Rehydration of Sugar Beet Plants after Water Stress: Effect of Cytokinins

The possibility to improve the recovery of sugar beet plants after water stress by application of synthetic cytokinins N⁶-benzyladenine (BA) or N⁶-(m-hydroxybenzyl)adenosine (HBA) was tested. Relative water content (RWC), net photosynthetic rate (P_N), transpiration rate (E), stomatal conductance (g_s), chlorophyll (Chl) a and Chl b contents, and photosystem 2 efficiency characterized by variable to maximal fluorescence ratio (F_v/F_m) were measured in control plants, in water-stressed plants, and after rehydration (4, 8, 24, and 48 h). Water stress markedly decreased parameters of gas exchange, but they started to recover soon after irrigation. Application of BA or HBA to the substrate or sprayed on leaves only slightly stimulated recovery of P_N, E, and g_s in rehydrated plants, especially during the first phases of recovery. Chl contents decreased only under severe water stress and F_v/F_m ratio was not significantly affected by water stress applied. Positive effects of BA or HBA application on Chl content and F_v/F_m ratio were mostly not observed.     

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 (P_N), transpiration rate (E), and stomatal conductance (g_s) 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 M abscisic acid (ABA), and/or 10 M N⁶-benzyladenine (BA). In all three species, application of ABA decreased g_s, E, and P_N already 1 h after application. However, during water stress g_s, E, and P_N 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.     

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 M ABA and 10 M BA were applied to plants sufficiently supplied with water. Spraying of leaves with ABA decreased stomatal conductance (g _s) and in consequence transpiration rate (E) and net photosynthetic rate (P _N) already 1 h after application, but 24 h after application the effect almost disappeared. 10 M 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 M BA, or 100 M ABA for 24 h and then the halves of split root system were dipped into different combinations of 1 M BA, 100 M 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 P _N were found when both halves of roots were immersed in ABA. Further, the effects of pre-treatment of plants with water, 1 M BA, 100 M 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 P _N 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.     

Cytokinins promotion of flowering in Cymbidium ensifolium var. misericors in vitro

Callus-derived rhizomes of Cymbidium ensifolium var. misericors produced flowers precociously on a defined basal medium (1/2MS) containing of NAA with thidiazuron (TDZ), N⁶-(2-isopentenyl) adenine (2iP) or N⁶-benzyladenine (BA) within 100 d of culturing. Among eight cytokinins tested, TDZ at 3.3–10 µM or 2iP at 10–33 µM combined with 1.5 µM NAA were the most effective combinations for achieving flower induction in vitro. These undersized flowers were physically normal and bloomed for two weeks in vitro.     

Effect of Foliar Application of Chitin and Chitosan Oligosaccharides on Photosynthesis of Maize and Soybean

On the first day after foliar application, chitosan pentamer (CH5) and chitin pentamer (CHIT5) decreased net photosynthetic rate (P _N) of soybean and maize, however, on subsequent days there was an increase in P _N in some treatments. CH5 caused an increase in maize P _N on day 3 at 10^–5 and 10^–7 M; the increases were 18 and 10 % over the control plants. This increase was correlated with increases in stomatal conductance (g _s) and transpiration rate (E), while the intercellular CO₂ concentration (C _i) was not different from the control plants. P _N of soybean plants did not differ from the control plants except for treatment CH5 (10^–7 M) which caused an 8 % increase on day 2, along with increased g _s, E, and C _i. On days 5 and 6 the CHIT5 treatment caused a 6–8 % increase in P _N of maize, which was accompanied by increases in g _s, E, and C _i. However, there was no such increase for soybean plants treated with CHIT5. In general, foliar application of high molecular mass chitin (CHH) resulted in decreased P _N, particularly for 0.010 % treated plants, both in maize and soybean. Foliar applications of chitosan and chitin oligomers did not affect (p > 0.05) maize or soybean height, root length, leaf area, shoot or root or total dry mass.     

