Dynamic Physiological Responses of Cinnamomum camphora with Monoterpene Protection under High Temperature Shock

Abstract

Monoterpenes can protect plants against high temperature, but the early events of protection are still unknown. In this study, the dynamic variations in reactive oxygen species metabolism, photosynthetic capacity, and related gene expression in linalool, eucalyptol, and camphor chemotypes of Cinnamomum camphora with and without monoterpene emission under 6 h high-temperature stress were investigated. With respect to the control (28 °C), 40 °C and Fos + 40 °C (fosmidomycin inhibited monoterpene biosynthesis under 40 °C) treatments increased H2O2 and thiobarbituric acid reactive substance levels in the three chemotypes, but without significant differences between the two treatments after 2 h. Compared with the 40 °C treatment, the Fos + 40 °C treatment further aggravated the increase after 4 h, with increases of 13.8%, 12.3%, and 12.3% in H2O2 levels as well as 16.5%, 17.4%, and 9.1% in thiobarbituric acid reactive substance levels, respectively, in linalool, eucalyptol, and camphor chemotypes. When the three chemotypes were treated with 40 °C and Fos + 40 °C, the ascorbic acid content was gradually decreased during the 2 h treatment. After 4 h, the Fos + 40 °C treatment further aggravated the decrease in ascorbic acid content, with decreases of 10.6%, 9.8%, and 20.1%, respectively, in the eucalyptol, linalool, and camphor chemotypes. This could be caused by the further down-regulation of the key gene GGP in antioxidant biosynthesis. Meanwhile, two genes (VTE3 and 4CL) in other non-enzymatic antioxidant formation were also further down-regulated in Fos + 40 °C treatment for 4 h. These might lead to the further increase in reactive oxygen species levels in Fos + 40 °C treatment lacking non-enzymatic antioxidants. The photosynthetic electron yield and transfer (φPo, Ψo and φEo) in the three chemotypes were significantly (p < 0.05) decreased under the 40 °C and Fos + 40 °C treatments for 0.5 h, and the photosynthetic rate was significantly (p < 0.05) decreased in the two treatments for 1 h. After 4 h, the Fos + 40 °C treatment aggravated the decrease, as the genes encoding the components of photosystem II (psbP and psbW) and ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcS and rbcL) were further down-regulated. These dynamic variations in the early events suggested that monoterpenes should act as signaling molecules to improve plant thermotolerance, as blocking monoterpene biosynthesis did not cause immediate effects on the physiological responses in contrast to the monoterpene-emitting plants during the 2 h high temperature stress, but resulted in serious damages after 4 h for suppressing related gene expression. This not only provides new proof for the isoprenoid thermotolerance mechanism by serving a signaling function, but also promotes the utilization of monoterpenes as anti-high-temperature agents, and the cultivation of high-temperature tolerance varieties with abundant monoterpene emission.