Oxidative Metabolism in ‘Valencia’ Sweet Orange (Citrus sinensis Osbeck) Abscission Zone Tissue Treated with the Abscission Agent 5-Chloro-3-Methyl-4-Nitro-1H-Pyrazole

Abstract

5-Chloro-3-methyl-4-nitro-1H-pyrazole (CMNP) is an abscission agent, standardized for the mechanical harvesting of late season ‘Valencia’ sweet oranges in Florida. This work was conducted to investigate the role of CMNP to induce oxidative stress in the abscission zone (AZ) of ‘Valencia’ sweet orange. Fully mature ‘Valencia’ sweet orange trees in a commercial grove were sprayed with 2.0 mm of CMNP. The experiment was repeated three times during the Apr.–May 2013 harvest season. Fruit were harvested at 0, 1, 2, and 3 days after CMNP application. Hydrogen peroxide (H2O2) concentration and malonic dialdehyde (MDA) concentration, as well as superoxide dismutase (SOD), ascorbate peroxidase (APOD), glutathione reductase (GR), peroxidase (POD), and lipoxygenase (LOX) specific activities were measured 0, 1, 2, and 3 days after CMNP treatment (DAT). Rate of lipid peroxidation remains unchanged throughout the abscission period. However, LOX activity increased 1 DAT in AZ of treated fruit, which might produce jasmonic acid (JA), known to promote abscission in citrus. Levels of H2O2 were similar in the AZ of control and treated fruit except at 3 DAT. The specific activity of SOD declined at 2 DAT, which showed compromised SOD defense against superoxide radicals (O·−). APOD activity declined sharply at 3 DAT. Interestingly, GR activity was 1.9-fold higher in CMNP-treated fruit at 3 DAT. Higher GR and low APOD activity reflects limited functioning of the APOD/GR cycle (e.g., APOD and GR) in scavenging of H2O2 at 3 DAT. Guaiacol POD activity transiently increased at 1 DAT and then declined. POD plays an important role in cell wall lignification and indole acetic acid (IAA) oxidation. The decline in POD activity may cause a decrease in lignification while higher activity made the AZ sensitive to ethylene and thus promote abscission in citrus fruit. This work also showed that CMNP-induced abscission is a collaborative effort of oxidative metabolism in flavedo tissue (FT) and AZ.