Gasometric measurement of sulfite oxidation

In the commonly used sulfite method the consumption of sulfite is determined by iodometry. Since however, the addition of organic substances may interfere with iodometry (e.g. due to chemical reactions with iodine) the gasometric measurement of sulfite oxidation has been developed for analysis of how different culture media may influence the oxygen transfer rate. The striking decrease of sulfite oxidation rate due to addition of culture media to the sulfite solution suggests that adsorption of orgnic components in the gas liquid interface may account for an additional diffusion barrier and thus for a decrease of the oxygen transfer coefficient which in addition gives an explanation for differences between values found by the sulfite method and by aerobic cultivations. Consequently identical values of oxygen transfer rate have been obtained for both systems whenever the sulfite system has been properly adjusted to the aerobic cultivation conditions. In so far, the gasometric sulfite method proved to be a unique tool for rapid determination of factors influencing oxygen transfer rate in fermentation processes which may give rise to a reappraisal as to the relevance of the sulfite method for oxygen transfer optimization.     

Adaptation of beta-ray gauge technique for measurement of water deficit of intact rice leaves

A linear relationship was obtained between the logarithm of count rate and absorber’s (Aluminum foil and leaf) effective thickness in the range of 4.25 to 18.00 mg cm^?2. Also relative water content (RWC) and absorber effective thickness was linearly related (Y = ?89.80 + + 15.84 X, and r = 0.99). From these relationships it was possible to determine the RWC of a rice leaf simply by measuring the radiation intensity of fresh, fully turgid and completely dry leaf by beta ray gauge technique without the added necessity of determining the mass of the leaf.     

NAD kinase activity in wheat leaves under water deficit

Total NAD kinase activity remained unaltered in the drought non-adapted wheat leaves under water deficit, but gradually decreased with water deficit in the adapted ones. The share of the calmodulin-dependent enzyme was significantly higher in the drought-hardened than in non-hardened plants; however, under severe water deficit the activity of the enzyme dropped by half. It seems, therefore, that NAD kinase activity does not limit phosphorylation of NAD in dehydrated plant tissues.     

Deciphering genetic variations of proteome responses to water deficit in maize leaves

The proteome of the basal part of growing Zea mays leaves was analyzed from 4 to 14 d after stopping watering and in well watered controls. The relative quantity of 46 proteins was found to increase in leaves of plants submitted to water deficit. Different types of responses were observed, some proteins showing a constant increase during water deficit, while others showed stabilization after a first increase or a transient increase. Isoforms encoded by the same gene showed different responses. The response to water deficit showed genetic variation. Some increased proteins were induced specifically in one of the two studied genotypes (e.g. ASR1) while others were significantly induced in both genotypes but to a different level or with different kinetics. Analyses of relations between protein quantities, relative water content (RWC) and abscisic acid (ABA) concentration allowed us to show that the quantitative variation of some proteins (e.g. ABA45 and OSR40 proteins) was linked to differences in ABA accumulation between the genotypes. Other proteins showed genetic variations that were not related to differences in water status or ABA concentration (e.g. a cystatin). Data obtained from these experiments, together with data from other experiments, contribute to the characterization of maize proteome response to drought in different conditions and in different genotypes. This characterization allows the search for candidate proteins, i.e. for protein whose genetic variation of expression could be partly responsible for the variability of plant responses to drought.     

Photochemical heat-shock response in common bean leaves as affected by previous water deficit

The heat sensitivity of photochemical processes was evaluated in the common bean (Phaseolus vulgaris) cultivars A222, A320, and Carioca grown under well-watered conditions during the entire plant cycle (control treatment) or subjected to a temporal moderate water deficit at the preflowering stage (PWD). The responses of chlorophyll fluorescence to temperature were evaluated in leaf discs excised from control and PWD plants seven days after the complete recovery of plant shoot hydration. Heat treatment was done in the dark (5 min) at the ambient CO₂ concentration. Chlorophyll fluorescence was assessed under both dark and light conditions at 25, 35, and 45°C. In the dark, a decline of the potential quantum efficiency of photosystem II (PSII) and an increase in minimum chlorophyll fluorescence were observed in all genotypes at 45°C, but these responses were affected by PWD. In the light, the apparent electron transport rate and the effective quantum efficiency of PSII were reduced by heat stress (45°C), but no change due to PWD was demonstrated. Interestingly, only the A222 cultivar subjected to PWD showed a significant increase in nonphotochemical fluorescence quenching at 45°C. The common bean cultivars had different photochemical sensitivities to heat stress altered by a previous water deficit period. Increased thermal tolerance due to PWD was genotype-dependent and associated with an increase in potential quantum efficiency of PSII at high temperature. Under such conditions, the genotype responsive to PWD treatment enhanced its protective capacity against excessive light energy via increased nonphotochemical quenching.     

