Low-frequency spectroscopy of four-photon scattering in aqueous solutions of biopolymers

The novel method of nonlinear laser spectroscopy — low frequency spectroscopy of four-photon scattering of laser radiation was applied to detect a considerable growth of ortho-H₂O spin isomer and also H₂O₂ molecule concentration in a hydrate layer at the interface between water and DNA, denatured DNA molecules and α-chymotrypsin. Spectra of rotational resonances of ortho/para-H₂O spin isomers were observed in aqueous solutions of different biopolymers and also in distilled water in the range from zero to 100 cm^?1 with the spectral resolution of 0.05–0.1 cm^?1. The fitting of four-wave mixing spectra shows notable growth of the H₂O₂ concentration and rotational line’s amplitude by a factor of ～3 in DNA solutions due to denaturizing. Besides, we studied the four-photon scattering spectra of α-chymotrypsin aqueous solutions at protein concentrations between 0 and 20 mg/cm³ in the range of ±7 cm^?1. We found that the velocity of sound in the protein aqueous solution measured by the shift of the Mandelstam-Brillouin scattering spectrum components was a cubic dependence on the protein concentration and reached the value of about 3000 m/s at 20 mg/cm³.     

Carbon dioxide gas exchange and the energy status of leaves of Primula palinuri under water stress

The photosynthetic rate of water stressed leaves of Primula palinuri was reduced drastically by stomatal closure, not by limitations imposed on the capacity of the photosynthetic apparatus, when water loss exceeded 20% of the water content of turgid leaves. The sudden decrease in phtosynthesis was not observed when the lower epidermis of the leaves had been removed. In these ‘stripped’ leaves, inhibition of photosynthesis increased only gradually during the wilting caused by increasing water stress and was complete when the relative water content was as low as 20% compared with the initial value. This corresponded to a water potential of about-40 bar. The light intensity at which half-maximum rates of photosynthesis were observed decreased as stress increased. In intact leaves photosynthesizing in the presence of CO₂, light scattering, which is a measure of thylakoid energization, increased steeply during stomatal closure. The observed increase corresponded to the light scattering level measured in the absence of CO₂. When the lower epidermis was removed, no sudden increase in thylakoid energization could be observed during dehydration. Thylakoid energization remained high even at low water potentials. It decreased drastically only below a relative water content of 20%. Irrespective, of the extent of water stress, CO₂ fixation of stripped leaves increased when the oxygen content of air was reduced from 21% to 2%. Usually the transition from 21 to 2% O₂ was accompanied by increased thylakoid energization. The increase in energization was more pronounced below than above a relative water content of 50%. The data show that energy-dissipating photorespiratory CO₂ turnover in the in tercellular space of water-stressed leaves whose stomata are closed decreases only slowly as water stress increases. Respiratory CO₂ production by leaves in the dark was even more resistant to water stress than photosynthesis. It was still significant at water potentials as low as-80 bar.     

Raman spectra and conformation of the glycerophosphorylcholine headgroup

The Raman and infrared spectra of L-α-glycerophosphylcholine (GPC) and its cadmium complex (GPC·CdCl₂) have been measured for both the anhydrous and hydrate crystals. The Raman spectra of GPC and GPC·CdCl₂ dissolved in water are also reported. All the spectra fall into three distinctive patterns, demonstrating the existence of at least three different kinds of rotational isomers; the first is stable in the anhydrous GPC crystal, the second is stable in the GPC hydrate and GPC·CdCl₂ hydrate crystals and the third is stable in the anhydrous GPC·CdCl₂ crystal and also in aqueous solutions of GPC·CdCl₂. The latter form is reported for the first time.Raman spectra of dimyristoyl-L-α-phosphatidylcholine (L-DMPC) and dipalmitoyl-(L and DL)-α-phosphatidylcholine (L and DL-DPPC) hydrates have been measured below, around and above the transition temperature T₁. The spectra below T₁ indicate that the conformation of the glycerophosphorylcholine (GPC) headgroup in the L-DMPC crystal is similar to that in the L-DPPC crystal. The headgroup conformation in the DL-DPPC crystal is quite different from that in the L-DPPC crystal, although they became the same aftera crystalline trasnformation of DL-DPPC. The changes of the Raman spectra through the T₁ transition for the above crystals show that the headgroup conformation remains unchanged at the beginning o the transition.     

