Objective:To identify the bioactive compounds in catechins isolation and its components from green tea GMB-4 clone.Methods:Green tea GMB-4 clones were extracted with distilled water at 90C.Samples were eluted into the...Objective:To identify the bioactive compounds in catechins isolation and its components from green tea GMB-4 clone.Methods:Green tea GMB-4 clones were extracted with distilled water at 90C.Samples were eluted into the column with 10%ethanol.Subsequently,the column was eluted with95%ethanol and evaporated separately.Green tea extract was identified by thin layer chromatography.Catechins were separated by the stationary phase in column chromatography using polyamide with 10%ethanol eluent and 95%ethanol.The results of isolations were analyzed by high performance liquid chromatographic(HPLC)and LCMS/MS.Analysis of catechins by HPLC was done by external standard.Results:Fraction from 10%ethanol showed that four major peaks at retention time of1.663,2.367,2.950 and 4.890,indicated the presence of four catechins components including catechin,epicatechins,gallocatechin and epigallocatechin.Whereas,fraction from 95%ethanol showed two main peaks at retention time of 5.167 and 9.82,which indicated the presence of epigallocatechin gallate(EGCG)and epicatechin gallate(ECG).EGCG(m/z 459),epigallocatechin(m/z 307),ECG(m/z 443),and epicatechin(m/z 291)were isolated and separated successfully using HPLC and LC-MS/MS.Conclusions:The HPLC and LC-MS/MS methods were successfully tuned for the qualitative analysis of green tea extract with EGCG and ECG.Four major catechins were separated and identified by LC-MS/MS,such as EGCG,epigallocatechin,ECG and epicatechin.The result of HPLC analysis showed that EGCG and ECG were main components from catechins isolation of green tea GMB-4 clone.展开更多
The research screening of adoptive elite tea clones was conducted at NTRI, Mansehra during 2011-2012. Nine clones 101Aa, 105aa, 108aa, 561aa, ll7aa, 219ab, 470bb and 180bd were evaluated for seedling performance. Rand...The research screening of adoptive elite tea clones was conducted at NTRI, Mansehra during 2011-2012. Nine clones 101Aa, 105aa, 108aa, 561aa, ll7aa, 219ab, 470bb and 180bd were evaluated for seedling performance. Randomized complete block design was used with three replications. Data was recorded on various morphological characters after 8 months. The results showed that high survival percentage, shoot length, number of roots plant-1, number of leaves plant1 and root length were observed in clone 105aa. While the highest fresh weight and dry weight of leaves were observed in clones 117aa and 105aa. The clone 105aa was drought resistant, high survival percentage and root growth. On the basis of the results, clone 105aa was recommended for cultivation through cuttings in the hilly areas of Pakistan where unequal rainfall distribution was a major hitch.展开更多
[ Objective ] This study aimed to verify the feasibility of in silico cloning of functional candidate genes in tea. [ Method ] Theobroma cacao caffeine syn- thase gene BCS1 was used as a probe to search the establishe...[ Objective ] This study aimed to verify the feasibility of in silico cloning of functional candidate genes in tea. [ Method ] Theobroma cacao caffeine syn- thase gene BCS1 was used as a probe to search the established tea EST database using BLAST; 26 tea ESTs highly homologous to BCS1 were obtained, which were assembled using CAP （contig assembly program） of BioEdit software; subsequently, two EST configs harboring ORF were obtained, which were named TCSnewl and TCSnew2, respectively. Nucleotide sequences and deduced amino acid sequences of theses two genes were compared with those of cDNA of tea caffeine synthase gene TCS in the GenBank database that was cloned with experimental biological method. A phylogenetic tree was constructed for homalogous analysis of the deduced amino acid sequences of theses three genes. [ Result] in silico cloning of functional candidate genes in tea using a homologous gene of distantly related species as a probe is a feasible technical means. [ Conclusion] This study provided the basis for in silico cloning of other functional genes in tea.展开更多
以4个茶树种质为试验材料,克隆得到茶叶中氨基酸合成转化关键酶基因:谷氨酸脱氢酶(GDH)、谷氨酰胺合成酶(GS)、谷氨酰胺α-酮戊二酸氨基转移酶(GOGAT)的保守区。在GenBank登录号分别为:JN602371、JN602372、JN602373。通过SYBR Green I...以4个茶树种质为试验材料,克隆得到茶叶中氨基酸合成转化关键酶基因:谷氨酸脱氢酶(GDH)、谷氨酰胺合成酶(GS)、谷氨酰胺α-酮戊二酸氨基转移酶(GOGAT)的保守区。在GenBank登录号分别为:JN602371、JN602372、JN602373。通过SYBR Green I实时荧光定量PCR检测,发现GDH酶基因在茶树种质0314C(相对高氨基酸种质)中的表达显著增强,而在0212-15种质(相对低氨基酸种质)中却显著下降。GS、GOGAT酶基因在种质0314C中表达显著下降,而在0212-15、0318D种质中显著增强。通过回归分析,GS基因表达与茶氨酸、赖氨酸、丙氨酸呈负相关,而GDH与茶氨酸呈正相关。展开更多