In vitro culture of adult and juvenile bud explants of Passiflora species

Cultivar E23, an F1 hybrid of P. edulis and P. edulis f. flavicarpa is usually propagated by shoot-tip grafting. Various media were tested to evaluate the potential of E23 for in vitro propagation. Adult tissue was difficult to culture and did not respond to media containing low (<10 µM) concentrations of growth regulators. Growth of adult buds on intact stem sections was promoted by 1 week of dark incubation on MS basal medium plus 150 µM 2iP, 200 µM adenine sulphate and 17.1 µM IAA (3 mg l^–1), and further developed into shoots on MS medium plus 4.9 µM 2iP (1 mg l^–1) and 5.7 µM IAA (1 mg l^–1). By contrast, juvenile shoots of E23, and Passiflora species: edulis f. flavicarpa, edulis, alata, caerulea, mollissima, coccinea, herbertiana and suberosa grew rapidly on MS medium plus 10 µM kinetin and 5 µM IAA. Rapid multiplication was achieved on MS plus 20 µM BA, 10 µM kinetin, 5 µM IAA, and roots initiated on MS plus 5 µM IAA.Abbreviations IAA indole-3-acetic acid - 2iP N6-iso pentenyl adenine - BA N6-benzyl adenine     

The Effects of Intermittent Flooding on Seedlings of Three Forest Species

Under greenhouse conditions, seedlings of three forest species, baldcypress (Taxodium distichum), nuttall oak (Quercus nuttallii), and swamp chestnut oak (Quercus michauxii) were subjected to an intermittent flooding and subsequent physiological and growth responses to such conditions were evaluated. Baldcypress showed no significant reductions in stomatal conductance (g _s) or net photosynthetic rate (P _N) in response to flood pulses. In nuttall oak seedlings g _s and P _N were significantly decreased during periods of inundation, but recovered rapidly following drainage. In contrast, in swamp chestnut oak g _s was reduced by 71.8 % while P _N was reduced by 57.2 % compared to controls. Baldcypress displayed no significant changes in total mass while oak species had significantly lower leaf and total mass compared to their respective controls. Thus baldcypress and nuttall oak showed superior performance under frequent intermittent flooding regimes due to several factors including the ability for rapid recovery of gas exchange soon after soil was drained. In contrast, swamp chestnut oak seedlings failed to resume gas exchange functions after the removal of flooding.     

Photosynthetic characteristics of dipterocarp species planted on degraded sandy soils in southern Thailand

To elucidate whether dipterocarp species, dominant late-successional species of tropical forests in Southeast Asia, actually have a disadvantage when planted on open site in terms of their photosynthetic characteristics, we investigated photosynthesis in dipterocarp seedlings planted in the open on degraded sandy soils in southern Thailand. These species were compared with seedlings of Acacia mangium Willd., a fast-growing tropical leguminous tree, which is often planted on degraded open site in Southeast Asia. The dipterocarp seedlings had an irradiance-saturated net photosynthetic rate (P _N), stomatal conductance (g _s), carboxylation efficiency, and photosynthetic capacity comparable to or superior to those of A. mangium. In particular, seedlings of Dipterocarpus obtusifolius Teijsm. ex Miq. showed an irradian-ce-saturated P _N of 21 μmol m⁻² s⁻¹, a value higher than any previously reported for a dipterocarp species, accompanied by high g _s (0.7 mol m⁻² s⁻¹) and high photosynthetic capacity. Thus dipterocarp species do not necessarily have a disadvantage in terms of their photosynthetic characteristics on open sites with degraded sandy soils.     

Accumulation of zeatin O-glycosyltransferase in Phaseolus vulgaris and Zea mays following cold stress

