I am in trouble with this contamination-like formation (please see the attached file). I transferred plants from the jars having this whitish formation into new ones containing different concentrations of Kanamycin in order to get rid of it. But, one day after, I observed it again. We made gram -/+ testing, suprisingly they are not bacteria. What could they be? It has not affected plant growth or development negatively yet. The cultivated plant in my experiment is Stevia. Note that, some jars do not have this formation on the media.
It looks like an anaerobe, because it looks like it is at the bottom of the container below the media. But if it is not bacteria based on the gram stain, and it isn't responding to kanamycin, it is more than likely fungal. I must caution you, low concentrations of antifungals (e.g. Nystatin, Amphotericin B) can be just as toxic to plants.
Why you think the contamination isn't caused by a bacteria? Do you have photos from microscopy observations?
I'll repeat the morphological test as to whether it is basteria or not. Then, I will share the results to you.
It looks like a bacterial contamination and you can see the bio film formation wherever the water on the surface of the semi solid medium moved, which carried the bacteria obviously. Usually it does not affect the shoot explant in that medium (if it is a callus the negative effect of the bacteria will be visible) in one subculture period say about 30 days, but when you do the subculture (transferring to the fresh medium) it will harm the whole batch contaminating through the manipulation if you are not careful. Single Antibiotic may or may not work on the organism unless otherwise it is specific.
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