CsCBF2 contributes to cold repression of chlorophyll and carotenoid biosynthesis in albino Camellia sinensis cv. Baiye 1

Abstract C-repeat binding factors (CBFs) play a pivotal role in regulating cold response in higher plants. Camellia sinensis cv. Baiye 1, a representative albino tea cultivar, has been identified as temperature-sensitive based on long-term observations by tea farmers. However, it remains unclear whether CsCBFs are involved in temperature-mediated albinism and seasonal greening in ‘Baiye 1’, and the mechanisms by which CBFs regulate cold responses in albino leaves are unknown. In this study, we demonstrate that CsCBF2 suppresses the seasonal greening of albino leaves by inhibiting chlorophyll and carotenoid biosynthesis under cold stress. In tea plantations, the accumulation of chlorophylls and carotenoids in the albino shoots of ‘Baiye 1’ is closely correlated with the effective accumulated temperature during its seasonal greening process. Weighted Gene Co-expression Network Analysis revealed negative associations between CsCBF expression and chlorophylls, carotenoids, as well as their biosynthetic genes REVEILLE 1 (CsRVE1) and Zeaxanthin epoxidase 1 (CsZEP1) under temperature fluctuations during seasonal greening. Cold-induced upregulation of CsCBF2 expression and decreased chlorophylls and carotenoids under controlled climate conditions. Transient suppression of CsCBF2 by antisense oligodeoxynucleotide elevated expressions of target genes, and increased chlorophylls and carotenoids. CBF-binding cis-elements were identified in CsRVE1, Protochlorophyllide oxidoreductase A (CsPORA), and CsZEP1 promoters. Luciferase assays suggested CsCBF2 binding to the CRT/DRE cis-elements and repressing expression of CsRVE1, CsPORA, and CsZEP1. These findings highlight CsCBF2 as a key transcriptional repressor involved in the seasonal greening of albino ‘Baiye 1’ under cold stress, by modulating cold responses and inhibiting genes associated with chlorophyll and carotenoid biosynthesis.