Abstract
Copper (Cu) interferes with numerous biological functions in plants, including plant growth, which is partly governed by plant hormones. In the present study, Cu stress effect on the roots of pre-emerging maize seedlings in terms of growth, nutrient composition, protein modifications, and root hormone homeostasis was investigated, focusing on possible metabolic differences between the root apex and the rest of the root tissues. Significant decreases in root length and root biomass after 72 h of Cu exposure (50 and 100 μM CuCl2), accompanied by reductions in Ca, Mg, and P root contents, were found. Cu also generated cell redox imbalance in both root tissues and revealed by altered enzymatic and non-enzymatic antioxidant defenses. Oxidative stress was evidenced by an increased protein carbonylation level in both tissues. Copper also induced protein ubiquitylation and SUMOylation and affected 20S proteasome peptidase activities in both tissues. Drastic reductions in ABA, IAA, JA (both free and conjugated), GA3, and GA4 levels in the root apex were detected under Cu stress. Our results show that Cu exposure generated oxidative damage and altered root hormonal homeostasis, mainly at the root apex, leading to a strong root growth inhibition. Severe protein post-translational modifications upon Cu exposure occurred in both tissues, suggesting that even when hormonal adjustments to cope with Cu stress occurred mainly at the root apex, the entire root is compromised in the protein turnover that seems to be necessary to trigger and/or to sustain defense mechanisms against Cu toxicity.
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Acknowledgments
We thank Dr. Myriam S. Zawoznik for her helpful criticism and for improving the English. CLM is a Research Fellow at the UBA (Argentina). LBP and SMG are Career Investigators from CONICET (Argentina). Hormone measurements were performed at Servei Central d’Instrumentació Científica (SCIC) of Universitat Jaume I (Spain).
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This work was supported by the Universidad de Buenos Aires (20020170100331BA UBACYT), and Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 0266).
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Supplemental Fig. 1
Hydroponic culture and maize seedlings. The left panel is a top view of the hydroponic system. The right panels show representative images of control maize seedlings and Cu-treated (100 μM of CuCl2) maize seedlings at 72 h of metal treatment (BMP 1349 kb).
Supplemental Fig. 2
Representative image of the dot blots obtained to determine carbonylated proteins. Root protein extracts obtained from the root apex (Ap) and the remaining root tissue (Rt) of control (C) and Cu-treated seedlings (50 and 100 μM of CuCl2) were derivatized with 2,4-DNPH. Equal amounts of total protein (7.5 μg) were blotted onto a PVDF, blocked in 5% (w/v) non-fat dried milk in PBS, and incubated at room temperature for 1 h with anti-DNP (Sigma, St Luis) as the primary antibody. Dots were subsequently visualized using a secondary rabbit antibody conjugated with horseradish peroxidase (HRP) and stained using 3,3′-diaminobenzidine (DAB) as substrate. The dot blots shown are representative of four replicates prepared in five independent experiments (PPT 136 kb).
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Matayoshi, C.L., Pena, L.B., Arbona, V. et al. Early responses of maize seedlings to Cu stress include sharp decreases in gibberellins and jasmonates in the root apex. Protoplasma 257, 1243–1256 (2020). https://doi.org/10.1007/s00709-020-01504-1
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DOI: https://doi.org/10.1007/s00709-020-01504-1