Genetic structure of Sclerotinia sclerotiorum populations from sunflower and cabbage in West Azarbaijan province of Iran.

Sclerotinia sclerotiorum is one of the most destructive fungal pathogens infecting a wide array of plant species worldwide. Management of this pathogen relies on the coordinated use of fungicides and resistant host cultivars with other control measures, but the effectiveness of these methods requires knowledge of the genetic variability and structure of the fungal populations. To provide insight into the genetic diversity and structure of this pathogen in West Azarbaijan province of Iran, a total of 136 isolates were collected from symptomatic sunflower and cabbage plants within fields in three regions and analysed using inter-simple sequence repeat (ISSR) markers and intergenic spacer (IGS) region of the rRNA gene sequences. A total of 83 ISSR multilocus genotypes (MLGs) were identified, some of which were shared among at least two regional or host populations but in a low frequency. High genotypic diversity, low levels of clonal fraction, and random association of ISSR loci in a region indicated a low level of clonal reproduction, and possibly a high level of sexually recombining life cycle for the pathogen in the province. Marker analyses revealed that the pathogen was spatially homogeneous among fields, and thus similar control measures, such as the choice of resistant cultivars and fungicides, may effectively manage S. sclerotiorum within the region. Four IGS haplotypes (IGS1-IGS4) were detected within populations with IGS3 being the most prevalent haplotype. The low IGS haplotype diversity, the absence of spatial structure, and shared MLGs among populations may suggest a single introduction and subsequent dispersal of S. sclerotiorum within West Azarbaijan province.

Taxonomic and pathogenic characterization of Phytopythium species from West Azarbaijan, Iran, and description of two new species.

Phytopythium is a globally distributed genus, species of which occur in various ecological niches. Despite their importance as plant pathogens, information on species diversity and their distribution in West Azarbaijan Province of Iran is limited. This study aimed to investigate the taxonomy and plant pathogenicity of Phytopythium isolates recovered from soils in the province. A polyphasic taxonomy based on morphological, cultural, and multilocus sequence data revealed two new species, described here as P. babaiaharii and P. longitubum. In addition, a putatively new species and five known species, P. boreale, P. carbonicum, P. mercuriale, P. ostracodes, and P. vexans, were found in the studied region. Phytopythium ostracodes, P. mercuriale, and P. boreale were the three most frequent species isolated from soil, although P. mercuriale was only found in one field. Among the studied species, P. mercuriale, P. ostracodes, and P. vexans exhibited various levels of pathogenicity on sugar beet, sunflower, and tomato, and P. boreale was documented for first time as a plant pathogen. Phytopythium ostracodes was highly aggressive on sugar beet and sunflower but lowly aggressive on tomato, whereas P. vexans exhibited high aggressiveness toward the three crop plants. Both P. boreale and P. mercuriale were found to be lowly aggressive on the host plants. The results indicated that members of Phytopythium, particularly plant-pathogenic species, are common in arable soils of West Azarbaijan Province and can pose a threat to agricultural crops in the region.

Molecular and morphological characterization of three new species of Pythium from Iran: P. ershadii, P. pyrioosporum, and P. urmianum.

Three new species of Pythium: P. ershadii, P. pyrioosporum, and P. urmianum from soils of various regions in Iran are described and illustrated. These species are morphologically distinct from all other known species. Pythium ershadii is morphologically characterized by pyriform ornamented oogonia and rarely production of pyriform oospores. Pythium pyrioosporum differs from other species of the genus by the production of pyriform oospores and smooth walled oogonia, oospores with a tapering elongation toward a hypogynous antheridium and intercalary hypogynous antheridia. Pythium urmianum is distinguished by the presence of intercalary hypogynous antheridia, smooth walled oogonia formed laterally on hyphae or on short side branches and peanut-shaped oospores. Phylogenetic relationships of these new taxa with other Pythium species were investigated using internal transcribed spacers rDNA and partial coxI sequence data. The three species reside in clade E1 and are separated from closely related species.

MAP kinase phosphorylation and cAMP assessment in fungi.

The cyclic AMP (cAMP) signaling and mitogen-activated protein (MAP) kinase pathways are the most important signal transduction pathways in eukaryotes. In many plant pathogenic fungi they play pivotal roles in virulence and development. Identification and understanding the role of signal transduction pathways in regulation of cellular responses require robust biochemical techniques. Determination of both the phosphorylation status of MAPKs and the intracellular levels of cAMP is required to unravel the function of these pathways during adaptation of fungi to environmental stress conditions or when particular fungal genes are disrupted or silenced. Here we describe protocols to determine the phosphorylation status of three different MAPKs including Fus3, Slt2 and Hog1 as well as a protocol to measure the intracellular levels of cAMP levels. These protocols can be adapted for a wide range of fungi.