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Measurement of Nucleic Acid Components in Soil¹

ABSTRACT

A procedure is described for the quantitative estimation of ribonucleic acid phosphorus in soil. Organic phosphorus was extracted from soil by digestion with 1 N NaOH for 19 hours at 30°C. Acidification of the alkaline extract resulted in precipitation of organic materials or humus. The soluble organic phosphorus in the acid extract was separated from excess salt and concentrated by adsorption on activated alumina. After removal from the alumina, the extract was resolved by anion-exchange chromatography. Samples of the eluants from chromatographic peaks were analyzed in a spectrophotometer. Techniques were evaluated by the addition of known quantities of nucleic acid to soils and the determination of the extent of nucleotide losses in the several steps involved.

The data showed that a Carrington soil containing 575 ppm. of organic phosphorus, and a Webster soil containing 327 ppm., contained not more than 1 ppm. and 6 ppm., respectively, of ribonucleic acid phosphorus by the above described technique. The large fraction of soil organic phosphorus was not identified, but the evidence was considered adequate that only a small portion of the soil organic phosphorus could be accounted for as nucleic acids. Purine bases were not detected in soil hydrolysates. Paper chromatograms were run on an acid hydrolysate of soil and the migration of the ultraviolet absorbing spots compared with the migration of known purine bases. Under ultraviolet light no migration of the soil hydrolysate could be detected. The absence of detectable quantities of purine bases also indicated that very little nucleic acid exists in the soil.

¹ Contribution from Department of Agronomy, Iowa Agr. Exp. Sta., and Division of Soil and Plant Relationships, U. S. Dept. of Agriculture, cooperating. Journal Paper J-2420, Project 1070, of the Iowa Agr. Exp. Sta. Presented before Division III, Soil Science Society of America, Dallas, Texas, Nov. 19, 1953.

² Graduate assistant, Associate Professor and Bacteriologist, respectively. The authors express their appreciation to Dr. F. Schlenk for his cooperation and helpful suggestions during the course of this investigation.

Received for publication October 7, 1953.