Comparative metabolic behavior and interrelationships of Tc and S in soybean plants.

The comparative behavior of sulfur (S) and technetium (Tc) in soybean seedlings shows gross subcellular distributions to be similar for these oxyanions. More than 75% of the tissue-deposited Tc remains soluble and extractable. Differences in Tc fixation/incorporation were noted for the nuclear and chloroplast fractions of leaf and root cells. Pulse studies showed that soluble protein and nitrate reductase levels rose in response to Tc accumulation by sink leaves but not source leaves. In vitro assay of chloroplast-based S reduction and incorporation systems showed Tc to be reduced and incorporated into amino nitrogen-containing products. A hypothesis related to the metabolic behavior of Tc in plants is presented.

Organic Constituents and Complexation of Nickel(II), Iron(III), Cadmium(II), and plutonium(IV) in Soybean Xylem Exudates.

The xylem exudates of soybean (Glycine max cv Williams), provided with fixed N, were characterized as to their organic constituents and in vivo and in vitro complexation of plutonium, iron, cadmium, and nickel. Ion exchange fractionation of whole exudates into their compound classes (organic acid, neutral, amino acid, and polyphosphate), followed by thinlayer electrophoresis, permitted evaluation of the types of ligands which stabilize each element. The polyvalent elements plutonium(IV) and iron(III) are found primarily as organic acid complexes, while the divalent elements nickel(II) and cadmium(II) are associated primarily with components of the amino acid/peptide fraction. For plutonium and cadmium, it was not possible to fully duplicate complexes formed in vivo by back reaction with whole exudates or individual class fractions, indicating the possible importance of plant induction processes, reaction kinetics, and/or the formation of mixed ligand complexes. The number and distribution of specific iron- and nickel-containing complexes varies with plant age and appears to be related to the relative concentration of organic acids and amino acids/peptides being produced and transported in the xylem as the plant matures.

Concentration of orally administered and chronically fed 95mTc in Japanese quail eggs.

A chronic feeding study using 95mTc incorporated into alfalfa and an acute study where 95mTc was amended to alfalfa showed that about 8.4% of ingested Tc was transferred to eggs. After 10 days of chronic feeding, 80% of the Tc was in yolk, 20% in albumin and less than 1% in shell and associated membranes. At necropsy, technetium concentrations in the three largest oocytes were nearly equal. The biological half-time for Tc was about one to two days in acute studies. Results from the chronic feeding study also indicated that Tc levels in albumin reach a maximum between three and five days while maximum yolk concentration is attained in about six to seven days. Albumin concentrations declined about 20-50% after Day 6.

Cadmium uptake kinetics in intact soybean plants.

The absorption characteristics of Cd(2+) by 10- to 12-day-old soybean plants (Glycine max cv Williams) were investigated with respect to influence of Cd concentration on adsorption to root surfaces, root absorption, transport kinetics and interaction with the nutrient cations Cu(2+), Fe(2+), Mn(2+), and Zn(2+). The fraction of nonexchangeable Cd bound to roots remained relatively constant at 20 to 25% of the absorbed fraction at solution concentration of 0.0025 to 0.5 micromolar, and increased to 45% at solution concentration in excess of 0.5 micromolar. The exchangeable fraction represented 1.4 to 32% of the absorbed fraction, and was concentration dependent. Using dinitrophenol as a metabolic inhibitor, the ;metabolically absorbed' fraction was shown to represent 75 to 80% of the absorbed fraction at concentration less than 0.5 micromolar, and decreased to 55% at 5 micromolar. At comparatively low Cd concentrations, 0.0025 to micromolar 0.3, root absorption exhibited two isotherms with K(2) values of 0.08 and 1.2 micromolar. Root absorption and transfer from root to shoot of Cd(2+) was inhibited by Cu(2+), Fe(2+), Mn(2+), and Zn(2+). Analyses of kinetic interaction of these nutrient cations with Cd(2+) indicated that Cu(2+), Fe(2+), Zn(2+), and possibly Mn(2+) inhibited Cd absorption competitively suggesting an involvement of a common transport site or process.

Root absorption and transport behavior of technetium in soybean.

The absorption characteristics and mechanisms of pertechnetate (TcO(4) (-)) uptake by hydroponically grown soybean seedlings (Glycine max cv Williams) were determined. Absorption from 10 micromolar solutions was linear for at least 6 hours, with 30% of the absorbed TcO(4) (-) being transferred to the shoot. Evaluation of concentration-dependent absorption rates from solutions containing 0.02 to 10 micromolar TcO(4) (-) shows the presence of multiphasic absorption isotherms with calculated K(s) values of 0.09, 8.9, and 54 micromolar for intact seedlings. The uptake of TcO(4) (-) was inhibited by a 4-fold concentration excess of sulfate, phosphate, selenate, molybdate, and permanganate; no reduction was noted with borate, nitrate, tungstate, perrhenate, iodate, or vanadate. Analyses of the kinetics of interaction between TcO(4) (-) and inhibiting anions show permanganate to be a noncompetitive inhibitor, while sulfate, phosphate, and selenate, and molybdate exhibit characteristics of competitive inhibitors of TcO(4) (-) transport suggesting involvement of a common transport process.

