Subcellular Localization of Galloylated Catechins in Tea Plants [Camellia sinensis (L.) O. Kuntze] Assessed via Immunohistochemistry.
Galloylated catechins, as the main secondary metabolites in the tea plant, including (-)-epigallocatechin-3-gallate and (-)-epicatechin-3-gallate, comprise approximately three-quarters of all the tea plant catechins and have stronger effects than non-galloylated catechins, both on the product quality in tea processing and the pharmacological efficacy to human beings. The subcellular localization of galloylated catechins has been the primary focus of studies that assess biosynthesis and physiological functions. Classical histochemical localization staining reagents can not specifically detect galloylated catechins; thus, their subcellular localization remains controversial. In the present study, we generated a monoclonal antibody (mAb) against galloylated catechins, which can be used for the subcellular localization of galloylated catechins in the tea plant by immunohistochemistry. Direct ELISA and ForteBio Octet Red 96 System assay indicated the mAb could recognize the galloylated catechins with high specificities and affinities. In addition, tea bud was ascertained as the optimal tissue for freezing microtomic sections for immunohistochemistry. What's more, the high quality mAbs which exhibited excellent binding capability to galloylated catechins were utilized for the visualization of them via immunohistochemistry. Our findings demonstrated that vacuoles were the primary sites of localization of galloylated catechins at the subcellular level.
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China; College of Food Science and Technology, Yunnan Agricultural UniversityKunming, China.
Huanhuan
H
Xu
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China; College of Food Science and Technology, Yunnan Agricultural UniversityKunming, China.
Ya
Y
Wang
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China.
Yana
Y
Chen
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China.
Pan
P
Zhang
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China.
Yi
Y
Zhao
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China; College of Food Science and Technology, Yunnan Agricultural UniversityKunming, China.
Yewei
Y
Huang
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China; College of Pu-er Tea, Yunnan Agricultural UniversityKunming, China; State Key Laboratory for Conservation and Utilization of Bio-Resources in YunnanKunming, China.
Xuanjun
X
Wang
Key Laboratory of Pu-er Tea Science, Ministry of Education, Yunnan Agricultural UniversityKunming, China; Tea Research Center of YunnanKunming, China; State Key Laboratory for Conservation and Utilization of Bio-Resources in YunnanKunming, China.
Jun
J
Sheng
Camellia sinensis; HPLC; catechins; galloylated catechins; immunohistochemistry; subcellular localization