How to eliminate this problem ?
I am using MS medium with PGR ,and working with orchid's
Contamination can occur any time in the media it depends upon the activation of the microorganism. Some times microbial spore may be dormant and might have started multiplying lately.Try sub-culturing immediately to save your results. Check if no one actually opens them and if anybody does it should be done between the burners and the mouth needs to be flamed.
Please check that (1) contamination is evenly distributed in the medium (may happen due to improper autoclave or dormant spore), (2) it starts from the explant and then grows in the medium (may happen due to endophytic microbes in the plant system)
Kindly check the laboratory condition first and then try to screen the endophytic fungi from the explant material. if found any endophytic organism , use appropriate antibiotic to avoid the problem...
Endophytic microbes may survive during initial sterilization treatment. They may be initially dormant and might have started multiplying lately. Use of antibiotics will be useful
First identify the microorganism and the principal causes of contamination that may be: Endogenous explant contamination; desktop due to inadequate asepsis; equipment with inefficient operation or instruments of work and outside of the container with inadequate sterilization beyond the operator and the presence of insects in the growth room. After this identification, use appropriate techniques cited in the literature as 0.5-2% NaOCl for 10-15minutes, rinse the explant in alcohol at 70% for 5 seconds.
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