Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudiana

Nanotechnology, particularly the use of iron oxide nanoparticles (IONPs), has gained significant attention in agricultural research due to its potential to enhance plant growth, development, and stress tolerance. However, the green synthesis of IONPs using plant extracts remains underexplored, espec...

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Main Authors: Sher Muhammad, Abid Ali Khan, Muhammad Rameez Khan, Sidra Mukhtar, Abeer Kazmi, Amir Ali, Ayesha Siddiqa, Kayley Aileen Hernández Ramírez, Juan Pedro Luna-Arias, Gabriela Medina-Pérez, Armando Pelaez-Acero, Silvia Armenta, Ajaz Ahmad
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Plant Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2025.1569613/full
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author Sher Muhammad
Abid Ali Khan
Muhammad Rameez Khan
Sidra Mukhtar
Abeer Kazmi
Abeer Kazmi
Amir Ali
Amir Ali
Ayesha Siddiqa
Kayley Aileen Hernández Ramírez
Juan Pedro Luna-Arias
Gabriela Medina-Pérez
Armando Pelaez-Acero
Silvia Armenta
Ajaz Ahmad
author_facet Sher Muhammad
Abid Ali Khan
Muhammad Rameez Khan
Sidra Mukhtar
Abeer Kazmi
Abeer Kazmi
Amir Ali
Amir Ali
Ayesha Siddiqa
Kayley Aileen Hernández Ramírez
Juan Pedro Luna-Arias
Gabriela Medina-Pérez
Armando Pelaez-Acero
Silvia Armenta
Ajaz Ahmad
author_sort Sher Muhammad
collection DOAJ
description Nanotechnology, particularly the use of iron oxide nanoparticles (IONPs), has gained significant attention in agricultural research due to its potential to enhance plant growth, development, and stress tolerance. However, the green synthesis of IONPs using plant extracts remains underexplored, especially in the context of agricultural applications. In this study, the green synthesis of IONPs using Moringa oleifera leaf extract is reported, with the extract serving as both a reducing and capping agent. The synthesized nanoparticles were characterized using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX), revealing spherical and polygonal shapes with an iron peak at 6.5-7.5 keV, consistent with the expected size and composition. These IONPs were incorporated into Murashige and Skoog (MS) medium to replace the conventional iron source and evaluate their effects on Stevia rebaudiana micropropagation. The results demonstrate that IONPs at lower concentrations (5.60 mg/L) significantly promoted early shoot and root initiation (5.2 and 5.3 days, respectively), while higher concentrations (11.20 mg/L and 22.40 mg/L) delayed growth initiation and inhibited development. Notably, 22.4 mg/L IONPs enhanced leaf growth (length: 3.20 cm, width: 1.90 cm), fresh weight (238.90 mg), and dry weight (20.67 mg), outperforming the positive control (FeSO4·7H2O). IONPs also increased the total phenolic content (TPC) and total flavonoid content (TFC) in plant tissues, with the highest values (4.54 mg GAE/g and 2.07 mg QAE/g) observed at 22.40 mg/L. The antioxidant capacity, measured by DPPH scavenging activity, was significantly enhanced, reaching 89.70%. Additionally, IONPs promoted the accumulation of diterpene glycosides, including stevioside (4.30 mg/g DW) and rebaudioside A (6.70 mg/g DW), especially at higher concentrations. These findings suggest that IONPs, particularly at 22.40 mg/L, are a promising and environmentally friendly alternative to traditional iron sources, offering enhanced plant growth, improved antioxidant defenses, and increased production of valuable secondary metabolites in S. rebaudiana.
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publisher Frontiers Media S.A.
