i got multiple shoots from cotyledonary node explants of cotton. basal callus formed at basal part of the explant. i did 2 subcultures along with the basal callus into fresh medium. after 2 subculture, i cut and removed the basal callus and transferred the multiple shoots to fresh medium. after 1 week i observed chlorosis and withering of the transferred shoots. i want to know whether the removal of basal callus is responsible for this or any other reasons?
In your case, plants need certain time for cutting. Go with callus for multiplication as much as you get enough clones. Then, let the shoots further growth and development, prior to cuting from basal callus. Finally place them into fresh MS medium containing 0.2 mg/L IAA in jars.
It is possible that the basal callus represents the source of growth regulators or other signals necessary for development of cotton regenerants. You should go through the literature and try to find protocols established for cotton regeneration from previous research. If no one says nothing about removing, or not removing callus then try to find it out for yourself. If you are sure to observe that shoots do better with callus than without, stick to that observation - it is a valid one - and try to keep them together with the callus for as long as possible.
If the callus eventually falls apart, then try making up for it by various combinations of cytokinins, or cytokinins and auxins. Try out first the combinations of plant growth regulators that other researchers have reported for shoot regeneration in cotton, then make lesser modifications if you find out that it doesn't work quite properly.
If you find good data in literature, then it should work quite easy for you. If there's no good data in literature it will be harder for you, but that will give you a good basis to publish your own regeneration protocol in the end, when you work it out.
Good luck!
Please, could you tell me about the type and concentration of the hormone that just added to the medium of culture so that I have the answer to your question. The reason callus formed in the base of the ex-plant is the type or concentration of the hormone.
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