Spent coffee grounds as an alternative fertilizer: impact on bioaccessibility of antioxidants and commercial quality of lettuce.

BACKGROUND: During the processing of spent coffee grounds (SCGs) several residues are obtained, which are mostly disposed of in landfills. There is an urgent need for a comprehensive waste management strategy for these residues. This study evaluates the potential of SCGs as a biofertilizer by assessing their effects on lettuce leaves and the release of antioxidants following in vitro digestion and fermentation. RESULTS: Lettuce plants were grown with different amounts of SCGs (0-150 g kg-1) in the substrate. High SCG concentrations in the soil generated lighter colored tissues, a decrease in the green color, less root development, and lower dry weight of leaves (P < 0.05). The SCG levels also affected the release of antioxidants by the final product. This effect was more pronounced in the digested fraction: applying the Ferric Reducing Antioxidant Power (FRAP) method, the addition of SCGs from 10 g kg-1 to 125 g kg-1 increased the amount of antioxidant from 43.88 ± 4.81 to 105.96 ± 29.09 µmol Trolox g-1 of dry weight (P < 0.05). The Indigo Carmine Reducing Capacity (ICRED) method also showed a similar trend, but in this case the highest value was obtained with 150 g kg-1 of SCGs (16.41 ± 3.93 mmol catechin g-1 of dry weight) (P < 0.05). Moreover, in the fermented fraction a significant increase in the antioxidant released was found with low levels of SCG(P<0.05), while lettuces fertilized with intermediate amounts of SCGs (25 and 50 g kg-1) presented the highest amount of insoluble antioxidant (P < 0.05). CONCLUSION: A compromise should be found in order to achieve a product with a high antioxidant capacity and an acceptable visual quality. © 2024 Society of Chemical Industry.

Biofortifying multiple micronutrients and decreasing arsenic accumulation in rice grain simultaneously by expressing a mutant allele of OAS-TL gene.

Rice grains typically contain relatively high levels of toxic arsenic (As) but low levels of essential micronutrients. Biofortification of essential micronutrients while decreasing As accumulation in rice would benefit human nutrition and health. We generated transgenic rice expressing a gain-of-function mutant allele astol1 driven by the OsGPX1 promoter. astol1 encodes a plastid-localized O-acetylserine (thiol) lyase (OAS-TL) with Ser189Asn substitution (OsASTOL1S189N), which enhances cysteine biosynthesis by forming an indissociable cysteine synthase complex with its partner serine acetyltransferase (SAT). The effects on growth, As tolerance, and nutrient and As accumulation in rice grain were evaluated in hydroponic, pot and field experiments. The expression of OsASTOL1S189N in pOsGPX1::astol1 transgenic lines enhanced SAT activity, sulphate uptake, biosynthesis of cysteine, glutathione, phytochelatins and nicotianamine, and enhanced tolerance to As. The expression of OsASTOL1S189N decreased As accumulation while increased the accumulation of multiple macronutrients (especially sulphur, nitrogen and potassium) and micronutrients (especially zinc and selenium) in rice grain in a pot experiment and two field experiments, and had little effect on plant growth and grain yield. Our study provides a new strategy to genetically engineer rice to biofortify multiple essential nutrients, reducing As accumulation in rice grain and enhancing As tolerance simultaneously.

Determination of elements in cereals, pseudocereals, and legumes by microwave plasma-atomic emission spectrometry.

A novel method for multi-element analysis in cereals, pseudocereals, and legumes was developed for principal (calcium, magnesium, potassium, and phosphorus) and trace (manganese, zinc, iron, copper, and aluminum) element determination using a microwave plasma-atomic emission spectrometer (MP-AES). The method was validated using certified reference analyte values from durum wheat (DUWF-1), corn bran (BRAN-1), quinoa (KINO-1), rice (SRM 1568b), and soy (SRM 3234). Spike recoveries were assessed using field-grown crops that represent staple and minor crops with variable matrix compositions. A closed-vessel microwave-assisted digestion method consisting of 12 mL of deionized water, 2 mL of HNO3, and 2 mL of H2O2 was efficient for the mineralization of all crops. Acceptable measurement agreement was achieved between certified and determined values for all reference materials with recovery ranges from 89 to 120 percent. Plant breeders can use the method to develop and screen crops for improved nutrient density.

