Rehydration Experiments Research Articles

Evidence for variations in cryogenic extraction deuterium biases of plant xylem water across foundational northeastern US trees

AbstractMeasurements of oxygen and hydrogen isotopes in plant xylem water (2H, 18O) have helped to redefine conceptual and numerical models of the hydrological cycle and understand how plants compete for subsurface water. Recent experiments have shown that Cryogenic Vacuum Extraction (CVE) of plant xylem water can result in a δ2H bias. We tested if CVE δ2H‐biases varied significantly across seven foundational northeastern US forest trees with a series of tree core rehydration experiments. Our analysis demonstrated that CVE δ2H‐biases were well predicted by sample gravimetric water content and varied significantly with tree species identity. We show that species‐level δ2H‐bias corrections can result in substantially different understandings of plant water uptake and transpiration versus uncorrected data or generic bias corrections. This research demonstrates an urgent need for the critical evaluation of CVE for plant water extraction. In the absence of a stronger understanding of CVE δ2H‐biases, we recommend that xylem water δ2H observations should not be used in plant water uptake studies.

Passive stomatal closure under extreme drought in an angiosperm species.

The phytohormone abscisic acid (ABA) plays a major role in closing the stomata of angiosperms. However, recent reports of some angiosperm species having a peaking-type ABA dynamic, in which under extreme drought ABA levels decline to pre-stressed levels, raises the possibility that passive stomatal closure by leaf water status alone can occur in species from this lineage. To test this hypothesis, we conducted instantaneous rehydration experiments in the peaking-type species Umbellularia californica through a long-term drought, in which ABA levels declined to pre-stress levels, yet stomata remain closed. We found that when ABA levels were lowest during extreme drought, stomata of U. californica were passively closed by leaf water status alone, with stomata reopening rapidly to maximum rates of gas exchange on instantaneous rehydration. This contrasts with leaves early in drought, in which ABA levels were highest and stomata did not reopen on instantaneous rehydration. The transition from ABA-driven stomatal closure to passively driven stomatal closure as drought progresses in this species occurs at very low water potentials facilitated by highly embolism-resistant xylem. These results have important implications for understanding stomatal control during drought in angiosperms.

The Life-History Traits of Soil-Dwelling Nematode (Acrobeloides sp.) Exhibit More Resilience to Water Restriction Than Caenorhabditis elegans.

In the context of climate warming, the intensity and frequency of drought occurrences are progressively increasing. However, current research on the impacts of drought on the life-history traits and physiological activities of animals rarely encompasses soil animals that play crucial roles within soil ecosystems. Therefore, this study focused on a soil nematode species (Acrobeloides sp.) and a model nematode (Caenorhabditis elegans) to investigate whether nematodes adjust the trade-off of their life-history traits to confront arid environments, utilizing a Petri dish experiment. Subsequently, we assessed the resilience of the two nematode species to moisture variations by comparing the extent of changes in various indicators (i.e., life-history traits, physiological traits, and oxidative stress) of nematodes before and after drought and rehydration. The results revealed that both nematode species are capable of adapting to arid environments by altering the trade-off between life-history traits. Specifically, they reduce reproductive investment and body mass while maintaining life span, thus responding to drought conditions. Follow-up rehydration experiments post-drought stress highlighted that the soil-dwelling nematode exhibits a superior recovery capacity in response to moisture fluctuations in comparison to the model nematode. To the best of our knowledge, this is the first investigation into life history of drought adaptation within soil-dwelling nematodes. Moreover, the findings hold significant implications for the exploration of drought adaptation and its mechanisms in soil-dwelling animals.

