Foliar selenium biofortification of soybean: the potential for transformation of mineral selenium into organic forms.

Introduction: Selenium (Se) deficiency, stemming from malnutrition in humans and animals, has the potential to disrupt many vital physiological processes, particularly those reliant on specific selenoproteins. Agronomic biofortification of crops through the application of Se-containing sprays provides an efficient method to enhance the Se content in the harvested biomass. An optimal candidate for systematic enrichment, guaranteeing a broad trophic impact, must meet several criteria: (i) efficient accumulation of Se without compromising crop yield, (ii) effective conversion of mineral Se fertilizer into usable organically bound Se forms (Seorg), (iii) acceptance of a Se-enriched crop as livestock feed, and (iv), interest from the food processing industry in utilization of Se-enriched outputs. Hence, priority should be given to high-protein leafy crops, such as soybean. Methods: A three-year study in the Czech Republic was conducted to investigate the response of field-grown soybean plants to foliar application of Na2SeO4 solutions (0, 15, 40, and 100 g/ha Se); measured outcomes included crop yield, Se distribution in aboveground biomass, and the chemical speciation of Se in seeds. Results and Discussion: Seed yield was unaffected by applied SeO4 2-, with Se content reaching levels as high as 16.2 mg/kg. The relationship between SeO4 2-dose and Se content in seeds followed a linear regression model. Notably, the soybeans demonstrated an impressive 73% average recovery of Se in seeds. Selenomethionine was identified as the predominant species of Se in enzymatic hydrolysates of soybean, constituting up to 95% of Seorg in seeds. Minor Se species, such as selenocystine, selenite, and selenate, were also detected. The timing of Se spraying influenced both plant SeO4 2- biotransformation and total content in seeds, emphasizing the critical importance of optimizing the biofortification protocol. Future research should explore the economic viability, long-term ecological sustainability, and the broad nutritional implications of incorporating Se-enriched soybeans into food for humans and animals.

Positive Selection of Specific Antibodies Produced against Fusion Proteins.

A method for the positive selection of specific antibodies for target proteins expressed as fusion proteins for the production of antiserum is presented. As proof of concept, the fusion protein FLAG::His::GFP::His::FLAG was expressed in Escherichia coli, purified, and used for the immunization of rabbits. The obtained serum was precleared via protein A affinity. A CusF::FLAG fusion protein was expressed in the periplasm of E. coli and purified. GFP without tags was also expressed in E. coli and purified via organic extraction. These proteins were then coupled to NHS-activated sepharose and used for the positive selection of Anti-GFP and Anti-FLAG antibodies. The obtained sera were tested for their specificity against different protein samples and fusion proteins in Western blots. A high specificity of the antibodies could be achieved by a single affinity chromatography step. In general, we advise to express the target protein with different tags and in different E. coli compartments for antibody production and affinity chromatography.

The effectiveness of various treatments in changing the nutrient status and bioavailability of risk elements in multi-element contaminated soil.

Potential changes in the mobility and bioavailability of risk and essential macro- and micro-elements achieved by adding various ameliorative materials were evaluated in a model pot experiment. Spring wheat (Triticum aestivum L.) was cultivated under controlled condition for 60 days in two soils, uncontaminated Chernozem and multi-element contaminated Fluvisol containing 4900 ± 200 mg/kg Zn, 35.4 ± 3.6 mg/kg Cd, and 3035 ± 26 mg/kg Pb. The treatments were all contained the same amount of sulfur and were as follows: (i) digestate from the anaerobic fermentation of biowaste, (ii) fly ash from wood chip combustion, and (iii) ammonium sulfate. Macro- and micro-nutrients Ca, Mg, K, Fe, Mn, Cu, P, and S, and risk elements Cd, Cr, Pb, and Zn were assayed in soil extracts with 0.11 mol/l solution of CH3COOH and in roots, shoots, and grain of wheat after 30 and 60 days of cultivation. Both digestate and fly ash increased levels of macro- and micro-nutrients as well as risk elements (especially Cd and Zn; the mobility of Pb decreased after 30 days of cultivation). The changes in element mobility in ammonium sulfate-treated soils appear to be due to both changes in soil pH level and inter-element interactions. Ammonium sulfate tended to be the most effective measure for increasing nutrient uptake by plants in Chernozem but with opposite pattern in Fluvisol. Changes in plant yield and element uptake in treated plants may have been associated with the higher proline content of wheat shoots cultivated in both soils compared to control. None of the treatments decreased uptake of risk elements by wheat plants in the extremely contaminated Fluvisol, and their accumulation in wheat grains significantly exceeded maximum permissible levels; these treatments cannot be used to enable cereal and other crop production in such soils. However, the combination of increased plant growth alongside unchanged element content in plant biomass in pots treated with digestate and fly ash suggests that these treatments have a beneficial impact on yield and may be effective treatments in crops grown for phytoremediation.

Applications of organic and inorganic amendments induce changes in the mobility of mercury and macro- and micronutrients of soils.

Both soil organic matter and sulfur (S) can reduce or even suppress mercury (Hg) mobility and bioavailability in soil. A batch incubation experiment was conducted with a Chernozem and a Luvisol artificially contaminated by 440 mg · kg(-1) Hg showing wide differences in their physicochemical properties and available nutrients. The individual treatments were (i) digestate from the anaerobic fermentation of biowaste; (ii) fly ash from wood chip combustion; and (iii) ammonium sulfate, and every treatment was added with the same amount of S. The mobile Hg portion in Chernozem was highly reduced by adding digestate, even after 1 day of incubation, compared to control. Meanwhile, the outcome of these treatments was a decrease of mobile Hg forms as a function of incubation time whereas the contents of magnesium (Mg), potassium (K), iron (Fe), manganese (Mn), copper (Cu), zinc (Zn), and phosphorus (P) were stimulated by the addition of digestate in both soils. The available calcium (Ca) contents were not affected by the digestate addition. The experiment proved digestate application as the efficient measure for fast reduction of mobile Hg at extremely contaminated soils. Moreover, the decrease of the mobile mercury portion was followed by improvement of the nutrient status of the soils.