Zeatin O-glycosides have been reported as inactive and stable storage forms of cytokinins whose concentrations increase in cold stressed plants. Zeatin O-glycosides accumulation in developing bean seeds has been correlated with an increase of zeatin O-glycosyltransferase , which is specific to trans-zeatin, and catalyzes the conjugation of zeatin O-glycosides. When Phaseolus vulgaris and Zea mays seedlings were grown for 3 days at 25 and then incubated at 4 or 10 for 6 days no further growth was observed in roots. Hypertrophy was observed in the root tips of both species. In shoot-hypocotyl complexes, in contrast, growth occurred when seedlings were incubated at 10 . Western analysis, with Mabs specific to zeatin O-glycosyltransferase, detected antigenically related proteins in roots, shoot tips and cotyledons after seedlings were cold stressed for 1–6 days at 4 or 10 . Immunolocalization, of both maize and bean root sections grown at 25 revealed antigenically related proteins that were detected at low levels in cortical cells. The signal intensified upon cold stress. The localization of zeatin O-glycosyltransferase in Z. mays root tips was directly comparable to the distribution of the zeatin O-glycosides. The enzyme was detected in the nucleus, cytoplasm, and closely associated with the plasma membrane and in the cell wall of Z. mays root cells. Southern analysis suggested that more than one gene in Z. mays that were homologous to zeatin O-glycosyltransferase in P. vulgaris. Zeatin O-glycosyltransferase may be involved in modulation of cytokinins under cold stress.     

Effect of Growth Regulators on Photosynthesis, Transpiration and Related Parameters in Water Stressed Cotton

Gas exchange in Gossypium hirsutum L. cv. H-777 as affected by water deficit and growth regulators (IAA, GA₃, BAP, ABA, ethrel) was examined. Sixty days after sowing, growth regulators in concentration 50 µM were applied as foliar spray and irrigation was withheld to get desired (moderate and severe) water deficit. All the parameters were measured on the third leaf from the top between 10:00 and 11:00. Net photosynthetic rate (P_N), transpiration rate (E), stomatal conductance (g_s), carboxylation efficiency (CE), and water potential (_w) decreased significantly with the increasing water stress, however, water use efficiency (WUE) was unaffected. Foliar spray with IAA, GA₃ and BAP partially counteracted the effect of water deficit on the above parameters except _w, which became more negative. ABA and up to some extent ethrel increased WUE and maintained higher _w, however, caused further decrease in P_N, E, and g_s.     

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 M 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 M BAP slightly delayed leaf senescence: in 17-d-old control plants, net photosynthetic rate (P_N) 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 M BAP slightly improved recovery of plants during rehydration after water stress in terms of increased g_ad, g_ab and P_N, 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 M BAP. 10 M BAP had mostly negative effects on the parameters measured.     

Gas Exchange and Chlorophyll Fluorescence in Symbiotic and Non-Symbiotic Ryegrass Under Water Stress

The symbiotic association of endophyte fungus, Neotyphodium lolii, and ryegrass improves the ryegrass resistance to drought. This is shown by a 30 % increase in the number of suckers in infected plants (E+), compared to plants lacking endophyte (E−), and by a higher water potential in the E+ than E− plants. The E+ plants have higher stomatal conductance (g _s), transpiration rate, net photosynthetic rate (P _N), and photorespiratory electron transport rate than the E− plants. The maximal photochemical efficiency (F_v/F_m) and the actual photochemical efficiency (Φ_PS2) are not affected by the endophyte fungus. The increase in P _N of the E+ plants subjected to water stress was independent from internal CO₂ concentration. An increased P _N was observed in E+ plants also in optimal water supply. Hence the drought resistance of E+ plants results in increased g _s, P _N, and photorespiratory electron transport rate. This revised version was published online in June 2006 with corrections to the Cover Date.     

Micropropagation of Two Australian Native Fruit Species,Davidsonia pruriens and Davidsonia jerseyana G. Harden & J.B. Williams

Nodal explants from in vitro grown seedlings of Davidsonia pruriens and D. jerseyana, established on MS media were treated with various concentrations of three cytokinins. D. pruriens developed optimum shoot growth in terms of shoot height and number of leaves per shoot when 1.0 µM BA was added to basal MS medium while optimum shoot growth for D. jerseyana was obtained when 0.01 µM 2iP was added to the medium. Optimum root initiation and development was obtained when actively growing axillary shoots were cultured on 1/2MS medium plus 32.2 µM IBA for 3–5 days for D. pruriens and 2–3 days for D. jerseyana before transfer to PGR-free medium containing 10 µM riboflavin. Root initiation of more than 80% was achieved with multiple genotypes of D. pruriens and three genotypes of D. jerseyana using juvenile material. The plantlets were transferred to pots and grown in the greenhouse with a success rate of 60% for D. pruriens and 75% for D. jerseyana. Adult D. jerseyana stem explants produced 2–5 shoots per nodal explant upon treatment with 0.1 µM BA. Side shoots from adult D. jerseyana produced similar results for shoot multiplication as for juvenile material. Protocol for multiplication of adult D. pruriens was achieved with much greater difficulty by using material from the green house. Axillary shoots were initiated when 100 µM TDZ was applied to the stem of an adult pot plant and the resultant side shoots were cultured on MS medium containing 1.0 µM BA and 1.0 µM GA₃.     

