Marwa H. E. Elnaiem et al / Int. J. Res. Ayurveda Pharm. 8 (Suppl 3), 2017 187 Research Article www.ijrap.net MORPHOLOGICAL AND MOLECULAR CHARACTERIZATION OF WILD MUSHROOMS IN KHARTOUM NORTH, SUDAN Marwa H. E. Elnaiem 1 , Ahmed A. Mahdi 1,3 , Ghazi H. Badawi 2 and Idress H. Attitalla 3 * 1 Department of Botany & Agric. Biotechnology, Faculty of Agriculture, University of Khartoum, Sudan 2 Department of Agronomy, Faculty of Agriculture, University of Khartoum, Sudan 3 Department of Microbiology, Faculty of Science, Omar Al-Mukhtar University, Beida, Libya Received on: 26/03/17 Accepted on: 20/05/17 *Corresponding author E-mail: idressattitalla2004@yahoo.cm DOI: 10.7897/2277-4343.083197 ABSTRACT In a study of the diversity of wild mushrooms in Sudan, fifty-six samples were collected from various locations in Sharq Elneel and Shambat areas of Khartoum North. Based on ten morphological characteristics, the samples were assigned to fifteen groups, each representing a distinct species. Eleven groups were identified to species level, while the remaining four could not, and it is suggested that they are Agaricales sensu Lato. The most predominant species was Chlorophylum molybdites (15 samples). The identified species belonged to three orders: Agaricales, Phallales and Polyporales. Agaricales was represented by four families (Psathyrellaceae, Lepiotaceae, Podaxaceae and Amanitaceae), but Phallales and Polyporales were represented by only one family each (Phallaceae and Hymenochaetaceae, respectively), each of which included a single species. The genetic diversity of the samples was studied by the RAPD-PCR technique, using six random 10-nucleotide primers. Three of the primers (OPL3, OPL8 and OPQ1) worked on fifty-two of the fifty-six samples and gave a total of 140 bands. The three primers showed 100% polymorphism among samples, regardless of their groups, while the groups showed polymorphisms ranging from 0% to 100% among their samples with the three primers. A dendrogram was constructed using the Jaccard’s similarity coefficient. The genetic similarities between samples ranged from 10% to 100%. Cluster analysis of the samples revealed distinct groupings. Both morphological characteristics and RAPD were useful in studying mushroom diversity, with some agreement between them. Accordingly, we believe that a hybrid system embodying both approaches can yield a more satisfactory identification of mushroom samples. Keywords: wild mushrooms, RAPD, mushroom identification, Khartoum North, Sudan INTRODUCTION Fungi play an important role in the changes that take place around us because of their ubiquity and their astonishingly large numbers 1 . They fulfil many roles beneficial to humans 2 . However, fungi have been relatively poorly collected and studied from most countries, regions and habitats. This is at least in comparison to plants and larger animals that are considerably easier to collect and identify 3 . One of the large groups of fungi are the Basidiomycota with over 30 000 species. They include many familiar mushrooms and toadstools, bracket fungi, puffballs, earth balls, earth stars, stinkhorns, false truffles, jelly fungi, smuts, rusts and some less familiar forms 4 . In modern times, the cultivation and annual production of mushrooms has steadily increased 5 . The world has witnessed dramatic acceleration in total worldwide cultivated mushroom production and mushroom scientists are making a great effort to bring wild species under domestication to increase production 6 , because growing awareness in the past two decades of the nutritional merits of mushrooms has increased consumption in an era in which people have become more concerned about human nutrition 5 . Sudan’s ecological diversity is reflected in the richness of biodiversity, as seen in mushrooms. Nevertheless, very few studies were conducted on mushroom biology, with no studies targeting their diversity. Therefore, more studies to investigate the mushroom flora of the Sudan, particularly from the genetic perspectives, should be welcomed. Molecular approaches have proven to be potent tools in the classification of complex fungal taxonomic groups, including mushrooms 7 , as DNA techniques are quick and reliable to establish the identities of wild collections and are helpful in mushroom taxonomy 8 . However, macroscopic features are still important today for recognizing fungi and making an initial identification. Indeed, the comparison of DNA sequences obtained from fungi is meaningful only if these fungi have previously been characterized and named by conventional methods 4 . Therefore, the objective of this study was to investigate the diversity of wild mushrooms in Khartoum North (Sudan) using both traditional (morphological characteristics) and molecular techniques. MATERIALS & METHODS Collection of samples Mushroom samples were collected from different areas in Khartoum North (Sudan), from February to September 2013. Intact button, medium-sized and mature specimens from each mushroom species were collected to represent different stages of mushroom’s development; to ease identification. All field notes and macroscopic characteristics were recorded. Photos of samples to illustrate different characteristics and the growth environment were also taken, and each specimen was given a serial number. The samples were transported to the laboratory and were kept aerated throughout the period of examination. Samples were cleaned with water and surface-sterilized with 70% ethanol. Small pieces of each fresh specimen were preserved in about 700 μl of CTAB (Cetyl trimethylammonium