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J K Cheesbrough

Publications and source records attributed to J K Cheesbrough.

4 recordsLinked to original sources

Photosynthesis by intact isolated chloroplasts on solid support.

A new approach to measurements of photosynthesis by isolated chloroplasts has been devised. Intact isolated chloroplasts were trapped in the cavities of membrane filters. The thin layers of chloroplasts so obtained were assayed for O(2) evolution and CO(2) assimilation in leaf-chambers. Photosynthetic gas exchange could be demonstrated to take place either in a closed or a flow-through system. The chloroplasts were morphologically intact as shown by light or scanning electron microscopy and displayed stable rates of photosynthesis in the presence of phosphate and alkaline phosphatase. The methods described open the way to in vitro measurement of photosynthesis, by chloroplasts under conditions more closely resembling those in leaves.

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Measurement of CO(2) and H(2)O Vapor Exchange in Spinach Leaf Discs : Effects of Orthophosphate.

A leaf chamber has been designed which allows the measurement of both CO(2) and water vapor exchange in Spinacia oleracea leaf discs. The center of the disc lies within a cylindrical gas chamber and its margins are enclosed within a cavity through which water or various metabolites can be pumped. In saturating light and normal atmospheres, the leaf discs have a relatively low resistance to H(2)O vapor transfer (r(w) = 1.87 seconds per centimeter) and can support high rates of photosynthesis for several hours. The abaxial surface of a disc had a higher resistance to water vapor transfer (r(w) = 3.22 seconds per centimeter) than the adaxial (r(w) = 2.45 seconds per centimeter) despite having a higher stomatal frequency (abaxial, 105/square millimeter; adaxial, 58/square millimeter). In 2% O(2), the discs required an internal concentration of CO(2) of 115 microliters per liter to support one-half of the maximal velocity of apparent photosynthesis (average value, 66 milligrams CO(2) per square decimeter per hour). In 20% O(2), the comparable values are 156 microliters per liter and 56 milligrams CO(2) per square decimeter per hour. In air, apparent photosynthesis saturated at intensities (750 microeinsteins per square meter per second) well below that of daylight but, when the internal CO(2) was raised to 700 to 900 microliters per liter, photosynthesis was not saturated even at daylight intensities (2025 microeinsteins per square meter per second). The distribution of Prussian blue crystals, formed after ferrocyanide feeding, showed that water entered the disc via the vasculature. When 25-minute pulses of orthophosphate were provided in the feeding solution, there were concentration-dependent increases in both r(w) and r(m) leading to inhibition of photosynthesis. The orthophosphate-dependent inhibitions were reversible.

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Effects of mannose on photosynthetic gas exchange in spinach leaf discs.

When mannose is provided in the transpiration stream to spinach (Spinacia oleracea) leaf discs, a series of specific and nonspecific changes occur in CO(2) and H(2)O vapor exchange as a function of feeding time. The initial increases in apparent photosynthesis and transpiration are nonspecific effects due to osmotic changes leading to passive stomatal opening. The mannose-specific effects are: (a) time-dependent changes in the CO(2) concentration required for saturation; (b) complex kinetics of the inhibition of CO(2) assimilation dependent on CO(2) and O(2) concentrations and the duration of feeding (high CO(2) and low O(2) lead to rapid inhibitions of photosynthesis); (c) elimination of the capacity of 2% O(2) to stimulate photosynthesis; and (d) oscillations in the CO(2) exchange rate following transitions from 20% to 2% O(2). The mannose-specific effects are reversible by orthophosphate. The mannose-dependent changes in gas exchange are attributed to altered [ATP]/[ADP] ratios.

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Simultaneous measurement of oscillations in oxygen evolution and chlorophyll a fluorescence in leaf pieces.

In spinach (Spinacia oleracea) and barley (Hordeum vulgare) leaves, chlorophyll a fluorescence and O(2) evolution have been measured simultaneously following re-illumination after a dark interval or when steady state photosynthesis has been perturbed by changes in the gas phase. In high CO(2) concentrations, both O(2) and fluorescence can display marked dampening oscillations that are antiparallel but slightly out of phase (a rise or fall in fluorescence anticipating a corresponding fall or rise in O(2) by about 10 to 15 seconds). Infrared gas analysis measurements showed that CO(2) uptake behaved like O(2) evolution both in the period of oscillation (about 1 minute) and in its relation to fluorescence. In the steady state, oscillations were initiated by increases in CO(2) or by increases or decreases in O(2). Oscillations in O(2) or CO(2) did not occur without associated oscillations in fluorescence and the latter were a sensitive indicator of the former. The relationship between such oscillations in photosynthetic carbon assimilation and chlorophyl a fluorescence is discussed in the context of the effect of ATP or NADPH consumption on known quenching mechanisms.

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