J. Pospíšilová

Participation of Phytohormones in the Stomatal Regulation of Gas Exchange During Water Stress

Almost all processes in the life of a plant are directly or indirectly affected by both stresses and phytohormones. Nevertheless, apart from abscisic acid, the role of phytohormones in plant response to water stress is far from being fully elucidated. This review tries to answer the question whether interactions between abscisic acid and some other phytohormones might be important in the regulation of stomatal opening during water stress and subsequent rehydration. Firstly, it describes the changes in the contents of individual endogenous phytohormones during water stress. Then, it deals with the effects of applied phytohormones on stomatal opening, and on transpiration and photosynthetic rates in different plants species. Finally, it focuses on the alleviation or stimulation of absicic acid-induced stomatal closure by application of other phytohormones.

Cytokinins and water stress

It is almost impossible to find a single process in plant life that is not affected by both stress and hormones directly or indirectly. This minireview is focused on the interactions between water stress and cytokinins (CKs). The attention was paid mainly to changes in endogenous CK content and composition under water stress, involvement of CK in plant responses to water stress mainly in stomatal regulation of gas exchange, water relations of transgenic plants with elevated CK content, and possibilities to ameliorate the negative effects of water stress by application of exogenous CKs.

Interactions between abscisic acid and cytokinins during water stress and subsequent rehydration

With the aim to contribute to elucidation of the role of phytohormones in plant responses to stresses the endogenous contents of abscisic acid (ABA) and cytokinins (CK) were followed in French bean, maize, sugar beet, and tobacco during water stress and subsequent rehydration. The effects of pre-treatments with exogenous ABA or benzyladenine (BA) before imposition of water stress were also evaluated. The content of ABA increased by water stress, and with the exception of bean plants increased content of ABA remained also after rehydration. In all plant species the ABA content was further increased by ABA pre-treatment, but in bean and maize it decreased by BA pre-treatment. The highest total content of CK was observed in bean and the lowest in maize during water stress. In their spectrum, the storage CK were dominant in bean, and inactive CK in tobacco while in sugar beet and maize all groups were present in comparable amounts. In all plant species, the contents of CK increased during water stress and with exception of bean they decreased back after rehydration. ABA pre-treatment further increased contents of CK in water-stressed bean and tobacco. BA pre-treatment increased contents of CK in sugar beet and tobacco after rehydration.

Production of reactive oxygen species and development of antioxidative systems during in vitro growth and ex vitro transfer

Ex vitro transfer is often stressful for in vitro grown plantlets. Water stress and photoinhibition, often accompanying the acclimatization of in vitro grown plantlets to ex vitro conditions, are probably the main factors promoting production of reactive oxygen species (ROS) and in consequence oxidative stress. The extent of the damaging effects of ROS depends on the effectiveness of the antioxidative systems which include low molecular mass antioxidants (ascorbate, glutathione, tocopherols, carotenoids, phenols) and antioxidative enzymes (superoxide dismutase, ascorbate peroxidase, catalase, glutathione reductase, monodehydroascorbate reductase, dehydroascorbate reductase). This review is focused on ROS production and development of antioxidative system during in vitro growth and their further changes during ex vitro transfer.

Photosynthesis of Transgenic Pssu-ipt Tobacco

Summary The elevated content of endogenous cytokinin (CK), as the result of the introduction of the chimeric P ssu-ipt gene in tobacco, induced changes in growth, water regime and photosynthesis of transgenic grafts and rooted plants grown in a greenhouse. Despite closed stomata remarkable water stress was detected in P ssu-ipt plants. The ABA content was lower than in wild type tobacco, indicative of disturbances in ABA transport or metabolism. The rates of net photosynthesis (measured as CO 2 uptake or O 2 evolution) decreased by more than 50% in P ssu-ipt grafts and by 20% in P ssu-ipt rooted plants, respectively. The partial reactions of the electron transport chain in transgenic P ssu-ipt tobacco were differently influenced: the activity of the reaction centre of PSII was hardly affected; the PS I activity and the intersystem electron transport chain was inhibited up to 70%, particularly in the P ssu-ipt grafts. Although the slight decrease in the potential photochemical efficiency of PSII, expressed as F v /F m , was found in transgenic plants, the actual quantum yields, photochemical efficiencies, and qP under steady-state conditions indicated that transgenic plants were not seriously limited by the redox state of QA. The reaction centre of PSII was well preserved. In transgenic grafts, photophosphorylation capacity was strongly reduced, which could correspond with lower non-photochemical quenching. The above mentioned changes are the results of elevated CK content per se rather than the effect of altered water relations in the plants, caused by the disproportion of shoots and root system in both P ssu-ipt grafts and plants.

Development of Water Stress under Increased Atmospheric CO2 Concentration

The increase in water use efficiency (the ratio of photosynthetic to transpiration rates) is likely to be the commonest positive effect of long-term elevation in CO2 concentration (CE). This may not necessarily lead to decrease in long-term water use owing to increased leaf area. However, some plant species seem to cope better with drought stress under CE, because increased production of photosynthates might enhance osmotic adjustment and decreased stomatal conductance and transpiration rate under CE enable plants to maintain a higher leaf water potential during drought. In addition, at the same stomatal conductance, internal CO2 concentration might be higher under CE which results in higher photosynthetic rate. Therefore plants under CE of the future atmosphere will probably survive eventual higher drought stress and some species may even be able to extend their biotope into less favourable sites.