Bridging agro-science and human nutrition: zinc nanoparticles and biochar as catalysts for enhanced crop productivity and biofortification.

The integration of zinc nanoparticles (Zn NPs) with biochar offers a transformative approach to sustainable agriculture by enhancing plant productivity and human nutrition. This combination improves soil health, optimizes nutrient uptake, and increases resilience to environmental stressors, leading to superior crop performance. Our literature review shows that combining Zn NPs with biochar significantly boosts the crop nutrient composition, including proteins, vitamins, sugars, and secondary metabolites. This enhancement improves the plant tolerance to environmental challenges, crop quality, and shelf life. This technique addresses the global issue of Zn deficiency by biofortifying food crops with increased Zn levels, such as mung beans, lettuce, tomatoes, wheat, maize, rice, citrus, apples, and microgreens. Additionally, Zn NPs and biochar improve soil properties by enhancing water retention, cation exchange capacity (CEC), and microbial activity, making soils more fertile and productive. The porous structure of biochar facilitates the slow and sustained release of Zn, ensuring its bioavailability over extended periods and reducing the need for frequent fertilizer applications. This synergy promotes sustainable agricultural practices and reduces the environmental footprint of the traditional farming methods. However, potential ecological risks such as biomagnification, nanoparticle accumulation, and toxicity require careful consideration. Comprehensive risk assessments and management strategies are essential to ensure that agricultural benefits do not compromise the environmental or human health. Future research should focus on sustainable practices for deploying Zn NPs in agriculture, balancing food security and ecological integrity and positioning this approach as a viable solution for nutrient-efficient and sustainable agriculture.

Prospects for mineral biofortification of wheat: classical breeding and agronomy.

Low intake of micro- and macroelements and vitamins in food negatively affects the health of more than two billion people around the world provoking chronic diseases. For the majority of the world's population, these are soft and durum wheats that provide beneficial nutrients, however their modern high-yielding varieties have a significantly depleted grain mineral composition that have reduced mineral intake through food. Biofortification is a new research trend, whose main goal is to improve the nutritional qualities of agricultural crops using a set of classical (hybridization and selection) methods as well and the modern ones employing gene/QTL mapping, bioinformatic analysis, transgenesis, mutagenesis and genome editing. Using the classical breeding methods, biofortified varieties have been bred as a part of various international programs funded by HarvestPlus, CIMMYT, ICARDA. Despite the promise of transgenesis and genome editing, these labor-intensive methods require significant investments, so these technologies, when applied to wheat, are still at the development stage and cannot be applied routinely. In recent years, the interest in wheat biofortification has increased due to the advances in mapping genes and QTLs for agronomically important traits. The new markers obtained from wheat genome sequencing and application of bioinformatic methods (GWAS, meta-QTL analysis) has expanded our knowledge on the traits that determine the grain mineral concentration and has identified the key gene candidates. This review describes the current research on genetic biofortification of wheat in the world and in Russia and provides information on the use of cultivated and wild-relative germplasms to expand the genetic diversity of modern wheat varieties.

Zinc oxide nanoparticles mediated salinity stress mitigation in Pisum sativum: a physio-biochemical perspective.

Salinity is the major abiotic stress among others that determines crop productivity. The primary goal is to examine the impact of Zinc Oxide Nanoparticles (ZnO NPs) on the growth, metabolism, and defense systems of pea plants in simulated stress conditions. The ZnO NPs were synthesized via a chemical process and characterized by UV, XRD, and SEM. The ZnO NPs application (50 and 100) ppm and salt (50 mM and 100 mM) concentrations were carried out individually and in combination. At 50 ppm ZnO NPs the results revealed both positive and negative effects, demonstrating an increase in the root length and other growth parameters, along with a decrease in Malondialdehyde (MDA) and hydrogen peroxide concentrations. However, different concentrations of salt (50 mM and 100 mM) had an overall negative impact on all assessed parameters. In exploring the combined effects of ZnO NPs and salt, various concentrations yielded different outcomes. Significantly, only 50 mM NaCl combined with 50 ppm ZnO NPs demonstrated positive effects on pea physiology, leading to a substantial increase in root length and improvement in other physiological parameters. Moreover, this treatment resulted in decreased levels of MAD, Glycine betaine, and hydrogen peroxide. Conversely, all other treatments exhibited negative effects on the assessed parameters, possibly due to the high concentrations of both stressors. The findings offered valuble reference data for research on the impact of salinity on growth parameters of future agriculture crop.