Effects of Angular Leaf Spot and Rust on Leaf Gas Exchange and Yield of Common Bean (Phaseolus Vulgaris)

Isolated and interactive effects of angular leaf spot (caused by Phaeoisariopsis griseola) and rust (caused by Uromyces appendiculatus) on leaf gas exchange and yield was studied in common bean (Phaseolus vulgaris L. cv. Carioca) plants. Gas exchange was measured on 37, 44, 51, and 58 d after planting using a portable photosynthesis system. The inoculation of plants with P. griseola (P), U. appendiculatus (U), and the combination of both pathogens (P+U) caused a significant reduction of net photosynthetic rate (P _N) and yield. The reduction of stomatal conductance (g _s), P _N, and yield was higher under P and combination of P+U than under U treatment. By effect of U, the reduction on yield was higher than the reductions on gas exchange parameters. On the treatment P+U, a reduction of 23 % in P _N and a correspondent reduction of 32 % in yield was observed. The interactive effects of the pathogens on yield could be explained in part by the decreases in g _s and in P _N of diseased bean leaves. The combined effect of both diseases on yield and gas exchange parameters suggests an antagonistic interaction.     

Effects of Cytokinins on In Vitro Seed Germination and Early Seedling Morphogenesis in Lotus corniculatus L.

We determined the effects of zeatin (ZEA), isopentenyl adenine (2iP), kinetin (KIN), benzyladenine (BA), and thidiazuron (TDZ) on seed germination, elongation of seedling shoots and roots, frequency of regeneration, and the number of regenerants per seedling in Lotus corniculatus L. Sterilized seeds were cultured in vitro on Murashige and Skoog (1962) medium containing 3% sucrose, 0.7% agar, and various cytokinins (0, 0.08, 0.22, 0.35, 0.80, 2.20, and 3.50 μM). After 30 days, seedlings were transferred to cytokinin-free medium for another 60 days. All cytokinins stimulated the rate and percentage of seed germination at least twofold in optimum concentrations; TDZ and ZEA were the most active, followed closely by BA, whereas KIN and 2iP stimulated germination in higher concentrations only. Elongation of shoots and roots was strongly inhibited at the lowest TDZ and BA concentrations, whereas ZEA, KIN, and 2iP exerted moderate, dose-dependent inhibition. The frequency of regenerant-producing seeds was highest on ZEA and BA, whereas the greatest number of regenerants per seedling was found on TDZ. It is concluded that the culture of seeds on cytokinin-containing media, followed by transfer to cytokinin-free medium, is a suitable procedure for rapid production of a large number of uniform regenerants. The presumed role of particular cytokinins is discussed.     

Physiological Effects of Plant Hormones in Cotton Under Drought

Effects of plant hormones indole-3-yl-acetic acid (IAA), gibberellic acid (GA), benzylaminopurine (BAP), abscisic acid (ABA) and ethrel (ETH) in 5 M concentration on gas exchange, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO, EC 4.1.1.39) activity, pigment content and yield in cotton (Gossypium hirsutum L. cv. H-777) under drought were studied. At reproductive stage (55 – 60 d after sowing) these hormones were sprayed on shoots one day prior to stress imposition by withholding irrigation. The soil moisture of control plants was kept at field capacity. Net photosynthetic rate (P_N), stomatal conductance (g_s), transpiration rate (E), carboxylation efficiency (CE), water use efficiency (WUE), RuBPCO activity, boll number per plant, seed number per plant and lint mass per plant significantly decreased at drought while chlorophyll (Chl) b content and flower number per plant increased. ABA and ETH significantly reduced gas exchange parameters, Chl a and Chl b content. Detrimental drought effect on P_N, g_s, E, CE, RuBPCO and lint mass per plant was significantly alleviated by BAP and also its effect on seed number and lint mass per plant was significantly alleviated with the ABA treatment.     