Interaction of Cytokinins and Abscisic Acid During Regulation of Stomatal Opening in Bean Leaves

Effects of benzyladenine (BA) and abscisic acid (ABA) applied separately or simultaneously on parameters of gas exchange of Phaseolus vulgaris L. leaves were studied. In the first two experimental sets) 100 μM ABA and 10 μM BA were applied to plants sufficiently supplied with water. Spraying of leaves with ABA decreased stomatal conductance (gs) and in consequence transpiration rate (E) and net photosynthetic rate (PN) already 1 h after application, but 24 h after application the effect almost disappeared. 10 μM BA slightly decreased gas exchange parameters, but in simultaneous application with ABA reversed the effect of ABA. Immersion of roots into the same solutions markedly decreased gas exchange parameters and 24 h after ABA application the stomata were completely closed. The effect of ABA was ameliorated by simultaneous BA application, particularly after 1-h treatment. In the third experimental set, plants were pre-treated by immersing roots into water, 1 μM BA, or 100 μM ABA for 24 h and then the halves of split root system were dipped into different combinations of 1 μM BA, 100 μM ABA, and water. In plants pre-treated with ABA all gas exchange parameters were small and they did not differ in plants treated with H2O+H2O, H2O+BA, or BA+BA. In plants pre-treated with BA or H2O, markedly lower values of PN were found when both halves of roots were immersed in ABA. Further, the effects of pre-treatment of plants with water, 1 μM BA, 100 μM ABA, or ABA+BA on the development of water stress induced by cessation of watering and on the recovery after rehydration were followed. ABA markedly decreased gas exchange parameters at the beginning of the experiment, but in its later phase the effect was compensated by delay in development of water stress. BA also delayed development of water stress and increased PN in water-stressed leaves. BA reversed the effect of ABA at mild water stress. Positive effects of BA and ABA pre-treatments were observed also after rehydration.

Effects of Pre-Treatments with Abscisic Acid and/or Benzyladenine on Gas Exchange of French Bean, Sugar Beet, and Maize Leaves During Water Stress and After Rehydration

Net photosynthetic rate (PN), transpiration rate (E), and stomatal conductance (gs) during water stress and after rehydration were measured in Phaseolus vulgaris, Beta vulgaris, and Zea mays. Immediately before imposition of water stress by cessation of watering, plants were irrigated with water (control), 100 μM abscisic acid (ABA), and/or 10 μM N6-benzyladenine (BA). In all three species, application of ABA decreased gs, E, and PN already 1 h after application. However, during water stress gs, E, and PN in plants pre-treated with ABA remained higher than in plants pre-treated with water. Positive effects of ABA application were observed also after rehydration. In contrast, the effects of pre-treatment with BA were species-specific. While in bean plants BA application ameliorated negative effect of water stress, only very slight effects were observed in maize, and in sugar beet BA even aggravated the effects of water stress.

Effect of benzylaminopurine on rehydration of bean plants after water stress

The possibility of improving the recovery of plant photosynthesis after water stress by cytokinin-induced stimulation of stomatal opening or delay of leaf senescence was tested. The 6-benzylaminopurine (BAP) in concentrations 1 and 10 μM was applied to the substrate (sand + nutrient solution) or sprayed on primary leaves of 14-d-old Phaseolus vulgaris L. plants sufficiently supplied with water or water-stressed for 4 d. The later ones having relative water content decreased to 69 % were fully rehydrated during the following three days. Parameters of photosynthesis and water relations were measured in primary leaves of 7-, 10-, 14-, and 17-d-old plants. Application of 1 μM BAP slightly delayed leaf senescence: in 17-d-old control plants, net photosynthetic rate (PN) and chlorophyll (Chl) content, and when sprayed on leaves also some of Chl a fluorescence kinetic parameters of BAP-treated leaves were slightly higher than those of untreated leaves. Both types of application of 1 μM BAP slightly improved recovery of plants during rehydration after water stress in terms of increased gad, gab and PN, i.e., parameters which were markedly decreased by mild water stress. However, contents of Chl a, Chl b and carotenoids and parameters of Chl a fluorescence kinetic were not markedly affected by mild water stress and after rehydration were not stimulated by 1 μM BAP. 10 μM BAP had mostly negative effects on the parameters measured.

Effect of carbon dioxide enrichment during in vitro cultivation and acclimation to ex vitro conditions

Tobacco and carnation plantlets were grown in vitro on Murashige and Skoogs medium with 2 % saccharose. Carnation plantlets were also grown fully photoautotrophically on a medium without saccharose. The ambient CO2 concentration was increased from 0.6 to 10 or 40 g m-3 during the last 3 weeks of in vitro cultivation or during the first 3 weeks of acclimation to ex vitro condition (plantlets transplanted to pots with sand and nutrient solution) or during both growth phases. CO2 enrichment during in vitro cultivation markedly stimulated growth of tobacco plantlets, and also of carnation plantlets, both with and without saccharose. CO2 enrichment during the acclimation period promoted plant growth more effectively in plantlets grown in vitro at a CO2 concentration of 0.6 g m-3 than in plantlets grown in either growth phase at higher CO2 concentrations.