RATES OF PHOTOADAPTATION IN SEA ICE DIATOMS FROM MCMURDO SOUND,ANTARCTICA1

Sea ice microalgae are released from their relatively stable light environment to the water column seasonally, and any subsequent growth in a vertically mixed water column may depend, in part, on their photoadaptation rates. In this study we followed the time course of photoadaptation in natural sea ice algal communities from bottom ice and surface ice by measuring their photophysiological response to an artificial shift in the ambient irradiance field. Microalgae from under-ice habitats, were incubated under full sunlight (LL-HL) and microalgae from surface ice habitats were incubated under artificial light to mimic under-ice irradiance (HL-LL). During 3- to 4-day time course studies, opposite shifts in chlorophyll: carbon, α, P^B_m, and I_k were observed, depending on the direction of the irradiance change. First-order rate constants (k) ranged from 0.0067 to 0.29 h^?1 for photosynthetic parameters, although P^B_m did not always show a clear change over time. Rates of photoadaptation for ice algae are comparable to k values reported for temperate phytoplankton, suggesting that sea ice algae may be equally capable of adapting to the light conditions experienced in a vertically mixed water column. This study presents the first evidence that sea ice microalgae are physiologically capable of adapting to a planktonic life and thus could serve as a seed population for polar marine phytoplankton blooms.     

Pushed to the limit: consequences of climate change for the Araucariaceae: a relictual rain forest family

Background and Aims

Under predicted climate change scenarios, increased temperatures are likely to predispose trees to leaf and other tissue damage, resulting in plant death and contraction of already narrow distribution ranges in many relictual species. The effects of predicted upward temperatures may be further exacerbated by changes in rainfall patterns and damage caused by frosts on trees that have been insufficiently cold-hardened. The Araucariaceae is a relictual family and the seven species found in Australia have limited natural distributions characterized by low frost intensity and frequency, and warm summer temperatures. The temperature limits for these species were determined in order to help understand how such species will fare in a changing climate.

Methods

Experiments were conducted using samples from representative trees of the Araucariaceae species occurring in Australia, Agathis (A. atropurpurea, A. microstachya and A. robusta), Arauacaria (A. bidwilli, A. cunninghamii and A. heterophylla) and Wollemia nobilis. Samples were collected from plants grown in a common garden environment. Lower and higher temperature limits were determined by subjecting detached winter-hardened leaves to temperatures from 0 to –17 °C and summer-exposed leaves to 25 to 63 °C, then measuring the efficiency of photosystem II (F_v/F_m) and visually rating leaf damage. The exotherm, a sharp rise in temperature indicating the point of ice nucleation within the cells of the leaf, was measured on detached leaves of winter-hardened and summer temperature-exposed leaves.

Key Results

Lower temperature limits (indicated by FT₅₀, the temperature at which PSII efficiency is 50 %, and LT₅₀ the temperature at which 50 % visual leaf damage occurred) were approx. –5·5 to –7·5 °C for A. atropurpurea, A. microstachya and A. heterophylla, approx. –7 to –9 °C for A. robusta, A. bidwillii and A. cunninghamii, and –10·5 to –11 °C for W. nobilis. High temperature damage began at 47·5 °C for W. nobilis, and occurred in the range 48·5–52 °C for A. bidwillii and A. cunninghamii, and in the range 50·5–53·5 °C for A. robusta, A. microstachya and A. heterophylla. Winter-hardened leaves had ice nucleation temperatures of –5·5 °C or lower, with W. nobilis the lowest at –6·8 °C. All species had significantly higher ice nucleation temperatures in summer, with A. atropurpurea and A. heterophylla forming ice in the leaf at temperatures >3 °C higher in summer than in winter. Wollemia nobilis had lower FT₅₀ and LT₅₀ values than its ice nucleation temperature, indicating that the species has a degree of ice tolerance.