record_format Article
series Frontiers in Plant Science
spelling doaj-art-2691e2b1f95b4d81a9aafd112af3e25b2025-08-20T03:14:19ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-04-011610.3389/fpls.2025.15696131569613Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudianaSher Muhammad0Abid Ali Khan1Muhammad Rameez Khan2Sidra Mukhtar3Abeer Kazmi4Abeer Kazmi5Amir Ali6Amir Ali7Ayesha Siddiqa8Kayley Aileen Hernández Ramírez9Juan Pedro Luna-Arias10Gabriela Medina-Pérez11Armando Pelaez-Acero12Silvia Armenta13Ajaz Ahmad14Biotechnology Laboratory, Agricultural Research Institute (ARI) Tarnab Peshawar, Peshawar, PakistanDepartment of Chemical Sciences, University of Laki Marwat, Khyber Pakhtunkhwa, PakistanBiotechnology Laboratory, Agricultural Research Institute (ARI) Tarnab Peshawar, Peshawar, PakistanBiotechnology Laboratory, Agricultural Research Institute (ARI) Tarnab Peshawar, Peshawar, PakistanThe State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, ChinaThe Key Laboratory of Aquatic Biodiversity and Conservation of Chinese Academy of Sciences, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei, ChinaDepartment of Botany, PMAS Arid Agriculture University, Rawalpindi, PakistanDepartment of Cell Biology, and Nanoscience and Nanotechnology Ph.D. Program, Center for Research and Advanced Studies of the National Polytechnic Institute (CINVESTAV), Mexico City, MexicoDepartment of Botany, PMAS Arid Agriculture University, Rawalpindi, PakistanInstitute of Agricultural Sciences, Autonomous University of the State of Hidalgo, Hidalgo, MexicoDepartment of Cell Biology, and Nanoscience and Nanotechnology Ph.D. Program, Center for Research and Advanced Studies of the National Polytechnic Institute (CINVESTAV), Mexico City, MexicoInstitute of Agricultural Sciences, Autonomous University of the State of Hidalgo, Hidalgo, MexicoInstitute of Agricultural Sciences, Autonomous University of the State of Hidalgo, Hidalgo, MexicoInstitute of Agricultural Sciences, Autonomous University of the State of Hidalgo, Hidalgo, MexicoDepartment of Clinical Pharmacy, College of Pharmacy, King Saud University, Riyadh, Saudi ArabiaNanotechnology, particularly the use of iron oxide nanoparticles (IONPs), has gained significant attention in agricultural research due to its potential to enhance plant growth, development, and stress tolerance. However, the green synthesis of IONPs using plant extracts remains underexplored, especially in the context of agricultural applications. In this study, the green synthesis of IONPs using Moringa oleifera leaf extract is reported, with the extract serving as both a reducing and capping agent. The synthesized nanoparticles were characterized using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX), revealing spherical and polygonal shapes with an iron peak at 6.5-7.5 keV, consistent with the expected size and composition. These IONPs were incorporated into Murashige and Skoog (MS) medium to replace the conventional iron source and evaluate their effects on Stevia rebaudiana micropropagation. The results demonstrate that IONPs at lower concentrations (5.60 mg/L) significantly promoted early shoot and root initiation (5.2 and 5.3 days, respectively), while higher concentrations (11.20 mg/L and 22.40 mg/L) delayed growth initiation and inhibited development. Notably, 22.4 mg/L IONPs enhanced leaf growth (length: 3.20 cm, width: 1.90 cm), fresh weight (238.90 mg), and dry weight (20.67 mg), outperforming the positive control (FeSO4·7H2O). IONPs also increased the total phenolic content (TPC) and total flavonoid content (TFC) in plant tissues, with the highest values (4.54 mg GAE/g and 2.07 mg QAE/g) observed at 22.40 mg/L. The antioxidant capacity, measured by DPPH scavenging activity, was significantly enhanced, reaching 89.70%. Additionally, IONPs promoted the accumulation of diterpene glycosides, including stevioside (4.30 mg/g DW) and rebaudioside A (6.70 mg/g DW), especially at higher concentrations. These findings suggest that IONPs, particularly at 22.40 mg/L, are a promising and environmentally friendly alternative to traditional iron sources, offering enhanced plant growth, improved antioxidant defenses, and increased production of valuable secondary metabolites in S. rebaudiana.https://www.frontiersin.org/articles/10.3389/fpls.2025.1569613/fullMoringa oleiferaStevia Rebaudianananotechnologyiron oxide nanoparticlessecondary metabolites
spellingShingle Sher Muhammad
Abid Ali Khan
Muhammad Rameez Khan
Sidra Mukhtar
Abeer Kazmi
Abeer Kazmi
Amir Ali
Amir Ali
Ayesha Siddiqa
Kayley Aileen Hernández Ramírez
Juan Pedro Luna-Arias
Gabriela Medina-Pérez
Armando Pelaez-Acero
Silvia Armenta
Ajaz Ahmad
Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudiana
Frontiers in Plant Science
Moringa oleifera
Stevia Rebaudiana
nanotechnology
iron oxide nanoparticles
secondary metabolites
title Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudiana
title_full Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudiana
title_fullStr Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudiana
title_full_unstemmed Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudiana
title_short Effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth, antioxidant activity, and stevioside accumulation in micro-propagated Stevia rebaudiana
title_sort effective substitution of ferrous sulfate with iron oxide nanoparticles enhances growth antioxidant activity and stevioside accumulation in micro propagated stevia rebaudiana
topic Moringa oleifera
Stevia Rebaudiana
nanotechnology
iron oxide nanoparticles
secondary metabolites
url https://www.frontiersin.org/articles/10.3389/fpls.2025.1569613/full
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