I have a problem to break dormancy seed in Arabidopsis wilt tipe and mutant aux 1. I'll grow it in 1/2 MS, so what the best treatment to break it?
Thanks before
In my experience, the best way to break dormancy is to place the seeds on 1/2X MS agar plates, wrap them in aluminum foil and place them at 4 degrees C (in a refrigerator) for 3-5 days. Then unwrap, place the seeds under the light and they should germinate after 24 hours. Good luck!
many kind of treatment can be applied. e.g. KNO3 , NaOH, GA3 in different dosage.
Dear Hajiloo
Thank you for your answer. Have you ever break the dormancy of Arabidopsis seed with GA3? Do you have a recipe or the method for it?
Actuallay i have search about it but i'm not sure if the method that i've found is the best
In my experience, the best way to break dormancy is to place the seeds on 1/2X MS agar plates, wrap them in aluminum foil and place them at 4 degrees C (in a refrigerator) for 3-5 days. Then unwrap, place the seeds under the light and they should germinate after 24 hours. Good luck!
I agree with Katie. Even shorter cold treatments (2 days) work nicely to break dormancy. If still your seeds do not germinate place them in MS agar + sucrose + 10 microM GA3, or even better GA4. With GA3 it may take more days to germinate but unless they are really old or recalcitrant most of them will germinate
Hi Nisrina,
I agree with Katie and Ana Elisa but I think that without sucrose in 1/2X MS it's better to germination in some conditions.
Sucrose stimulate ABA accumulation and that may delay germination in Arabidopsis.
Maybe it's better a 1/2x MS + GA3 only.
Best regards,
Vitor
Article Mechanisms of glucose signaling during germination of Arabidopsis
Dear Nisrina,
if you want to grow Arabidopsis on 1/2 MS agar plates, you can simply put sterile seeds on them and then keep plates in 4 C degrees for 2-3 days. You could also take as many sterile seeds as you need, put them into eppendorf tube and keep in 4 C as above - after that transfer them to plates or to the liquid medium in sterile conditions.
Best regards,
Julia
Hi Nisrina, we mostly use a few days cold (4 C) treatment (in the dark) to break dormancy in Arabidopsis (moist filter paper will do, agar-medium is not needed, especially not when later sowing on soil). For more recalcitrant Brassicaceae, 10 uM GA4 on moist filter paper has worked well in our hands.
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