Dear M. Alizadeh

To prepare the leaf discs around 30 ml of (1.5% agar solution was poured into a 7 cm Petri dish) after cooling the agar (simply agar)  leaf in this media and condition was able to survive 10 days, all tool used in cutting the leaves were sterilized in 90% ethanol to reduce contamination. I was able to keep the leaf for at least 10 days simply. I add parafilm covering over the dishes.

Hoping this will be helpful            

Regards

Prof. Houda Kawas

97

Hi Alizadeh

I agree with Prof.Houda

Dear Alizadeh

You can use benzimidazole alone or with water agar, it is useful to keep detached leaf of plant for more than a month by  wrapping  the neck of cucumber leaf (cotyledon) with cotton and wet the cotton with a very low concentration of benzimidazole about 15-25 µg ml, or float the leaf discs over the surface of benzimidazole solution.

I attached a relative reference

Good luck

Agar used in plant tissue culture medium ( e.g., MS medium)  should be good for your proposed experiment.

Dear Alizadeh,

1. You listed 3 kinds of agars for us to choose: "Which Agar is better? 1-Agar agar, 2-plant agar, 3-bacteriologic agar". I did not see the detailed descriptions of these 3 products. But, for now, I definitely will choose 'plant agar', because you are dealing with plant leaves.

2. Just a reminder: I don't know your research goal, but I am not sure whether the leaves will remain real 'fresh' (like those on a plant) when they are placed on 'plain' agar for 48h, without nutrients such as carbon source (macro-elements) and micro-elements.

The media to use will depend upon why you are wanting to keep the leaf discs fresh. I assume you want to use them for an experiment after 48 hours perhaps? If so, I would be placing them on a tissue culture agar media with no hormones - just a simple water:agar media would d, or you could use a 1/4 MS media. It won't matter too much as long as the leaf pieces do not become contaminated.  Surface sterilsie the leaf and place on the medai in petri dishes and tape up. Hope this helps.

Dear all,

I want to assay an insecticide toxicity in a 48h period using leaf-dip method and I didn't know which type of agar is best for this goal.

Thank you all very much. Your comments and ideas were very useful.

you most welcome

Houda

Sometimes I need to send leaf pieces to PCR. Sometimes even to another city or after even 30 days.

Therefor I use small eppendorfs as they close totally. Inseref is 0.5 MS with plant agar. They can even kept by RT! 

As they are taken from tissue culture they need no sterilisation. But I use leaves from the greenhouse too. After surface sterilisation.

Maybe you keep bigger leaves like this in Falcons. And cut the didcs only when you are starying the experiment. As the discs are open afresh.

i read so much researches all the methods are based of a rooting detached leaves on agar and no one explained the method in details >>>> i tried it once but the leaves dried quickly >>>> finally i intended to try a tissue culture media 

hope not to fail