I'm thinking 2 hours. This is my first qPCR experiment on plants and I'm hoping to catch the earliest changes in root gene expression when iron is applied to leaf tissue.
I've found a similar study which considers "1 day later" after foliar application of iron. Gene expression in the roots must occur before that however. Do you think 2 hours is too short?
If 2 hours is not long enough to effect root gene expression, this would be a surprise to me and I would consider this an interesting result anyway! Gene expression in leaf tissue should be effected immediately after iron application, these will be subjected to qPCR as well...
Any comments are appreciated, I anticipate treating/processing plants soon. Such a tense and anxious time, wish me luck :)
Since you couldn't find the response in any bibliography, I think it would be more interesting if you try obtaining the answer through a kinetic study at different time points. You could then realize a QRT-PCR analysis to compare the expression over time, I'm sure that you'll get interesting results.
Not to be too flippant, but how big are your plants at the time of treatment? If they are little seedlings (
Matthew Wheal , The plants I am working on are 100 - 200 mm root to shoot tip.
What I mean by "must", is referring to the time it takes for metabolites/micro-nutrients to move in vivo. I can't find a straight forward answer, but some studies which consider the uptake/transport of isotope labelled substrates suggest that transport can occur in hours rather than days. However, gene expression studies I find consider days, not hours.
If iron can be absorbed and mobilized in hours, for example in a small (10-20mm) plant, then iron-regulated gene expression should be affected pretty quickly. Or maybe I'm wrong in my understanding and iron isn't absorbed as quickly or gene expression doesn't occur as quickly.
There is the possibility that a sudden increase of iron in plant leaf tissue will quickly deplete the concentration of metal-chelating metabolites, thus the iron itself may not even have to reach root tissue in order to elicit a root response. My intuition says gene expression "must" occur rapidly because I am under the impression that gene expression is under constant regulation. Typically, gene expression can be effected almost immediately in response to sudden changes, such as that from a sudden increase in iron. But not all stimuli result in immediate changes..
Ended up spraying/watering and harvesting plants the next day.
So 16h treatment time..
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