We are trying to develop an explant-to-plant regeneration protocol for marshmallow (Althaea Officinalis) and have tried every auxin/cytokinin combination we can think of on leaf and cotyledon explants, but so far just callus and roots formed. Any ideas or tips would be very gratefully received.
Try other explants, like shoot tip. However, please note that the success of plant regeneration via tissue culture is highly genotype-dependent. Thus, if possible, try multiple genotypes within this species on similar media.
Hey Pascal
which type of callus u have found from the leaf and cotyledon explants. You have not mentioned the age of explant which u have tried. I think you should try pulse treatment of auxins and also combination with low cytokinins, it will be better for the callus. First u have to identify the nature of callus, organogenic or embryogenic afterthat set the experiments. I suggest you to do some more experiments and some more additives in the media. Good luck.........
In spite of the other explant sources as Mr. Mathe adviced; you need to check the hormonal balance, because you develop root and I am assuming that were from the callus, if that is the case; your callus can be consider organogenic callus (rhizogenic), and some time when one obtain this answer, mainly depended from the amount of auxin accumulated inside the tissue of the callus due to the repetitive subcultures and of course the external concentration supplemented in the culture medium to induce the callus on the differentiation process.
Additional suggestions: try to use embryos, increase cytokinin concentrations, delete auxins.
Thank you for all your recommendations. It is difficult to make any kind of predictions with this species - we have seen root formation for example with high cytokinin to auxin ratio. We will certainly try shoot tips and although we had hoped to avoid zygotic embryos, due to the extra work involved in excision from seed, we will probably try them too. Thanks again.
hello pascal,
you have tried with an explant and have raised callus along with that. you can treat one part of your grown material for shooting by raising the auxins slowly with that of cytokinins in lesser concentration (example. NAA 1.0 mg/L + .05mg/L). this will help in giving you shoot formation too. hence you can go for hardening also.
all the best.
Hello Aparna -Thanks for your message, but I'm not sure what planta regeneration system is.
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