Quantitative contribution of cryogenic vacuum extraction and radial water transport to xylem-source water deuterium offsets

The positions and magnitudes of deuterium offsets between bulk xylem and corresponding source waters are under debate and quantifying them is essential for isotope-based ecohydrological investigations. In this study, stable isotopes (δ2H, δ18O, and δ13C), iteration method, and rehydration experiments were combined to quantitatively determine the magnitude of cryogenic vacuum extraction (CVE)- and radial water transport (RWT)-induced deuterium offsets using one riparian tree species Salix babylonica L. A modified potential water source line (MPWL) was proposed to identify the total δ2H offsets between bulk xylem and source waters. The relationships between δ2H offsets induced by CVE or RWT and plant water content, leaf δ13C values, soil water content (SWC), and the depth to the water table (WTD) were investigated. Results showed that the bulk xylem waters in different tissue positions of S. babylonica showed −7.0 ‰ to −4.0 ‰ deuterium depletion relative to MPWL at four different sites (p < 0.01). The isotopic compositions of sap water coincided well with MPWL on the dual-isotope plot at the four sites. The CVE- and RWT-induced δ2H offsets accounted for 75.1 % and 24.9 % of the total δ2H offsets, respectively. The CVE-induced δ2H offsets were significantly negatively correlated with plant water content. In comparison, the RWT-induced δ2H offsets were negatively related to plant leaf δ13C values, trunk water content, and SWC, but positively correlated with WTD. This study provides a quantitative contribution of two major sources of deuterium offsets. The results provide critical insights into isotope-based plant water source identification and evapotranspiration partitioning.

Optimization of In Vitro Germination, Viability Tests and Storage of Paeonia ostii Pollen.

Paeonia ostii is an important woody oil crop mainly cross-pollinated. However, the low yield has become an important factor restricting the industrial development of P. ostii. Cross-pollination has become one of the important measures to increase the seed yield. Therefore, conservation of pollen with high vitality is crucial to ensure successful pollination of P. ostii. In this study, we found an effective methodological system to assess the viability, ability to germinate, and optimal storage conditions of P. ostii pollen grains. The optimal medium in vitro was 50 g/L sucrose, 100 mg/L boric acid, 50 g/L PEG6000, 100 mg/L potassium nitrate, 300 mg/L calcium nitrate, and 200 mg/L magnesium sulfate at pH 5.4. Optimal germination condition in vitro was achieved at 25 °C for 120 min, allowing easy observation of the germination percentage and length of the pollen tubes. In addition, the viability of pollen grains was assessed by comparing nine staining methods. Among them, MTT, TTC, benzidine-H2O2, and FDA were effective to distinguish between viable and non-viable pollen, and the results of the FDA staining method were similar to the pollen germination percentage in vitro. After evaluation of pollen storage, thawing and rehydration experiments showed that thawing at 4 °C for 30 min and rehydration at 25 °C for 30 min increased the germination percentage of pollen grains stored at low temperatures. The low-temperature storage experiments showed that 4 °C was suitable for short-term storage of P. ostii pollen grains, while -80 °C was suitable for long-term storage. This is the first report on the in vitro germination, viability tests, and storage of P. ostii pollen grains, which will provide useful information for P. ostii germplasm conservation and artificial pollination.

Resistant xylem from roots to peduncles sustains reproductive water supply after drought-induced cavitation of wheat leaves.

Many annual grasses exhibit drought avoiding life cycles where rapid reproduction must be completed before soil water is exhausted. This strategy would seem to require a hydraulic system capable of sustaining reproduction at all costs to the rest of the plant, yet little is known about the whole-plant structure of hydraulic vulnerability in grasses. We examine vulnerability to water-stress induced xylem cavitation in roots, flag leaves, basal and apical regions of peduncles of wheat (Triticum aestivum var Krichauff) to understand the staged failure of xylem function in severe drought. The functionality of segmented vulnerabilities is tested by conducting rehydration experiments after acute dehydration. We show the water supply to peduncles is more drought resistant than in leaves due to greater xylem cavitation resistance, ensuring a pathway of water can be maintained from the roots to the reproductive tissues even after severe water deficit. Differential rehydration of peduncles compared to leaves following drought confirmed the functionality of xylem supply from roots to seed after water stress sufficient to completely cavitate flag leaf vessels. These results demonstrate that a proportion of the hydraulic pathway between roots and seeds remains functional under extreme dehydration, suggesting that vulnerability traits in this key grass species reflect its reproductive strategy.

