,461 bp in length, with a G+C content of 50.85%.

The interaction between plants and plant pathogens can have significant effects on ecosystem performance. For their growth and development, both bionts rely on amino acids. While amino acids are key transport forms of nitrogen and can be directly absorbed from the soil through specific root amino acid transporters, various pathogenic microbes can invade plant tissues to feed on different plant amino acid pools. In parallel, plants may initiate an immune response program to restrict this invasion, employing various amino acid transporters to modify the amino acid pool at the site of pathogen attack. The interaction between pathogens and plants is sophisticated and responses are dynamic. Both avail themselves of multiple tools to increase their chance of survival. In this review, we highlight the role of amino acid transporters during pathogen infection. Having control over the expression of those transporters can be decisive for the fate of both bionts but the underlying mechanism that regulates the expression of amino acid transporters is not understood to date. We provide an overview of the regulation of a variety of amino acid transporters, depending on interaction with biotrophic, hemibiotrophic or necrotrophic pathogens. In addition, we aim to highlight the interplay of different physiological processes on amino acid transporter regulation during pathogen attack and chose the LYSINE HISTIDINE TRANSPORTER1 (LHT1) as an example.

Alternaria species have often been reported as plant pathogens and are commonly isolated from diseased plant hosts. During an investigation of this genus in Algeria, seven Embellisia -like isolates were collected from Apiaceae. Phylogenetic analysis using sequences at four loci, the internal transcribed spacer of the rDNA region (ITS), glyceraldehyde-3-phosphate dehydrogenase ( gpd ), translation elongation factor 1-alpha (tef1), and RNA polymerase second largest subunit (rpb2), revealed that these isolates grouped into three sections, namely Embellisia , Embellisioides , and Eureka . Four isolates had significant differences with their closest species and were determined to be new species, namely Alternaria longiformis and A. radicicola spp. nov. The three other isolates of section Eureka were identified as A. eureka and A. hungarica , the latter species being described as a new record in Algeria. Detailed descriptions of new species are provided based on colony color, aspect, diameter, conidial size, septa, sporulation patterns and compared with other relevant Alternaria species within same sections. All these species were weakly pathogenic on carrot, coriander, and fennel under greenhouse experiments. Apiaceae may constitute a reservoir of Alternaria species that could represent potential pathogens for other plant families.