Empowering agriculture: The promise of zinc biofortification in rice.

Zinc (Zn) plays a crucial role in metabolism in both plant and animal life. Zn deficiency is a worldwide problem that has recently gotten worse. This micronutrient shortage can be largely attributed to eating foods that are poor in zinc. If biofortification methods were widely used, Zn enrichment of the organ or tissue of interest would increase dramatically. However, Zn absorption mechanisms in rice plants must be understood on a fundamental level before these methods can be used effectively. Plant systems' Zn transporters and metal chelators play a major role in regulating this intricate physiological characteristic. The Zn efficiency of specific species is affected by a variety of factors, including the plant's growth stage, edaphic conditions, the time of year, and more. Both old and new ways of breeding plants can be used for biofortification. We have highlighted the significance of recombinant and genetic approaches to biofortifying in rice. In this review, we have the metabolic role of zinc in rice, and the different transporter families involved in the transportation of zinc in rice. We have also discussed the combined approaches of agronomic and genetic in zinc biofortification in rice and potential outcomes and future predictions.

A short stature allele enhances tolerance to zinc deficiency and translocation of zinc in barley.

Background: Zinc (Zn) content is of great importance in healthy human diet, crop productivity and stress tolerance in soils with zinc deficiency. The genes used to increase yield per unit area such as semi-dwarf 1 (sdw1) is commonly considered to reduce mineral content of grain. Methods: In the present study, influence of sdw1.d, a widely used allele for short plant height in barley breeding, on zinc accumulation and tolerance to zinc deficiency were investigated. A near isogenic line of sdw1.d allele, its recurrent parent Tokak 157/37 and donor parent Triumph were grown in zinc-deficient and-sufficient hydroponic cultures. Two experiments were conducted until heading stage and physiological maturity. Results: In zinc-deficient conditions, sdw1.d allele increased shoot dry weight by 112.4 mg plant-1, shoot Zn concentration by 0.9 ppm, but decreased root Zn concentration by 6.6 ppm. It did not affect grain characteristics, but increased grain Zn content. In zinc-sufficient conditions, sdw1.d allele increased shoot Zn content, and decreased root Zn content. sdw1.d did not affect grain weight but increased grain Zn concentration by about 30% under zinc-sufficient conditions. The results showed that sdw1.d allele has no negative effect on tolerance to zinc deficiency, and even promotes tolerance to zinc deficiency by more Zn translocation. It was revealed that sdw1.d allele improves Zn accumulation under both zinc-deficient and zinc-sufficient condition. The sdw1.d allele could contribute to solving the problems in plant growth and development caused by zinc-deficiency via improving tolerance to zinc-deficiency. It could also provide a better Zn biofortification.

Functional characterization of the riboflavin-overproducing and dextran-producing Weissella cibaria BAL3C-5 C120T strain for the development of biofortified plant-based beverages.

Riboflavin (vitamin B2) is essential for human beings and it has to be provided by healthy nutrition. The use of fermentation with riboflavin-overproducing lactic acid bacteria (LAB) represents an ideal strategy to generate, by in situ biofortification, functional drinks. These beverages can positively contribute to consumer health and address nutritional deficiencies. In the present work, the functional capabilities of Weissella cibaria BAL3C-5 C120T for riboflavin-overproduction and dextran-production during fermentation of oat-, rice-, soybean- and almond-based drinks have been evaluated. It was confirmed that the strain was capable of producing riboflavin and dextran in the analysed drinks. This property was especially pronounced in the oat-based drink, where after 24 h of fermentation the strain was able to increase riboflavin and dextran levels up to 3.4 mg/L and 3.2 g/L, respectively. Moreover, under optimized conditions the strain was able to enrich the fermented oat-based drinks with the prebiotic oligosaccharide panose (up to 6.6 g/L). In addition, in the oat-based drinks BAL3C-5 C120T showed a good pH-lowering ability (from 7.0 to 3.8) as well as a high 80 % cell viability after one month of storage. Rheological analysis of the resulting fermented oat-based beverages revealed a thixotropic structure related to a gel-like behaviour which was not observed in the non-fermented control drinks. In summary, these results confirmed the unique characteristics of W. cibaria BAL3C-5 C120T strain for the development of biofortified and functional plant-based beverages with improved nutritional and rheological properties. Analysis of the BAL3C-5 C120T strain survival under gastrointestinal conditions and its autoaggregation properties, also indicated its potential use as a probiotic delivered in an oat-based fermented beverage. In this context, this study also promotes the utilization of W. cibaria species in health and food industries where it has not yet been used as a starter or adjunct culture.

