Immunohistochemical localization of caffeine in young Camellia sinensis (L.) O. Kuntze (tea) leaves

Breda, Shane Vontelin van; Merwe, C.F. van der; Robbertse, Hannes; Apostolides, Zeno

doi:10.1007/s00425-012-1804-x

Cited by 15 publications

(7 citation statements)

References 54 publications

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“…These enzymes have substrate (Table S1). Additionally, although xanthine alkaloids may not be homogeneously distributed at the subcellular and tissue level (48), reported concentrations of the relevant intermediates in Theobroma, Paullinia, and Citrus may be conservatively estimated to be in the 10-1,000 μM range (34,38,41), which are comparable to the K M values we obtained (Table S1). …”

Section: Resultssupporting

confidence: 83%

Convergent evolution of caffeine in plants by co-option of exapted ancestral enzymes

Huang

O’Donnell

Barboline

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

114

156

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Convergent evolution is a process that has occurred throughout the tree of life, but the historical genetic and biochemical context promoting the repeated independent origins of a trait is rarely understood. The well-known stimulant caffeine, and its xanthine alkaloid precursors, has evolved multiple times in flowering plant history for various roles in plant defense and pollination. We have shown that convergent caffeine production, surprisingly, has evolved by two previously unknown biochemical pathways in chocolate, citrus, and guaraná plants using either caffeine synthaseor xanthine methyltransferase-like enzymes. However, the pathway and enzyme lineage used by any given plant species is not predictable from phylogenetic relatedness alone. Ancestral sequence resurrection reveals that this convergence was facilitated by co-option of genes maintained over 100 million y for alternative biochemical roles. The ancient enzymes of the Citrus lineage were exapted for reactions currently used for various steps of caffeine biosynthesis and required very few mutations to acquire modern-day enzymatic characteristics, allowing for the evolution of a complete pathway. Future studies aimed at manipulating caffeine content of plants will require the use of different approaches given the metabolic and genetic diversity revealed by this study. convergent evolution | caffeine biosynthesis | enzyme evolution | paleomolecular biology

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Section: Resultssupporting

confidence: 83%

Convergent evolution of caffeine in plants by co-option of exapted ancestral enzymes

Huang

O’Donnell

Barboline

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

114

156

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“…They concluded that it might be possible that caffeine is bound with the cell wall phenolic compounds. During immunolocalization, caffeine washed out easily and the tissues that were poor in phenolic cell wall components produced a weaker signal due to a lower caffeine content [59]. A similar phenomenon to that described above was observed in the case of wood impregnation.…”

Section: Computational Studiessupporting

confidence: 64%

“…The presence of this alkaloid in beans and young leaves explains the possible role of caffeine as a chemical defense. Breda et al (2013) visually determined that caffeine was on the periphery of cells of xylem and phloem. They concluded that it might be possible that caffeine is bound with the cell wall phenolic compounds.…”

Section: Computational Studiesmentioning

confidence: 99%

Chemical Changes of Wood Treated with Caffeine

et al. 2021

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Earlier studies have revealed that wood treated with caffeine was effectively protected against decay fungi and molds. However, there is a need to establish how the caffeine molecule behaves after wood impregnation and how it can protect wood. The objective of the research was to characterize the interaction between caffeine and Scots pine (Pinus sylvestris L.) wood as well as to assess the stability of the alkaloid molecule in lignocellulosic material. For this purpose, an elementary analyzer was used to assess the nitrogen concentration in the treated wood. The results showed that caffeine is easily removed from the wood structure through large amounts of water. The changes occurring in the wood structure after impregnation were evaluated with regard to the results obtained by Fourier transform infrared (FTIR) spectroscopy of two model mixtures with caffeine and cellulose or lignin for the purpose of conducting a comparison with the spectrum of impregnated and non-impregnated samples. The observed changes in FTIR spectra involve the intensity of the C=O(6) caffeine carbonyl group and signals from guaiacyl units. It might indicate favorable interactions between caffeine and lignin. Additionally, molecular simulation of the caffeine’s interaction with the guaiacyl β-O-4 lignin model compound characteristic for the lignin structure using computational studies was performed. Consequently, all analyses confirmed that caffeine may interact with the methylene group derived from the aromatic rings of the guaiacyl group of lignin. In summary, scanning electron microscope (SEM) observations suggest that caffeine was accumulated in the lignin-rich areas of the primary walls.

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“…In addition, immunohistochemistry is advantageous for the localization of small molecules because the technique is substantially more specific than other standard histochemical methods using stains, such as vanillin-hydrochloric acid, which are not unique for a distinct small molecule, but rather an entire class of compounds, and false positives or side reactions may occur. Furthermore, immunohistochemistry has been used to determine effectively the localization of caffeine in young tea leaves ( van Breda et al, 2013 ). Monoclonal antibodies are a superior choice for immunohistochemistry because they detect only one epitope in a particular antigen.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, this method has been successfully used to localize some secondary metabolites in plant. Immunohistochemical localization with primary anti-caffeine antibodies and conjugated secondary antibodies on tea leaf sections proved at the tissue level that caffeine was localized and accumulated within vascular bundles, mainly the precursor phloem ( van Breda et al, 2013 ). Similarly, delta 9-tetrahydrocannabinol (THC) localization in glandular trichomes and bracteal tissues of Cannabis was examined with a mAb, which can specifically recognize THC ( Kim and Mahlberg, 1997 ).…”

Section: Introductionmentioning

confidence: 99%

Subcellular Localization of Galloylated Catechins in Tea Plants [Camellia sinensis (L.) O. Kuntze] Assessed via Immunohistochemistry

Chen

et al. 2016

Front. Plant Sci.

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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.

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Immunohistochemical localization of caffeine in young Camellia sinensis (L.) O. Kuntze (tea) leaves

Cited by 15 publications

References 54 publications

Convergent evolution of caffeine in plants by co-option of exapted ancestral enzymes

Convergent evolution of caffeine in plants by co-option of exapted ancestral enzymes

Chemical Changes of Wood Treated with Caffeine

Subcellular Localization of Galloylated Catechins in Tea Plants [Camellia sinensis (L.) O. Kuntze] Assessed via Immunohistochemistry

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