شناسایی یک گونه از Xylaria sp. بعنوان قارچ اندوفیت روی پرتقال تامسون ناول

Citrus endophyte fungi are unknown organism that living within tissues of fruit trees in various environment due to the drought stress, frost damage and physiological effects. They have special role in survival of these trees by producing specific secondry metabolites. In this research one isolate of endophytic fungi was separated from healthy leaves of orange tree (Citrus sinensis) Tompson cultivar in Lahijan city from Guilan provience. For fungal isolation samples were washed with tap water and then their inner side were cut along the main vein. For surface sterilization samples were soaked in 96% ethanol for 1‌min, 3.5% sodium hypochlorite for 5‌‌min and 96% ethanol for 30‌ sec respectively. Then rinsed with sterile distilled water twice and dried on sterile filter paper. Finally those samples were cultured on PDA medium and incubated at 25°C for ten days until observation of the fungal growth. According to morphological characteristics of this isolate formed shiny white mycelium on PDA. After 7-10 days black spots observed on colonies that became milky color stroma with
2-3mm thickness at base and 1.2-2.7 cm length was rised from each one. In molecular assessment genomic DNA of the endophytic fungi was extracted using Fast Prep method. 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 Xylaria venosula with 99% similarity and its NCBI, Gen Bank accession number was MF919405.