International Journal of Agriculture Sciences ISSN: 0975-3710&E-ISSN: 0975-9107, Volume 9, Issue 20, 2017, pp.-4209-4210. Available online at http://www.bioinfopublication.org/jouarchive.php?opt=&jouid=BPJ0000217 Research Article FIRST REPORT OF EPICHLOË SYLVATICA ON Brachypodium pinnatum GRASS FROM IRAN KOMAKI AMANMOHAMMAD1, AGHAJANI MOHAMMAD ALİ1 AND HASSAN ZOHAİB UL2 1Department of Plant Protection, Research Center of Agricultural Sciences and Natural Resources, Golestan province, Gorgan, Iran. of Plant Protection, Faculty of Agriculture, Ataturk University, Erzurum, Turkey. *Corresponding Author: Email-amkomaki@gmail.com 2Department Received: April 03, 2017; Revised: April 12, 2017; Accepted: April 13, 2017; Published: April 30, 2017 Abstract- The aim of this study was identification of the endophytic fungus on Brachypodium pinnatum plants from forests of Northern part of Iran. According to the investigation, the grass was infected with Epichloë sylvatica Leuchtm. & Schardl 1998 and this fungus is a new record for Iranian mycoflora and reports for first time. Keywords- Epichloë, Brachypodium, Endophyte, Biodiversity, Iran Citation: Komaki Amanmohammad, et al., (2017) First Report of Epichloë Sylvatica on Brachypodium pinnatum Grass from Iran. International Journal of Agriculture Sciences, ISSN: 0975-3710 & E-ISSN: 0975-9107, Volume 9, Issue 20, pp.-4209-4210. Copyright: Copyright©2017 Komaki Amanmohammad, et al., This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited. Academic Editor / Reviewer: Ansari Rizwan Ali Introduction Genus Epichloë (Fr.) Tul. & C. Tul. is an endophytic fungus Classified in Clavicipitaceae family (Order Hypocreales: Ascomycota) [1]. The asexual form of Epichloë fungus placed in Neotyphodium A.F. Glenn, C.W. Bacon & Hanlin (Anamorph) form systemic infections in a wide range of grasses of temperate areas [2-4]. The 30% of all cool-season grass species (Poaceae subfamily Poöideae), including forage, turf grasses, wild grasses of meadows, woodlands and rangelands in worldwide are associated with Epichloë spp. [5]. These fungi form systemic and asystemic associations throughout the aerial parts of the host plants. The hyphae of these fungi colonize in intercellular apoplastic spaces of culms, leaf sheaths and seeds [6-9]. To identify this fungus, different Epichloë species description have been used [1,5,10-12]. The aim of this study was description the morphological characteristics of fungus and report as a new fungal species for Iranian mycoflora. Materials and Methods The Brachypodium pinnatum plants infected with Epichloë sylvatica were collected from the forests of Northern part of Iran (Forest of Shast-Kola, Gorgan, Golestan Province, 13.05.2010; Forest of Babol, Mazandaran Province, 07.05.2010). The infected plants then transferred to the laboratory and the fungus was inoculated to the PDA medium and incubated at 21 oC. The Koch’s test has been done on their host (B. pinnatum) by cultivating their seeds in the pots. After emerging the stem of plant the spores of fungus sprayed on them and kept in plastic bag for 24 h under greenhouse conditions. The microscopic slides were investigated by microscope (Olympus BH2). The fungal morphology has described according to the collected samples from the natural places. Results The description of samples are given in the following text. Epichloë sylvatica Leuchtm. & Schardl 1998 The fleck leaf of B. pinnatum was colonized by the endophytic fungus and produced their stomata around the sheath and fleck leaf. First of all, the colonized sheath by fungus had white and metallic color and gradually changed to yellowish orange by maturation. Stromata length on the sheath found around 45-65 mm. The surface of young stroma was smooth but the perithecia emerged in mature stroma. In each millimeter square (mm 2) an average of 32 (20-50) perithecia have been observed. The shapes of perithecia were piriform with short neck and the color was