Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt Stress

Nanoparticles play an important role in plant response to abiotic stresses including salt stress. In this study, the physiological and histological responses of CuO, ZnO, and CaO nanoparticle (NP) applications on callus tissues developed from two <i>alfalfa</i> lines (Erzurum and Muş) ex...

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Main Authors: Mustafa Akçay, Merve Simsek Geyik, Busra Yazicilar, Fatma Boke, Hayrunnisa Nadaroglu, Okkes Atıcı, İsmail Bezirganoğlu
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Agronomy
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Online Access:https://www.mdpi.com/2073-4395/15/1/180
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author Mustafa Akçay
Merve Simsek Geyik
Busra Yazicilar
Fatma Boke
Hayrunnisa Nadaroglu
Okkes Atıcı
İsmail Bezirganoğlu
author_facet Mustafa Akçay
Merve Simsek Geyik
Busra Yazicilar
Fatma Boke
Hayrunnisa Nadaroglu
Okkes Atıcı
İsmail Bezirganoğlu
author_sort Mustafa Akçay
collection DOAJ
description Nanoparticles play an important role in plant response to abiotic stresses including salt stress. In this study, the physiological and histological responses of CuO, ZnO, and CaO nanoparticle (NP) applications on callus tissues developed from two <i>alfalfa</i> lines (Erzurum and Muş) exposed to salt (NaCl) stress were evaluated. The NPs were synthesized from the extracts obtained from healthy walnut shells using the green synthesis approach and then characterized by Scanning Electron Microscopy (SEM) and X-ray diffraction analysis (XRD). The leaf explants were placed in an MS medium containing 4 mg L<sup>−1</sup> 2,4-D (2,4-dichlorophenoxyacetic acid), 50 mM NaCl, and 0.8 ppm of NPs for 1 month in the dark. CaO NP is determined to be more effective than CuO and ZnO in callus induction from leaf explants. Malondialdehyde (MDA) content was higher in the callus treated with 0.8 ppm CuO NP + 50 mM NaCl compared to other treatments. The callus induction stage, without salt treatments, showed the best results with 0.8 ppm CaO NPs for both H<sub>2</sub>O<sub>2</sub> levels and peroxidase (POX) activity compared to the other NPs. The highest protein rate was obtained from the callus induction stage and callus formation stage after 50 mM treatment NaCl with 0.8 ppm CuO. The LCSM results displayed, under in vitro conditions, that the treatment of NPs can greatly suppress the negative effects of salt stress on calli samples. SEM analysis supported the results obtained by laser scanning confocal microscopy (LSCM) analysis. Our findings suggest that CuO, CaO, and ZnO NPs can offer a simple and effective method to protect <i>alfalfa</i> callus from NaCl stress severity. Furthermore, these NPs, particularly CaO, hold potential for broader application and should be evaluated under various abiotic conditions beyond salt stress.
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spelling doaj-art-df34e88f1d5b4cac85ba1900ce6a09412025-01-24T13:17:04ZengMDPI AGAgronomy2073-43952025-01-0115118010.3390/agronomy15010180Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt StressMustafa Akçay0Merve Simsek Geyik1Busra Yazicilar2Fatma Boke3Hayrunnisa Nadaroglu4Okkes Atıcı5İsmail Bezirganoğlu6Department of Chemistry and Chemical Processing Technologies, Kars Vocational School, Kafkas University, Kars 36000, TürkiyeDepartment of Molecular Biology and Genetics, Erzurum Technical University, Erzurum 25050, TürkiyeDepartment of Molecular Biology and Genetics, Erzurum Technical University, Erzurum 25050, TürkiyeDepartment of Molecular Biology and Genetics, Erzurum Technical University, Erzurum 25050, TürkiyeDepartment of Food Technology, Vocational College of Technical Sciences, Ataturk University, Erzurum 25240, TürkiyeDepartment of Biology, Faculty of Science and Arts, Atatürk University, Erzurum 25240, TürkiyeDepartment of Molecular Biology and Genetics, Erzurum Technical University, Erzurum 25050, TürkiyeNanoparticles play an important role in plant response to abiotic stresses including salt stress. In this study, the physiological and histological responses of CuO, ZnO, and CaO nanoparticle (NP) applications on callus tissues developed from two <i>alfalfa</i> lines (Erzurum and Muş) exposed to salt (NaCl) stress were evaluated. The NPs were synthesized from the extracts obtained from healthy walnut shells using the green synthesis approach and then characterized by Scanning Electron Microscopy (SEM) and X-ray diffraction analysis (XRD). The leaf explants were placed in an MS medium containing 4 mg L<sup>−1</sup> 2,4-D (2,4-dichlorophenoxyacetic acid), 50 mM NaCl, and 0.8 ppm of NPs for 1 month in the dark. CaO NP is determined to be more effective than CuO and ZnO in callus induction from leaf explants. Malondialdehyde (MDA) content was higher in the callus treated with 0.8 ppm CuO NP + 50 mM NaCl compared to other treatments. The callus induction stage, without salt treatments, showed the best results with 0.8 ppm CaO NPs for both H<sub>2</sub>O<sub>2</sub> levels and peroxidase (POX) activity compared to the other NPs. The highest protein rate was obtained from the callus induction stage and callus formation stage after 50 mM treatment NaCl with 0.8 ppm CuO. The LCSM results displayed, under in vitro conditions, that the treatment of NPs can greatly suppress the negative effects of salt stress on calli samples. SEM analysis supported the results obtained by laser scanning confocal microscopy (LSCM) analysis. Our findings suggest that CuO, CaO, and ZnO NPs can offer a simple and effective method to protect <i>alfalfa</i> callus from NaCl stress severity. Furthermore, these NPs, particularly CaO, hold potential for broader application and should be evaluated under various abiotic conditions beyond salt stress.https://www.mdpi.com/2073-4395/15/1/180callus<i>alfalfa</i>nanoparticleconfocal laser scanning analysisin vitro assay
spellingShingle Mustafa Akçay
Merve Simsek Geyik
Busra Yazicilar
Fatma Boke
Hayrunnisa Nadaroglu
Okkes Atıcı
İsmail Bezirganoğlu
Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt Stress
Agronomy
callus
<i>alfalfa</i>
nanoparticle
confocal laser scanning analysis
in vitro assay
title Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt Stress
title_full Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt Stress
title_fullStr Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt Stress
title_full_unstemmed Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt Stress
title_short Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintenance of <i>Alfalfa</i> (<i>Medicago sativa</i> L.) Under In Vitro Salt Stress
title_sort assessment of the effects of newly fabricated cao cuo zno nanoparticles on callus formation maintenance of i alfalfa i i medicago sativa i l under in vitro salt stress
topic callus
<i>alfalfa</i>
nanoparticle
confocal laser scanning analysis
in vitro assay
url https://www.mdpi.com/2073-4395/15/1/180
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