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Abstract This research was designed to study the biological effects of actinomycetes and essential oils on microbial contaminants in banana plants tissue culture in vitro. Microbial contaminations are considered to be of the most serious constraints facing banana plant in tissue culture proliferation in vitro.Four antibiotics, namely rifampicin, gentamicin, chloramphenicol and vancomycin each at 50, 100 and 200 mg /litre and three antifungal agents, namely ketoconazole, fluconazole and nystatin each at 50, 100 and 200 mg/litre were used in the culture susceptibility tests of the identified bacteria and fungi, respectively. To control banana plants contaminated by bacteria in vitro, actinomycetes (Streptomyces bobilii or S. grisiobrunneus) were tested. Actinomycetes as antimicrobial agent were tested in two ways, first in Petri dishes to test the antagonism between antimicrobial agents from actinomycetes and bacteria as well as fungi contamination, second in culture medium from the beginning at concentration of ( 0, 5, 10 and 15%) in different stages of banana indirect stages (shooting and rooting). Data obtained showed that, after twelve weeks, S. grisiobrunneus was more effective than S. bobilii at any concentration tested and at different stages. Addition of S. grisiobrunneus at the concentration of 10 % to the culture media during all studied stages was the prefer treatment which caused the best number of explants free contamination and the best number of survived explants |