I’m working on callus induction of different genotypes of Sugar beet; I’ve a problem one of this genotype produced hard callus. I don’t know how to avoid this phenomenon, can you help me?
MS+1mg/L BAP+0.5mg/L NAA,1% sucrose and 0.8% Plant Agar. I've used leaf, stems, cotyledons and hypocotyls as starting material.
Possible suggestions,
Just try to
1. Increase sucrose concentration to 3 %
2. Use little bit lower concentration of Agar from 0.8% to 0.75 to 0.7%
It will probably produce soft callus
Thanks for your suggestions dot. Magdum, unfortunately I cannot increase the sucrose concentration to 3% because it improves the production of phenolic compounds in sugar beet and recently I’ve tried to lower the percentage of Plant agar to 0.7%, I’m waiting for seeing what happens.
i had read in the tobacco callus induction, that if you culture in the light, will produce more tight structure, regular shape. while culturing in the dark will produce less compact and irregular callus. u can try to apply the technique. http://nhjy.hzau.edu.cn/kech/xbgc/sy/PDF/ChaoYanjie.pdf
Dear doc. Morris,
Thanks for your answer.
I need soft calli for initiation of cell suspensions cultures, perform a mutagenesis, obtain again the calli and then regenerate them. Yes, the hard callus primary callus and does it grow but remain hard after subculture.
I think that the 5x5 matrix could be a solution but at the moment I would try to increase auxin content and decrease the cytokinin one. I wish to obtain calli and regenerate them.
I do not know if I solved the browning problem since I tried the ascorbate treatment and the AgNO3 medium a short time ago. I have to wait and see what happens. I have tried the charcoal but it seems to reduce the callus induction without decreasing the browning problem.
I am working on another plant very similar to Miscanthus, Arundo donax (giant reed). My colleague obtained calli of A.donax from buds and immature inflorescences, but they had some problems. The calli from buds are not easy to obtain and difficult to regenerate, while the immature inflorescences are difficult to find and they are produced only during a short period of time.
The calli induction from rhizome could resolve the problem of periodicity and moreover the meristem produced by the rhizome should be the most undifferentiated since it can generate rhizome, shoot and roots.
Dear doc. Drogba,
thanks for your suggestion. I induce calli in darkness, but i does not help anyway.
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