A DTPA soil test was developed to identify near‐neutral and calcareous soils with insufficient available Zn, Fe, Mn, or Cu for maximum yields of crops. The extractant consists of 0.005M DTPA (diethylenetriaminepentaacetic acid), 0.1M triethanolamine, and 0.01M CaCl2, with a pH of 7.3. The soil test consists of shaking 10 g of air‐dry soil with 20 ml of extractant for 2 hours. The leachate is filtered, and Zn, Fe, Mn, and Cu are measured in the filtrate by atomic absorption spectrophotometry.The soil test successfully separated 77 Colorado soils on the basis of crop response to Zn, Fe, and Mn fertilizers. Critical nutrient levels must be determined separately for each crop using standardized procedures for soil preparation, grinding, and extraction. The critical levels for corn using the procedures reported herein were: 0.8 ppm for Zn, 4.5 ppm for Fe, and tentatively 1.0 ppm for Mn, and 0.2 ppm for Cu.Development of the soil test was based, in part, on theoretical considerations. The extractant is buffered at pH 7.30 and contains CaCl2 so that equilibrium with CaCO3 is established at a CO2 level about 10 times that of the atmosphere. Thus, the extractant precludes dissolution of CaCO3 and the release of occluded nutrients which are normally not available to plants. DTPA was selected as the chelating agent because it can effectively extract all four micronutrient metals. Factors such as pH, concentration of chelating agent, time of shaking, and temperature of extraction affect the amount of micronutrients extracted and were adjusted for maximum overall effectiveness.

We investigated the capability of the plant-growth-promoting and biocontrol fungus Trichoderma harzianum Rifai 1295-22 (T-22) to solubilize in vitro some insoluble or sparingly soluble minerals via three possible mechanisms: acidification of the medium, production of chelating metabolites, and redox activity. T-22 was able to solubilize MnO2, metallic zinc, and rock phosphate (mostly calcium phosphate) in a liquid sucrose-yeast extract medium, as determined by inductively coupled plasma emission spectroscopy. Acidification was not the major mechanism of solubilization since the pH of cultures never fell below 5.0 and in cultures containing MnO2 the pH rose from 6.8 to 7.4. Organic acids were not detected by high-performance thin-layer chromatography in the culture filtrates. Fe2O3, MnO2, Zn, and rock phosphate were also solubilized by cell-free culture filtrates. The chelating activity of T-22 culture filtrates was determined by a method based on measurement of the equilibrium concentration of the chrome azurol S complex in the presence of other chelating substances. A size exclusion chromatographic separation of the components of the culture filtrates indicated the presence of a complexed form of Fe but no chelation of Mn. In liquid culture, T. harzianum T-22 also produced diffusible metabolites capable of reducing Fe(III) and Cu(II), as determined by the formation of Fe(II)-Na2-bathophenanthrolinedisulfonic acid and Cu(I)-Na2-2,9-dimethyl-4,7-diphenyl-1,10-phenanthrolinedisulfonic acid complexes. This is the first report of the ability of aTrichoderma strain to solubilize insoluble or sparingly soluble minerals. This activity may explain, at least partially, the ability of T-22 to increase plant growth. Solubilization of metal oxides by Trichoderma involves both chelation and reduction. Both of these mechanisms also play a role in biocontrol of plant pathogens, and they may be part of a multiple-component action exerted by T-22 to achieve effective biocontrol under a variety of environmental conditions.

High Cd content in durum wheat (Triticum turgidum L. var durum) grain grown in the United States and Canada presents potential health and economic problems for consumers and growers. In an effort to understand the biological processes that result in excess Cd accumulation, root Cd uptake and xylem translocation to shoots in seedlings of bread wheat (Triticum aestivum L.) and durum wheat cultivars were studied. Whole-plant Cd accumulation was somewhat greater in the bread wheat cultivar, but this was probably because of increased apoplastic Cd binding. Concentrationdependent 109 Cd 2؉ -influx kinetics in both cultivars were characterized by smooth, nonsaturating curves that could be dissected into linear and saturable components. The saturable component likely represented carrier-mediated Cd influx across root-cell plasma membranes (Michaelis constant, 20-40 nM; maximum initial velocity, 26-29 nmol g ؊1 fresh weight h ؊1 ), whereas linear Cd uptake represented cell wall binding of 109 Cd. Cd translocation to shoots was greater in the bread wheat cultivar than in the durum cultivar because a larger proportion of root-absorbed Cd moved to shoots. Our results indicate that excess Cd accumulation in durum wheat grain is not correlated with seedling-root influx rates or root-toshoot translocation, but may be related to phloem-mediated Cd transport to the grain.