Discovery of a conserved translationally repressive upstream open reading frame within the iron-deficiency response regulator IDEF2.

BACKGROUND: Iron (Fe) deficiency affects 30-50% of the world's population. Genetic biofortification of staple crops is a promising strategy for improving human nutrition, but the number of effective precision breeding targets for Fe biofortification is small. Upstream open reading frames (uORFs) are cis-regulatory elements within the 5' leader sequence (LS) of genes that generally repress translation of the main open reading frame (mORF). RESULTS: We aligned publicly available rice (Oryza sativa L.) ribo-seq datasets and transcriptomes to identify putative uORFs within important Fe homeostasis genes. A dual luciferase assay (DLA) was used to determine whether these uORFs cause repression of mORF translation and pinpoint LS regions that can be mutated for mORF derepression. A translationally repressive uORF region was identified in two positive regulators of the Fe-deficiency response: IDEF1 and IDEF2. The IDEF2-uORF peptide was highly conserved among monocots and a mutation series in the 5' LS of the wheat (Triticum aestivum L.) TaIDEF2-A1 gene demonstrated variable mORF derepression. CONCLUSIONS: Together these results reveal a possible regulatory mechanism by which IDEF2 transcription factors modulate the Fe deficiency response in monocots, and highlight novel precision breeding targets to improve crop nutrition and abiotic stress tolerance.

The use of propionic and lactic acid bacteria to produce cobalamin and folate in injera, an Ethiopian cereal-based fermented food.

Like in many developing countries, the traditional Ethiopian diet relies mainly on starchy staple foods and often lacks sufficient animal-sourced foods which are crucial for cobalamin intake. Furthermore, the concentration of folate in traditionally prepared injera, an Ethiopian cereal-based fermented staple food, is highly variable and injera contains biologically inactive corrinoids. This study aimed to improve the cobalamin and folate content of injera by using cobalamin-producing Propionibacterium freudenreichii and folate-producing Lactiplantibacillus plantarum strains, both individually and combined. Since injera is fermented using backslopping, we also assessed the ability of these strains to produce cobalamin and folate consistently across successive fermentation batches. Changes in the microbial ecosystem were monitored using real-time PCR. The theoretical contribution of the injera prepared using the selected strains to the cobalamin and folate intake of children and women of reproductive age was also calculated. Results showed that using the selected bacterial strains individually increased cobalamin (up to 19.2 µg/100 g of dry matter) and folate (up to 180.2 µg/100 g of dry matter) levels in the injera dough over several backslopping fermentation batches. Regular consumption of the injera with enhanced vitamin content produced using each strain alone would be capable of fulfilling the entire recommended nutrient intake for cobalamin and up to 29 % of the recommended intake for folate for children and women of reproductive age. However, when the strains were used together, the production of both vitamins was reduced. The presence of certain common endogenous bacterial species and genera exhibited significant variability, highlighting the complex response of the native microbiota to the different inoculation strategies employed. Future experiments should consider selecting a microbial consortium comprising non-competing microorganisms to ensure the simultaneous production of cobalamin and folate in fermented foods.

Foliar Sprays of Multi-Nutrient Fertilizer Containing Selenium Produce Functional Tomato Fruits with Higher Shelf Life.

