Tea plants grown in high-latitude areas are often damaged by extreme freezing temperatures in winter, leading to huge economic losses. Here, the physiological and gene expression characteristics of two tea cultivars (Xinyang No. 10 (XY10), a freezing-tolerant cultivar and Fudingdabaicha (FDDB), a freezing-sensitive cultivar) during overwintering in northern China were studied to better understand the regulation mechanisms of tea plants in response to natural freezing stress. Samples were collected at a chill (D1), freezing (D2) and recovery (D3) temperature in winter. TEM analysis of integrated leaf ultrastructure at D2 revealed lower malondialdehyde and relative electrical conductivity in XY10 than in FDDB, with serious cell structure damage in the latter, indicating XY10 was more resistant to freezing stress. Differential gene expression analysis among the different samples over winter time highlighted the following gene functions in cell wall metabolism (CesAs, COBLs, XTHs, PGs, PMEs), transcription factors (ERF1B and MYC2), and signal transduction (CDPKs and CMLs). The expression pattern of cellulose and pectin-related genes suggested higher accumulation of cellulosic and pectic materials in the cell wall of XY10, agreeing with the results of cell wall and its components. These results indicated that under the regulation of cell wall genes, the freezing-resistant tea cultivar can better maintain a well-knit cell wall structure with sufficient substances to survive natural freezing damage. This study demonstrated the crucial role of cell wall in tea plant resistance to natural freezing stress and provided important candidate genes for breeding of freezing-resistant tea cultivars.

Tea plant can enrich a large amount of fluorine (F) in the cell wall of its mature leaves, thus posing the risk of excessive intake of F for tea consumers. This study investigated the effect of foliar calcium (Ca) application (0.05-1 mM) on F accumulation in tea plant leaves by analyzing the association of F with cell wall materials, pectin methylesterification structure, and cell wall genes. Ca spray could effectively reduce the F level, the content of wall carbohydrates (excluding the chelate pectin fraction) and the pectin methylesterification degree in tea plant leaves. Correlation analysis further revealed that the tea leaf F content was positively correlated with most of the cell wall materials. Transcriptomics analysis indicated that the key genes involved in reorganization of cell wall polysaccharides (such as CslE6, XTH32, PG, PGIP, PME18, PMEU1, and PMEI9) were associated with the variations in F content and cell wall components. All the results suggest that Ca may lessen the tea leaf F level by reducing the accumulation of cell wall materials and intervening the cell wall structure and gene expression. This study provided useful information for reducing the F level and solving the F safety problem in tea plant.