Even after taking ultra care in Plant Tissue Culture, we used to get contamination in plant cultures. some times it may be due to the endophytes present in the tissue. Few of them can be visible at the cuture initiation level and a few of them can be multiplication level. (depending on the plant).
I request you to help me to avoid these endophytic microbial contaminants in both levels.
You may consider using some antibiotic to treat the plant tissue before starting the culture. I believe some protocols should be available if you search for it online. You may need to set up a preliminary experiment test the different antibiotics to find out the appropriate one(s) and the proper concentration(s).
Otherwise, you could also try separate the plant tissue to single-cell level, either by microscopic operation or isolation techniques similar to the ones used in microbial culture. Pick up some isolates and try to culture them in isolated containers. You should be able to find some one without endocellular microbes, unless you are culturing root tissue of some highly symbiotic microbe-plant duos.
I cannot give you any more detailed information since I have leaved wet lab for a while, but I wish this will help. And perhaps someone else can answer your question better.
If you are using the tip of apical meristem portion for culturing , every likely , it doesnt have phloem and , hence , unlikely to have any such endophytic bacterium...
avoid using drugs, and try to generate new callus from meristematic cells, so that you will obtain new free cells without endophytic bacteria.
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