Journal of Advances in Biology & Biotechnology 10(4): 1-8, 2016; Article no.JABB.29724 ISSN: 2394-1081 SCIENCEDOMAIN international www.sciencedomain.org Isolation of Indigenous Brown Rot Fungi from Rotten Wood from Selected Areas of Pakistan Raja Tahir Mahmood 1*, Muhammad Javaid Asad 2, Muhammad Asgher 3, Muhammad Gulfraz 4, Dawood Ahmed 5, Pervez Anwar 6 and Nasib Zaman 7 1Department of Biotechnology, Mirpur University of Science and Technology (MUST), Mirpur-10250, AJK, Pakistan. 2Department of Biochemistry, PMAS-Arid Agriculture University, Rawalpindi, Pakistan. 3Department of Biochemistry, University of Agriculture, Faisalabad, Pakistan. 4Department of Chemistry, COMSATS Institute of Information Technology, Abbottabad, Pakistan. 5Department of Medical Laboratory Technology, University of Haripur, KPK, Pakistan. 6Department of Biochemistry and Molecular Biology, University of Gujrat, Sailkot Campus, Pakistan. 7Centre of Biotechnology and Microbiology, University of Swat, KPK, Pakistan. Authors’ contributions This work was carried out in collaboration between all authors. Author RTM designed the study, performed the experiments, wrote the protocol and wrote the first draft of the manuscript. Authors MJA, MG and MA managed the analyses of the study. Authors DA, PA and NZ managed the literature searches. All authors read and approved the final manuscript. Article Information DOI: 10.9734/JABB/2016/29724 Editor(s): (1) James W. Lee, Department of Chemistry and Biochemistry, Old Dominion University, USA. (2) Reviewers: (1) Anonymous, Botanical Institute, Brasil. (2) Asemoloye Michael Dare, Department of Botany and Microbiology, University of Ibadan, Nigeria. (3) Aline Elesbão do Nascimento, Universidade Católica de Pernambuco, Brazil. (4) Mohamed M .Y . Deabes, Food Toxicology and Contaminants Dept., National Research Centre, Egypt. Complete Peer review History: http://www.sciencedomain.org/review-history/17147 Received 26 th September 2016 Accepted 28th November 2016 Original Research Article Published 6th December 2016 ABSTRACT Aims: Wood decaying fungi are very good candidate for industrial processes like; enzymes productions, dyes biodegradation and wastewater decolorization. Many studies have been conducted to explore their potential for successful industrialization. There is still need to isolate more and more new species so that these can be used for beneficial processes. Methodology: The current study was conducted to collect Brown Rot Fungi (BRF) species from _____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected]; Mahmood et al.; JABB, 10(4): 1-8, 2016; Article no.JABB.29724 rotten wood and their morphological identification. The collection was made from Islamabad, Rawalpindi and Murree areas of Pakistan. Collected fungi species were cultured on Malt Extract Agar media at pH 5.5 and pure culture plates were submitted in Mycology Laboratory, Department of Plant Pathology, PMAS-Arid Agriculture University Rawalpindi Pakistan for morphological identification by mycologist. Results: More than 20 species were collected, out of these 05 were cultured on Malt Extract Agar media and identified on morphological basis. Two of these are common brown rot fungi belong to basidomycota and three are un-common brown rot fungi belong to ascomycota. Conclusion: Rotten wood is a very good source of brown rot fungi. Isolated BRF species can be used to explore their potential of biodegradation of textile dyes. Keywords: Wood decaying; brown rot fungi; wastewater; decolorization; malt extract agar. 1. INTRODUCTION Daedalea is a genus of fungi which was circumscribed, it is restricted to species produce Brown rot is a kind of wood decay caused by brown rots and have different morphology as Basidiomycota, namely Agaricomycetes and described by different scientists [11-12]. some species of ascomycota. This class Daedalea dickinsii belong to Polyporaceae family encompasses many orders and families, though which is an annual red rot fungus [13]. the overwhelming majority of the BRF belongs C. puteana, brown rot fungi belong to Boletaceae to the Agaricales, Hymenochaetales, family also cause rot in wood [14-15]. It is mostly Gloeophyllales and Polyporales . Interestingly, present on timber, moisture places and where only 6% of all the known wood decay fungi are poor ventilation. It produces cellular enzymes now known to cause a brown rot and are almost which has role for decomposition of classic exclusively associated with conifers [1]. cycles of nature [16]. Brown Rot Fungi (BRF) includes wood decaying From Pakistan many BRF species has been basidiomycetes, some uncommon BRF belongs reported but not widely applied for beneficial to ascomycota like Aspergillus fumigatus and