Conclusions

While lower temperature limits in the Australian Araucariaceae are generally unlikely to affect their survival in wild populations during normal winters, unseasonal frosts may have devastating effects on tree survival. Extreme high temperatures are not common in the areas of natural occurrence, but upward temperature shifts, in combination with localized radiant heating, may increase the heat experienced within a canopy by at least 10 °C and impact on tree survival, and may contribute to range contraction. Heat stress may explain why many landscape plantings of W. nobilis have failed in hotter areas of Australia.     

Regional Variability and Drivers of Below Ice CO<Subscript>2</Subscript> in Boreal and Subarctic Lakes

Northern lakes are ice-covered for considerable portions of the year, where carbon dioxide (CO₂) can accumulate below ice, subsequently leading to high CO₂ emissions at ice-melt. Current knowledge on the regional control and variability of below ice partial pressure of carbon dioxide (pCO₂) is lacking, creating a gap in our understanding of how ice cover dynamics affect the CO₂ accumulation below ice and therefore CO₂ emissions from inland waters during the ice-melt period. To narrow this gap, we identified the drivers of below ice pCO₂ variation across 506 Swedish and Finnish lakes using water chemistry, lake morphometry, catchment characteristics, lake position, and climate variables. We found that lake depth and trophic status were the most important variables explaining variations in below ice pCO₂ across the 506 lakes_. Together, lake morphometry and water chemistry explained 53% of the site-to-site variation in below ice pCO₂. Regional climate (including ice cover duration) and latitude only explained 7% of the variation in below ice pCO₂. Thus, our results suggest that on a regional scale a shortening of the ice cover period on lakes may not directly affect the accumulation of CO₂ below ice but rather indirectly through increased mobility of nutrients and carbon loading to lakes. Thus, given that climate-induced changes are most evident in northern ecosystems, adequately predicting the consequences of a changing climate on future CO₂ emission estimates from northern lakes involves monitoring changes not only to ice cover but also to changes in the trophic status of lakes.     

Adaptation of the Common Ice Plant to High Copper and Zinc Concentrations and Their Potential Using for Phytoremediation

A facultative halophite Mesembryanthemum crystallinum L. (the common ice plant) was shown to grow successively at the high concentrations of Cu and Zn. Although 25 μM CuSO₄ or 800 μM ZnSO₄ retarded markedly plant growth, they did not interfere with the completion of plant development and the formation of viable seeds. In such plants, leaves accumulated more than 200 μg of Cu and 1700 μg of Zn per 1 g of dry weight. A damaging effect of heavy metals (HMs) was manifested in a reduced content of water in leaves and proline accumulation in them. As copper is a metal with transient valence, copper salts are more toxic than zinc salts, which was manifested in a stronger inhibition of the chlorophyll synthesis. Both HMs induced oxidative stress, as evident from increased activities of guaiacol peroxidase and lipoxygenase. Moderate Cu and Zn concentrations did not damage cell membranes in leaves, as evident from the absence of their action on electrolyte leakage either under optimum conditions or after heat treatment. A capability of a substantial HM accumulation by the common ice plant and their considerable transport to shoots (up to 50 μg of Cu and 560 μg of Zn per plant) make it possible to consider the common ice plant as a promising phytoremediator. __________ Translated from Fiziologiya Rastenii, Vol. 52, No. 6, 2005, pp. 848–858. Original Russian Text Copyright ? 2005 by Kholodova, Volkov, Kuznetsov.     

