00b25489f8289362c11f18443c03702a agronomy-15-00580.pdf 9ea662df2fb5bd61d8645d478dedee6ab7297921 agronomy-15-00580.pdf db546145535212354e252f3bbd6d4395396a40a41e3b4b1fe8f91d2e35c994ae agronomy-15-00580.pdf Title: Enhancing Organic Selenium Forms in Alfalfa Forage Through Inorganic Selenium Foliar Application: Insights from Laboratory and Field Studies Using X-Ray Spectroscopy Subject: Selenium (Se) is an essential micronutrient, yet its deficiency remains a global concern. This study investigates the biofortification of alfalfa (Medicago sativa cv. ProINTA Super Monarca GR9) via foliar Se application to enhance Se accumulation and transformation into bioavailable organic forms. A controlled environment experiment in a plant growth chamber and a one-season open-field trial (January 2023, Argentina) were conducted. Treatments included sodium selenate (Se(VI)), sodium selenite (Se(IV)), and a 1:1 mixture, applied at 45 and 90 g Se ha-1, with and without the biostimulant BIOFORGE®. Treated plants exhibited increased Se content, correlating with the applied doses. X-ray absorption spectroscopy (XAS) confirmed that most inorganic Se was transformed into organic Se forms, with Se(IV) treatments yielding the highest concentrations of organic Se species such as selenocysteine (SeCys) and selenomethionine (SeMet). Open-field trials showed a complete conversion of Se, though total Se accumulation was lower than in controlled conditions. Se treatments did not affect forage quality or biomass production. The biostimulant slightly reduced Se uptake but did not compromise biofortification. These results highlight Se(IV) as the optimal treatment for alfalfa biofortification, presenting a sustainable strategy to enhance dietary Se intake through functional foods. Keywords: biofortification; selenate; selenite; direct speciation; synchrotron Author: María-Jesús Sánchez-Martín, Mónica Gaggiotti, Laura Simonelli, Carlo Marini, Federico Marini, Roberto Boada, Mercè Llugany, Manuel Valiente, Gabriel Céccoli, María Micaela Stoffel, Marcia Viltres-Portales, Juan Manuel Picco, María Gabriela Guevara and Fernando Felipe Muñoz Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Wed Feb 26 17:14:28 2025 CET ModDate: Wed Feb 26 17:19:23 2025 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 22 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2365260 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- WYEKQU+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 QKZKVH+VnURWPalladioL Type 1 Custom yes yes yes 16 0 NHKUHV+URWPalladioL-Roma Type 1 Custom yes yes yes 21 0 HDUGBN+URWPalladioL-Bold Type 1 Custom yes yes yes 27 0 VCESIG+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 QMDDXF+CMSY10 Type 1 Builtin yes yes yes 37 0 ZXDKIP+EURM10 Type 1 Builtin yes yes yes 63 0 HYYHEA+CMBSY10 Type 1 Builtin yes yes yes 129 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-04-09 02:08:52 CEST RepresentationInformation: agronomy-15-00580.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-04-08 08:39:38 CEST Size: 2365260 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 381 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Se-Biofortified Alfalfa at Laboratory Scale Destination: subsection.2.1 Item: Title: Application of Se Foliar Treatments Destination: subsection.2.2 Item: Title: Gas Exchange Measurements and Chlorophyll Index Determination Destination: subsection.2.3 Item: Title: Forage Quality Assessment Destination: subsection.2.4 Item: Title: Open-Field Se-Biofortification of Alfalfa Destination: subsection.2.5 Item: Title: Determination of Total Selenium, Macro- and Micronutrient Concentration Destination: subsection.2.6 Item: Title: Selenium Speciation Analysis by XAS Destination: subsection.2.7 Item: Title: Statistical Analysis Destination: subsection.2.8 Item: Title: Results and Discussion Destination: section.3 Children: Item: Title: Se-Biofortified Alfalfa at Laboratory Scale Destination: subsection.3.1 Children: Item: Title: Physiological Parameters Destination: subsubsection.3.1.1 Item: Title: Effect of Foliar Selenium Application on Yield Parameters and Selenium Content in Alfalfa Plants Grown Under Controlled Conditions Destination: subsubsection.3.1.2 Item: Title: Effects of Selenium on Forage Quality in Alfalfa Plants Grown Under Controlled Conditions Destination: subsubsection.3.1.3 Item: Title: Effects of Selenium on Mineral Elements in Alfalfa Plants Grown Under Controlled Conditions Destination: subsubsection.3.1.4 Item: Title: Selenium Speciation Analysis by XAS in Alfalfa Plants Grown Under Controlled Conditions Destination: subsubsection.3.1.5 Item: Title: Se-Biofortified Alfalfa at Open-Field Destination: subsection.3.2 Children: Item: Title: Effects of Selenium on Yield and Forage Quality Destination: subsubsection.3.2.1 Item: Title: Effect of Foliar Selenite Application on Selenium Content in Alfalfa Plants Grown Under Open-Field Conditions Destination: subsubsection.3.2.2 Item: Title: Effects of Selenium on Mineral Elements in Alfalfa Plants Grown Under Open-Field Conditions Destination: subsubsection.3.2.3 Item: Title: Selenium Speciation Analysis by XAS in Alfalfa Plants Grown Under Open-Field Conditions Destination: subsubsection.3.2.4 Item: Title: Conclusions Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: Enhancing Organic Selenium Forms in Alfalfa Forage Through Inorganic Selenium Foliar Application: Insights from Laboratory and Field Studies Using X-Ray Spectroscopy Author: María-Jesús Sánchez-Martín, Mónica Gaggiotti, Laura Simonelli, Carlo Marini, Federico Marini, Roberto Boada, Mercè Llugany, Manuel Valiente, Gabriel Céccoli, María Micaela Stoffel, Marcia Viltres-Portales, Juan Manuel Picco, María Gabriela Guevara and Fernando Felipe Muñoz Subject: Selenium (Se) is an essential micronutrient, yet its deficiency remains a global concern. This study investigates the biofortification of alfalfa (Medicago sativa cv. ProINTA Super Monarca GR9) via foliar Se application to enhance Se accumulation and transformation into bioavailable organic forms. A controlled environment experiment in a plant growth chamber and a one-season open-field trial (January 2023, Argentina) were conducted. Treatments included sodium selenate (Se(VI)), sodium selenite (Se(IV)), and a 1:1 mixture, applied at 45 and 90 g Se ha-1, with and without the biostimulant BIOFORGE®. Treated plants exhibited increased Se content, correlating with the applied doses. X-ray absorption spectroscopy (XAS) confirmed that most inorganic Se was transformed into organic Se forms, with Se(IV) treatments yielding the highest concentrations of organic Se species such as selenocysteine (SeCys) and selenomethionine (SeMet). Open-field trials showed a complete conversion of Se, though total Se accumulation was lower than in controlled conditions. Se treatments did not affect forage quality or biomass production. The biostimulant slightly reduced Se uptake but did not compromise biofortification. These results highlight Se(IV) as the optimal treatment for alfalfa biofortification, presenting a sustainable strategy to enhance dietary Se intake through functional foods. 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