Water potential,water saturation deficit and their relationship in leaves of different insertion levels

The water potential (Ψ _w ) and the water saturation deficit (δW _sat) in leaves of different insertion levels of potted kale plants were simultaneously measured. In non-wilting plantsδW _sat gradually decreased andΨ _w slightly increased from the upper to the lower leaves. During the wilting of the plants induced by decreasing of soil moistureΨw practically decreased paralelly in all the leaves but the same decrease ofΨ _w was connected with the lowest increase ofδW _sat in upper leaves and the highest increase ofδW _sat in lower leaves. Not only the values ofΨ _w andδW _sat but also their relationship varied considerably with the leaf insertion levels.     

Respiratory energy demand for protein turnover and ion transport in wheat leaves upon water deficit

The effect of water stress on the respiratory energy demand for the main biosynthetic and transport processes was estimated in the leaves of spring wheat ( Triticum aestivum L. cv. San Pastore) acclimated and non-acclimated to drought. ATP-consuming processes were assessed from the effects of selective inhibitors of RNA synthesis, protein synthesis and proteolysis, Ca²⁺-ATPase and P-type ATPases on respiration. The proportions of energy consumed by these processes were compared with the theoretical ATP production calculated from the rate of oxygen consumption measured manometrically. Respiratory energy production increased significantly in both acclimated leaves and in leaves stressed by drought. In the fully grown wheat leaves, Ca²⁺-dependent reactions and protein turnover consumed about 37% and 34% of the total respiratory energy, respectively. The costs of ion transport constituted another 15% of the total ATP production. Both acclimation and drought stress in non-acclimated leaves resulted in a decrease of leaf sensitivity towards inhibitors of RNA and protein syntheses as well as a decrease in Ca²⁺-mediated processes; but also in an increase of leaf sensitivity towards inhibitors of proteolysis and ouabain-sensitive ATPase in non-acclimated plants. This indicates a shift in ATP input into the energy-requiring processes towards greater expenses for ion transport upon water deficit. However, in acclimated leaves under drought stress, distribution of respiratory energy became almost the same as in control plants.     

Temperature effects in the measurement of water saturation deficit (relative water content) in Tobacco and Kale

An expressive effect of temperature on water saturation deficit determined by “leaf-disc extrapolation method” was found. This error may reach about 15 to 40% of measured value with the temperature gradient of 10° between tissuein situ and under conditions of W.S.D. determination.     

Effects of plant water deficit on the daily carbon balance of leaves of cacao seedlings

The daily carbon balance of individual source leaves of Theobroma cacao L. seedlings was measured at 2- to 3-day intervals during a 19-day period of increasing plant water deficit and during an 8-day period of recovery following rewatering. In each case, responses of stressed seedlings were compared to those of irrigated controls. Leaves of irrigated cacao seedlings assimilated approximately 41 mg carbohydrate dm^-2 during 12-h photoperiods, and exported an average of 34 mg carbohydrate dm^-2 during 24-h measurement cycles. The rate of carbon export from cacao leaves was sharply reduced as leaf water potential (ψ) declined between -0.8 and -2.0 MPa. Further, the rate of export was closely associated with the net assimilation rate (A), with export capacity being severely reduced as A fell to near zero. Net accumulation of dry matter occurred as long as A remained greater than approximately 20 mg carbohydrate dm^-2 over the 12-h photoperiod, but at lower assimilation rates, export exceeded concomitant assimilation. Carbon export continued at the expense of leaf carbon reserves as photoassimilation fell to near zero during periods of severe water stress (ψ < -2.0 MPa). Night respiration rate was independent of plant water status.     