基金supported by a grant of Directorate of General Higher Education,Ministry of Education and Culture of Indonesia for the"BPPDN"scholarship
文摘Objective:To identify the bioactive compounds in catechins isolation and its components from green tea GMB-4 clone.Methods:Green tea GMB-4 clones were extracted with distilled water at 90C.Samples were eluted into the column with 10%ethanol.Subsequently,the column was eluted with95%ethanol and evaporated separately.Green tea extract was identified by thin layer chromatography.Catechins were separated by the stationary phase in column chromatography using polyamide with 10%ethanol eluent and 95%ethanol.The results of isolations were analyzed by high performance liquid chromatographic(HPLC)and LCMS/MS.Analysis of catechins by HPLC was done by external standard.Results:Fraction from 10%ethanol showed that four major peaks at retention time of1.663,2.367,2.950 and 4.890,indicated the presence of four catechins components including catechin,epicatechins,gallocatechin and epigallocatechin.Whereas,fraction from 95%ethanol showed two main peaks at retention time of 5.167 and 9.82,which indicated the presence of epigallocatechin gallate(EGCG)and epicatechin gallate(ECG).EGCG(m/z 459),epigallocatechin(m/z 307),ECG(m/z 443),and epicatechin(m/z 291)were isolated and separated successfully using HPLC and LC-MS/MS.Conclusions:The HPLC and LC-MS/MS methods were successfully tuned for the qualitative analysis of green tea extract with EGCG and ECG.Four major catechins were separated and identified by LC-MS/MS,such as EGCG,epigallocatechin,ECG and epicatechin.The result of HPLC analysis showed that EGCG and ECG were main components from catechins isolation of green tea GMB-4 clone.
文摘The research screening of adoptive elite tea clones was conducted at NTRI, Mansehra during 2011-2012. Nine clones 101Aa, 105aa, 108aa, 561aa, ll7aa, 219ab, 470bb and 180bd were evaluated for seedling performance. Randomized complete block design was used with three replications. Data was recorded on various morphological characters after 8 months. The results showed that high survival percentage, shoot length, number of roots plant-1, number of leaves plant1 and root length were observed in clone 105aa. While the highest fresh weight and dry weight of leaves were observed in clones 117aa and 105aa. The clone 105aa was drought resistant, high survival percentage and root growth. On the basis of the results, clone 105aa was recommended for cultivation through cuttings in the hilly areas of Pakistan where unequal rainfall distribution was a major hitch.
基金Supported by National Science and Technology Support Program of China(2011BAD01B01)Youth Talent Innovation Fund of Fujian Academy of Agricultural Sciences(2011QC-2)Special Fund for"Double Hundred Plan"of Fujian Academy of Agricultural Sciences(sbmx1303-1)
文摘[ Objective ] This study aimed to verify the feasibility of in silico cloning of functional candidate genes in tea. [ Method ] Theobroma cacao caffeine syn- thase gene BCS1 was used as a probe to search the established tea EST database using BLAST; 26 tea ESTs highly homologous to BCS1 were obtained, which were assembled using CAP （contig assembly program） of BioEdit software; subsequently, two EST configs harboring ORF were obtained, which were named TCSnewl and TCSnew2, respectively. Nucleotide sequences and deduced amino acid sequences of theses two genes were compared with those of cDNA of tea caffeine synthase gene TCS in the GenBank database that was cloned with experimental biological method. A phylogenetic tree was constructed for homalogous analysis of the deduced amino acid sequences of theses three genes. [ Result] in silico cloning of functional candidate genes in tea using a homologous gene of distantly related species as a probe is a feasible technical means. [ Conclusion] This study provided the basis for in silico cloning of other functional genes in tea.
文摘以4个茶树种质为试验材料,克隆得到茶叶中氨基酸合成转化关键酶基因:谷氨酸脱氢酶(GDH)、谷氨酰胺合成酶(GS)、谷氨酰胺α-酮戊二酸氨基转移酶(GOGAT)的保守区。在GenBank登录号分别为:JN602371、JN602372、JN602373。通过SYBR Green I实时荧光定量PCR检测,发现GDH酶基因在茶树种质0314C(相对高氨基酸种质)中的表达显著增强,而在0212-15种质(相对低氨基酸种质)中却显著下降。GS、GOGAT酶基因在种质0314C中表达显著下降,而在0212-15、0318D种质中显著增强。通过回归分析,GS基因表达与茶氨酸、赖氨酸、丙氨酸呈负相关,而GDH与茶氨酸呈正相关。