Technical note: On uncertainties in plant water isotopic composition following extraction by cryogenic vacuum distillation

Abstract. Recent studies have challenged the interpretation of plant water isotopes obtained through cryogenic vacuum distillation (CVD) based on observations of a large 2H fractionation. These studies have hypothesized the existence of an H-atom exchange between water and organic tissue during CVD extraction with the magnitude of H exchange related to relative water content of the sample; however, clear evidence is lacking. Here, we systematically tested the uncertainties in the isotopic composition of CVD-extracted water by conducting a series of incubation and rehydration experiments using isotopically depleted water, water at natural isotope abundance, woody materials with exchangeable H, and organic materials without exchangeable H (cellulose triacetate and caffeine). We show that the offsets between hydrogen and oxygen isotope ratios and expected reference values (Δ2H and Δ18O) have inversely proportional relationships with the absolute amount of water being extracted, i.e. the lower the water amount, the higher the Δ2H and Δ18O. However, neither Δ2H nor Δ18O values, were related to sample relative water content. The Δ2H pattern was more pronounced for materials with exchangeable H atoms than with non-exchangeable H atoms. This is caused by the combined effect of H exchange during the incubation of materials in water and isotopic enrichments during evaporation and sublimation that depend on absolute water amount. The H exchange during CVD extraction itself was negligible. Despite these technical issues, we observed that the water amount-dependent patterns were much less pronounced for samples at natural isotope abundance and particularly low when sufficiently high amounts of water were extracted (&gt;600 µL). Our study provides new insights into the mechanisms causing isotope fractionation during CVD extraction of water. The methodological uncertainties can be controlled if large samples of natural isotope abundance are used in ecohydrological studies.

Effect of Temperature on Rehydration of Cornelian Cherry Fruit: A Kinetics Study

Cornelian cherry is an important resource for human health and nutrition. Cornelian cherry is very rich in contains many mineral substances such as vitamin C. For this reason, it is important that cornelian cherry can be dried and used later. This was studied investigated the rehydration capacity and the kinetics of dried cornelian cherry fruit under certain conditions. Drying was carried out in a thin layer using a convective dryer. The drying conditions were determined as the different temperatures of air (50, 60, and 70°C) and the constant velocity of air of 1 m/s. The rehydration experiments of dried cornelian cherry were actualized in a hot water bath with a water circulator. The experiments were made with rehydration water temperatures of 25, 50, and 75°C. The rehydration characterization of dried cornelian cherry was evaluated in terms of rehydration curves, rehydration rate curves, final moisture content, and rehydration coefficient (COR). Fick’s 2nd law was employed to determine the coefficients of effective diffusivity of the rehydration process. The kinetics models of Peleg, the Weibull, the first-order, and the exponential were used to determine the kinetics of rehydration. The model of Weibull was determined to be the most suitable model for investigating the kinetics of rehydration of cornelian cherry samples. The Model fit was evaluated by considering statistical parameters such as R2, χ2, RMSE, and P%. In addition, the energy of activation of rehydration was figured with the rate constants of the most suitable models.

Comparative cryogenic extraction rehydration experiments reveal isotope fractionation during root water uptake in Gramineae.

Determining whether isotope fractionation occurs during root water uptake is a prerequisite for using stem or xylem water isotopes to trace water sources. However, it is unclear whether isotope fractionation occurs during root water uptake in gramineous crops. We conducted prevalidation experiments to estimate the isotope measurement bias associated with cryogenic vacuum distillation (CVD). Next, we assessed isotope fractionation during root water uptake in two common agronomic crops, wheat (Triticum aestivum L.) and maize (Zea mays L.), under flooding after postdrought stress conditions. Cryogenic vacuum distillation caused significant depletion of 2 H but negligible effects on 18 O for both soil and stem water. Surprisingly CVD caused depletion of 2 H and enrichment of 18 O in root water. Stem and root water δ18 O were more than soil water δ18 O, even considering the uncertainty of CVD. Soil water 18 O was depleted compared with irrigation water 18 O in the pots with plants but enriched relative to irrigation water 18 O in the pots without plants. These results indicate that isotope fractionation occurred during wheat and maize root water uptake after full irrigation and led to a heavy isotope enrichment in stem water. Therefore, the xylem/stem water isotope approach widely used to trace water sources should be carefully evaluated.

