Dear Eugene Sosnovsky!

It is possible (I have made some experiments using wheat) to increase the leaf thickness using for plants cultivation high levels of light intensity (see Russian Botanical Journal, 1978) and to decrease the leaf thickness using for plants cultivation artificial drougth or low concentrations of crude oil.

Best regards,

Professor Vadim Polonskiy,

Krasnoyarsk State Agrarian University,

Head of Dept. of Botany and Plant Physiology

Some MYB domain proteins are known to control cell shape and tricome development. Also there are several genes known to control stomata patterns (ie TOO MANY MOUTHS). You could do a transient genetic manipulation with some candidate genes. This kind of work is done in tomato and Arabidopsis as routine.

Try to contact directly the working group of

http://www.bot1.biozentrum.uni-wuerzburg.de/wissenschaftforschung/hedrich/

My first question is: Do you have any mathod for ficus transformation?

There is plenty of ways to manipulate structural features of leaves, however most of them involve modification of gene expression and for this you need to have efficient transformation method. Of course that is not all but before writing something more I need that answer.

Regards

Robert Malinowski

Dear Eugene

For stomate number and density I use nail polish (colorless) which I spread thinly on the leaves, after desiccation I take off gently polish then I put it on the glass and I observed and photographed under the microscope or binocular.

Measurement of anatomical features

You fixe mature leaves in formalin–70% alcohol–glacial acetic acid (5:90:5). You dehydrated leaf samples in an alcohol series (70–100%), cleared in xylene and embedded in paraffin (56– 58 8C). Transverse sections of 10 mmthickness for leaf obtained using a rotary microtome. These slides were deparaffinized, rehydrated and stained with safranin– fast green (1% aqueous safranin and 0.5% fast green in 95% alcohol). Sections of samples were photographed under a light microscope (Olympus BX41, Japan) and digitized (Olysia BioReport software) to determine anatomical parameters.

Best regards

Dear Eugene,

I agree with the proposal sent by Nima Tachbibi in using anatomical techniques.

My suggestion is depending on how much do yo want to spend in time, effort and cost.

The use of fixatives and embed the samples in parafin is quite good, and you can obtain many information, depending on the dyes you use. Safranin-fast green stain is widely used for geneal pourposes. the time you spend is higher compared with other techniques and the cost is a bit higher if you have not facilities.

Using fixative but in spite of obtaining parafin sections you use a freezing microtome you can obtaion 15-20 micron thickness. If your aim is to see structural changes maybe you will be interested in remove the cell content. Using sodium hypochlorite during 10 minutes and then neutralizing the bleach agent you can stain them with the same dyes and see modification in the cellwall. It is a cheap and give general good results. If you remove the cell content you will loose part of information (modification in cell content produced by the host).

Another approach you can select, is to obtain demithin sections (1-1.5 microns) using plant samples embebdded in resins (Spurr, LrWhite,JB4,...). Here you can use several aqueous dies (I prefer Tolouidine Blue O, because it is metachromatic). But you must select wich area of the sample is interesting for you. I.E. I have been working analyzing the impact of aracnids in leaf domatia looking for the mechanism used to fix eggs on stomata in this structures.

If you want to see surfaces there are other techniques I can suggest.

You can take a look to the link I attach. There are some pictures I included long time ago with several kinds of vegetal material and different dyes, techniques, etc. If you need other kind of pictures I can show you some pictures from my PhD. There I studied the effect of ozone in leaf histology in open top chambers and in the field in several populations.

Best regards,

David Lázaro-Gimeno

http://www.uv.es/lagida/Laboratory_eng.htm

Dear Eugene

I agree with David Lazaro-Gimeno in anatomical techniques, it's expensive and lengthy but for the density and nomber of somate my technique is very easy cheaper and gives good results.

Best regards

I thought He wants to manipulate some features of leaf anatomy not just record it.

You can manipulate some components of cell cycle machinery or factors influencing growth or patterning. In case of stomata substantial progress has been made and many factors were characterised. Paper below is a nice example:

http://www.sciencemag.org/content/333/6048/1436.short

For trichome manipulation you can also find some nice papers (best from Hülskamp lab) and in case of the cell cycle I have some experience. One major problem: YOU NEED METHOD FOR TRANSFORMING PLANTS.

Regards

Robert

Dear Eugene,

I found strong effect of soil water salinity on especially stomatal density but in some species also on stomatal size. An easy way to manipulate leaf characters would thus be to water your plants with different salinity concentrations. But of course be aware to use salinity levels your plants can stand, we don't want them to die right.

Cheers,

Nele

Dear Eugene

The anatomical features vary according to the climatic features,trichome density increases as temp is low like how u use a sweater in cool season,stomata density is genetical but under polluted and dusty conditions its number and size reduces.For trichome and stomatal studies temporory preparations stained with dil.iodine is best,But your host plant highly resisting type,choose plants without waxy coating.Contact me for further details if you wish.

Best of luck

Dr.Shankaran.

Exposure to abiotic stress factors such as Ultraviolet-B radiation would cause thickening of leaf and the cell walls.

There are reports that plants grown under Ultraviolet-B radiation stress have developed resistance to herbivory due the development of thick walls and also changes in the biochemistry of the tissues.

If someone doing research on forest tree growth rings ?