Abstract:
In quest to ameliorate the issues of recurring incidences of multidrug-resistant organisms and over-exploration of
plants, researchers have delved into researches involving endosymbiotic microorganisms, known as endophytes. These
are microorganisms inhabiting the internal part of plants, which have been verified to possess great potentials of
bioengineering novel products for therapeutic purposes. The aim of this study was to isolate and identify endophytic
fungal species present in Mitracarpus scaber, molecularly characterize the pure isolates, and test for phytochemical
compounds present. Freshly collected non-diseased leaves of M. scaber were subjected to a four-step surface
sterilization. Thereafter, they were sliced into fragments of 1 cm-length, exposing the leaf blades and the midribs, and
were aseptically inoculated on sterilized malt extract agar, containing chloramphenicol (500 mg/l), in Petri dishes.
Hyphal tips of actively growing fungi from the plant material were harvested and further sub-cultured for purification
and isolation. Segments were aseptically cut from the actively growing pure isolates, on malt extract agar and inoculated
onto the fermentation medium (sterilized local rice), in 500 ml Erlenmeyer flasks, which were properly sealed with
sterile cotton and kept at static condition at 25 oC for 21 days. Ethyl acetate was used to extract the metabolites from
the end products of the fermentation processes. Four isolates obtained from the endophytic fungi (EDF), present in the
leaf of M. scaber, were subjected to molecular identification. The DNA extraction was done using Zr fungal/bacterial
DNA miniprep. The extracted DNA was amplified through PCR, and sequenced. The resultant sequences were compared
with GenBank database, using Basic Local Alignment Search Tool. The result of phytochemical screening of the extracts
revealed the presence of flavonoids, tannins and phenols, in large amounts; terpenoids, alkaloids and cardiac glycosides,
in moderate amounts; steroids, hydrogen cyanide and saponins in very low quantities, while that of molecular
characterization identified four organisms: Penicillium sclerotiorum, Clasporium cladosporiodies, Cryptococcus
nemorosus, and Phyllosticta capitalensis.
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