Causes and Factors of Cryogenic Extraction Biases on Isotopes of Xylem Water

AbstractCryogenic vacuum extraction (CVE) has been considered as the standard technique for the analysis of plant water stable isotopes in ecohydrological research. Recent studies reported that CVE can introduce significant bias in stable isotope analyses, yet the causes and influencing factors of the CVE‐induced deuterium offsets remain poorly understood. Here, we performed rehydration experiments on plant samples from two species and three organs with two distinct‐isotopic spiking waters. Centrifugation and high‐pressure mechanical squeezing were used to separate sap water and tissue water for stable isotope analyses. Plant waters extracted by CVE differed significantly from reference waters in δ2H, but not in δ18O. The δ2H bias was linearly correlated to the xylem water content, and this relationship is affected significantly by plant organs/species and the isotopic signature of the spiking water. Moreover, the δ2H bias induced by CVE (−8.52 ± 0.90‰) was significantly greater than the δ2H difference between the tissue and sap waters (−3.33 ± 0.76‰) for apple stems possessing similar water contents. Thus, hydrogen‐exchange between plant organics and water, and isotopic heterogeneity within plants both contribute to the negative δ2H bias, but the former is the dominant cause. The multiple factors governing the CVE‐induced δ2H bias, make it difficult to establish a unified bias correction equation. Our results question the usefulness of cryogenic extraction as a standard for plant water extraction.

Optimization of In Vitro Germination and storage of Armeniaca sibirica Pollen

Armeniaca sibirica is an important nut species with ecological and economic value. However, low fruit yield has become a significant obstacle to industrial production of A. sibirica. High and stable yield breeding via cross-pollination has become a solution for improving production. Therefore, pollen with high vitality is essential to ensure successful hybridization in the A. sibirica breeding program. In this study, we used pollen from a A. sibirica clone to optimize the composition of germination medium. The orthogonal assay test strategy (OATS) indicated an agar-solidified medium containing 15 g/100 mL sucrose, 0.01 g/100 mL boric acid, and 0.01 g/100 mL calcium nitrate at pH 6.0 was optimal for pollen germination. The pollen germination rate and tube length in the optimal medium were checked at seven different temperatures and times. The results indicated that 25 °C was the optimal temperature. Controlled unfreezing and rehydration experiments showed that the germination rate of cryopreserved pollen reached its highest value when pollen was thawed for 24 h at 4 °C and rehydrated for 4 h at 25 °C, demonstrating that unfreezing and rehydration greatly improved the vitality of pollen. A. sibirica pollen could remain high viability at 25 °C (4 weeks) or 4 °C (8 weeks) for short-term storage, and -20 °C and -80 °C were suitable temperatures for long-term storage (52 weeks). Our results can provide helpful information for pollen sources in the A. sibirica cross breeding program.

Withdrawn: Similar Isotopic Biases of Plant Stem Bulk Water From Different Water Sources by Cryogenic Vacuum Distillation Demonstrated Through Rehydration Experiments

AbstractZhao, P., Sprenger, M., Barzegar, R., Tang, X., &amp; Adamowski, J. (2022). Similar isotopic biases of plant stem bulk water from different water sources by cryogenic vacuum distillation demonstrated through rehydration experiments. Geophysical Research Letters, 49, e2021GL096474. The above article from Geophysical Research Letters, published online on 30 March 2022 in Wiley Online Library (http://wileyonlinelibrary.com), has been withdrawn by agreement among the authors, the Editor‐in‐Chief Harihar Rajaram, the American Geophysical Union, and Wiley Periodicals, LLC. The withdrawal has been agreed because the authors discovered, after acceptance of the article but before final publication, some accidental transcription/typo errors in the data (for some δ2H and δ18O values) which would affect some of the results and cannot be sufficiently corrected to the authors' satisfaction.

