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Concentrations of inorganic and organic solutes in extracts from individual epidermal,mesophyll and bundle-sheath cells of barley leaves
Authors:Wieland Fricke  Roger A Leigh  A Deri Tomos
Institution:(1) School of Biological Sciences, University of Wales, LL57 2UW Bangor, Gwynedd, UK;(2) Biochemistry and Physiology Department, AFRC Institute of Arable Crops Research, Rothamsted Experimental Station, AL5 2JQ Harpenden, Hertfordshire, UK
Abstract:The distribution of solutes between epidermal, mesophyll and bundle-sheath cells in barley (Hordeum vulgare L. cv. Klaxon) leaves was studied by analysing extracts obtained from single cells with a modified pressure probe. Activity of the cytoplasmic marker enzyme, malate dehydrogenase, revealed that epidermal cell extracts were completely vacuolar in origin, but extracts from mesophyll cells also contained cytoplasmic constituents. The extracts were analysed for osmolality and the concentrations of K, Na, Ca, Cl, P, S, NO 3 , sugars and total amino acids. Epidermal and mesophyll cell extracts had similar osmolalities but these varied between 420 and 565 mosmol, kg 1 depending on the leaf developmental stage; the osmolality of bundle-sheath extracts was approximately 100 mosmol, kg–1 lower. Under the growth conditions used, K and NO 3 were found in all three cell types and their concentrations generally ranged between 180 and 230 mM. In contrast, Ca was almost restricted to epidermal cells, where it increased to 70 mM during leaf ageing. Phosphorus was only detectable (ge 5 mM) in extracts from mesophyll and bundle-sheath cells, while Cl concentrations were highest in epidermal and lowest in mesophyll cell extracts. The concentrations of sugars and amino acids were close to the detection limit (approx. 2 mM) in epidermal cells but mesophyll cells contained total sugar (glucose, fructose and sucrose) of up to 78 mM and total amino-acid concentrations of up to 13.5 mM. Concentrations in bundle-sheath cells were intermediate between those in the epidermis and mesophyll.Abbreviations EDX analysis energy dispersive X-ray analysis - MDH malate dehydrogenase We wish to thank Paul Richardson, Jeremy Pritchard, Peter Hinde and Andrew Davies (Banger) for their helpfull discussion and technical advice. This work was financed by a grant (LR5/521) from the Agricultural and Food Research Council.
Keywords:Epidermis (leaf)  Hordeum  Mesophyll  Osmolality  Single-cell sampling  Solute compartmentation
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