I used Chloramphenicol, Streptomycin, Gentamicin and Cefoxitin to remove bacteria from trichoderma spore. After sterilization microscopic observation showed bacateria free. but when i subcultured it on PDA plate or on broth i got reappearance of bacteria again. So which antibiotic including other factors i may use or follow to get bacteria free culture of Trichoderma?
well Moosa thanks. Yes I was also thinking about technical problems but i repeated it several times and got same results. I am not sure about resistant nature of that bacteria. but i tried as for example if i used Chloramphenicol (CP) than i tried several protocol such 2%,1% CP for 5/10/30 min then washed. but result same. after sterilization i got bacteria free only on eppendorf tube but later on same as was in before.
Amend your antibiotic agar with a physical technique. A sterile glass rod is placed in/on the antibiotic agar medium. Inoculate fungal specimen on one side and incubate until fungal hyphae have grown under and emerge on other side, subculturing the emerging hyphae to antibiotic agar.
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