Summary
1. In 0.1 ml of bleeding sap of uninoculated plants aspartic acid was found to occur as the sole amino-acid usually in a small concentration. With larger quantities of sap we also found a spot in the chromatograms and in the electropherogram, that most probably is identical with threonine.
2. Under the same conditions with a bleeding period of 4–7 hrs. aspartic acid was the only amino-acid detectable in the chromatogram of the bleeding sap of plants inoculated with ineffective strains.
3. Sap of plants inoculated with effective strains contains aspartic acid asparagine, glutamine, hydroxyproline and ‘threonine’. Sometimes in addition to these substances 1 or 2 unknown substances were found, havingR f-phenol values about identical with those of valine and leucine.
4. Glutamic acid never was detected in the bleeding sap. We suggest, that glutamic acid formed in the nodules might be rapidly transformed into glutamine and transported in this form.
5. The uptake of water is significantly higher with effective strains than with ineffective ones or with plants that have not been inoculated.
6. With this technique it is possible to select effective strains within 3 weeks after planting.
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References
Boissonnas, R. A., Séparation rapide des acides aminés par chromatographie ascendante bidimensionelle sur papier. Helv. Chim. Acta33, 1966 (1950).
Burris, R. H. and Wilson, P. W., Comparison of the metabolism of ammonia and molecular nitrogen in Azotobacter. J. Biol. Chem.165, 595 (1946).
Consden, L., Gordon, A. H. and Martin, A. J. P., Qualitative analysis of proteins: a partitition chromatographic method, using paper. Bioch. J.38, 224 (1944).
Dent, C. E., A study of the behaviour of some sixty amino-acids and other ninhydrinreacting substances on phenol-collidine filter paper chromatograms, with notes as to the occurrence of some of them in biological fluids. Bioch. J.43, 169 (1948).
Draper, O. J. and Pollard, A. L., The purification of phenol for paper partitition chromatography, Science109, 448 (1949).
Hunt, G. E., A comparative chromatographic survey of the amino-acids in five species of legume roots. Am. J. Botany38, 452 (1951).
Thomson, J. F., Zacharias, R. M. and Stewart, F. C., Investigations on nitrogen compounds and nitrogen metabolism in plants. I. The reaction of nitrogen compounds with ninhydrin on paper: a quantitative procedure. Plant Physiol.26, 375 (1951).
Wilson, P. W., The comparative biochemistry of nitrogen fixation. Advances Enzymol.13, 345 (1952).
Wilson, P. W. and Burris, R. H., The mechanisms of biological nitrogen fixation Bact. Rev.11, 41 (1947).
Woiwod, A. J., A technique for examining large numbers of bacterial culture filtrates by partitition chromatography. J. Gen. Microbiol.3, 312 (1949).
Zelitsch, I., Rosenblum, E. D., Burris, R. H. and Wilson, P. W., Isolation of the key intermediate in biological nitrogen fixation by Clostridium. J. Biol. Chem.191, 295 (1951).
Zelitsch, I., Rosenblum, E. D., Burris, R. H. and Wilson, P. W., Comparison of the metabolism of ammonia and molecular nitrogen in Clostridium. J. Bacteriol.62, 747 (1951).
Zelitsch, I., Wilson, P. W. and Burris, R. H., The amino acid composition and distribution of N 152 in soybean root nodules supplied N 152 -enriched N2. Plant Physiol.27, 1 (1952).
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Wieringa, K.T., Bakhuis, J.A. Chromatography as a means of selecting effective strains ofRhizobia . Plant Soil 8, 254–262 (1957). https://doi.org/10.1007/BF01666160
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DOI: https://doi.org/10.1007/BF01666160