External application of brassinolide enhances cold resistance of tea plants (Camellia sinensis L.) by integrating calcium signals.

ABSTRACT

MAIN CONCLUSION: Exogenous brassinolide can activate the expression of key genes in the calcium signalling pathway to enhance cold resistance of tea plants. Brassinolide is an endogenous sterol phytohormone containing multiple hydroxyl groups that has the important function of improving plant cold resistance and alleviating freeze damage. To explore the molecular mechanism of how brassinolide improves the cold resistance of tea plants, "Qiancha 1" was used as the material, and the method of spraying brassinolide on the leaves was adopted to explore its effects on the tea plants under 4 °C low-temperature treatment. The results showed that brassinolide can significantly increase the protective enzyme activity of tea plants under cold stress and reduce cold damage. At the transcriptome level, brassinolide significantly enhanced the expression of key genes involved in calcium signal transduction, Calmodulin (CaM), Calcium-dependent protein kinase (CDPK), calcineurin B-like protein (CBL) and calmodulin-binding transcriptional activators (CAMTA), which then activated the downstream key genes transcriptional regulator CBF1 (CBF1) and transcription factor ICE1 (ICE1) during cold induction. Quantitative real-time PCR (qRT‒PCR) results showed that the expression of these genes was significantly induced after treatment with brassinolide, especially CaM and CBF1. When calcium signalling was inhibited, the upregulated expression of CBF1 and ICE1 disappeared, and when CAMTA was knocked down, the expression of other genes under cold stress was also significantly reduced. The above results indicate that brassinolide combined with the calcium signalling pathway can improve the cold resistance of tea plants. This study provides a new theoretical basis for the study of the cold resistance mechanism of brassinolide.