We have got a good callus culture after optimization of auxin/cytokinin hormones concentration .The callus cultures were subcultured on shooting induction media
We have got green and compact cultures which are surviving for long time in the same media without any subculture.No shoots were induced on callus.
We could not realize the underlying challenge for shoot induction in callus culture.
Swati Sharan the required types and concentrations of PGRs for shoot induction from callus vary from plant to plant. Therefore, go through the literature and find out a good ratio of cyt-auxin PGRs. If you are maintaining callus on 2,4-D or any other auxin, just subculture calli on cytokinin medium. If you can discuss in details about the use of PGRs and plant name, I may help you better in this regard. All the best...
Hello! from our experience this can happen when callus has been subcultured for a long time in the presence of high cytokinin concentration. Probably cytokinins are accumulated in the tissue and inhibit further regeneration. To overcome this problem you can transfer the calluses to the media without any PGRs for about 1-2 month (depending on plant species). After that transfer your calluses to regeneration media. Sometimes it helps
Swati Sharan, I agree with the colleagues above about the specificity of the aux/cytokinin ratio for each plant species, but I would also suggest you to in one of these subcultures suddenly shift for a media with high cytokinin level, can be a way to retrigger SAM and induce shoot
Lidia Samarina Thanks a lot for your invaluable inputs.These cultures were maintained for a long time in the presence of auxin/cytokinins because of lockdown.
You should transfer callus in a new plant growth regulators-free medium (half-strength) supplement with 1 g /L activated charcoal.
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