► Tea products from green and purple leaf coloured cultivars were analysed. ► Unaerated (green) tea products contained higher levels of TP, catechins and AA. ► Purple tea cultivars also contain ...catechins and have lower caffeine levels. ► Antioxidant activities of the purple teas were attributed to the anthocyanins. ► Anthocyanins are highly soluble in water than methanol.
Black (aerated) and green (unaerated) tea products, processed from 10 green and 18 purple leaf coloured cultivars of Kenyan origin, and two tea products, from the Japanese cultivars, Yabukita and Yutakamidori, were assayed for total polyphenols (TP) content, individual catechin profiles and in vitro antioxidant capacity (AA). In addition, the phenolic content of the tea products was determined using the Folin–Ciocalteu phenol reagent. Catechin fractions were identified using reverse phase high performance liquid chromatography (HPLC) with a binary gradient elution system.
The AA% of the tea products was determined using a 2,2′-diphenyl picrylhydrazyl (DPPH) radical assay method. The results showed that TPs, catechin profiles and antioxidant activities were significantly (p⩽0.05) higher in unaerated than in aerated teas. Tea products from the purple leaf coloured tea cultivars had levels of TPs, total catechin (TC) and antioxidant activities similar to those from the green leaf coloured cultivars, except for teas from the Japanese cultivars that were very low in the assayed parameters. Caffeine content was significantly (p⩽0.05) lower in products from the purple leaf coloured cultivars than in those from the green leaf coloured tea cultivars. Antioxidant activity (%) was higher in tea products from the Kenyan germplasm than in those from the Japanese cultivars. Antioxidant potency of tea products was significantly (r=0.789∗∗, p⩽0.01) influenced by the total anthocyanin content of the purple leaf coloured cultivars. Cyanidin-3-O-glucoside was the anthocyanin most highly correlated with AA% (r=0.843∗∗, p⩽0.01 in unaerated tea). Total catechins in the unaerated products from the green leaf coloured tea cultivars were also significantly correlated with antioxidant capacity (r=0.818∗∗, p⩽0.01). Results from this study suggest that the antioxidant potency of teas is dependent on the predominant flavonoid compound, the type of tea cultivar and the processing method.
► Anthocyanins were characterised in selected Kenyan tea germplasm. ► The common anthocyanins found in nature were identified in addition to the catechins. ► The major anthocyanidin is malvidin. ► ...Anthocyanidins correlated negatively with the catechins. ► Unaerated (green) tea products retained higher anthocyanins than black (aerated) tea.
Characterization and quantification of anthocyanins in selected tea cultivars processed into black (aerated) and green (unaerated) tea products was carried out in this study. The anthocyanins were extracted from tea products processed from a number of newly bred purple leaf coloured Kenyan tea cultivars (Camellia sinensis) using acidified methanol/HCl (99:1 v/v). Extracted anthocyanins were purified by C18 solid phase extraction (SPE) catridges and characterised by HPLC-UV–Visible. They were identified according to their HPLC retention times, elution order and comparison with authentic standards that were available. Total monomeric anthocyanins were determined by the pH-differential method. Although the tea cultivars gave different yields of anthocyanins, the unaerated (green) teas had significantly (p⩽0.05) higher anthocyanin content than the aerated (black) teas. This was attributed to the degradation of anthocyanins by polyphenol oxidase products (catechin O-quinones) formed during the auto-oxidation (fermentation) process of black tea manufacture. Of the six most common natural anthocyanidins, five were identified in the purified extracts from purple leaf coloured tea, in both aerated (black) and unaerated (green) teas namely; delphinidin, cyanidin, pelargonidin, peonidin and malvidin. The most predominant anthocyanidin was malvidin in both tea products. In addition, two anthocyanins namely, cyanidin-3-O-galactoside and cyanidin-3-O-glucoside were also identified. Tea catechins were also identified in the tea products derived from the purple coloured tea cultivars namely, epigallocatechin (EGC), catechin (+C), epicatechin (EC), epigallocatechin gallate (EGCG), and epicatechin gallate (ECG). Correlation between the total catechins versus the total anthocyanins and anthocyanin concentration in unaerated teas revealed significant negative correlations (r=−0.723∗ and r=−0.743∗∗, p⩽0.05 and p⩽0.01, respectively). However, in aerated (black) tea the correlations were insignificant (r=−0.182 and r=−0.241, p>0.05).
