Xiaolan Jiang; Yajun Liu; Weiwei Li; Lei Zhao; Fei Meng; Yunsheng Wang; Huarong Tan; Hua Yang; Chaoling Wei; Xiaochun Wan; Liping Gao; Tao Xia

“ In our experiment (data not shown) , however, utilizing partly purified ANR enzyme from tea plant, only EC and EGC were detected, without any trace of C and GC. The results suggested the function of recombinant proteins may be not consistent with the enzyme in tea plant, or there may be another ANR gene in tea plant. In our transgenic experiment (data not shown) , the level of ANR expression is consistent well with the content of DMACA-stained compounds. ”

“ Phenolic compounds, an important secondary metabolite in tea, account for 18% to 36% dry weight in the fresh leaves and tender stem. More than 96 phenolic compounds have been identified from 41 green teas and 25 fermented teas, which included the following: 28 acylated glycosylated flavonols, 19 O-glycosylated flavonols, 15 phenolic acid derivatives, 12 catechins, 7 C-glycosylated flavones, 6 PAs, and 3 flavonols [13] . ”

“ Analyses of enzymatic activity involved in catechins galloylated and hydrolysis biosynthesis in different organs and leaves at different developmental stages.The enzyme activity unit was defined as the amount of product formed by one milligram of enzyme catalyzed in one minute (μg min−1 mg−1 protein) ”

“ ANR, anthocyanidin reductase; UGT, uridine diphosphate Glycosyltransferase; GCH, galloylated catechins hydrolase; ECGT, epicatechins 1-O-galloyl-β-D-glucose O-galloyltransferase. Extensive studies on model organisms like tobacco, Arabidopsis thaliana, and Medicago truncatula facilitated the understanding of regulation system and subcellular location in the flavonoid pathway [17] – [18] . In the recent years, more number of researches were conducted on the synthetic regulation, transport and galloylation of phenolic compounds, and polymerization of PAs synthesis [5] , [18] – [19] . ”

“ The DAD was set at 280 nm and 340 nm for real-time monitoring of the peak intensities. Ultraviolet (UV) spectra were continuously recorded from 200 nm to 600 nm for the identification of plant components. Mass spectra were acquired simultaneously using the electrospray ionization in the positive and negative ionization modes at the fragmentation voltages of 175 V over the range of m/z 100 to 2000. A drying gas flow of 12 L min−1, drying gas temperature of 325°C, nebulizer pressure of 35 psi, and capillary voltages of 3500 V were used. ”

“ Ectopic expression of VvMybPA2 promotes proanthocyanidin biosynthesis in grapevine and suggests additional targets in the pathway. Plant Physiol 149: 1028–1041 pmid:19098092↑ Akagi T, Ikegami A, Tsujimoto T, Kobayashi S, Sato A, et al. (2009) DkMyb4 Is a Myb Transcription Factor Involved in Proanthocyanidin Biosynthesis in Persimmon Fruit. Plant Physiol 151: 2028–2045 pmid:19783643↑ Xie DY, Sharma SB, Dixon RA (2004) Anthocyanidin reductases from Medicago truncatula and Arabidopsis thaliana . ”

“ Separation of catechin constituents from five tea cultivars using high-speed counter-current chromatography. J Chromatogr A 1083: 223–228 pmid:160787121 2 Chen HD, Parks TA, Chen XX, Gillitt ND, Jobin C, et al. (2011) Structural identification of mouse fecal metabolites of theaflavin 3,3 '-digallate using liquid chromatography tandem mass spectrometry. ”

“ Table S1 Sequences of primers used to amplify genes involved in phenolic compounds biosynthesis in '''Camellia sinensis''' (L.) . (DOC) pone.0062315.s007.docWe thank Prof. Fenglin Hu for assistance with LC-MS analysis.
References ↑ Sueoka N, Suganuma M, Sueoka E, Okabe S, Matsuyama S, et al. (2001) A new function of green tea: prevention of lifestyle-related diseases. Ann N Y Acad Sci 928: 274–280 pmid:11795518↑ Thielecke F, Boschmann M (2009) The potential role of green tea catechins in the prevention of the metabolic syndrome – A review. ”