Tea is the world's oldest and most popular caffeine-containing beverage with immense economic, medicinal, and cultural importance. Here, we present the first high-quality nucleotide sequence of the repeat-rich (80.9%), 3.02-Gb genome of the cultivated tea tree Camellia sinensis. We show that an extraordinarily large genome size of tea tree is resulted from the slow, steady, and long-term amplification of a few LTR retrotransposon families. In addition to a recent whole-genome duplication event, lineage-specific expansions of genes associated with flavonoid metabolic biosynthesis were discovered, which enhance catechin production, terpene enzyme activation, and stress tolerance, important features for tea flavor and adaptation. We demonstrate an independent and rapid evolution of the tea caffeine synthesis pathway relative to cacao and coffee. A comparative study among 25 Camellia species revealed that higher expression levels of most flavonoid- and caffeine- but not theanine-related genes contribute to the increased production of catechins and caffeine and thus enhance tea-processing suitability and tea quality. These novel findings pave the way for further metabolomic and functional genomic refinement of characteristic biosynthesis pathways and will help develop a more diversified set of tea flavors that would eventually satisfy and attract more tea drinkers worldwide.
Wild teas are valuable genetic resources for studying domestication and breeding. Here we report the assembly of a high-quality chromosome-scale reference genome for an ancient tea tree. The further RNA sequencing of 217 diverse tea accessions clarifies the pedigree of tea cultivars and reveals key contributors in the breeding of Chinese tea. Candidate genes associated with flavonoid biosynthesis are identified by genome-wide association study. Specifically, diverse allelic function of
CsANR
,
CsF3’5’H
and
CsMYB5
is verified by transient overexpression and enzymatic assays, providing comprehensive insights into the biosynthesis of catechins, the most important bioactive compounds in tea plants. The inconspicuous differentiation between ancient trees and cultivars at both genetic and metabolic levels implies that tea may not have undergone long-term artificial directional selection in terms of flavor-related metabolites. These genomic resources provide evolutionary insight into tea plants and lay the foundation for better understanding the biosynthesis of beneficial natural compounds.
Colletotrichum, the causative agent of anthracnose, is an important pathogen that invades the tea plant (Camellia sinensis). In this study, 38 isolates were obtained from the diseased leaves of tea plants collected in different areas of Zhejiang Province, China. A combination of multigene (ITS, ACT, GAPDH, TUB2, CAL, and GS) and morphology analyses showed that the 38 strains belonged to two different species, namely, C. camelliae (CC), and C. fructicola (CF). Pathogenicity tests revealed that CC was more invasive than CF. In vitro inoculation experiments demonstrated that CC formed acervuli at 72 hpi and developed appressoria on wound edges, but CF did not develop these structures. Under treatment with catechins and caffeine, the growth inhibition rates of CF were remarkably higher than those of CC, indicating that the nonpathogenic species CF was more vulnerable to catechins and caffeine. Growth condition testing indicated that CF grew at a wide temperature range of 15–35°C and that the optimum temperature for CC growth was 25°C. Growth of both CC and CF did not differ between acidic and weakly alkaline environments (pH 5–8), but the growth of CC was significantly reduced at pH values of 9 and 10. Furthermore, the PacC/RIM101 gene, which associated with pathogenicity, was identified from CC and CF genomes, and its expression was suppressed in the hyphae of both species under pH value of 5 and 10, and much lower expression level was detected in CC than that in CF at pH 6. These results indicated that temperature has more important effect than pH for the growth of two Colletotrichum species. In conclusion, the inhibition by secondary metabolite is an important reason why the pathogenicity by CC and CF are different to tea plant, although the environmental factors including pH and temperature effect the growth of two Colletotrichum species.
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