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A prerequisite for successful in vitro culture is the establishment of an aseptic technique, thus the experiment was to investigate suitable sterilization regimes for the leaf explants of Gnetum africanum , an endangered green leafy vegetable. Three sterilization regimes were tested to establish the best regime using three to four days old leaves. The surface sterilized explants were later aseptically introduced onto the surfaces of sterile Murashige and Skoog agar media, incubated at 25°C for three weeks in the growth chamber. 100% sterility was observed from the regime which was significantly different (P<0.05) from the other two regimes thus the best regime adopted for further experiments was; washing in two drops of Tween 20/100 mls of sterile water, soaking in 70% ethanol for 2 min and later in 1% sodium hypochlorite for 20 min. Fungal contaminants responsible for in vitro contaminations was also investigated and possible isolates were identified as Asperigilus niger (28.71%); A. flavus (26.73%); Rhyziopus spp. (24.75%) and Mucor Spp (19.81%) respectively. Keywords : Gnetum africanum, A. niger, in vitro culture, green leafy vegetable African Journal of Biotechnology Vol. 12(28), pp. 4473-4476

Explant establishment for callus initiation of a Nigerian endangered leafy vegetable, Gnetum africanum (WILLD)