Selenium (Se) is a nutrient whose daily intake is often below the recommended levels in people. Biofortification with Se is a method to increase this intake by raising the Se concentration in tomato fruits, an effect dependent on sources and modes of application. Additionally, Se application can promote the enhancement of other compounds in tomato fruits, altering their metabolism, which may increase the fruit's shelf life. This study aimed to determine how different strategies of applying a multi-nutrient fertilizer containing Se (SeMNF) can increase the Se content and other bioactive compounds and enhance the shelf life of tomato (Solanum lycopersicum L.) fruits. Different foliar fertilization strategies involving the use of SeMNF were evaluated in field trials conducted on commercial tomato crops. Indeterminate-growth tomatoes were used, and different Se doses and application strategies were tested. Harvesting was conducted in three phases according to fruit ripening. Each harvested fruit was assessed for the Se content, macro and micronutrients, total phenolic compounds, vitamin C, antioxidant activity, carotenoids, pH, total titratable acidity, and total soluble solids in tomato fruits. Doses of 15 g ha-1 of Se, split into three applications, increased the Se content in the fruits at 1 and 2 harvests. The application of SeMNF at Se doses above 10 g of Se ha-1 increased firmness, days of ripening, and the nutritional quality of the tomatoes (higher contents of carotenoids (+39%), lycopene (+33%), antioxidant activity (+16%), total phenolic compounds (+38%), and vitamin C (+14%) in a dose-dependent effect of the application strategy used. These results contributed to an increase in the shelf life of tomatoes, consequently reducing food waste.

Agronomic Biofortification of Plants with Iodine and Selenium: A Potential Solution for Iodine and Selenium Deficiencies.

Iodine and selenium deficiencies are widespread both in developed countries and developing countries. The soil is the fundamental source of iodine and selenium for plants, and iodine and/or selenium-depleted soil restrains the cultivation of crops to cover recommended daily intakes of iodine and selenium. Although food fortification strategies, including salt iodization, increase the dietary intake of these minerals, their global deficiencies have not been eliminated. Therefore, new strategies have been developed to prevent iodine and selenium deficiencies, and biofortification is one of them. The aim of this review is to assert the outcomes of the studies that investigate the optimum conditions for biofortification with iodine and selenium and to recognize the role of biofortification practices as a potential solution for preventing iodine and selenium deficiencies. The findings of studies show that biofortification with iodine and selenium can be a solution for iodine and selenium deficiencies. Agronomic biofortification is currently a more convenient method to increase selenium and iodine contents in plants. However, the most effective agronomic biofortification conditions are crucial to acquire biofortified food. Moreover, increasing the awareness of the producers and consumers on biofortification has a determinative role in the achievement of biofortification practices for human health. Although research about iodine and selenium biofortification has been increased, the effectiveness of biofortified foods to meet recommended daily intakes is still unknown. More research is needed to understand most effective biofortification conditions for plants and bioavailability of biofortified foods for humans.

Boosting pro-vitamin A content and bioaccessibility in leaves by combining engineered biosynthesis and storage pathways with high-light treatments.

Biofortification of green leafy vegetables with pro-vitamin A carotenoids, such as ß-carotene, has remained challenging to date. Here, we combined two strategies to achieve this goal. One of them involves producing ß-carotene in the cytosol of leaf cells to avoid the negative impacts on photosynthesis derived from changing the balance of carotenoids and chlorophylls in chloroplasts. The second approach involves the conversion of chloroplasts into non-photosynthetic, carotenoid-overaccumulating chromoplasts in leaves agroinfiltrated or infected with constructs encoding the bacterial phytoene synthase crtB, leaving other non-engineered leaves of the plant to sustain normal growth. A combination of these two strategies, referred to as strategy C (for cytosolic production) and strategy P (for plastid conversion mediated by crtB), resulted in a 5-fold increase in the amount of ß-carotene in Nicotiana benthamiana leaves. Following several attempts to further improve ß-carotene leaf contents by metabolic engineering, hormone treatments and genetic screenings, it was found that promoting the proliferation of plastoglobules with increased light-intensity treatments not only improved ß-carotene accumulation but it also resulted in a much higher bioaccessibility. The combination of strategies C and P together with a more intense light treatment increased the levels of accessible ß-carotene 30-fold compared to controls. We further demonstrated that stimulating plastoglobule proliferation with strategy P, but also with a higher-light treatment alone, also improved ß-carotene contents and bioaccessibility in edible lettuce (Lactuca sativa) leaves.

Assessing the carotenoid profiles and allelic diversity of yellow maize inbred lines adapted to mid-altitude subhumid maize agroecology in Ethiopia.

