Summary
Guard cells and epidermal cells of the abaxial (lower) and adaxial (upper) epidermis ofPisum sativum L., mutant Argenteum, are the predominant sites of flavonoid accumulation within the leaf. This was demonstrated by the use of a new method of simultaneous isolation and separation of intact, highly-purified guard cell and epidermal cell protoplasts from both epidermal layers and of protoplasts from the mesophyll. Isolated guard and epidermal protoplasts retained flavonoid patterns of the parent epidermal tissue; quercetin 3-triglucoside and its p-coumaric acid ester as major constituents, kaempferol 3-triglucoside and its p-coumaric acid ester as minor compounds. Total flavonoid content in the lower epidermis was estimated to be ca. 80 fmol per guard cell protoplast and 500 fmol per epidermal cell protoplast. Protoplasts isolated from the upper epidermis had about 20–30% as much of these flavonoids. Mesophyll protoplasts retained only about 25 fmol total flavonoid per protoplast.
By fluorescence microscopy, using the alkaline-induced yellow-green fluorescence characteristics of flavonols, we suggest that these flavonol glycosides are present in cell vacuoles. There was no indication for the presence of flavine-like compounds.
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Abbreviations
- uE:
-
adaxial (upper) epidermis
- IE:
-
abaxial (lower) epidermis
- GCP:
-
guard cell protoplasts
- ECP:
-
epidermal cell protoplasts
- MCP:
-
mesophyll cell protoplasts
- PP:
-
protoplasts
- HPLC:
-
high performance liquid chromatography
- TLC:
-
thin layer chromatography
- CC:
-
column chromatography
- HOAc:
-
acetic acid
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Weissenböck, G., Hedrich, R. & Sachs, G. Secondary phenolic products in isolated guard cell, epidermal cell and mesophyll cell protoplasts from pea (Pisum sativum L.) leaves: Distribution and determination. Protoplasma 134, 141–148 (1986). https://doi.org/10.1007/BF01275712
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DOI: https://doi.org/10.1007/BF01275712