The histone-like protein H1-S and the response of tomato leaves to water deficit

Linker histone protein variants are expressed in different tissues, at various developmental stages or induced by specific environmental conditions in many plant species. In most cases, the function of these proteins remains unknown. In the work presented here an antisense strategy has been used to study the function of the drought-induced linker histone, H1-S of tomato. Three independent H1-S antisense tomato mutants, selected for their inability to accumulate H1-S in response to water stress, were studied. These mutants have been characterized at the physiological and morphological levels. Histone H1-S antisense transgenic plants developed normally indicating that H1-S does not play an important role in the basal functions of tomato development. No differences were detected in chromatin organization, excluding a structural role for H1-S in chromatin organization. However, differences between the wild-type and antisense plants were observed in leaf anatomy and physiological activities. This analysis indicates that H1-S has more than one function, at different times, in controlling plant water status, highlighting the complexity of the water stress response.     

Water saturation deficit in the wilting plant. The preference of young leaves and the translocation of water from old into young leaves

U 100 a? 120denných rostlin krmné kapusty a ?epky byly sledovány rozdíly v dynamice vzniku a dal?ího vývoje momentálního vodního deficitu (VD) u r?zně starých list?. VD byl stanovován ter?íkovou metodou s extrapolaí dosycovací k?ivky do po?átku (?atský 1962b). U list?, oddělených od rostliny a vadnoucích bez p?ísunu vody, je VD nejvy??í u mladých a nejni??í u starých list?, tedy v podstatě odpovídá rozdíl?m v intensité transpirace. P?i od?íznutí celé rostliny vadnou listy r?zného stá?í v podstatě stejnou intensitou. V pozděj?ích fázích vadnutí byly v některých pokusech stanoveny mírně vy??í hodnoty VD starých list?. P?i pomalém vadnutí rostliny in situ, indukovaném sni?ováním p?dní vlhkosti, byla po?ínaje st?edními hodnotami VD, tj. pr?měrně od 8 a? 20 % stanovena velmi z?etelná preference mladých list? v zásobování vodou. P?i celkovém nedostatku vody v rostlině nejprve silně vadnou a později odumírají starí a dospělé listy; VD mladých list? se dlouho udr?uje na poměrně nízkých hodnotách. Tento pr?běh vadnutí rostliny in situ byl stanoven jak p?ímým mě?ením VD, tak i nep?ímo stanovením poklesu procentuálního obsahu vody v listech. Na zji?těné preferenci mladých list? v zásobování vodou se uplatňuje i translokace vody do mladých list? z vadnoucích list? star?ích.     

Photosynthetic responses and proline content of mature and young leaves of sunflower plants under water deficit

In mature and young leaves of sunflower (Helianthus annuus L. cv. Catissol-01) plants grown in the greenhouse, photosynthetic rate, stomatal conductance, and transpiration rate declined during water stress independently of leaf age and recovered after 24-h rehydration. The intercellular CO₂ concentration, chlorophyll (Chl) content, and photochemical activity were not affected by water stress. However, non-photochemical quenching increased in mature stressed leaves. Rehydration recovered the levels of non-photochemical quenching and increased the F_v/F_m in young leaves. Drought did not alter the total Chl content. However, the accumulation of proline under drought was dependent on leaf age: higher content of proline was found in young leaves. After 24 h of rehydration the content of proline returned to the same contents as in control plants.     

The interactive effect of water deficit and UV-B radiation on salicylic acid accumulation in barley roots and leaves

The aim of the paper was to determine the effect of water deficit (WD) and UV-B radiation acting individually and in combination on salicylic acid (SA) accumulation as well as on the activity of phenylalanine ammonia-lyase (PAL) and benzoic acid hydroxylase (BA2H) that control its biosynthetic route from phenylalanine. An additional aim was to test whether the interaction of these stresses limits the negative effect of a single stress on tissue hydration and membrane injury. Two-week-old seedlings were subjected to water deficit (WD), UV-B irradiation (UV-B) and three different combinations of WD and UV-B: (I) WD and UV-B applied at the same time, (II) UV-B applied before WD, and (III) WD applied before UV-B. Water deficit was imposed by immersing the root system in aerated nutrient solution with polyethylene glycol (PEG 6000) of water potential – 0.5 MPa. UV-B dosage was 24 kJ m⁻² day⁻¹ (0.84 W m⁻²) at the canopy level. UV-B and WD imposed individually and jointly, caused, in a time-dependent manner, an increase in SA content in both organs. Increased levels of SA in WD stressed plants were accompanied by an increase in the activity of PAL and BA2H. However, in plants exposed to UV-B were accompanied only by an increase in the activity of BA2H. Under WD conditions, an earlier increase of SA content was observed in roots than in leaves, which may indicate the involvement of SA in the signal transduction between roots and leaves. In plants exposed to sequential action of WD and UV-B, regardless of the order of its imposition, the effect of each single factor on SA accumulation in leaves was strengthened. WD had a greater effect on water loss and membrane injury than UV-B radiation. In plants exposed to WD after pre-treatment with UV-B radiation, a cross-tolerance mechanism was observed. Leaves of these plants did not show increased lipid peroxidation, measured in terms of malondialdehyde content, and a decrease in water content. This protective action was probably caused by the increase of the SA level in leaves of the UV-B treated plants prior to WD imposition.     