Desiccation-rehydration measurements in bryophytes: current status and future insights.

Desiccation-rehydration experiments have been employed over the years to evaluate desiccation tolerance of bryophytes (Bryophyta, Marchantiophyta, and Anthocerotophyta). Researchers have applied a spectrum of protocols to induce desiccation and subsequent rehydration, and a wide variety of techniques have been used to study desiccation-dependent changes in bryophyte molecular, cellular, physiological, and structural traits, resulting in a multifaceted assortment of information that is challenging to synthesize. We analysed 337 desiccation-rehydration studies, providing information for 351 species, to identify the most frequent methods used, analyse the advances in desiccation studies over the years, and characterize the taxonomic representation of the species assessed. We observed certain similarities across methodologies, but the degree of convergence among the experimental protocols was surprisingly low. Out of 52 bryophyte orders, 40% have not been studied, and data are lacking for multiple remote or difficult to access locations. We conclude that for quantitative interspecific comparisons of desiccation tolerance, rigorous standardization of experimental protocols and measurement techniques, and simultaneous use of an array of experimental techniques are required for a mechanistic insight into the different traits modified in response to desiccation. New studies should also aim to fill gaps in taxonomic, ecological, and spatial coverage of bryophytes.

Cloning and Drought Resistance Analysis of Soybean GmHsps_p23-like Gene

Soybean (Glycine max (L.) Merr.) is an important cultivated crop, which requires much water during its growth, and drought seriously affects soybean yields. Studies have shown that the expression of small heat shock proteins can enhance drought resistance, cold resistance and salt resistance of plants. In this experiment, soybean GmHsps_p23-like gene was successfully cloned by RT-PCR, the protein encoded by the GmHsps_p23-like gene was subjected to bioinformatics analysis, and the pCAMBIA3301-GmHsps_p23-like overexpression vector and pCBSG015-GmHsps_p23-like gene editing vector were constructed. Agrobacterium-mediated method was used to transform soybeans to obtain positive plants. RT-PCR detection, rehydration experiment and drought resistance physiological and biochemical index detection were performed on the T2 generation positive transgenic soybean plants identified by PCR and Southern hybridization. The results showed that the overexpression vector plant GmHsps_p23-like gene expression increased. After rehydration, the transgenic overexpression plants returned to normal growth, and the damage to the plants was low. After drought stress, the SOD and POD activities and the PRO content of the transgenic overexpression plants increased, while the MDA content decreased. The reverse was true for soybean plants with genetically modified editing vectors. The drought resistance of the overexpressed soybeans under drought stress was higher than that of the control group, and had a stronger drought resistance. It showed that the expression of soybean GmHsps_p23-like gene can improve the drought resistance of soybean. The cloning and functional verification of soybean GmHsps_p23-like gene had not been reported yet. This is the first time that PCR technology has been used to amplify the soybean GmHsps_p23-like gene and construct an expression vector for this gene. This research has laid the foundation for transgenic technology to improve plant drought resistance and cultivate new drought-resistant transgenic soybean varieties.

Vegetative desiccation tolerance in Eragrostiella brachyphylla: biochemical and physiological responses

