Genome-wide identification of CsCCD gene family in tea plant (Camellia sinensis) and expression analysis of the oolong tea processing with supplementary LED light / 生物工程学报

Carotenoid cleavage dioxygenase (CCD) family is important for production of volatile aromatic compounds and synthesis of plant hormones. To explore the biological functions and gene expression patterns of CsCCD gene family in tea plant, genome-wide identification of CsCCD gene family was performed. The gene structures, conserved motifs, chromosome locations, protein physicochemical properties, evolutionary characteristics, interaction network and cis-acting regulatory elements were predicted and analyzed. Real time-quantitative reverse transcription PCR (RT-qPCR) was used to detect the relative expression level of CsCCD gene family members under different leaf positions and light treatments during processing. A total of 11 CsCCD gene family members, each containing exons ranging from 1 to 11 and introns ranging from 0 to 10, were identified. The average number of amino acids and molecular weight were 519 aa and 57 643.35 Da, respectively. Phylogenetic analysis showed the CsCCD gene family was clustered into 5 major groups (CCD1, CCD4, CCD7, CCD8 and NCED). The CsCCD gene family mainly contained stress response elements, hormone response elements, light response elements and multi-factor response elements, and light response elements was the most abundant (142 elements). Expression analysis showed that the expression levels of CsCCD1 and CsCCD4 in elder leaves were higher than those in younger leaves and stems. With the increase of turning over times, the expression levels of CsCCD1 and CsCCD4 decreased, while supplementary LED light strongly promoted their expression levels in the early stage. The expression level of NCED in younger leaves was higher than that in elder leaves and stems on average, and the expression trend varied in the process of turning over. NCED3 first increased and then decreased, with an expression level 15 times higher than that in fresh leaves. In the late stage of turning over, supplementary LED light significantly promoted its gene expression. In conclusion, CsCCD gene family member expressions were regulated by mechanical force and light. These understandings may help to optimize tea processing techniques and improve tea quality.

A predicted protein functional network aids in novel gene mining for characteristic secondary metabolites in tea plant (Camellia sinensis)

Modeling a protein functional network in concerned species is an efficient approach for identifying novel genesin certain biological pathways. Tea plant (Camellia sinensis) is an important commercial crop abundant innumerous characteristic secondary metabolites (e.g., polyphenols, alkaloids, alkaloids) that confer tea qualityand health benefits. Decoding novel genes responsible for tea characteristic components is an important basisfor applied genetic improvement and metabolic engineering. Herein, a high-quality protein functional networkfor tea plant (TeaPoN) was predicted using cross-species protein functional associations transferring andintegration combined with a stringent biological network criterion control. TeaPoN contained 31,273 nonredundant functional interactions among 6,634 tea proteins (or genes), with general network topologicalproperties such as scale-free and small-world. We revealed the modular organization of genes related to themajor three tea characteristic components (theanine, caffeine, catechin) in TeaPoN, which served as strongevidence for the utility of TeaPoN in novel gene mining. Importantly, several case studies regarding geneidentification for tea characteristic components were presented. To aid in the use of TeaPoN, a concise webinterface for data deposit and novel gene screening was developed (http://teapon.wchoda.com). We believe thatTeaPoN will serve as a useful platform for functional genomics studies associated with characteristic secondarymetabolites in tea plant.

Comparison of volatile profiles and bioactive components of sun-dried Pu-erh tea leaves from ancient tea plants on Bulang Mountain measured by GC-MS and HPLC / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

To explore the volatile profiles and the contents of ten bioactive components (polyphenols and caffeine) of sun-dried Pu-erh tea leaves from ancient tea plants on Bulang Mountain, 17 samples of three tea varieties were analyzed by headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and high-performance liquid chromatography (HPLC). A total of 75 volatile components were tentatively identified. Laomaner (LME), Laobanzhang (LBZ), and other teas on Bulang Mountain (BL) contained 70, 53, and 71 volatile compounds, respectively. Among the volatile compounds, alcohols (30.2%-45.8%), hydrocarbons (13.7%-17.5%), and ketones (12.4%-23.4%) were qualitatively the most dominant volatile compounds in the different tea varieties. The average content of polyphenol was highest in LME (102.1 mg/g), followed by BL (98.7 mg/g) and LBZ (88.0 mg/g), while caffeine showed the opposite trend, 27.3 mg/g in LME, 33.5 mg/g in BL, and 38.1 mg/g in LBZ. Principal component analysis applied to both the volatile compounds and ten bioactive components showed a poor separation of samples according to varieties, while partial least squares-discriminant analysis (PLS-DA) showed satisfactory discrimination. Thirty-four volatile components and five bioactive compounds were selected as major discriminators (variable importance in projection (VIP) >1) among the tea varieties. These results suggest that chromatographic data combined with multivariate analysis could provide a useful technique to characterize and distinguish the sun-dried Pu-erh tea leaves from ancient tea varieties on Bulang Mountain.

