جداسازی و شناسایی دو گونه Amesia از حشرات دو راسته ‏Coleoptera و Dermaptera

With the aim of isolation and identification of insect-associated fungi, different life stages of insects from different orders, including adults and larvae of Coleoptera and adults of Dermaptera, were collected from habitats in the downtown regions and suburbs of Ahvaz. To isolate fungal agents, full or part body of larva or adult were surface-sterilized by 2% sodium hypochlorite (2 min), followed by washing with sterile distilled water. Full or part bodies were plated into dishes containing potato dextrose agar (PDA) supplemented with streptomycin and the dishes were incubated at 28 °C. The grown colonies were purified by the single-spore method. In Primary morphological study, both isolates were diagnosed that they belong to the family Chaetomiaceae (SCUA-02 and SCUA-05) and used for morphological and molecular identification.The isolates were cultivated in potato-dextrose-broth (PDB) and mycelial biomass was collected using sterile filter paperand then freeze-dried. Genomic DNA was extracted by an organic method based on phenol and chloroform. Partial region of nrRNA (around 1200 bp from ITS and 28s-D1/D2) was amplified using universal primer pair ITS1 and NL4, and sequenced. The sequences were edited and used in phylogenetic analysis and BLASTn search algorithm. In BLASTn search, isolates SCUA-02 and SCUA-05 showed 100% and 98% sequence identity with type strains of Amesiaatrobrunnea CBS 379.66and Amesiagelasinospora CBS 673.80, respectively. In phylogenetic tree constructed based on ITS region, both isolates clustered with strains of the same species, but failed to form sufficiently supported clades. Proper morphology of the isolates, particularly the shape and size of ascoma and ascospores, in compare to original description of the type strains of both mentioned and closely related species showed that the SCUA-02 is Amesiaatrobrunnea species while the SCUA-05 is not the same with any one and considerate as Amesia sp. This firstly reports the association of these fungi with mentioned insects.