•We examine the antioxidant potency of green and black teas fortified with milk and sweeteners.•We study the effect of tea with milk and sweeteners on glutathione an innate body antioxidant.•Added ...milk lowered the antioxidant capacity of green and black tea in a dose dependent manner.•Common sweeteners lowered antioxidant capacity of tea except for Stevia.•Tea consumption enhanced tissue glutathione levels in mice.
Several studies have demonstrated that tea flavonoids protect cells and tissues against free radicals which have been implicated in the etiology of oxidative stress-related disease disorders. However, black tea is commonly consumed with additives that could otherwise affect the bioavailability of the active tea molecules. In this study, the biochemical parameters of Kenyan teas were determined and the effect of added milk and sweeteners on the antioxidant activity of Kenyan teas was investigated. The effect of tea antioxidants on glutathione (GSH) was also evaluated in vivo in a time series study using Swiss mice. Green teas had the highest levels of total polyphenols, total and individual catechins, while black teas had high levels of total thearubigins, total theaflavins and theaflavin fractions. The antioxidant activity was high in green teas though some of the black teas were as efficacious as the green teas. The addition of milk, sugar and honey significantly (p<0.05) decreased the antioxidant activity of tea in a concentration-dependent manner. Addition of the sweetener, stevia (Stevia rebaudiana Bertoni), showed no significant (p>0.05) influence on the antioxidant activity of tea and therefore can be recommended as a preferred sweetener for tea. Significantly (p<0.001) higher levels of GSH were observed in plasma than in other tissues. GSH levels were generally highest 2h after tea consumption, which indicates the need to repeatedly take tea every 2h to maximise its potential health benefits.
•Kenyan purple clones has more total polyphenols than the Japanese Yabukita variety.•K-purple and TRFK 91/1 can make good green because of higher catechin quality index.•TRFK 73/5, a purple clone has ...the lowest caffeine content among the studied clones.•Kenyan purple teas has more theanine than the Japanese and Chinese varieties studied.•TRFK 73/4, a purple clone has higher EGCG levels than the Japanese variety, Yabukita.
The Kenyan tea industry wishes to diversify its tea products, and in line with this, anthocyanin – rich teas were developed at the Tea Research Foundation of Kenya. These teas have purple-coloured leaves and the green colour is masked. In total, 12 accessions of the purple leaf coloured teas and 2 standard tea varieties were studied. Clones Hanlu and Yabukita are Chinese and Japanese tea varieties, respectively, known for good green tea, and they were used as reference standards. Little if any research had been done to characterize the quality of these purple leaf coloured teas and this study investigated their total polyphenols (TPP), catechins, caffeine, gallic acid and theanine. These are the major green tea quality parameters. Results showed that the new Kenyan tea clones had higher total polyphenols than had the reference standard tea varieties, which had 17.2% and 19.7% while the lowest among the Kenyan clones was 20.8%. On catechin quality index, K-purple and TRFK 91/1 showed high index values of 15.9 and 13.3, respectively, while clones TRFK 83/1 and 73/5 showed low index values of 0.74 and 1.0, respectively. Hanlu had the highest caffeine level with 2.42% while clones TRFK KS 3, TRFK KS 2 and TRFK 83/1 had relatively high caffeine levels among the purple leaf coloured teas, with 2.33%, 2.22% and 2.21%, respectively. Clone TRFK 73/5 had the lowest caffeine content, with 1.16%. Theanine analysis showed that most purple leaf coloured teas had more theanine than had the reference standard clones, except TRFK 83/1 and K-purple, which were lower than the reference standard clones. The implication of the green tea chemical quality parameters is also discussed. It is concluded that all the studied clones/varieties have above the minimum 14% of total polyphenols. Clones K-purple and TRFK 91/1 showed high green tea quality indices with the latter doubling with high levels of theanine; hence its highly recommended for green tea manufacture.
