[Arabidopsis] replies to "How to check ROS levels in plants"

Y Jiang via arab-gen%40net.bio.net (by jiangyqcn from hotmail.com)
Thu Mar 5 11:54:25 EST 2009


Hi there,

 

a few days ago, I posted here to ask "how to check ROS levels in plants" and received a few replies. Thanks them. For those who also want to see the answers, posted as follows:

 

 

Hi,
maybe you did alredy got your answer otherwise you could use DAB stainig.
is a Diaminobenidine sensitive to H2O2.
Di
 
Diana Molino
Laboratoire de Biologie Cellulaire
INRA Versailles
Route de St-Cyr (RD10)
F-78026 VERSAILLES Cedex
Tel: +33 1 30 83 31 13
Fax: +33 1 30 83 30 99
 
 
you can try with d399 of molecular probs
 
Yuval Kaye
Department of Plant & environmental Sciences
Institute of Life Sciences 
The Hebrew University of Jerusalem
Givat Ram, Jerusalem, 91904, Israel
E-Mail:yuval.kaye from mail.huji.ac.il
 
 
 
I can suggest you few ways:

-Fluorescence using a ROS-specific dye. I use H2DCFDA from invitrogen, in conjuction with fluorescence microscope or microplate fluorescence reader.
 
           Krause, M., and Durner, J. (2004). Harpin inactivates mitochondria in Arabidopsis suspension cells. Mol Plant Microbe Interact 17, 131-139.-- Cristina


-Cerium deposits + electron microscopy

     Yao, N., Tada, Y., Sakamoto, M., Nakayashiki, H., Park, P., Tosa, Y., and Mayama, S. (2002). Mitochondrial oxidative burst involved in apoptotic response in oats. Plant J 30, 567-579.


Have fun, Cristina

Palmieri M. Cristina, phD student
Helmholtz Zentrum München
Institute of Biochemical Plant Pathology (BIOP) 
Ingolstädter Landstraße 1, D-85764 Munich
Tel.      +49 (0)8931873293
e-mail  cristina.palmieri from helmholtz-muenchen.de
 
 
Hi Y Jiang, 
There are few common staining methods that are use for several type ROS. H2O2 is normally the usual suspect in all stress conditions since it has the longest half life of all ROS types. Another ROS type is superoxide ions which are quickly dismutase to H2O2.
I attach here for you methods for both types  - for H2O2: DCF-DA and DAB staining, for superoxid: NBT staining. I suggest you try to stain H2O2 because it is the easiest and usually it is sufficient.  
 
Good luck,
Gadi
Gad Miller, Ph.D
Biochemistry Department
Mail Stop 200
University of Nevada
Reno NV 89557
775-784-4071

 

 

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