762 



Hall, Miller, and Herrero 



the radioactive spots on the chromatograms. Another aliquot of 

 the purified unknown was chromatographed in two dimensions with 

 the same solvents (3) and gave a single, molybdenum blue spot; its 

 autoradiogram showed only one spot (Figure 5D, Sh 44b) overlap- 

 ping the molybdenum blue area of the chromatogram. The Rf values 

 of the phosphorylated metabolite in six solvents are given in Table 1. 

 Comparison of the Rf values of the two phosphorylated, radioactive 

 spots with those of known phosphorylated compounds gave incon- 

 clusive residts. 



Table 1. Rf values of phosphorylated cotton metabolite. 



* Developed at 2° C. 



Radiochromatograms made with the crude water extract from 

 cotton showed at least seven radioactive compounds, but only two 

 important ones coinciding with two fluorescent bands in the paper 

 (Figure 5C, Ch 61-a and Ch 61-b). The principal metabolite exhibited 

 a brilliant blue fluorescence and the less active one a dark blue flu- 

 orescence. 



Two water soluble, phosphorylated radioactive metabolites were 

 also demonstrated in Coleus tissue processed in the same manner. The 

 second active fraction, after purification on the anion exchange col- 

 umn, contained more radioactivity than found in cotton. The radio- 

 chromatograms in six solvent systems (Figure 6) showed two radio- 

 active spots on each chromatogram. Rf values of the two metabolites, 



Tabic 2. Rf values of Coleus metabolites. 



