In order to identify the endophytic fungi of some plants of the family Asteraceae, samples of Achillea filipendulina Lam. were collected from Golestan province (Iran) in spring 2016. The plants were rinsed gently with running water, and then were cut into 0.5-1 cm pieces. The surface sterilization was done by sodium hypochlorite (1.5-2.5% NaOCl), followed by 75% ethanol. The surface sterilized samples were placed on Potato-Dextrose-Agar (PDA, Merck, Germany) containing 50 ppm tetracycline, and incubated at 25±2 C for 30 days. After purification, endophytic fungi was cultured on PDA and OMA (Oat Meal Agar) and exposed to 12 hr cycles of UV light and daylight for 14 days to stimulate sporulation. Their identification was based on morphological studies (Ahmed & Cain 1972, Arenal et al. 2005, Asgari & Zare 2010, Kornerup & Wanscher 1989). The method for measuring asci and ascospores and the terminology used for the descriptions are those used by Ahmed & Cain (1972). Based on morphological characteristics, Preussia africana Arenal, Platas & Peláez (Sporomiaceae, Pleosporales) was identified. A description of the fungus is given bellow: Colony reaching 70 mm diam. on PDA in 14 days at 25 °C, dark brown to black. Ascomata 205-288 μm diam., scattered, immersed, pear-shaped, smooth-walled, light brown to dark brown with a narrow, cylindrical neck, 52-60 × 20-40 μm, covered with ornamental hyphae measuring 12-20 × 2-4 μm. Peridium brown, pseudoparenchymatous in surface view, and 10-15 μm thick. Asci 90-110 × 16-17 μm, eight-spored, hyaline, cylindrical, gradually to abruptly tapering into a short stipe up to 4 × 13 μm. Pseudoparaphyses 1-2 μm diam., filiform, septate. Ascospores 42-43 × 3-4 μm, cylindrical, four-celled, middle cells of equal length and broader than terminal cells, germ slit parallel to slightly sinuous; gelatinous sheath hyaline and narrow (Fig. 1). In order to confirm the identification of this fungus based on molecular data, DNA extraction of examined isolate was performed by CTAB method, molecular analysis based on the ITS1-5.8S-ITS2 region, 28S rRNA gene and a fragment of the translation elongation factor 1α gene amplified using primers ITS4/ITS5, LROR/LR5 and EF1-983F/Efgr. BLAST searches were conducted in GenBank. All sequences generated in this study are deposited in GenBank (Accession numbers: LSU = MH255557, ITS = MH250006, EF-1α = MK952151). Living culture of this fungus is deposited at the Iranian Fungal Culture Collection (Iran 3262C) of the "Iran" herbarium. Preussia africana was reported for the first time from Canary Islands and Spain on leaves of Viburnum tinus subsp. rigidum (Arenal et al. 2005) and in this study, it is reported for the first time from Iran. Specimen examined: Iran: Golestan province, Chaharbagh, Isolated from A. filipendulina, 14.5.2016, coll. Sareh Hatamzadeh (Iran 3262C).
All Science Journal Classification (ASJC) codes
- Plant Science
- Ecology, Evolution, Behavior and Systematics
- Achillea filipendulina
- Golestan province