I am teaching phytohormones to MSc students. For the practical part of the course we conducted some experiments related to the effect of ABA on germination, stomatal closure and leaf senescence. I would like to see whether ABA can be quantified from the extract of salt or drought-stressed leaves using a bioassay protocol. The extract can be the rude extract of the leaf or leaf extract in a buffer. I would be grateful if somebody provide me such protocols.
I think William Davis Set up a sensitive but laborious assay on Commelina communis leaf strips. It requires translucent varieties with easy to remove epidermis, http://www.plantphysiol.org/content/102/2/497.full.pdf. Enid macRobbie wrote fundamental works on Commelina. Also Vicia faba strips can be assayed
Dear Guido
Thanks for your answer and also many thanks for the suggested paper. Actually, during last three years I was busy with finding the sensitivity of different methods for stomatal ABA responses, and we found some interesting result about ABA sensitivity in different tissues. However what I need now, is finding a protocol for quantifying ABA amount in the tissues without using other methods such as using HPLC, ELISA or GC MS. I what to see if possible a bioassay protocol using plant responses. As in the paper that you suggest they used stomatal response from epidermis of Commelia. I would try that method to see if it is work.
Thanks
Sasan
I used ELISA method (using Phytodetek ABA, Agdia, IN, USA) to quantify ABA concentration of drought-stressed leaves of petunia, and it worked very nicely. You can check my previous article which had very interesting correlation data between stomatal conductance and leaf ABA level. Hope this helps.
Article Physiological and molecular responses to drought in Petunia:...
Hi Jongyun
Thanks for the paper, that's interesting because I also found some interesting results for ABA level in the leaf and stomatal conductance also the relationships between them using different closing stimuli such as ABA, desiccation, low RH and darkness. It seems ABA somehow is involved in rhytmic response of stomata. However, here I am looking for a protocol for bioassay measurement of ABA.
Cheers
Dear Dongping
Thanks for your suggession. Could you explain me a bit about this method?
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