I am using tobacco callus for suspension culture in MS basal and its color is brown. I did not find any contamination in suspension when plated in nutrient agar. callus was friable and white in color.
can anyone give me some paper on plant regeneration from suspension culture.
Are those cells dead?? You said you cultured it in MS "basal' medium. Does this MS basal powder you used to make medium contains other important ingredients such as vitamins...etc? There are many types of MS powder mix sold from companies. Some contains only macro-elements, some contains both macro-elements and vitamins,......etc.
What kind of tobacco cells are you doing suspension culture? BY-2 cells?
According to this protocol (for BY-2) cells: "Brown cells should not be used because they are damaged." (see attached article, yellow highlight). Actually, not only for BY-2 cells, other cells turn brown are usually not a good sign.
Cause of browning in cultures cultured in vitro is phenolic compounds.
you can addition activated charcoal or PVP to media Or re culture of cultures during few periods , Immersion of explants in antioxidant solution consisting of 150 mg L −1 citric acid plus 100 mg L −1 ascorbic acid before culture or its addition to media
Thank you Yuan-Yeu Yau and Ahmed Al mayahi for your answer. I am working on two variety of tobacco from the root we developed callus and did suspension culture from the same callus. I am using MS media sold from company containing micro, macro nutrient and vitamins without sucrose and agar. We added sucrose during preparation. We didn't find microbial growth when plated on nutrient agar. We didn't check the cell viability, but the cell number is increasing when we take 1 ml suspension and centrifuge we found mass is increased.
Is it possible color of suspension is because of the variety of tobacco which I am using.
For your question: "Is it possible color of suspension is because of the variety of tobacco which I am using?"
I am not sure about your variety. But, for BY-2 variety, the name 'BY' is from their 'bright yellow' color. They are not brown when grown in suspension culture.
Attached paper is a story of BY-2 tobacco cells.
PS. Could you please attach a picture of your brown color tissue culture?
The attached paper described 'Soybean Suspension Cultures'. They also observed 'brown color' cultures. They add 2% PVP to the culture and found good result.
This is from their results:
- "Significant differences in coloration between control and PVP-treated soybean suspension cultures were apparent after 1 week by visual inspection. Control cultures grown in 2B5 medium, as well as cultures grown with the addition of 1% PVP, became dark brown in color. The cells from cultures grown in 2B5 medium plus 2% PVP, however, remained white in color throughout 1 week of growth."
They also suggested that the brown color substance is phenolic compounds.
- "The 1% PVP-treated cells had a much reduced amount of total phenolics, while the amount of phenolics was negligible in the 2% PVPtreated cultures."
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