Comparison of volatile profiles and bioactive components of sun-dried Pu-erh tea leaves from ancient tea plants on Bulang Mountain measured by GC-MS and HPLC / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

To explore the volatile profiles and the contents of ten bioactive components (polyphenols and caffeine) of sun-dried Pu-erh tea leaves from ancient tea plants on Bulang Mountain, 17 samples of three tea varieties were analyzed by headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and high-performance liquid chromatography (HPLC). A total of 75 volatile components were tentatively identified. Laomaner (LME), Laobanzhang (LBZ), and other teas on Bulang Mountain (BL) contained 70, 53, and 71 volatile compounds, respectively. Among the volatile compounds, alcohols (30.2%–45.8%), hydrocarbons (13.7%–17.5%), and ketones (12.4%–23.4%) were qualitatively the most dominant volatile compounds in the different tea varieties. The average content of polyphenol was highest in LME (102.1 mg/g), followed by BL (98.7 mg/g) and LBZ (88.0 mg/g), while caffeine showed the opposite trend, 27.3 mg/g in LME, 33.5 mg/g in BL, and 38.1 mg/g in LBZ. Principal component analysis applied to both the volatile compounds and ten bioactive components showed a poor separation of samples according to varieties, while partial least squares-discriminant analysis (PLS-DA) showed satisfactory discrimination. Thirty-four volatile components and five bioactive compounds were selected as major discriminators (variable importance in projection (VIP) >1) among the tea varieties. These results suggest that chromatographic data combined with multivariate analysis could provide a useful technique to characterize and distinguish the sun-dried Pu-erh tea leaves from ancient tea varieties on Bulang Mountain.

Enraizamento de estacas de genótipos de Camellia sinensis L. em meio ácido, presença de alumínio e ácido indolbutírico / Rooting of Camellia sinensis L. in acid medium, the presence of aluminum and indolebutyric acid

RESUMO Experimentos foram conduzidos para avaliar o efeito do meio ácido e do alumínio, assim como para determinar a concentração mais apropriada de ácido indolbutírico (AIB) para o enraizamento de estacas de diferentes genótipos de Camellia sinensis L (planta de chá). Para tal, foram coletados ramos de plantas-matrizes em Pariquera-Açu-SP, Brasil, no inverno de 2012 e preparadas estacas semi-lenhosas, contendo uma gema e uma folha, que foram mantidas em viveiro com 70% de sombreamento. A irrigação do substrato foi feita com água e soluções contendo ácido fosfórico e sulfato de alumínio a pH 5,5, 4,5; 3,5. No primeiro experimento, o delineamento experimental foi o inteiramente casualizado em esquema fatorial 3 x 7, três genótipos (F 15, IAC 259 e Comum) e sete condições diferentes de enraizamento (vermiculita a pH 6,5, vermiculita acidificada com ácido fosfórico a pH 3,5; 4,5 e 5,5 ou vermiculita acidificada com sulfato de alumínio a pH 3,5; 4,5 e 5,5). No segundo experimento, o tratamento que promoveu o maior enraizamento no primeiro experimento (vermiculita acidificada com sulfato de alumínio a pH 3,5) foi combinado ao tratamento com AIB. O delineamento experimental utilizado foi o inteiramente casualizado em esquema fatorial 3 x 6, sendo três genótipos e seis concentrações de AIB (0, 2000, 4000, 6000, 8000 e 1000 mg L-1). A vermiculita acidificada com sulfato de alumínio a pH 3,5, combinada a aplicação de 10000 mg L-1 de AIB por 30 segundos, foi o tratamento mais adequado para o enraizamento de estacas semi-lenhosas dos genótipos F15, IAC 259 e Comum.

ABSTRACT The experiments were carried out to evaluate the effect of medium acidic and aluminum, as well as determine the most suitable concentration of indolebutyric acid (IBA) for rooting cuttings of different genotypes of Camellia sinensis L (tea plant). Such, stems were collected from mother plants in Pariquera-Açu, São Paulo state, Brazil, in winter 2012 and prepared semi-hardwood cuttings, with one bud and one leaf, which were kept in a nursery with 70% of shading. Irrigation substrate was taken with water and solutions containing phosphoric acid and aluminum sulfate at pH 5.5, 4.5; 3,5. In the first experiment, the experimental design was completely randomized in a factorial scheme 3x7, three genotypes (F 15, IAC 259 and Comum) and seven different conditions of rooting (vermiculite at pH 6.5, acidified with phosphoric acid or aluminum sulfate vermiculite at pH 3.5, 4.5 and 5.5). In the second experiment, the treatment that promoted the highest rooting in the first experiment (vermiculite acidified with aluminum sulfate at pH 3.5) was combined with treatment with IBA The experimental design was completely randomized in a factorial scheme 3x6, three genotypes and six concentrations of IBA (0, 2.000, 4.000, 6.000, 8.000 and 10.000 mg L-1). Vermiculite acidified with aluminum sulphate to pH 3.5, combined application of 10.000 mg L-1 IBA for 30 seconds was the most appropriate treatment for cutting propagation of genotypes F15, IAC 259 and Comum. Vermiculite acidified with aluminum sulfate at pH 3.5, combined treatment with 10,000 mg L-1 IBA for 30 seconds, was the most suitable conditions for the rooting of cuttings Camellia sinensis L.