Diurnal changes in water deficit and recovery patterns of intact rice leaves under natural and controlled environment conditions

In the natural monsoon environment, the leaves of Bala and Jaya cultivars experienced more than 10 per cent LWD for about 8 and 6 1/2 h in the diurnal cycle, respectively. The maximum LWD of around 14 per cent occurred between 14.30 to 15.30 at the time of minimum relative humidity (EH) and maximum air temperature (Ta). On the other hand, under controlled environment conditions, LWD values of both the cultivars remained below 10 per cent in the entire diurnal cycle. There was a close relationship between LWD and Ta (r = 0.90 for Bala and 0.91 for Jaya) as well as RH (r = -0.95 for Bala and -0.85 for Jaya). The results suggested that RWC values obtained on an interval of 30 min were not enough to observe more than the general trend as a complete recovery of stress achieved within 25 min of rewatering the stressed leaves. Calculations indicated that the determination of LWD with the techniques using excised leaf are likely to incur error as rice leaf may loose water at a rate of 0.3 MPa min⁻¹.     

Changes in water status and osmolyte contents in leaves and roots of olive plants (Olea europaea L.) subjected to water deficit

Two-year-old olive trees (Olea europaea L., cv. Coratina) were subjected to a 15-day period of water deficit, followed by 12 days of rewatering. Water deficit caused decreases in predawn leaf water potential (Ψ_w), relative water content and osmotic potential at full turgor (Ψ _π100) of leaves and roots, which were normally restored upon the subsequent rewatering. Extracts of leaves and roots of well-watered olive plants revealed that the most predominant sugars are mannitol and glucose, which account for more than 80% of non-structural carbohydrates and polyols. A marked increase in mannitol content occurred in tissues of water-stressed plants. During water deficit, the levels of glucose, sucrose and stachyose decreased in thin roots (with a diameter <1 mm), whereas medium roots (diameter of 1–5 mm) exhibited no differences. Inorganic cations largely contribute to Ψ _π100 and remained stable during the period of water deficit, except for the level of Ca²⁺, which increased of 25% in water-stressed plants. The amount of malate increased in both leaves and roots during the dry period, whereas citrate and oxalate decreased. Thin roots seem to be more sensitive to water deficit and its consequent effects, while medium roots present more reactivity and a higher osmotic adjustment. The results support the hypothesis that the observed decreases in Ψ_w and active osmotic adjustment in leaves and roots of water-stressed olive plants may be physiological responses to tolerate water deficit.     

Cyclitols and carbohydrates in leaves and roots of 13 Eucalyptus species suggest contrasting physiological responses to water deficit

In many tree species, physiological adaptations to drought include the accumulation of osmotically active substances and/or the presence of particular compatible solutes, among them cyclitols. Recently, the cyclitol quercitol was identified in species of Eucalyptus, a diverse genus whose speciation is probably driven by adaptation to water availability. We subjected seedlings of 13 Eucalyptus species from different ecosystems ('mesic' and 'xeric') and different sub-generic taxonomic groups to 10 weeks of water deficit (WD) treatment. Pre-dawn water potentials (psi(pdwn)) and relative water content (RWC) were determined in shoots, and total osmolality, soluble low-molecular-weight carbohydrates and cyclitols were measured in leaves and roots. Responses to water deficit followed two distinct patterns: Eucalyptus species from 'mesic' environments adjusted concentrations of sucrose (through increased levels of sucrose and decreases in RWC) in response to water deficit, whereas 'xeric' species increased concentrations of quercitol (through reductions in RWC). In root tissues, only species from xeric environments contained high levels of quercitol and mannitol, increasing under WD conditions. We suggest that the former (mesic) strategy may be beneficial to respond to short-lasting drought conditions, because sucrose is easily metabolized, whereas the latter (xeric) strategy may relate to an effective acclimation to longer-lasting drought. These physiological response groups are also related to taxonomic groups within the genus.