معرفی و شناسایی Cladosporium uredinicola بعنوان یکی از قارچ های اندوفیت‌ چای (Camellia sinensis)

Endophytic fungi are organisms inhabiting the living plant organs at some time during their life, without causing apparent harm to the host. In this research one isolate of endophytic fungi was separated from one-year-old and asymptomatic leaves of tea plant from tea-garden which was located in siyahkal city from Gilan province. For fungal isolation, leaf samples were immersed in water with saturated detergent for 30 min, then washed with tap water. Leaf samples soaked with %75 ethanol for 30 sec and rinsed three times with sterile distilled water, finally after disinfecting with %1.5 sodium hypochlorite for 3 min and rinsing with sterile distilled water 5 times then the samples dried on sterile filter paper. After these stages a leaf square was cut from each sample with sterile scalpel and plated on PDA medium then incubated at 25°C for ten days until observation of the fungal growth. According to morphological and cultural characteristics this isolate formed compacted shiny olivaceous mycelium on PDA. Conidiophores were tall and dark and their size were 87-90 × 2.1-2.3 µm. Conidia were dark, 1-celled and variable in shape from cylindrical to irregular and some typically lemon-shaped usually in branched chains, 2-3.5 × 2.1-7.8 µm. In molecular assessment genomic DNA of the endophytic fungi was extracted using CTAB method with some amendation. Amplification of ITS-rDNA, was done in a thermocycler using primer pair ITS1‌(TCCGTAGGTGAACCTGCGG) and ITS4‌‌(CCTCCGCTTATTGATATGC). Then for identification, PCR products sequencing was evaluated in BLAST. With analysis of rDNA internal transcribed spacer (ITS) sequence this isolate was further confirmed Cladosporium uredinicola with 99% similarity and its NCBI, Gene Bank accession number was KY488352.