Ortho/para spin-isomers of H2O molecules as a factor responsible for formation of two structural motifs in water

According to the latest results obtained by small-angle X-ray scattering and X-ray spectroscopy, it was suggested that water on a nanometer scale represents a fluctuating mixture of clusters with tetrahedral structure and a subphase with partially broken hydrogen bonds, whereas the nuclear configuration of the H₂O molecule corresponds to single tetrahedral coordination. The basic reason of such structural partition is not clear until now. Here we show that it can be associated with existence of two nuclear H₂O spin isomers that have different probability to be in one or the other subphase. The para molecule can transfer an excess of its rotational energy to the environment, up to complete stopping of rotation because its rotational quantum number J = 0 in the basic state. This property is favorable for formation of clusters with closed H-bonds. Ortho molecules with odd-numbered J states lack this property and thus should be predominantly present in the locations with impaired bonds.     

Structure of cellulose II hydrate

A hydrate of cellulose II can be formed by swelling Fortisan fibers in hydrazine and then washing in water. The hydrate is stable at 93% relative humidity and has a monoclinic unit cell with dimensions a = 9.02 Å, b = 9.63 Å, c = 10.34 Å, and γ = 116.0°; the space group is P2₁. The unit cell contains disaccharide sections of two chains and approximately four water molecules. The structure was refined using the LALS method, based on 10 observed and 10 unobserved reflections. An antiparallel arrangement of adjacent chains was assumed, since this occurs in cellulose II (the starting material), and the hydrate also reverts to cellulose II on dehydration. Refinement of the positions and side-chain conformations of the chains shows that the chains are stacked in the same way as in cellulose II, and the hydrate is formed by insertion of water molecules between the stacks. However, all efforts to arrange the water molecules in crystallographically regular positions led to unsatisfactory agreement between the observed and calculated intensities. These results suggest an irregular arrangement of the water molecules, which was modeled using water-weighted atomic scattering factors. The analysis resulted in two refined models with relative chain staggers of ～ +c/4 and ～ -c/4, which are indistinguishable in terms of the x-ray agreement. Our preference is for the +c/4 model, for which the stacks of chains are analogous to those in cellulose II.     

Light Scattering as an Indicator of the Energy State in Leaves of the Crassulacean Acid Metabolism Plant Kalanchoë pinnata

Both transmittance changes in a weak beam of green light (light scattering) and the slow decay of chlorophyll a fluorescence were used as indicators of the energy state of leaves of a Crassulacean acid metabolism plant, Kalanchoë pinnata, at frequent intervals during 12-hour light/12-hour dark cycles. To induce light scattering and fluorescence changes, leaves were exposed to red light for 6 minutes. When measurements were made during the light period, the leaves were kept in darkness for 6 minutes before illumination. In the middle of the light period, when malic acid decarboxylation was very active and stomatal conductance was low, light scattering changes were small and indicated that the energy state of leaves was low. This result was supported by determination of adenylate levels. Light scattering and ATP/ADP ratios increased during the late light period when the tissue was deacidified. Illumination produced maximum light scattering changes between the 2nd and 5th hour of the dark period, when rates of dark CO₂ fixation were highest. Light scattering and fluorescence measurements taken from leaves, which were illuminated with red or far-red light in the presence or absence of O₂ showed that, in addition to linear electron transport, K. pinnata has the potential for both cyclic and pseudocyclic electron transport. The results are relevant with regard to the high ATP demand during Crassulacean acid metabolism.     

The Response of Leaf Water Potential and Crassulacean Acid Metabolism to Prolonged Drought in Sedum rubrotinctum

Plants of Sedum rubrotinctum R. T. Clausen were studied in a green-house over a 2-year period without watering. Only the apical leaves survived and were turgid at the end of the experiment. The midday leaf water potential of these apical leaves was −1.20 megapascals, while the leaf water potential of comparable leaves on well-watered control plants was −0.20 megapascals. The unwatered plants appear to have maintained turgor by means of an osmotic adjustment. After 2 years without water the plants no longer exhibited a nocturnal accumulation of titratable acidity. However, the daytime levels of titratable acidity of the unwatered plants were more than 2-fold greater than the levels in well-watered control plants. Well-watered plants of S. rubrotinctum exhibited seasonal shifts in biomass stble carbon isotope ratios, indicating a greater proportion of day versus night CO₂ uptake in the winter than in the summer. The imposition of water stress prevented the expression of this seasonal rhythm and restricted the plants to dark CO₂ uptake.     