In the present study, twelve different types of commercial tea samples were assayed to determine their phenolic composition and antioxidant activity. Reverse phase high performance liquid ...chromatography using a binary gradient system was used for the identification and quantification of individual catechins. Subsequently, total phenolic content was determined spectrophotometrically according to the Folin-ciocalteus method. Total theaflavins and thearubigins were also determined. The radical scavenging behavior of the polyphenols on 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) was also studied spectrophotometrically. The results showed that total polyphenols, total catechins and antioxi-dant activity were significantly (P<0.05) different in the commercial tea samples. Green tea had the highest levels of catechins, total polyphenols and total antioxidant activity. White tea (silvery tip) a rare specialty type of tea was not significantly different from green tea. Statistical analysis showed an essential catechin content influence of the tea extracts on antioxidant activity. Epigallocatechin gallate (EGCG) was the most potent catechin and the most potent in antioxidant activity (r = 0.989***). Epigallo-catechin (EGC) (r = 0.787, P<0.001), epicatechin (EC) + catechin (+C) and epicatechigallate (ECG) also showed significant (P<0.05) antioxidant activity. Black tea contained high levels of theaflavins and thearubigins, which accounted for most of the antioxidant potential in this type of tea product (r = 0.930*** and r = 0.930*** respectively). These results suggest that conversion of catechins during black tea processing did not affect the free-radical potency of black tea. Gallic acid (GA) also showed significant(r = 0.530*) contribution to the antioxidant activity in black tea. Green, black and white tea products processed from Kenyan tea cultivars originally selected for black tea had significantly (P<0.05) higher antioxidant activity than green tea processed from tea cultivars from Japan and China. These results seem to suggest that the cultivar type is critical in determining the antioxidant potency of tea product and that black teas processed from suitable cultivars could be potent in antioxidant activity when compared to green teas.
A study to determine the physiological and biochemical responses of eight tea Camellia sinensis (L.) O. Kuntze cultivars to water-deficit stress was conducted in a ‘rain-out shelter’ using potted ...plants. Three levels of soil moisture content 34, 26, or 18% (v/v) water were applied to three plants of each cultivar in a complete randomised design, and the whole experiment was replicated three-times. The treatments were applied for 12 weeks, during which time plant water status, shoot extension rates, changes in gas exchange parameters, and leaf proline and glycinebetaine concentrations were determined. The imposition of severe water-deficit conditions 18% (v/v) soil water content caused a significant (P ≤ 0.05) decline in the relative water content of leaves, shoot water potentials, and shoot extension rates from mean values of 84.8% to 50.6%, –0.80 to –1.15MPa, and 1.87 to 0.29 mm d–1, respectively, compared to plants grown in a well-watered soil 34% (v/v) soil water content. The three gas exchange parameters measured (stomatal conductance, evapotranspiration rate, and rate of net photosynthesis) also declined significantly (P ≤ 0.05) with decreasing soil moisture content. In contrast, water-deficit stress increased the accumulation of leaf proline and glycinebetaine from mean values of 0.104 to 0.244 μmol g–1 FW, and from 1.567 to 2.025 μmol g–1 DW, respectively. The eight tea cultivars differed significantly (P ≤ 0.05) in their responses to water-deficit stress. Proline accumulation was significantly (P ≤ 0.05) higher in the drought-tolerant cultivars, ‘TRFK 306’, ‘TRFCA SFS150’, and ‘EPK TN14/3’, suggesting that proline concentration could be used as a marker for drought-tolerance in tea.
