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A new protocol on Prometheus Wiki “Making leaf surface imprints” does just what you need! Have a look at: http://prometheuswiki.publish.csiro.au/tiki-index.php?page=Making+leaf+surface+imprints
Great to hear you’ve found a protocol Pablo – it’d be great if you could share it on PrometheusWiki, perhaps in combination with the suggestions below. Here are some suggestions from a few of our Editors:
From Andr�s Baresch of the Smithsonian Tropical Research Institute (via editor Lou Santiago).
“as a starting point and to help the person asking the question a short description of what I have done:
On fresh leaves:
Peeling hairs with scotch tape (can take several attempts until hairs start to detach) then using nail polish to generate a first imprint that will eliminate remaining hairs (sometimes this nail polish gets firmly attached, it often comes out after several tries with scotch tape), this procedure is followed by the final peel that should come out clean and easy.
In some cases there is a direction by which hairs fall easier so is important to try from several angles to pull of the scotch tape.I have been able to eliminate the cuticle (bearing hairs) of Cecropia peltata (after treated with NaOH 10%) leaving it clean for venation images, I could try to see if is still possible to do an imprint with polish over that surface.”
From Lawren Sack:
“Recommend trying to shave the leaf and use nailpolish first. Then, could try dental epoxy impressions and using nailpolish on that. Failing that, can try epidermal peels from semi-macerated leaves and looking at the stomata from the inside. That can be a real pain. At that stage, looking for an SEM and looking at shaved and unshaved leaves is the way forward…”Hope this is helpful!
> Something which discusses the nature of Neutron Probe Calibration.
> Best shape calibration curves etc.
> Mathematical models for dealing with Probe readings from near the surface.Hi Simon
There are many protocols published, but most are just hard copy. We are trying to dig out an electronic version that will have information on calibrations and relations to lower limits for water extraction etc. Rana> Something which discusses the nature of Neutron Probe Calibration.
> Best shape calibration curves etc.
> Mathematical models for dealing with Probe readings from near the surface.From Rana Munns:
Dear Dr Habib
That is a very good idea to include protocols on extracting antioxidant enzymes. I will contact the editor in charge of Tissue Chemistry and we will invite some protocols.
I would like to add that in the Munns and Tester (2008) review, we cautioned about the physiological interpretation of changes in antioxidants, which can be secondary responses to the stress. These measurements need to be accompanied by measurements of growth rate and of photosynthesis. Otherwise we do not know whether the changes in antioxidants indicate tolerance to the stress, or sensitivity to it.
Rana Munns
Dear Habib,
You might find the protocol on [http://prometheuswiki.publish.csiro.au/tiki-index.php?page=Imaging-PAM+Chlorophyll+Fluorometer|Imaging PAM Chlorophyll Fluorometer] useful. This is the only protocol on chlorophyll fluorescence available at this stage, so any further contributions will be very welcome!
Emma
PrometheusWiki Editorial Assistant> I am planning to measure the relative water content as an indication of drougth stress resistance. I have stressed and not stressed plants to compare. My question is: should I water stressed and not stressed plants before measuring RWC?
No, dont water the plants.
You could start by doing the traditional RWC measurement, by rehydrating either a detached leaf or a section of a leaf for 3-4 h etc. If the RWC is lower than the control, the plant has responded to the drought treatment. This does not tell you whether or not it has lost turgor; it may have lost turgor, or it may have osmotically adjusted and not lost turgor. This is explained in the Boyer et al article, FPB, 2008. If you cannot access it, write to me and I will send you a pdf (rana.munns@csiro.au).
