[Objective] This study aimed to analyze the difference in the contents of gallic acid and catechins of tea resources from Yunnan Province. [Method] By using high performance liquid chromatography （HPLC）, the content...[Objective] This study aimed to analyze the difference in the contents of gallic acid and catechins of tea resources from Yunnan Province. [Method] By using high performance liquid chromatography （HPLC）, the contents of gallic acid （GA）, catechins （C）, epicatechin （EC）, epigallocatechin （EGC）, epicatechin gallate （ECG） and epigallocatechin gallate （EGCG） of 121 tea germplasms from the China National Germplasm Tea Repositories （CNGTR） at the Tea Research Institute of Yunnan Academy of Agricultural Sciences （TRIYAAS） were measured. [Result] The content of GA ranged from 0.210% to 1.902%, with an average of 0.834%, explaining rela- tively low GA content among tea germplasms. The content of C ranged from 0.069% to 8.865%, with an average of 1.916%. The content of EC ranged from 0.126% to 2.865%, with an average of 1.112%. The content of EGC ranged from 0.00% to 3.709%, with an average of 0.954%. The content of ECG ranged from 0.739% to 8.957%, with an average of 4.063%. The content of EGCG ranged from 0.819% to 11.77%, with an average of 5.939%. The content of total C ranged from 6.354% to 22.654%, with an average of 14.042%. [Conclusion] There was relatively big difference of catechin contents among different tea resources, indicating that there was plentiful biodiversity of Yunnan tea germplasms. At the same time, three tea germplasms with high epigallocatechin gallate content （≥10%） was selected preliminarily, which would provide important materials for breeding tea cultivars with high EGCG content in the future.展开更多
A bacterial artificial chromosome （BAC） library is an invaluable resource tool to initiate tea plant genomics research, and the preparation of high molecular weight （HMW） genomic DNA is a crucial first step for co...A bacterial artificial chromosome （BAC） library is an invaluable resource tool to initiate tea plant genomics research, and the preparation of high molecular weight （HMW） genomic DNA is a crucial first step for constructing a BAC Library. In order to construct a BAC library for enhancing tea plant genomics research, a new method for the preparation of tea pant high molecular weight （HMW） genomic DNA must be developed due to young tea plant leaves and shoots are notably rich in both tea polyphenols and tea polysaccharides. In this paper, a modified method for preparing high quality tea plant HMW genomi~ DNA was optimized, and the quality of tea plant genomic DNA was evaluated. The results were as follows： Critical indicators of HMW DNA preparation were the appearance of the smooth nuclei in solution （as opposed to sticky-gummy） before agarose plug solidification, non-dark colored nuclei plugs after lysis with an SDS/proteinase K solution, and the quality and quantity of HMW DNA fragments after restriction enzyme digestion. Importantly, 1% dissolved PVP-40 and 1% un-dissolved PVP-40 during the nuclei extraction steps, in conjunction with the removal of PVP-40 from the plug washing and nuclei lysis steps, were critical for achieving HWM tea plant DNA suitable for BAC library construction. Additionally, a third PFGE fraction selection step to eliminate contaminating small DNA fragments. The modifications provided parameters that may have prevented deleterious interactions from tea polyphenols and tea polysaccharides. The HMW genomic DNA produced by this new modified method has been used to successfully construct a large-insert tea plant BAC library, and thus may be suitable for BAC library construction from other plant species that contain similarly interfering compounds.展开更多
To investigate the protective effect of epigallocatechin gallate （EGCG） on the immune function of dendritic cells （DCs） after ultraviolet B irradiation （UVB） and its underlying mechanisms, the monocytes were iso...To investigate the protective effect of epigallocatechin gallate （EGCG） on the immune function of dendritic cells （DCs） after ultraviolet B irradiation （UVB） and its underlying mechanisms, the monocytes were isolated from peripheral blood and cultivated into DCs with cytokines, such as GM-CSF and IL-4. DCs were harvested after cultivation for 7 d and subjected to irradiation with different dosages of UVB. Then, 200 μg/ml of EGCG were added in certain groups immediately after irradiation. DCs simply treated with UVB or treated with both UVB and EGCG were co-cultured with lymphocytes, and MTT assay was used to detect the ability of DCs to stimulate proliferation of lymphocytes. Surface markers CDS0, CD86, HLA-DR and CD40 were detected by flow cytometry, and the levels of IL-10 and IL-12 secreted from DCs 2d h after cultivation were measured by ELISA. It was demonstrated that UVB irradiation could inhibit the ability of DCs to stimulate the proliferation of lymphocytes and surface expressions of CDS0, CD86, HLA-DR and CD40 on DCs in a dose-dependent manner. The inhibition rate of DCs was improved to some extent after treatment with 200μg/ml of EGCG. When the concentra- tion of EGCG exceeded 100 μg/ml, the enhancing effect of EGCG on the expression of the co-stimulating molecules on DCs could be demonstrated in a dose-dependent manner. UVB showed no significant influence on the secretion of IL-10 and IL-12 from DCs, while EGCG could down-regulate the secretion level of IL-12 and up-regulate that of IL-10. It is concluded that EGCG can antagonize the inhibitory effect on DCs induced by UVB irradiation. This function has some relationship with its protecting effect of the expression of the co-stimulating molecule on the surface of DCs and the secretion level of IL-10 and IL-12.展开更多
