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Intact mesophyll cells can be rapidly isolated from the cladophylls ofAsparagus officinalis by gentle scraping with a plastic card, the yield being higher than 80% on a chlorophyll basis. The cells can be stored for at least 24h without loss of photosynthetic capacity and were found to be stable under a variety of conditions. In contrast to cell preparations from other plant species, photosynthetic activity was little affected by the presence of sorbitol as an osmoticum up to a concentration of 1.5 M. Similarly, the pH value of the medium influenced photosynthesis to only a small extent at a constant [CO2] of 200 μM. The response of the cells' photosynthetic capacity to light, temperature and CO2 concentration was similar to those reported for isolated cells from other plant species. Isolated cells ofA. officinalis can be used under a large range of conditions which gives them a measure of flexibility not possible with most plant cells which have sharply defined optimal conditions for photosynthesis. The isolated cells have a photosynthetic capacity of 40-60% of that of the intact cladophyll. The loss of photosynthetic activity observed upon isolation could not be accounted for by breakage of the cells. Virtually all of the cells were shown to be intact on the basis of Evans Blue exclusion and more than 80% of the cells contained intact chloroplasts and vacuoles. The entire loss of photosynthetic activity could be accounted for by a decrease in sucrose synthesis rather than by an equal decrease in the synthesis in all products. A six- to seven fold increase in the level of(14)C in hexose phosphates in the isolated cells supports the notion of inhibition of the sucrose-synthesis pathway. |
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