Fungal and Bac contaminations are common in vitro culture. However, Ppm works alright with some of the contaminated plants . But bleach and ethanol doesn't work at all. Any recommended solutions or media types?
I think you can change the concentration of your bleach and ethanol. You can also use fungicides and substances that kill bacteria in the culture medium, such as hydrogen peroxide, nanosilver, etc. Of course, if your contamination is endogenous, you have less chance of complete control of the contamination.
Hello Maduka Wehella
You may refer to the link below. It may be helpful.
https://www.researchgate.net/post/How-to-prevent-the-microbial-contamination-in-plant-tissue-and-cell-culture#:~:text=The%20best%20way%20to%20get,bleach%20by%20using%20autoclaved%20water
Best.
The best way to avoid contamination (at least by virus) is ti cut the meristems (apical). You avoid most of them. To avoid fungus or bacteria you may try antibiotics, bacteriostatic compounds of antimicotic substances. Normally there is a protocol for each, but most of them would say it will be on the same place. cut the leaves, place them in ...
I think you are asking about the surface sterilization of the explants. There is a series of procedure you have to follow that. Just go through some of our recently published articles you will understand the procedure. It is not a difficult task as you think but yes, it is very important to initiate the cultures.
Dear Maduka
I agree with Saeed and add that, the type of explants and the concentration of bleaching are directly proportional. Thus, if the possibility of contamination of the explants were high , the higher concentrations of bleaching is required and the extra attention should be taken for handling procedure . Also, I agree with Mahipal if the question was intended Surface sterilization of explants. Therefore, to obtain cultures free of contamination from fungi and bacteria, wash the excised plants under running water for 30 minutes with soap, then under sterile conditions, soak them for 20 minutes in chlorine (NaOCl or KOCl solution), and then wash them 3 times with sterile distilled water. Finally, plant it inside the Laminar Air Flow Cabinet.............Good luck
regards
Metwally Bekhit
Meristem cells in shoot apex are devoid of pathogens like bacteria, fungi or viruses.
Suggest meristem tip culture to obtain pathogen free plants you are working with.
For details of this procedure read plant tissue culture book.
Or...start with seeds that have been treated with diluted bleach. Germinate the seeds in sterile containers on a sterile salts solution. The plants that grow might be free of any organism.