Ice nucleation temperature of individual leaves in relation to population sizes of ice nucleation active bacteria and frost injury

Ice nucleation temperatures of individual leaves were determined by a tube nucleation test. With this assay, a direct quantitative relationship was obtained between the temperatures at which ice nucleation occurred on individual oat (Avena sativa L.) leaves and the population sizes of ice nucleation active (INA) bacteria present on those leaves. In the absence of INA bacteria, nucleation of supercooled growth-chamber grown oat leaves did not occur until temperatures were below approximately −5°C. Both nucleation temperature and population size of INA bacteria were determined on the same individual, field-grown oat leaves. Leaves with higher ice nucleation temperatures harbored larger populations of INA bacteria than did leaves with lower nucleation temperatures. Log₁₀ mean populations of INA bacteria per leaf were 5.14 and 3.51 for leaves with nucleation temperatures of −2.5°C and −3.0°C, respectively. Nucleation frequencies (the ratio of ice nuclei to viable cells) of INA bacteria on leaves were lognormally distributed. Strains from two very different collections of Pseudomonas syringae and one of Erwinia herbicola were cultured on nutrient glycerol agar and tested for nucleation frequency at −5°C. Nucleation frequencies of these bacterial strains were also lognormally distributed within each of the three sets. The tube nucleation test was used to determine the frequency with which individual leaves in an oat canopy harbored large populations of INA bacteria throughout the growing season. This test also predicted relative frost hazard to tomato (Lycopersicon esculentum Mill) plants.     

Neutron scattering from water in DNA and muscle tissue

Neutron scattering studies of rotational modes of water in DNA and muscle tissue are reported. Spectra from wet DNA and rat muscle are compared with those from ice and water. No evidence is found for an ice-like water component in biological systems. The relevance of neutron scattering for such a study is examined and possible discrepancies with other measurements are discussed. The similarity of the neutron spectra for wet DNA and muscle tissue is taken as evidence that water-nucleotide and water-peptide interactions are similar to water-water interactions when probed on a short time scale.     

Stress tolerance and stress-induced injury in crop plants measured by chlorophyll fluorescence in vivo: chilling, freezing, ice cover, heat, and high light

The proposition is examined that measurements of chlorophyll fluorescence in vivo can be used to monitor cellular injury caused by environmental stresses rapidly and nondestructively and to determine the relative stress tolerances of different species. Stress responses of leaf tissue were measured by F_R, the maximal rate of the induced rise in chlorophyll fluorescence. The time taken for F_R to decrease by 50% in leaves at 0°C was used as a measure of chilling tolerance. This value was 4.3 hours for chilling-sensitive cucumber. In contrast, F_R decreased very slowly in cucumber leaves at 10°C or in chilling-tolerant cabbage leaves at 0°C. Long-term changes in F_R of barley, wheat, and rye leaves kept at 0°C were different in frost-hardened and unhardened material and in the latter appeared to be correlated to plant frost tolerance. To simulate damage caused by a thick ice cover, wheat leaves were placed at 0°C under N₂. Kharkov wheat, a variety tolerant of ice encapsulation, showed a slower decrease in F_R than Gatcher, a spring wheat. Relative heat tolerance was also indicated by the decrease in F_R in heated leaves while changes in vivo resulting from photoinhibition, ultraviolet radiation, and photobleaching can also be measured.     

