The ability of apoplastic ascorbate to protect poplar leaves against ambient ozone concentrations: a quantitative approach.

Shoots of a sensitive (Populus nigra 'Brandaris') and a more tolerant (Populus euramericana 'Robusta') poplar clones were exposed for 30 days to Filtered Air or ambient O3-concentrations in fumigation cabinets. At regular intervals were determined: gas exchange of the leaves, the internal air space (Vair) and apoplastic water volume (Vapo) and the reduced (ASA) and oxidized (DHA) ascorbate concentration in the apoplast and in the mesophyll cells. The apoplastic ASA-concentration was 0.2 mM at the start of the experiment for both cultivars, while the effective cell wall thickness, estimated from Vapo, varied from 0.3 to 0.6 micron. Model calculations revealed that only 30% of the O3 molecules entering the apoplast was intercepted at these values. The O3-treatment induced a decline in stomatal conductance, an increase in Vapo and in the apoplastic ASA-concentration. As a result the estimated O3-flux to the cell membrane strongly declined. However, these responses occurred after the O3-induced reduction in photosynthesis. Moreover, they did not prevent early senescence of the leaves at a prolonged exposure. Therefore, it is concluded that the increase in apoplastic ASA-concentration was rather a general stress reaction of the affected poplar leaf than a (specific) defence reaction induced by O3. Our results suggest that other factors than the scavenging efficiency of apoplastic ASA were responsible for the difference in O3 sensitivity between both poplar cultivars.

Influence of acetylcholine agonists and antagonists on the swelling of etiolated wheat (Triticum aestivum L.) mesophyll protoplasts.

Etiolated wheat (Triticum aestivum L.) mesophyll protoplasts swell within 30 min in darkness after a red light (R) pulse or addition of acetylcholine (ACh), if 0.5 mM CaCl2 is present in the medium. In addition, ACh is also able to induce swelling in the presence of both 0.1 mM KCl or NaCl. Besides ACh, only carbamylcholine out of the choline derivatives tested was active in induction of swelling in the presence of K(+) or Na(+). The K(+)/Na(+)-dependent ACh-induced protoplast swelling was nullified by a 'calmodulin inhibitor', but not by Ca(2+)-channel blockers, Li(+) or VO 4 (3-) . The antagonists atropine (of muscarine-sensitive ACh receptors, mAChRs) andD-tubocurarine (of nicotine-sensitive ACh receptors, nAChRs) nullified the Ca(2+) - and the K(+)/Na(+)-dependent protoplast swelling responses, respectively, while having no effect on the Ca(2+)-dependent R-induced swelling response. Moreover, muscarine and nicotine mimicked ACh in the Ca(2+)- and K(+)/Na(+)-dependent swelling responses respectively. Just as is the case in animal cells, the proposed mAChRs appear to be associated with a phosphatidylinositol-dependent pathway, whereas the proposed nAChRs are phosphatidylinositol independent. Similarity between the action of ACh via the proposed mChRs and R via phytochrome in protoplast swelling indicates they share in common signal-transduction pathway.

The role of calcium ions in phytochrome-controlled swelling of etiolated wheat (Triticum aestivum L.) protoplasts.

Protoplasts from dark-grown wheat (Triticum aestivum L.) maintained at a constant osmotic potential at 22°C, were found to swell upon red irradiation (R) and the effect was negated by subsequent far-red light (FR), indicating phytochrome involvement. Swelling only occurred when Ca(2+) ions were present in the surrounding medium, or were added within 10 min after R. Furthermore, Mg(2+), Ba(2+) or K(+) could not replace this requirement for Ca(2+). The presence of K(+) did not enhance the Ca(2+)-dependent swelling response. When the Ca(2+)-ionophore A 23187 was added to the medium, protoplasts swelled in the dark to the same extent as after R. Both the Ca(2+)-channelblocker Verapamil and La(3+) inhibited R-induced swelling. It is proposed that R causes the opening of Ca(2+)-channels in the plasma membrane. Boyle-van't Hoff analyses of protoplast volume after R and FR are consistent with the conclusion that R irradiation causes changes in membrane properties.

TL-DNA from Agrobacterium rhizogenes plasmid pRi1855 reduces the osmotic pressure in transformed plants grown in vitro.

Growth, water content, osmotic pressure and solute content were examined for normal potato (Solanum tuberosum L. cv. Desiree) and a derivative (line D9X8a), which was genetically transformed with TL-DNA from Agrobacterium rhizogenes. Plants were grown (i) in vitro, (ii) in a growth chamber and (iii) in the field. In vitro, the transformed potato plants produced more biomass than the untransformed plants, partly because they had a higher water content. Potassium concentration and osmotic pressure were lower in cell sap extracted from the transformed potato shoots. In some cases the difference was as much as 50%. These differences were less clear, absent or reversed in plants from a growth chamber or from the field. In the field, however, transformed potato senesced early. It is suggested that a cellular basis for these observations may be changes induced by Ri TL-DNA expression products in plant membrane properties.

Developmental regulation of RI TL-DNA gene expression in roots, shoots and tubers of transformed potato (Solanum tuberosum cv. Desiree).

Expression of TL-DNA from Agrobacterium rhizogenes plasmid pRi 1855 was examined in a transformed derivative of Solanum tuberosum cv. Desiree, D9X8a. Northern blot analysis identified at least nine TL-DNA coded transcripts in roots, shoots and tubers but their relative abundance differed within and between organs. This revealed a distinctive pattern of organ specified differential expression. Grafting experiments showed that the abnormal shape of tubers of transformed potato was probably determined by TL-DNA products synthesised within the tuber and not by diffusable products synthesised in other parts of the plant. The abundance of at least one transcript, tr5, was probably determined by culture conditions. Implications for functions and control of expression of Ri TL-DNA genes are discussed. It is suggested that Ri TL-DNA provides a convenient and extensive set of model genes to study variation and stability of expression of linked foreign genes introduced into plants.