yellowish orange at maturity. The height of perithecia were about 250412.5 µm and width was 112.5-237.5 µm. The wall of perithecia was thick with multilayered and transparent hyphae. There were numerous asci in each perithecia and the ascus was hyaline. Ascus had apical pore and their apical part was thick. Also, the ascus stalk was long and the asci size were about 165-375 × 12.5-8.75 µm. There were eight filliform ascospores in each ascus. Ascospores were hyaline and their size were about 160-282.5 × 1-1.5 µm. According to the morphological characters the fungus name was identified as Epichloë sylvatica Leuchtm. & Schardl 1998 [1]. In past the E. typhina have been reported in Iran [13]. E. typhina has ascus size about 163 ± 17.8 × 6.9 ± 0.2 µm and their ascospore sizes are about 176 ± 34.3 × 1.6 ± 0.2 µm [1]. Also, their asexual stage Neotyphodium have been reported from different parts of Iran [14,15]. E. sylvatica is a new record for Iraninan mycoflora and this species report for first time from B. pinnatum. The herbarium specimens are kept in the mycological herbarium at Agricultural Science and Natural Resources Research center of Gorgan, Golestan Province. Fig-1 Epichloë sylvatica stromata in infected plant Brachypodium pinnatum (a, b). International Journal of Agriculture Sciences ISSN: 0975-3710&E-ISSN: 0975-9107, Volume 9, Issue 20, 2017 || Bioinfo Publications || 4209 First Report of Epichloë Sylvatica on Brachypodium pinnatum Grass from Iran Fungal Genet Biol., 45, 84-93. [10] Mc Cargo P. D., Iannone L. J., Vignale M. V., Schardl C. L. and Rossi M. S. (2014) Mycologia, 339-352. [11] Schardl C.L. & Leuchtmann A. (1999) Mycologia, 91, 95–107. [12] Yan K., Yanling J., Kunran Z., Hui W., Huimin M. and Zhiwei W. (2011) Mycologia, 103 (6), 1341-1350. [13] Ershad D. (2009) Fungi of Iran. Agricultural Research, Education and Extension Organization, Iranian Research Inistitute of Plant Protection, Ministry of Jihad-e-Agriculture, 531pp. [14] Dehghanpour Farashah S., Sarifnabi B. & Mirlohi A.F. (2005) Rostaniha, 6, 131-150 (In Persian with English Summery). [15] Karimi S., Mirlohi A., Sabzalian M.R. and Seyed Tabatabaei B.E. (2012) Mycologia, 104 (6), 1281-1290. Fig-2 Epichloë sylvatica stromata; immature stromata (a) and mature stromata (b, c), cross section of infected plant stem (d). Fig-3 Epichloë sylvatica; Perithecia (a), Ascus (b), Ascospores (c). Conclusion According to the morphological study and comparison of their characteristics with different identification keys this fungal species is a new record for Iranian mycoflora. Author Contributions: All author equally contributed. Abbreviations: oC: Centigrade; mm: Millimeter; mm2: Millimeter square; µm: Micrometer Ethical approval: This article does not contain any studies with human participants or animals performed by any of the authors. Conflict of Interest: None declared References [1] Li W., Ji Y., Yu H. & Wang Z. (2006) Mycologia, 98(4), 560-570. [2] Schardl C. (1996) Annuals Review of Phytopsthology, 34, 109-130. [3] Schardl C., Leuchtmann A. & Spiering M.J. (2004) Annuals Review of Plant Biology, 55, 315-340. [4] Schardl C.L., Scott B., Florea S. and Zhang D. (2009) Epichloë endophytes: clavicipitaceous symbionts of grasses. In: Deising HB, ed. The Mycota V: plant relationships. 2nd ed. Berlin: Springer-Verlag. P 275-306. [5] Leuchtmann A., Schardl C.L. & Siegel M.R. (1994) Mycologia, 86, 802–812. [6] Christensen M.J. and Voisey C.R. (2009) Agron Monogr., 53, ASA, CSSA, SSSA. P 251-272. [7] Christensen M.J., Ball O.J.P., Bennett R. and Schardl C.L. (1997) Mycol Res., 101, 493-501. [8] Christensen M.J., Bennett R.J. and Schmid J. (2002) Mycol Res., 106, 93106. [9] Christensen M.J., Bennett R.J., Ansari H.A., Koga H., Johnson R.D., Bryan G.T., Simpson W.R., Koolaard J.P., Nickless E.M. and Voisery C.R. (2008) International Journal of Agriculture Sciences ISSN: 0975-3710&E-ISSN: 0975-9107, Volume 9, Issue 20, 2017 || Bioinfo Publications || 4210