Cytokinin quantification

dreimers at msuvx1.memphis.edu dreimers at msuvx1.memphis.edu
Sun Mar 20 15:05:57 EST 1994


In article <2mar03$r33 at nnrp.ucs.ubc.ca>, murali at unixg.ubc.ca (Murali Srinivasan) writes:
> 
> Does anybody have an easy method for qualitative and quantitative estimation
> of cytokinins from gram positive bacteria, preferably Bacillus spp?
> Thank you! 

I don't know of an "easy" way to do that.  If your sample is "clean" you can
use immunoassays that are fairly quick and easy. Normally I concentrate the
cytokinins using XAD-2 chromatography and then fractionate the cytokinins
in the sample by matching elution times of known standards using Sephadex
LH-20 chromatography in 35% ETOH.  You then use the tobacco callus bioassay
to determine cytokinin activity (Kinetin equivalents per liter) of your 
fractions-35 day incubation.  The easier and faster stuff follows.  With
a fairly clean sample at this point, you can run them through HPLC in
a 70% ETOH or 20% acetonitrile gradient and again match up elution times
with standards.  You can collect these fractions and do immunoassays or
do GC-Mass Spec.  This is probably more than you wanted so let me give
you a reference to check out.

Sturtevant, Dawn B. and Barbara J. Taller 1989
Cytokinin Production by Bradyrhizobium japonicum
Plant Physiology Vol. 89 pps. 1247-1252

I would be interested in hearing about your research.  I am looking at
cytokinin production in cyanobacteria. 



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