Eragrostiella brachyphylla is an angiosperm desiccation-tolerant resurrection plant, which can survive during desiccation in the air-dry state and recover completely on availability of water. The present study was conducted to understand the vegetative desiccation tolerance of Eragrostiella brachyphylla by evaluating its ability to recover the physiological, biochemical and morphological functions post desiccation. In order to understand the responses of Eragrostiella brachyphylla to desiccation and subsequent rehydration experiments were conducted in the hydrated state (HS), desiccated state (DS) and rehydrated state (RS). Scanning electron microscopy revealed significant changes between the three stages in the internal ultra-structures of leaves and stems. Compared to the other states, photosynthetic parameters such as chlorophyll a, chlorophyll b, total chlorophylland total carotenoid contents decreased significantly in the desiccated state. Superoxide radical (O2•−) content also increased, resulting in an oxidative burst during desiccation. Consequently, antioxidant enzymes such as catalase (CAT) superoxide dismutase (SOD) peroxidase (APX) and glutathione reductase (GR) activities were found to be significantly elevated in the desiccated state to avoid oxidative damage. Increased malondialdehyde (MDA) content and relative electrolyte leakage (REL) during desiccation provide evidence for membrane damage and loss of cell-wall integrity. During desiccation, the contents of osmolytes represented by sucrose and proline were found to increase to maintain cell structure integrity. After rehydration, all physiological, biochemical and morphological properties remain unchanged or slightly changed when compared to the hydrated state. Hence, we believe that these unique adaptations contribute to the remarkable desiccation-tolerance property of this plant.

Evidence for regional aeolian transport of freshwater micrometazoans in arid regions.

While separated by large expanses of dry terrain unsuitable for aquatic biota, aridland waters possess high biodiversity. How aquatic micrometazoans disperse to, and colonize, these isolated ephemeral habitats are not well understood. We used a multi-faceted approach including wind tunnel and rehydration experiments, and next-generation sequencing to assess potential movement of diapausing propagules of aquatic invertebrates by anemochory across regional scales (102-105 km). Wind tunnel experiments using dry playa sediments with added micrometazoan propagules demonstrated that after entrainment by saltation and downwind transport were subsequently recoverable as viable animals when rehydrated. Further, rehydration of fallen natural dust yielded micrometazoans, including rotifers, gastrotrichs, microcrustaceans, and nematodes. Using conserved DNA primers, we identified >3,300 eukaryotic Operational Taxonomic Units (excluding fungi) in the dust including some taxa found in rehydration experiments. Thus, we provide strong evidence that anemochory can disperse micrometazoans among isolated, ephemeral ecosystems in North American deserts and likely elsewhere.

How water availability influences morphological and biomechanical properties in the one-leaf plant Monophyllaea horsfieldii.

In its natural habitat, the one-leaf plant Monophyllaea horsfieldii (Gesneriaceae) shows striking postural changes and dramatic loss of stability in response to intermittently occurring droughts. As the morphological, anatomical and biomechanical bases of these alterations are as yet unclear, we examined the influence of varying water contents on M. horsfieldii by conducting dehydration–rehydration experiments together with various imaging techniques as well as quantitative bending and turgor pressure measurements. As long as only moderate water stress was applied, gradual reductions in hypocotyl diameters and structural bending moduli during dehydration were almost always rapidly recovered in acropetal direction upon rehydration. On an anatomical scale, M. horsfieldii hypocotyls revealed substantial water stress-induced alterations in parenchymatous tissues, whereas the cell form and structure of epidermal and vascular tissues hardly changed. In summary, the functional morphology and biomechanics of M. horsfieldii hypocotyls directly correlated with water status alterations and associated physiological parameters (i.e. turgor pressure). Moreover, M. horsfieldii showed only little passive structural–functional adaptations to dehydration in comparison with poikilohydrous Ramonda myconi.

Water content in the Martian mantle: A Nakhla perspective

Water contents of the Martian mantle have previously been investigated using Martian meteorites, with several comprehensive studies estimating the water content in the parental melts and mantle source regions of the shergottites and Chassigny. However, no detailed studies have been performed on the Nakhla meteorite. One possible way to determine the water content of a crystallizing melt is to use the water content in nominally anhydrous minerals (NAMs) such as clinopyroxene and olivine. During or after eruption on the surface of a planetary body and during residence in a degassing magma, these minerals may dehydrate. By reversing this process experimentally, original (pre-dehydration) water concentrations can be quantified. In this study, hydrothermal rehydration experiments were performed at 2kbar and 700°C on potentially dehydrated Nakhla clinopyroxene crystals. Rehydrated clinopyroxene crystals exhibit water contents of 130±26 (2σ) ppm and are thus similar to values observed in similar phenocrysts from terrestrial basalts. Utilizing clinopyroxene/melt partition coefficients, both the water content of the Nakhla parent melt and mantle source region were estimated. Despite previous assumptions of a relatively dry melt, the basaltic magma crystallizing Nakhla may have had up to 1.42±0.28 (2σ) wt.% H2O. Based on an assumed low degree of partial melting, this estimate can be used to calculate a minimum estimate of the water content for Nakhla’s mantle source region of 72±16ppm. Combining this value with values determined for other SNC mantle sources, by alternative methods, gives an average mantle value of 102±9 (2σ) ppm H2O for the Martian upper mantle throughout geologic time. This value is lower than the bulk water content of Earth’s upper mantle (∼250ppm H2O) but similar to Earth’s MORB source (54–330ppm, average ∼130ppm H2O).

