Tea is an economically important plant characterized by a large genome, high heterozygosity, and high species diversity. In this study, we assemble a 3.26-Gb high-quality chromosome-scale genome for the ‘Longjing 43’ cultivar of Camellia sinensis var. sinensis. Genomic resequencing of 139 tea accessions from around the world is used to investigate the evolution and phylogenetic relationships of tea accessions. We find that hybridization has increased the heterozygosity and wide-ranging gene flow among tea populations with the spread of tea cultivation. Population genetic and transcriptomic analyses reveal that during domestication, selection for disease resistance and flavor in C. sinensis var. sinensis populations has been stronger than that in C. sinensis var. assamica populations. This study provides resources for marker-assisted breeding of tea and sets the foundation for further research on tea genetics and evolution.

The tea plant (Camellia sinensis (L.) O. Kuntze) is one of the most economically important woody crops. Recently, many leaf color genotypes have been developed during tea plant breeding and have become valuable materials in the processing of green tea. Although the physiological characteristics of some leaf color mutants of tea plants have been partially revealed, little is known about the molecular mechanisms leading to the chlorina phenotype in tea plants. The yellow-leaf tea cultivar Zhonghuang 2 (ZH2) was selected during tea plant breeding. In comparison with Longjing 43 (LJ43), a widely planted green tea cultivar, ZH2 exhibited the chlorina phenotype and displayed significantly decreased chlorophyll contents. Transmission electron microscopy analysis revealed that the ultrastructure of the chloroplasts was disrupted, and the grana were poorly stacked in ZH2. Moreover, the contents of theanine and free amino acids were significantly higher, whereas the contents of carotenoids, catechins and anthocyanin were lower in ZH2 than in LJ43. Microarray analysis showed that the expression of 259 genes related to amino acid metabolism, photosynthesis and pigment metabolism was significantly altered in ZH2 shoots compared with those of LJ43 plants. Pathway analysis of 4,902 differentially expressed genes identified 24 pathways as being significantly regulated, including ‘cysteine and methionine metabolism’, ‘glycine, serine and threonine metabolism’, ‘flavonoid biosynthesis’, ‘porphyrin and chlorophyll metabolism’ and ‘carotenoid biosynthesis’. Furthermore, a number of differentially expressed genes could be mapped to the ‘theanine biosynthesis’, ‘chlorophyll biosynthesis’ and ‘flavonoid biosynthesis’ pathways. Changes in the expression of genes involved in these pathways might be responsible for the different phenotype of ZH2. A novel chlorophyll-deficient chlorina tea plant cultivar was identified. Biochemical characteristics were analyzed and gene expression profiling was performed using a custom oligonucleotide-based microarray. This study provides further insights into the molecular mechanisms underlying the phenotype of the chlorina cultivar of Camellia sinensis.

Sugars Will Eventually be Exported Transporters (SWEETs) are important in plant biological processes. Expression levels of CsSWEET1a and CsSWEET17 are induced by cold acclimation (CA) and cold stress in Camellia sinensis. Here, we found that CsSWEET17 was alternatively spliced, and its exclusion (Ex) transcript was associated with the CA process. Both plasma membrane-localized CsSWEET1a and CsSWEET17 transport hexoses, but cytoplasm-localized CsSWEET17-Ex does not. These results indicate that alternative splicing may be involved in regulating the function of SWEET transporters in response to low temperature in plants. The extra C-terminal of CsSWEET17, which is not found in the tonoplast fructose transporter AtSWEET17, did not affect its plasma membrane localization but promoted its sugar transport activities. The overexpression (OE) of CsSWEET1a and CsSWEET17 genes resulted in an increased sugar uptake in Arabidopsis, affecting plant germination and growth. The leaf and seed sizes of the CsSWEET17-OE lines were significantly larger than those of the wild type. Moreover, the OE of CsSWEET1a and CsSWEET17 significantly reduced the relative electrolyte leakage levels under freezing stress. Compared with the wild type, the expression of AtCWINV genes was suppressed in both CsSWEET1a-OE and CsSWEET17-OE lines, indicating the alteration in sugar contents in the cell walls of the OE lines. Furthermore, the interaction between CsSWEET1a and CsSWEET17 was confirmed using yeast two-hybrid and bimolecular fluorescence complementation assays. We showed that CsSWEET1a and CsSWEET17 form homo-/heterodimers in the plasma membrane and mediate the partitioning of sugars between the cytoplasm and the apoplast, thereby regulating plant growth and freezing tolerance.