Biofortification of provitamin A in maize is an attractive and sustainable remedy to the problem of vitamin A deficiency in developing countries. The utilization of molecular markers represents a promising avenue to facilitate the development of provitamin A (PVA)-enriched maize varieties. We screened 752 diverse tropical yellow/orange maize lines using kompetitive allele-specific PCR (KASP) makers to validate the use of KASP markers in PVA maize breeding. To this end, a total of 161 yellow/orange inbred lines, selected from among the 752 lines, were evaluated for their endosperm PVA and other carotenoid compounds levels in two separate trials composed of 63 and 98 inbred lines in 2020 and 2021, respectively. Significant differences (p < 0.001) were observed among the yellow maize inbred lines studied for all carotenoid profiles. An inbred line TZMI1017, introduced by the International Institute of Tropical Agriculture (IITA) showed the highest level of PVA (12.99 µg/g) and ß-carotene (12.08 µg/g). The molecular screening showed 43 yellow maize inbred lines carrying at least three of the favorable alleles of the KASP markers. TZMI1017 inbred line also carried the favorable alleles of almost all markers. In addition, nine locally developed inbred lines had medium to high PVA concentrations varying from 5.11 µg/g to 10.76 µg/g and harbored the favorable alleles of all the KASP PVA markers. Association analysis between molecular markers and PVA content variation in the yellow/orange maize inbred lines did not reveal a significant, predictable correlation. Further investigation is warranted to elucidate the underlying genetic architecture of the PVA content in this germplasm. However, we recommend strategic utilization of the maize-inbred lines with higher PVA content to enhance the PVA profile of the breeding program's germplasm.

The 4T and 7T introgressions from Amblyopyrum muticum and the 5Au introgression from Triticum urartu increases grain zinc and iron concentrations in Malawian wheat backgrounds.

Micronutrient deficiencies (MNDs) particularly zinc (Zn) and iron (Fe) remain widespread in sub-Saharan Africa (SSA) due to low dietary intake. Wheat is an important source of energy globally, although cultivated wheat is inherently low in grain micronutrient concentrations. Malawian wheat/Am. muticum and Malawian wheat/T. urartu BC1F3 introgression lines, developed by crossing three Malawian wheat varieties (Kenya nyati, Nduna and Kadzibonga) with DH-348 (wheat/Am. muticum) and DH-254 (wheat/T. urartu), were phenotyped for grain Zn and Fe, and associated agronomic traits in Zn-deficient soils, in Malawi. 98% (47) of the BC1F3 introgression lines showed higher Zn above the checks Paragon, Chinese Spring, Kadzibonga, Kenya Nyati and Nduna. 23% (11) of the introgression lines showed a combination of high yields and an increase in grain Zn by 16-30 mg kg -1 above Nduna and Kadzibonga, and 11-25 mg kg -1 above Kenya nyati, Paragon and Chinese Spring. Among the 23%, 64% (7) also showed 8-12 mg kg -1 improvement in grain Fe compared to Nduna and Kenya nyati. Grain Zn concentrations showed a significant positive correlation with grain Fe, whilst grain Zn and Fe negatively and significantly correlated with TKW and grain yield. This work will contribute to the efforts of increasing mineral nutrient density in wheat, specifically targeting countries in the SSA.

Assessing a Fermented Whey Beverage Biofortified with Folate as a Potential Folate Source for Humans.

Folate, a vital water-soluble vitamin (B9), requires specific attention as its recommended daily intake frequently is not reached in countries without mandatory fortification. In this regard, biofortification with microorganisms like Bifidobacterium and Streptococcus offers a compelling approach for enhancing food with natural folates. A randomized, nonblinded, and monocentric human pilot study is conducted to assess the bioavailability of a folate-biofortified fermented whey beverage, comprising 3 intervention days and a controlled replenishment phase before and during the assay. Folate plasma concentration (5-CH3-H4folate) is determined using a stable isotope dilution assay and LC-MS/MS detection. Biokinetic parameters (cmax and tmax) are determined, and areas under the curve (AUC) normalized to the basal folate plasma concentration are calculated. An average bioavailability of 17.1% in relation to the 5-CH3-H4folate supplement, ranging from 0% to 39.8%, is obtained. These results reiterate the significance of additional research into folate bioavailability in general and dairy products. Further investigations are warranted into folate-binding proteins (FBP) and other potential limiting factors within the food and individual factors. In summary, biofortification via fermentation emerges as a promising avenue for enhancing the natural folate content in dairy and other food products.