While soybean is an exotic crop introduced in Kenya early last century, promiscuous (TGx) varieties which nodulate with indigenous rhizobia have only recently been introduced. Since farmers in Kenya ...generally cannot afford or access fertilizer or inoculants, the identification of effective indigenous Bradyrhizobium strains which nodulate promiscuous soybean could be useful in the development of inoculant strains. Genetic diversity and phylogeny of indigenous Bradyrhizobium strains nodulating seven introduced promiscuous soybean varieties grown in two different sites in Kenya was assayed using the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) of the 16S-23S rDNA intergenic spacer region and 16S rRNA gene sequencing. PCR-RFLP analysis directly applied on 289 nodules using Msp I distinguished 18 intergenic spacer groups (IGS) I-XVIII. Predominant IGS groups were I, III, II, IV and VI which constituted 43.9%, 24.6%, 8.3% 7.6% and 6.9% respectively of all the analyzed nodules from the two sites while IGS group VII, IX, X, XI, XII, XIV, XVI, XVII, XVIII each constituted 1% or less. The IGS groups were specific to sites and treatments but not varieties. Phylogenetic analysis of the 16S rRNA gene sequences showed that all indigenous strains belong to the genus Bradyrhizobium. Bradyrhizobium elkanii, Bradyrhizobium spp and Bradyrhizobium japonicum related strains were the most predominant and accounted for 37.9%, 34.5%, and 20.7% respectively while B. yuanmigense related accounted for 6.9% of all strains identified in the two combined sites. The diversity identified in Bradyrhizobium populations in the two sites represent a valuable genetic resource that has potential utility for the selection of more competitive and effective strains to improve biological nitrogen fixation and thus increase soybean yields at low cost.
AFLP markers were successfully employed to detect diversity and genetic differentiation among Indian and Kenyan populations of tea (Camellia sinensis (L.) O. Kuntze). Shannon's index of diversity was ...used to partition the total phenotypic variation into between and within population components. On average, most of the diversity was detected within populations, with 79% of the variation being within and 21% being between populations of Indian and Kenyan tea. A dendrogram constructed on the basis of band sharing distinctly separated the three populations of tea into China type (sinensis), Assam type (assamica) and Cambod type (assamica ssp. lasiocalyx) in a manner consistent with the present taxonomy of tea, the known pedigree of some of the genotypes and their geographical origin. Principal coordinate (PCO) analysis grouped Assam genotypes both from India and Kenya supporting the suggestion that the Kenyan clones have been derived from collections made in this region. The China types were more dispersed on the PCO plot which is a reflection of wider genetic variation. As would be expected, clones collected from the same region exhibited less overall genetic variation. AFLP analysis discriminated all of the tested genotypes from India and Kenya, even those which cannot be distinguished on the basis of morphological and phenotypic traits.PUBLICATION ABSTRACT
The tea industry is significant in the economies of tea-growing countries. Prospects of improving yield of made tea genomic information were explored using clones from a cross between clones TRFCA ...SFS150 and AHP S15/10. The 42 clones were tested in two distinct tea-growing regions in Kenya. Bulk segregant analysis was performed followed by complete genotyping. Out of 260 informative markers, 100 markers that showed 1:1 segregation were used to construct a linkage map. The map contained 30 (19 maternal and 11 paternal) linkage groups that spanned 1,411.5 cM with mean interval of 14.1 cM between loci. Based on the map, quantitative trait loci (QTL) analysis was done on yield data over 2003-2007 across the two sites, Timbilil and Kangaita. Twenty-three putative QTLs were detected, 16 in five different linkage groups for Timbilil, two in two groups for Kangaita, and the rest were associated with unassigned markers. No QTL was detected at both sites, which showed strong genotype × site interaction (G × E) but highly effective within-site heritability ( graphic removed generally > 0.7). Problems of overestimated and spurious QTL effects arising from the smallness of the population should be mitigated by generally high within-site heritability. At least two unassigned markers associated with yield at Kangaita over the whole study period, suggesting potential as candidate markers for site-specific marker-assisted selections. Implications of the results with respect to mapping population, G × E, and marker-assisted selection are discussed.
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