基金Supported by National Natural Science Foundation of China(31160175)Technology Innovation Talents Project of Yunnan Province(2011CI068)+1 种基金Special Fund for National Modern Agricultural Industrial Technology System Construction(nycytx-23)Seed Preservation Project of Ministry of Agriculture(NB2012-2130135)~~
文摘[Objective] This study aimed to analyze the difference in the contents of gallic acid and catechins of tea resources from Yunnan Province. [Method] By using high performance liquid chromatography （HPLC）, the contents of gallic acid （GA）, catechins （C）, epicatechin （EC）, epigallocatechin （EGC）, epicatechin gallate （ECG） and epigallocatechin gallate （EGCG） of 121 tea germplasms from the China National Germplasm Tea Repositories （CNGTR） at the Tea Research Institute of Yunnan Academy of Agricultural Sciences （TRIYAAS） were measured. [Result] The content of GA ranged from 0.210% to 1.902%, with an average of 0.834%, explaining rela- tively low GA content among tea germplasms. The content of C ranged from 0.069% to 8.865%, with an average of 1.916%. The content of EC ranged from 0.126% to 2.865%, with an average of 1.112%. The content of EGC ranged from 0.00% to 3.709%, with an average of 0.954%. The content of ECG ranged from 0.739% to 8.957%, with an average of 4.063%. The content of EGCG ranged from 0.819% to 11.77%, with an average of 5.939%. The content of total C ranged from 6.354% to 22.654%, with an average of 14.042%. [Conclusion] There was relatively big difference of catechin contents among different tea resources, indicating that there was plentiful biodiversity of Yunnan tea germplasms. At the same time, three tea germplasms with high epigallocatechin gallate content （≥10%） was selected preliminarily, which would provide important materials for breeding tea cultivars with high EGCG content in the future.
文摘A bacterial artificial chromosome （BAC） library is an invaluable resource tool to initiate tea plant genomics research, and the preparation of high molecular weight （HMW） genomic DNA is a crucial first step for constructing a BAC Library. In order to construct a BAC library for enhancing tea plant genomics research, a new method for the preparation of tea pant high molecular weight （HMW） genomic DNA must be developed due to young tea plant leaves and shoots are notably rich in both tea polyphenols and tea polysaccharides. In this paper, a modified method for preparing high quality tea plant HMW genomi~ DNA was optimized, and the quality of tea plant genomic DNA was evaluated. The results were as follows： Critical indicators of HMW DNA preparation were the appearance of the smooth nuclei in solution （as opposed to sticky-gummy） before agarose plug solidification, non-dark colored nuclei plugs after lysis with an SDS/proteinase K solution, and the quality and quantity of HMW DNA fragments after restriction enzyme digestion. Importantly, 1% dissolved PVP-40 and 1% un-dissolved PVP-40 during the nuclei extraction steps, in conjunction with the removal of PVP-40 from the plug washing and nuclei lysis steps, were critical for achieving HWM tea plant DNA suitable for BAC library construction. Additionally, a third PFGE fraction selection step to eliminate contaminating small DNA fragments. The modifications provided parameters that may have prevented deleterious interactions from tea polyphenols and tea polysaccharides. The HMW genomic DNA produced by this new modified method has been used to successfully construct a large-insert tea plant BAC library, and thus may be suitable for BAC library construction from other plant species that contain similarly interfering compounds.
文摘To investigate the protective effect of epigallocatechin gallate （EGCG） on the immune function of dendritic cells （DCs） after ultraviolet B irradiation （UVB） and its underlying mechanisms, the monocytes were isolated from peripheral blood and cultivated into DCs with cytokines, such as GM-CSF and IL-4. DCs were harvested after cultivation for 7 d and subjected to irradiation with different dosages of UVB. Then, 200 μg/ml of EGCG were added in certain groups immediately after irradiation. DCs simply treated with UVB or treated with both UVB and EGCG were co-cultured with lymphocytes, and MTT assay was used to detect the ability of DCs to stimulate proliferation of lymphocytes. Surface markers CDS0, CD86, HLA-DR and CD40 were detected by flow cytometry, and the levels of IL-10 and IL-12 secreted from DCs 2d h after cultivation were measured by ELISA. It was demonstrated that UVB irradiation could inhibit the ability of DCs to stimulate the proliferation of lymphocytes and surface expressions of CDS0, CD86, HLA-DR and CD40 on DCs in a dose-dependent manner. The inhibition rate of DCs was improved to some extent after treatment with 200μg/ml of EGCG. When the concentra- tion of EGCG exceeded 100 μg/ml, the enhancing effect of EGCG on the expression of the co-stimulating molecules on DCs could be demonstrated in a dose-dependent manner. UVB showed no significant influence on the secretion of IL-10 and IL-12 from DCs, while EGCG could down-regulate the secretion level of IL-12 and up-regulate that of IL-10. It is concluded that EGCG can antagonize the inhibitory effect on DCs induced by UVB irradiation. This function has some relationship with its protecting effect of the expression of the co-stimulating molecule on the surface of DCs and the secretion level of IL-10 and IL-12.
