Protein Aggregation, Radical Scavenging Capacity, and Stability of Hydrogen Peroxide Defense Systems in Heat-stressed Vinca and Sweet Pea Leaves

Abstract

Although heat stress injury is known to be associated with membrane dysfunctions, protein structural changes, and reactions of activated forms of oxygen, the underlying mechanisms involved are poorly understood. In this study, the relationships between thermotolerance and hydrogen peroxide (H2O2) defense systems, radical scavenging capacity [based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) reduction], and protein aggregation were examined in vinca [Catharanthus roseus (L.) G. Don `Little Bright Eye'], a heat tolerant plant, and sweet pea (Lathyrus odoratus L. `Explorer Mix'), a heat susceptible plant. Vinca leaves were 5.5 °C more thermotolerant than sweet pea leaves based on electrolyte leakage analysis. Vinca leaf extracts were more resistant to protein aggregation at high temperatures than sweet pea leaf extracts, with precipitates forming at ≥40 °C in sweet pea and at ≥46 °C in vinca. Vinca leaves also had nearly three times greater DPPH radical scavenging capacity than sweet pea leaf extracts. Two enzymatic detoxifiers of H2O2, catalase (CAT) and ascorbate peroxidase (APOX), demonstrated greater activities in vinca leaves than in sweet pea leaves. In addition, CAT and APOX were more thermostable in vinca, compared with sweet pea leaves. However, tissue H2O2 levels did not differ between controls and tissues injured or killed by heat stress in either species, suggesting that H2O2 did not play a direct role in acute heat stress injury in vinca or sweet pea leaves. Greater thermotolerance in vinca, compared with sweet pea, was associated with greater DPPH radical scavenging capacity, indicating that AOS other than H2O2 may be involved in acute heat stress injury.