Hi guys,
I wonder whether you guys have included SA (salicylic acid) in your study or not? If yes, what are the methods of its application? will you apply it as a solution through roots or will you just spray it on the leaves? and what are you planning about the concentration of SA?
My apology in advance if SA isn't a part of the project.
Certainly in our lab hormones have been applied using lanolin or delivered in the agar/media, but I think it depends on the solubility of the particular hormone (auxin is certainly fat soluble, and as far as I can tell from a quick google, so is SA.) Anyway, some good information on various techniques in the 'methods' section here: http://oer.nios.ac.in/wiki/index.php/Growth_Hormones
When you apply plant hormones, the 1st thing to know how to dissolve it. Different hormones come in different formulations (Easter, Salt, Acid..). Sodium salt is easy to dissolve in H2O. If it doesn't dissolve try DMSO, ethanol or ethanol and water. Sometimes the ethanol dissolved one may re-solidified when you add it in large quantities of water (to prepare working solution). Once I had this problem with SA and 2,4-D. My advise is:
1. Try to dissolve with DMSO (the stock solution) and then check if it works when diluted
2. Increase the pH (>7.5) of your working solution at very low buffer strength (K phosphate can be tried)
3. Add an appropriate attractant for foliar application
Thank you all for your valuable comments
I prepared 1M stock solution in ethanol and then diluted it into working solution. The problem isn't related to SA itself actually, the problem is that the ethanol itself affect the expression level of my gene. I just want to replace ethanol as a solvent...
Any suggestion will be highly appreciated
Try DMSO or the other option is you can add equal amount of ethanol that you used for dissolving the SA into your control treatment. If you spray water as a control, then you can calculate the amount of ethanol in your SA working solution so that you'll be able to see the effect of Ethanol on the level of expression of your gene. In any case you need to add equal amount of any solvent you used for the control treatments.
Plants are sprayed with a final concentration of 4 mM SA in 1% ethanol, and covered with a transparent plastic hood during the treatment period. Also, see sewelam et al., 2013.
Thank you Nasir but I don't wanna spray it, I want to apply it as aqueous solution to the plant roots
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