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. 2023 Aug 25;24(17):13198.
doi: 10.3390/ijms241713198.

The Chemical Composition and Transcriptome Analysis Reveal the Mechanism of Color Formation in Tea (Camellia sinensis) Pericarp

Yueyang Du  1 , Yongen Lin  1 , Kaikai Zhang  1 , Dylan O'Neill Rothenberg  1 , Huan Zhang  1 , Hui Zhou  1 , Hongfeng Su  1 , Lingyun Zhang  1
Affiliations

The Chemical Composition and Transcriptome Analysis Reveal the Mechanism of Color Formation in Tea (Camellia sinensis) Pericarp

Yueyang Du et al. Int J Mol Sci. .

Abstract

To elucidate the molecular mechanisms underlying the differential metabolism of albino (white), green, and purple pericarp coloration, biochemical profiling and transcriptome sequencing analyses were performed on three different tea pericarps, Zhongbaiyihao (Camellia sinensis L. var. Zhongbai), Jinxuan (Camellia sinensis L. var. Jinxuan), and Baitangziya (Camellia sinensis L. var. Baitang). Results of biochemical analysis revealed that low chlorophyll content and low chlorophyll/carotene ratio may be the biochemical basis for albino characteristics in the 'Zhongbaiyihao' pericarp. The differentially expressed genes (DEGs) involved in anthocyanin biosynthesis, including DFR, F3'5'H, CCoAOMT, and 4-coumaroyl-CoA, were highly expressed in the purple 'Baitangziya' pericarp. In the chlorophyll synthesis of white pericarp, GUN5 (Genome Uncoupled 5) and 8-vinyl-reductase both showed high expression levels compared to the green one, which indicated that albino 'Zhongbaiyihao' pericarp had a higher chlorophyll synthesis capacity than 'Jinxuan'. Meanwhile, chlorophyllase (CLH, CSS0004684) was lower in 'Baitang' than in 'Jinxuan' and 'Zhongbaiyihao' pericarp. Among the differentially expressed transcription factors, MYB59, WRKY41-like2 (CS ng17509), bHLH62 like1 (CS ng6804), and bHLH62-like3 (CSS0039948) were downregulated in Jinxuan pericarp, suggesting that transcription factors played a role in regulating tea pericarp coloration. These findings provide a better understanding of the molecular mechanisms and theoretical basis for utilizing functional components of tea pericarp.

Keywords: Camellia sinensis; albino; anthocyanin; coloration; pericarp.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Phenotypic characteristics of Zhongbaiyihao (white), Jinxuan (green), and Baitangziya (purple) pericarp (A) and pigment contents of Zhongbaiyihao (ZB), Jinxuan (JX), and Baitangziya (BT) (B–E). Errors bar represent standard deviation of three independent replicates. Bars showing different lower-case letters indicate significant differences between groups (p < 0.05, one-way ANOVA, Student’s t-test). BT is ‘Baitang’ (purple pericarp), JX is ‘Jinxuan’ (green pericarp), ZB is ‘Zhongbaiyihao’ (white pericarp). C, catechin; EC, epicatechin; ECG, epicatechin gallate; GC, gallocatechin; EGC, epigallocatechin; GCG, gallocatechin gallate; ECGC, epigallocatechin gallate.
Figure 2
Figure 2
Differentially expressed genes (DEGs) in different tea pericarps. (A) Principle component analysis of pericarp samples indicates a high degree of clustering among intra-group samples. (B) DEGs among BT vs. JX, ZB vs. BT, ZB vs. JX. (C) Venn diagram of DEGs between three groups. BT is ‘Baitang’ (purple pericarp), JX is ‘Jinxuan’ (green pericarp), ZB is ‘Zhongbaiyihao’ (white pericarp).
Figure 3
Figure 3
KEGG enrichment analysis of DEGs in different tea pericarp. JX vs. BT (A), ZB vs. BT (B), and ZB vs. JX (C). BT is ‘Baitang’ (purple pericarp), JX is ‘Jinxuan’ (green pericarp), ZB is ‘Zhongbaiyihao’ (white pericarp).
Figure 4
Figure 4
DEGs involved in flavonoid and phenylpropanoid biosynthesis in different tea pericarps. JX vs. ZB (A), ZB vs. BT (B), and BT vs. JX (C). BT is ‘Baitang’ (purple pericarp), JX is ‘Jinxuan’ (green pericarp), ZB is ‘Zhongbaiyihao’ (white pericarp).
Figure 5
Figure 5
Expression analysis for key genes and transcription factors involved in pigment biosynthesis. (A) Expression level analysis of key genes and transcription factors in different pericarp. Errors bar represent standard deviation of three independent replicates. Bars showing different lower-case letters indicate significant differences between groups (p < 0.05, one-way ANOVA, Student’s t-test). (B) Correlation analysis based on RNA-seq and qRT-PCR data. CLH, Chlorophyllase; HY5, ELONGATED HYPOCOTYL5; VR, Vestitone reductase; CHS, Chalcone synthase; DFR, Dihydroflavonol reductase; F3′5′H, Flavonoid 3′,5′-hydroxylase; CoAOMT, Caffeoyl coenzyme A O-methyltransferase; HEME, Uroporphyrinogen III decarboxylase; GUN5, Genome Uncoupled 5; ANR, Anthocyanidin reductase; LAR, Leucoanthocyanidin reductase. BT is ‘Baitang’ (purple pericarp), JX is ‘Jinxuan’ (green pericarp), ZB is ‘Zhongbaiyihao’ (white pericarp).
Figure 6
Figure 6
DEGs involved in the flavonoids and chlorophyll biosynthetic pathway. (A) DEGs involved in the flavonoids biosynthetic pathway; (B) DEGs involved in the chlorophyll biosynthetic pathway. Heat maps were created based on average expression levels (FPKM values). The color scale represents the FPKM value. Red indicates high expression, and green indicates low expression. BT is ‘Baitang’ (purple pericarp), JX is ‘Jinxuan’ (green pericarp), ZB is ‘Zhongbaiyihao’ (white pericarp). PAL, Phenylalanine ammonia-lyase; C4H, Cinnamate 4-hydroxylase; 4CL, 4-coumarate-CoAligase; HCT, Hydroxycinnamoyl-CoA shikimate/quinatehydroxy-cinnamoyltransferase; CCR, Cinnamoyl Co-A reductase; CoAOMT, Caffeoyl coenzyme A O-methyltransferase; CHS, Chalcone synthase; CHI, Chalcone isomerase; FLS, Flavonol synthase; F3’H, Flavonoid 3′-hydroxylase; F3’5’H, Flavonoid 3′5′-hydroxylase; DFR, Dihydroflavonol 4-reductase; LAR, Leucoanthocyanidin reductase; ANS, Anthocyanidin synthase; HEMA, Glutamyl-tRNA reductase; HEML, glutamate 1-semialdehyde aminotransferase; HEMB, porphobilinogen synthase; HEME, Uroporphyrinogen III decarboxylase; GUN5, Genome Uncoupled 5.
Figure 7
Figure 7
Differential expression of transcription factors involved in anthocyanidin and flavonoid pathways in different pericarps. The heatmap was created according to the expression levels of related transcription factors based on the FPKM value. The color scale represents the FPKM value. Red indicates high expression, and green indicates low expression. BT is ‘Baitang’ (purple pericarp), JX is ‘Jinxuan’ (green pericarp), ZB is ‘Zhongbaiyihao’ (white pericarp).
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