Toxicity of copper hydroxide nanoparticles, bulk copper hydroxide, and ionic copper to alfalfa plants: A spectroscopic and gene expression study
2018
Cota-Ruiz, Keni | Hernández-Viezcas, José A. | Varela-Ramírez, Armando | Valdés, Carolina | Núñez-Gastélum, José A. | Martínez-Martínez, Alejandro | Delgado-Rios, Marcos | Peralta-Videa, Jose R. | Gardea-Torresdey, Jorge L.
Bulk Cu compounds such as Cu(OH)₂ are extensively used as pesticides in agriculture. Recent investigations suggest that Cu-based nanomaterials can replace bulk materials reducing the environmental impacts of Cu. In this study, stress responses of alfalfa (Medicago sativa L.) seedlings to Cu(OH)₂ nanoparticle or compounds were evaluated. Seeds were immersed in suspension/solutions of a Cu(OH)₂ nanoform, bulk Cu(OH)₂, CuSO₄, and Cu(NO₃)₂ at 25 and 75 mg/L. Six days later, the germination, seedling growth, and the physiological and biochemical responses of sprouts were evaluated. All Cu treatments significantly reduced root elongation (average = 63%). The ionic compounds at 25 and 75 mg/L caused a reduction in all elements analyzed (Ca, K, Mg, P, Zn, and Mn), excepting for S, Fe and Mo. The bulk-Cu(OH)₂ treatment reduced K (48%) and P (52%) at 75 mg/L, but increased Zn at 25 (18%) and 75 (21%) mg/L. The nano-Cu(OH)₂ reduced K (46%) and P (48%) at 75 mg/L, and also P (37%) at 25 mg/L, compared with control. Confocal microscopy images showed that all Cu compounds, at 75 mg/L, significantly reduced nitric oxide, concurring with the reduction in root growth. Nano Cu(OH)₂ at 25 mg/L upregulated the expression of the Cu/Zn superoxide dismutase gene (1.92-fold), while ionic treatments at 75 mg/L upregulated (∼10-fold) metallothionein (MT) transcripts. Results demonstrated that nano and bulk Cu(OH)₂ compounds caused less physiological impairments in comparison to the ionic ones in alfalfa seedlings.
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