Growth rates of arctic juvenile Scolelepis squamata (Polychaeta: Spionidae) isolated from Chukchi Sea fast ice

In spring, Arctic coastal fast ice is inhabited by high densities of sea ice algae and, among other fauna, juveniles of benthic polychaetes. This paper investigates the hypothesis that growth rates of juveniles of the common sympagic polychaete, Scolelepis squamata (Polychaeta: Spionidae), are significantly faster at sea ice algal bloom concentrations compared to concurrent phytoplankton concentrations. Juvenile S. squamata from fast ice off Barrow, Alaska, were fed with different algal concentrations at 0 and 5?°C, simulating ambient high sea ice algal concentrations, concurrent low phytoplankton concentrations, and an intermediate concentration. Growth rates, calculated using a simple linear regression equation, were significantly higher (up to 115?times) at the highest algal concentration compared to the lowest. At the highest algal concentration, juveniles grew faster at 5?°C compared to those feeding at 0?°C with a Q ₁₀ of 2.0. We conclude that highly concentrated sea ice algae can sustain faster growth rates of polychaete juveniles compared to the less dense spring phytoplankton concentrations. The earlier melt of Arctic sea ice predicted with climate change might cause a mismatch between occurrence of polychaete juveniles and food availability in the near future. Our data indicate that this reduction in food availability might counteract any faster growth of a pelagic juvenile stage based on forecasted increased water temperatures.     

Frost injury and heterogeneous ice nucleation in leaves of tuber-bearing solanum species : ice nucleation activity of external source of nucleants

The heterogeneous ice nucleation characteristics and frost injury in supercooled leaves upon ice formation were studied in nonhardened and cold-hardened species and crosses of tuber-bearing Solanum. The ice nucleation activity of the leaves was low at temperatures just below 0°C and further decreased as a result of cold acclimation. In the absence of supercooling, the nonhardened and cold-hardened leaves tolerated extracellular freezing between −3.5° and −8.5°C. However, if ice initiation in the supercooled leaves occurred at any temperature below −2.6°C, the leaves were lethally injured.

To prevent supercooling in these leaves, various nucleants were tested for their ice nucleating ability. One% aqueous suspensions of fluorophlogopite and acetoacetanilide were found to be effective in ice nucleation of the Solanum leaves above −1°C. They had threshold temperatures of −0.7° and −0.8°C, respectively, for freezing in distilled H₂O. Although freezing could be initiated in the Solanum leaves above −1°C with both the nucleants, 1% aqueous fluorophlogopite suspension showed overall higher ice nucleation activity than acetoacetanilide and was nontoxic to the leaves. The cold-hardened leaves survived between −2.5° and −6.5° using 1% aqueous fluorophlogopite suspension as a nucleant. The killing temperatures in the cold-hardened leaves were similar to those determined using ice as a nucleant. However, in the nonhardened leaves, use of fluorophlogopite as a nucleant resulted in lethal injury at higher temperatures than those estimated using ice as a nucleant.

     

Shape shifting predicts ontogenetic changes in metabolic scaling in diverse aquatic invertebrates

Metabolism fuels all biological activities, and thus understanding its variation is fundamentally important. Much of this variation is related to body size, which is commonly believed to follow a 3/4-power scaling law. However, during ontogeny, many kinds of animals and plants show marked shifts in metabolic scaling that deviate from 3/4-power scaling predicted by general models. Here, we show that in diverse aquatic invertebrates, ontogenetic shifts in the scaling of routine metabolic rate from near isometry (b_R = scaling exponent approx. 1) to negative allometry (b_R < 1), or the reverse, are associated with significant changes in body shape (indexed by b_L = the scaling exponent of the relationship between body mass and body length). The observed inverse correlations between b_R and b_L are predicted by metabolic scaling theory that emphasizes resource/waste fluxes across external body surfaces, but contradict theory that emphasizes resource transport through internal networks. Geometric estimates of the scaling of surface area (SA) with body mass (b_A) further show that ontogenetic shifts in b_R and b_A are positively correlated. These results support new metabolic scaling theory based on SA influences that may be applied to ontogenetic shifts in b_R shown by many kinds of animals and plants.     

