Hello all
I've some explants grown in vitro conditions, in elongation step and acclimatization step of Eucalyptus sp. I had no problem with two step, but a few days ago I observed the presence of small callus in the leaves of shoots and after some days these fell off (photo attach).
The first thing I did was decreasing the hormone concentration (BAP), but there are any factor?
thanks for your answer Subba, you are referring all macronutrients or nitrate only?
You may find the solution to control the callus formation when reading the article " Plant Callus: Mechanisms of Induction and Repression" published on Plant Cell.,25(9): 3159–3173 by Ikeuchi et al. (2013).
How about to transfer the shoots to a new medium with low concentration of sucrose concentration and without plant growth substances?
use MS/2 or I think may be adding the agar concentration(example: 8 g/l to 8.8 g/l)
Many thanks to all for your comments. I'll read the articule recommended by Quynh Nguyen and try with other strength medium.
Nasrin, Why inscreasing agar concentration?
Best regards and thank you again
Auxin may be responsible. Are you using it? If so, try to lower concentration or replace for another one if you cannot eliminate it. For instance, 2-4D by IAA... or NAA...
Good luck
When you obtain the suitable growth on the medium that contained BAP, immediately transfer the shoots to medium free of hormones.
Dear Mauricio
First of all, How much 6-BAP are you using during elongation of Eucalyptus?. It is important, because one of the principal causses of callus induction during propagation via organogenesis in woody plants is the level of cytokin, this concentration must be reduced in the last subculture in the multiplication phase. Some time reducing thiamine levels, and the addition of B12 or fluroglucinol limited callus formation and improve shoots quality. For elongation of Eucalytus shoots I recomend you eliminate the callus, individualized shoots with 2 cm size or more and culture in liquid culture medium with 0.2-0.3 mg/L 6-BAP+0.1 mg/L ANA, without thiamine; you must obtain a vigorous plants. Good look!
Best regards,
Elisa
Muchas Gracias Elisa, Nunca he usado la estrategia de dejar fuera la tiamina, te cuento como me fue
Saludos
Nota: Note que hablas español, por eso he respondido asi
Ha sido un placer , prueba el medio de cultivo líquido es efectivo y económico, se producen miles de plantas de Eucalipto grandis en biofábrica con una estrategia similar.
Saludos,
Elisa
Increasing agar concentration causes to decrease uptake the nutrient and water and so callus can't growth and produce.
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