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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
agricultural and biological sciences
Immunohistochemical localization of caffeine in young Camellia sinensis (L.) O. Kuntze (tea) leaves
Planta, Volume 237, No. 3, Year 2013
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Description
The anatomical localization of caffeine within young Camellia sinensis leaves was investigated using immunohistochemical methods and confocal scanning laser microscopy. Preliminary fixation experiments were conducted with young C. sinensis leaves to determine which fixation procedure retained caffeine the best as determined by high-performance liquid chromatography analysis. High pressure freezing, freeze substitution, and embedding in resin was deemed the best protocol as it retained most of the caffeine and allowed for the samples to be sectioned with ease. Immunohistochemical localization with primary anti-caffeine antibodies and conjugated secondary antibodies on leaf sections proved at the tissue level that caffeine was localized and accumulated within vascular bundles, mainly the precursor phloem. With the use of a pressure bomb, xylem sap was collected using a micro syringe. The xylem sap was analyzed by thin-layer chromatography and the presence of caffeine was determined. We hypothesize that caffeine is synthesized in the chloroplasts of photosynthetic cells and transported to vascular bundles where it acts as a chemical defense against various pathogens and predators. Complex formation of caffeine with chlorogenic acid is also discussed as this may also help explain caffeine's localization. © 2012 Springer-Verlag Berlin Heidelberg.
Authors & Co-Authors
van Breda, Shane Vontelin
South Africa, Pretoria
University of Pretoria
van der Merwe, Christiaan Frederick
South Africa, Pretoria
University of Pretoria
Robbertse, Hannes
South Africa, Pretoria
University of Pretoria
Apostolides, Zeno
South Africa, Pretoria
University of Pretoria
Statistics
Citations: 16
Authors: 4
Affiliations: 1
Identifiers
Doi:
10.1007/s00425-012-1804-x
ISSN:
00320935
e-ISSN:
14322048