Hydrogen peroxide spraying alleviates drought stress in soybean plants

To ascertain the effect of exogenously applied hydrogen peroxide (H₂O₂) on drought stress, we examined whether the spraying of soybean leaves with H₂O₂ would alleviate the symptoms of drought stress. Pre-treatment by spraying leaves with H₂O₂ delayed foliar wilting caused by drought stress compared to leaves sprayed with distilled water (DW). Additionally, the relative water content of drought-stressed leaves pre-treated with H₂O₂ was higher than that of leaves pre-treated with DW. Therefore, we analyzed the effect of H₂O₂ spraying on photosynthetic parameters and on the biosynthesis of oligosaccharides related to water retention in leaves during drought stress. Under conditions of drought stress, the net photosynthetic rate and stomatal conductance of leaves pre-treated with H₂O₂ were higher than those of leaves pre-treated with DW. In contrast to DW spraying, H₂O₂ spraying immediately caused an increase in the mRNA levels of d-myo-inositol 3-phosphate synthase 2 (GmMIPS2) and galactinol synthase (GolS), which encode key enzymes for the biosynthesis of oligosaccharides known to help plants tolerate drought stress. In addition, the levels of myo-inositol and galactinol were higher in H₂O₂-treated leaves than in DW-treated leaves. These results indicated that H₂O₂ spraying enabled the soybean plant to avoid drought stress through the maintenance of leaf water content, and that this water retention was caused by the promotion of oligosaccharide biosynthesis rather than by rapid stomatal closure.     

Tolerance and physiological responses of Phragmites australis to water deficit

The water stress tolerance of Phragmites australis (Cav.) Trin ex. Steud. grown in the laboratory were investigated by examining effects of different levels of imposed water deficits on growth, photosynthesis and various physiological traits related to water stress. Individual plants were grown under conditions of unrestricted water supply and compared with groups of plants receiving 60, 30, 15 or 5% of previous daily water requirements, respectively.Water deficit was found to reduce the leaf area and the leaf biomass per plant due to decreased production of new leaves, increased leaf shedding and reduced average leaf size. Leaf production and leaf expansion growth were very sensitive to water availability and were reduced when plants were subjected to fairly mild water deficit. Osmolality in sap expressed from leaves and the concentration of proline in leaves were only significantly increased in severely stressed plants, indicating that osmotic adjustment was of minor importance until a critical stress level was reached. Photosynthetic parameters were rather unaffected until the water availability was very low and led to the assertion that reduced CO₂ assimilation was mainly due to stomatal closure and not biochemical changes. Water stress had no effect on the activity of Rubisco. The CO₂ assimilation rate and stomatal conductance decreased in such a way that the intrinsic water use efficiency (A/g_s) increased, indicating efficient CO₂ utilization in water stressed plants. The apparent quantum yield (φ_i) was reduced in leaves of the most stressed plants, probably due to a decrease in the CO₂ molar fraction in the chloroplasts following stomatal closure.The initial response of P. australis to water deficit is a reduction in leaf area, the remaining leaves staying physiological rather well functioning until they are severely stressed. A high intrinsic water use efficiency and the ability to maintain some capacity for photosynthesis under severe water stress can undoubtedly contribute to the survival of P. australis under dry conditions. Taken together with its well-developed adaptations to flooding, P. australis seems very well adapted to grow in wetland areas with a widely fluctuating hydroperiod. P. australis grows very well in rather deep water, but can also tolerate extensive periods of drought with reduced availability of water.     

The V amylose–H2O system: Structural changes resulting from hydration

X-ray diffraction and stereochemical analyses were used to study the hydrated structure of the helical amylose polymorph having a fiber repeat spacing of 8 Å. Intensity calculations using computer models confirmed six as the number of residues per turn and supported the space group P2₁2₁2₁. Both diffraction intensity and stereochemical methods indicate the suitability of residue G3 from the potassium acetate complex of cyclohexaamylose as opposed to residues with longer O(4)–O(1) vectors. Comparison of the present results with those obtained for V amylose dehydrate indicated no major conformational differences between the two helix skeletons. A net helical rotation of approximately 30° accompanied the hydrate–dehydrate transition and the rotational position in the hydrate allowed packing that was less close. Hydration water molecules were not located; noncarbohydrate peaks on the electron density maps were primarily due to Fourier series termination errors.