Validation of SNP Markers for Diversity Analysis, Quality Control, and Trait Selection in a Biofortified Cassava Population.

A validated marker system is crucial to running an effective genomics-assisted breeding program. We used 36 Kompetitive Allele-Specific PCR (KASP) markers to genotype 376 clones from the biofortified cassava pipeline, and fingerprinted 93 of these clones with DArTseq markers to characterize breeding materials and evaluate their relationships. The discriminating ability of the 36-quality control (QC) KASP and 6602 DArTseq markers was assessed using 92 clones genotyped in both assays. In addition, trait-specific markers were used to determine the presence or absence of target genomic regions. Hierarchical clustering identified two major groups, and the clusters were consistent with the breeding program origins. There was moderate genetic differentiation and a low degree of variation between the identified groups. The general structure of the population was similar using both assays. Nevertheless, KASP markers had poor resolution when it came to differentiating the genotypes by seed sources and overestimated the prevalence of duplicates. The trait-linked markers did not achieve optimal performance as all markers displayed variable levels of false positive and/or false negative. These findings represent the initial step in the application of genomics-assisted breeding for the biofortified cassava pipeline, and will guide the use of genomic selection in the future.

OsWNK9 regulates cadmium concentration in brown rice by restraining cadmium transport from straw to brown rice.

Selecting and breeding rice cultivars that enable strong cadmium (Cd) accumulation in rice straw but low accumulation in brown rice is a promising way to achieve Cd phytoremediation as well as to ensure the food safety of rice. Herein, we isolated a gene OsWNK9 from the quantitative trait locus associated with reducing Cd translocation from rice straw to brown rice and decreasing the Cd concentration in brown rice (BRCdC). Continuous strong expression of OsWNK9 was observed in nodes and internode and was induced after Cd supply. OsWNK9 was localized in the rice cell nucleus and participated in the regulation of Cd transport in yeast. Two independent oswnk9 rice mutants were generated via CRISPR/Cas9 gene-editing and showed significantly higher BRCdC than that of the wild type (WT). The BRCdC of knockout oswnk9 mutants was 0.227 mg kg-1and 0.238 mg kg-1, increased by 14 % and 19 % compared with that of the WT due to the lower Cd allocation in the basal stem, internode, and node III, which was unrelated to Cd uptake. Interestingly, OsWNK9 could promote iron (Fe) accumulation in rice under Cd-contaminated conditions, suggesting that OsWNK9 is an ideal gene for Cd phytoremediation and Fe biofortification in rice to support safe food production.

Soil and Foliar Zinc Biofortification of Triticale (x Triticosecale) under Mediterranean Conditions: Effects on Forage Yield and Quality.

Zinc (Zn) deficiency represents a significant global concern, affecting both plant and human health, particularly in regions with Zn-depleted soils. Agronomic biofortification strategies, such as the application of Zn fertilizers, offer a cost-effective approach to increase Zn levels in crops. This study aimed to assess the efficacy of soil and foliar Zn biofortification, applied as an aqueous solution of 0.5% zinc sulphate (ZnSO4·7H2O), on triticale (x Triticosecale) grown under Mediterranean conditions. The study was conducted over two growing seasons (2017/18 and 2018/19) in southern Spain, evaluating the effects on biomass yield; forage quality, including crude protein, Van Soest detergent fiber, organic matter digestibility, and relative forage value; and nutrient accumulation. Soil treatment consisted in the application of 50 kg of ZnSO4·7H2O ha-1 solely at the beginning of the first campaign to assess the residual effect on the second year. In contrast, the foliar treatment consisted of two applications of 4 kg of ZnSO4·7H2O ha-1 per campaign, one at the beginning of tillering and the other at the appearance of the first node. The foliar application increased the Zn content of the forage to adequate levels, while the soil application resulted in a 33% increase in biomass production, which is particularly beneficial for farmers. Overall quality was favored by the combined soil + foliar application, and no adverse antagonistic effects on other nutrients were detected. Instead, a synergistic interaction between Se and Zn was observed, which improved the efficacy of this important micronutrient for livestock and human wellbeing.