Effects of plutonium on soil microorganisms.

As a first phase in an investigation of the role of the soil microflora in Pu complex formation and solubilization in soil, the effects of Pu concentration, form, and specific activity on microbial types, colony-forming units, and CO(2) evolution rate were determined in soils amended with C and N sources to optimize microbial activity. The effects of Pu differed with organism type and incubation time. After 30 days of incubation, aerobic sporeforming and anaerobic bacteria were significantly affected by soil Pu levels as low as 1 mug/g when Pu was added as the hydrolyzable Pu(NO(3))(4) (solubility, <0.1% in soil). Other classes of organisms, except the fungi, were significantly affected at soil Pu levels of 10 mug/g. Fungi were affected only at soil Pu levels of 180 mug/g. Soil CO(2) evolution rate and total accumulated CO(2) were affected by Pu only at the 180 mug/g level. Because of the possible role of resistant organisms in complex formation, the mechanisms of effects of Pu on the soil fungi were further evaluated. The effect of Pu on soil fungal colony-forming units was a function of Pu solubility in soil and Pu specific activity. When Pu was added in a soluble, complexed form [Pu(2)(diethylenetriaminepentaacetate)(3)], effects occurred at Pu levels of 1 mug/g and persisted for at least 95 days. Toxicity was due primarily to radiation effects rather than to chemical effects, suggesting that, at least in the case of the fungi, formation of Pu complexes would result primarily from ligands associated with normal (in contrast to chemically-induced) biochemical pathways.

Cadmium distribution and chemical fate in soybean plants.

The distribution and chemical behavior of Cd(2+) in tissues and its chemical form in xylem water of soybean plants (cv. Williams) were investigated. Following root absorption, Cd is strongly retained by roots, with only 2% of the accumulated Cd being transported to leaves; as much as 8% was transported to seeds during seed filling. In vivo xylem exudates contained two anionic Cd complexes in addition to inorganic forms of Cd. Once accumulated in root and leaf tissues, Cd rapidly equilibrated between the insoluble, soluble, and organelle fractions. Of the solubles, which contain 50% of the Cd, >50% was associated with components of >10,000 molecular weight, and <8% was associated with <500 molecular weight components. Cadmium accumulated in soybean seeds was primarily associated with cotyledons. Fractionation of seeds showed the soy proteinate and soy whey to contain 32 and 50% of the accumulated Cd, respectively.

Nickel in plants: I. Uptake kinetics using intact soybean seedlings.

The absorption of Ni(2+) by 21-day-old soybean plants (Glycine max cv. Williams) was investigated with respect to its concentration dependence, transport kinetics, and interactions with various nutrient cations. Nickel absorption, measured as a function of concentration (0.02 to 100 mum), demonstrated the presence of multiple absorption isotherms. Each of the three isotherms conforms to Michaelis-Menten kinetics; kinetic constants are reported for uptake by the intact plant and for transfer from root to shoot tissues. The absorption of Ni(2+) by the intact plant and its transfer from root to shoot were inhibited by the presence of Cu(2+), Zn(2+), Fe(2+), and Co(2+). Competition kinetic studies showed Cu(2+) and Zn(2+) to inhibit Ni(2+) absorption competitively, suggesting that Ni(2+), Cu(2+), and Zn(2+) are absorbed using the same carrier site. Calculated K(m) and K(i) constants for Ni(2+) in the presence and absence of Cu(2+) were 6.1 and 9.2 mum, respectively, whereas K(m) and K(i) constants were calculated to be 6.7 and 24.4 mum, respectively, for Ni(2+) in the presence and absence of Zn(2+). The mechanism of inhibition of Ni(2+) in the presence of Fe(2+) and Co(2+) was not resolved by classical kinetic relationships.

Nickel in Plants: II. Distribution and Chemical Form in Soybean Plants.

The gross tissue distribution, intracellular fate, and chemical behavior of Ni(2+) in soybean plants (Glycine max cv. Williams) were investigated. Following root absorption, Ni was highly mobile in the plant, with leaves being the major sink in the shoots for Ni during vegetative growth. A senescence >70% of the Ni present in the shoot was remobilized to seeds. Fractionation of root and leaf tissues showed >90% of the Ni to be associated with the soluble fraction of tissues; ultrafiltration of the solubles showed >77% of the Ni to be associated with the 10,000 to 500 molecular weight components of both roots and leaves. Chemical characterization of the soluble components (10,000 to 500 and >500 molecular weight) by thin layer chromatography and electrophoresis resolved a number of Ni-containing organic complexes. Major Ni-containing components formed in the root are transported in the xylem stream, and undergo partial modification on deposition in leaves. Nickel accumulated in seeds is primarily associated with the cotyledons. Chemical fractionation of cotyledon components showed 80% of the Ni to be associated with the soluble whey fraction, while 70% of this fraction was composed of Ni-containing components with molecular weight <10,000.