I have to perform a pot experiment in which I have to separately visualize and collect root of different, intercropped herbaceous species and I need a stain to be injected in the above ground part and that can be transferred to the roots. This stain should allow me to discriminate roots from the stain-injected species from roots of the non injected species.
Can you suggest me some stain and, if available, examples of its application?
Thanks in advance.
This is a quite challenging question as most of the studies on roots using histochemistry are based on dyes which are cytotoxic - and you are asking for a peri-vital one.There is a publication by Dubrovsky et al., published in 2006 http://aob.oxfordjournals.org/content/97/6/1127.full.pdf+html, which uses Neutral Red for staining roots. They performed a cytotoxicity test fort this dye - and if you use the lowest concentration (possibly doing this in incremental doses) it could possibly serve your purpose. Bear in mind that it is a suggestion from a molecular pathologist trained as human pathologist - so all plant specialists out there should comment on this ( I have used NR as a counterstain for human tissue - but obviously not as a perivital stain - it works well). Good luck with your project.
Well, I am not sure injection approach will work. Park Nobel used a different approach. He was using NR but in adiffrent way. Specifically they did the following (I copy extract from his M&M of the DRENNAN and NOBEL paper (American Journal of Botany 83(2): 133-139. 1996):
"To allow identification of newly produced roots and of the extension of existing roots, the entire root systems were initially excavated, stained by dipping into a 0.05% (w/w) aqueous so- lution of neutral red (pH 6.5) for 10 min (Schumacher et al., 1983), and then replanted in the containers; subsequently produced root ma- terial was unstained and thus readily discernible."
May be in this way you can pre-stain one individuals but no the other. Then it may work for you.
Dear Joseph, That what I thought would happen when a medic stirs the pot. Fantastic luck that you are on ResearchGate. The idea of staining by drawing in a solution of dye into the roots is along the physiological route, simple yet elegant. I was considering that injection would draw the dye only upwards. I hope that Segio tries NR with your suggestion.
This not the case, but for external staining I used Congo Red to differentiate living roots from those that were recently died, and found a tendency of darker color for the latter
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