Gas Exchange of Spring Barley and Wheat Grown under Mild Water Shortage

During mild water stress (decrease of full water capacity from 60 to 35 %) net photosynthetic rate (P _N) of four spring barley and wheat genotypes was about twice lower than that for unstressed plants and was mainly limited by non-stomatal factors. Availability of CO₂ from intercellular spaces did not change significantly when stomatal conductance (g _s) decreased from 0.25-0.35 to 0.15-0.20 mol(H₂O) m⁻² s⁻¹. There may be two main processes leading to similar intercellular CO₂ concentration (c _i) in stressed and unstressed seedlings despite of twice lower P _N under mild water stress: (a) lower diffusion of CO₂ through stomata represented by lower g _s, (b) lower consumption of CO₂ by photosynthetic apparatus of stressed plants. Last factor is partially pronounced by lower response of P _N to c _i observed for stressed than for control plants. This revised version was published online in August 2006 with corrections to the Cover Date.     

Stomatal and non-stomatal limitations to photosynthesis in field-grown grapevine cultivars

Diurnal changes of photosynthesis in the leaves of grapevine (Vitis vinifera × V. labrusca) cultivars Campbell Early and Kyoho grown in the field were compared with respect to gas exchanges and actual quantum yield of photosystem 2 (Φ_PS2) in late May. Net photosynthetic rate (P_N) of the two cultivars rapidly increased in the morning, saturated at photosynthetic photon flux density (PPFD) from 1200 to 1500 μmol m⁻² s⁻¹ between 10:00 and 12:00 and slowly decreased after midday. Maximum P_N was 13.7 and 12.5 μmol m⁻² s⁻¹ in Campbell Early and Kyoho, respectively. The stomatal conductance (g_s) and transpiration rate changed in parallel with P_N, indicating that P_N was greatly affected by g_s. However, the decrease in P_N after midday under saturating PPFD was also associated with the observed depression of Φ_PS2 at high PPFD. The substantial increase in the leaf to air vapour pressure deficit after midday might also contribute to decline of g_s and P_N.     

Stimulation of adventitious root formation in non-woody stem cuttings by uridine

Uridine strongly stimulated adventitious root formation in stem cuttings of sunflower (Helianthus annuus L.), mung bean (Vigna radiata L.) and common bean (Phaseolus vulgaris L.). A dose response curve of uridine induced rooting showed that the optimum concentration of uridine was 0.1 µM. At all concentrations employed, uridine had no significant effect on root elongation. The rooting response of stem cuttings to the optimal concentration of indole-3-butyric acid (10 µM) in combination with 0.1 µM uridine did not significantly differ from their response to either of these compounds when applied alone. However, the rooting response of the cuttings to sub-optimal IBA (0.01 µM) was significantly stimulated by uridine. These findings suggested that uridine may have stimulated rooting by increasing the sensitivity of the rooting tissue to auxin.     

Nitrogen and phosphorus uptake by the Brazilian kelp Laminaria abyssalis (Phaeophyta) in culture

The uptake of ammonium, nitrate and phosphate by laboratory-grown young sporophytes of Laminaria abyssalis was measured in a perturbed system (batch mode) at 18 °C and 35 ± 5 µE m^–2 s^–1 photon flux density. Uptake of all appeared to follow saturation-type nutrient uptake kinetics. The NO _inf3 ^sup– (K _s = 14.0 µM, V _max = 5.0 µmol h^–1 g^–1 dry wt) and NH _inf4 ^sup+ (K _s = 4.6 µM, V _max= 2.0 µmol h^–1 g^–1 dry wt) were taken up simultaneously, although NH _inf4 ^sup+ was taken up more rapidly. Values of K ₃ and V _max for phosphate were, respectively, 2.21 µM and 0.83 µmol h^–1 g^–1 dry wt. Nitrate and phosphate were both consumed in similar rates (V _max /Ks 0.37) at low concentrations. NH _inf4 ^sup+ , thus, might be a more efficient form of N fertilizer if artificial enrichment of seawater is used.