processes [17-18]. Aspergillus sydowii, that are widely distributed in the world [2,3]. These fungi have the ability to There is further need to explore the potential remain active in extreme environmental areas of these microorganisms and their conditions regarding the presence of chemicals. application for various beneficial processes. The These can used to solve various environmental aim of current study was to isolate potential BRF issues like, wastewater treatment, toxic species from Rawalpindi, Islamabad and Azad dyes decolorization and industrial enzymes Kashmir and identify these on morphological production. Isolation of fungal species form new basis. Isolated species will be further used for area will play important role for reporting new bioremediation process in future to get maximum species and support the ongoing research [4-6]. benefits. Brown Rot Fungi has non-specific enzymatic 2. MATERIALS AND METHODS system comprising of maganase peroxidase, lignin peroxidase and laccase, having the ability 2.1 Study Area of cell wall component modification and degradation [7]. Brown Rot Fungi degrade Timber Market of Rawalpindi and Islamabad, cellulose and hemicellulose while cause the surrounding forest of Rawalpindi, Islamabad and modification of lignin and gave brown color to Murree. Islamabad is located at 33°26 ′N rotten wood [8]. Due to non-specific enzymatic 73°02 ′E/33.43°N 73.04°E at the edge of the system BRF can be actively used for the Pothohar Plateau at the foot of the Margalla Hills . bioconversion of textile dyes having similar While Rawalpindi and Murree are located in the structure like cell wall components. Isolated BRF surrounding of Islamabad. Further hilly areas and species can also be used for the production of high altitude forest of Bagh Azad Kashmir were extracellular enzymes which are responsible for also included. The collection of BRF from natural the wood degradation, because these are conditions will help in the isolation of fungal industrially important enzymes [9-10]. species that can survive in harsh conditions. 2 Mahmood et al.; JABB, 10(4): 1-8, 2016; Article no.JABB.29724 2.2 Sample Collection of Ficus carica (Fig. plant) on 24-11-2013. It was initially whitish in color with little shade of brown Macroscopic BRF growing on the brown rotten growing margins (Figs. 1a and b). Wood was wood were collected, tagged, wrapped in paper converted into brown patches where fungus and packed in plastic bags. Whole growing part showed active growth. It was cultured on malt of fungi collected without damage its structure, extract agar media at 28°C having pH 5, growth because it also help mycologist for identification. was clear after 96 hours (Fig. 1c). The culture These were brought to laboratory for further produce white mycelia which turn brownish at processing. Further brown rotten wood samples margins after 5 days of growth and spread on were also collected for the isolation of fungi from whole petri plate (Table 1). Coniophora is a the spores in the rotten wood that fall on wood genus of fungi of Boletaceae family with 20 during growth of fungi. known species with wide distribution. One notable member of the cellular fungus is 2.3 Culturing of Fungi Coniophora puteana which causes brown rot in wood [15]. This causes a darkening of the BRF were culture on Malt Extract Agar (MEA) timber (brown rot) and rarely found in building, media having composition (malt extract 20 g/L, cause decay with high moisture content. agar 15 g/L, dextrose 20 g/L, peptone 3 g/L, pH The fungi secrete extracellular enzymes 5.5) at 28°C temperature in incubator. For which break down potential food sources, which culturing, fungi sample were surface sterilized are then absorbed back into the fungal colony (70% ethanol/ 10% chlorox) and 1 mm piece [16]. These have the potential to damage shifted to media aseptically in Leminar Flow standing timber, finished wood products, fibers, Hood [19-20]. and wide range of non-cellulosic products such 2.4 Identification of Fungi as plastics, fuels, paints, glues and drugs. These enzymes include amylases, glucoamylases, Most of the fungus were macroscopic and could oxidases, lipases, pectinases, and proteases be identified by specialist mycologist. Those [22]. species culture on media plates, their slides were prepared and their identification was confirmed Daedalea dickinsii IEBL-2 was collected from on basis of spores shape, size, texture and Chakshahzad (CS) and Lake View Park shape of fungal hyphae by mycologist [21]. Islamabad on the dead trunk of Ficus carica on 14-11-2013. When collected, it was light white 3. RESULTS from lower side and brownish from upper surface (Figs. 2a and 2b) and was labeled as CS-2 and 3.1 Fungal Species