Osmolyte regulation by TonEBP/NFAT5 during anoxia-recovery and dehydration-rehydration stresses in the freeze-tolerant wood frog (Rana sylvatica).

BackgroundThe wood frog, Rana sylvatica, tolerates freezing as a means of winter survival. Freezing is considered to be an ischemic/anoxic event in which oxygen delivery is significantly impaired. In addition, cellular dehydration occurs during freezing because water is lost to extracellular compartments in order to promote freezing. In order to prevent severe cell shrinkage and cell death, it is important for the wood frog to have adaptive mechanisms for osmoregulation. One important mechanism of cellular osmoregulation occurs through the cellular uptake/production of organic osmolytes like sorbitol, betaine, and myo-inositol. Betaine and myo-inositol are transported by the proteins BGT-1 and SMIT, respectively. Sorbitol on the other hand, is synthesized inside the cell by the enzyme aldose reductase. These three proteins are regulated at the transcriptional level by the transcription factor, NFAT5/TonEBP. Therefore, the objective of this study was to elucidate the role of NFAT5/TonEBP in regulating BGT-1, SMIT, and aldose reductase, during dehydration and anoxia in the wood frog muscle, liver, and kidney tissues.MethodsWood frogs were subjected to 24 h anoxia-4 h recovery and 40% dehydration-full rehydration experiments. Protein levels of NFAT5, BGT-1, SMIT, and aldose reductase were studied using immunoblotting in muscle, liver, and kidney tissues.ResultsImmunoblotting results demonstrated downregulations in NFAT5 protein levels in both liver and kidney tissues during anoxia (decreases by 41% and 44% relative to control for liver and kidney, respectively). Aldose reductase protein levels also decreased in both muscle and kidney tissues during anoxia (by 37% and 30% for muscle and kidney, respectively). On the other hand, BGT-1 levels increased during anoxia in muscle (0.9-fold compared to control) and kidney (1.1-fold). Under 40% dehydration, NFAT5 levels decreased in liver by 53%. Aldose reductase levels also decreased by 42% in dehydrated muscle, and by 35% in dehydrated liver. In contrast, BGT-1 levels increased by 1.4-fold in dehydrated liver. SMIT levels also increased in both dehydrated muscle and liver (both by 0.8-fold).DiscussionOverall, we observed that osmoregulation through an NFAT5-mediated pathway is both tissue- and stress-specific. In both anoxia and dehydration, there appears to be a general reduction in NFAT5 levels resulting in decreased aldose reductase levels, however BGT-1 and SMIT levels still increase in certain tissues. Therefore, the regulation of osmoregulatory genes during dehydration and anoxia occurs beyond the transcriptional level, and it possibly involves RNA processing as well. These novel findings on the osmoregulatory mechanisms utilized by the wood frog advances our knowledge of osmoregulation during anoxia and dehydration. In addition, these findings highlight the importance of using this model to study molecular adaptations during stress.

Investigation of rehydration kinetics of open-sun dried okra samples

In this study rehydration kinetics of open-sun dried okra, which dried naturally and two pre-treatment, was investigated at 25 and 50 °C. By the obtained data, parameters with respect to rehydration kinetics and diffusion mechanism were calculated. In dehydration experiments, it was determined that blanching pre-treatment has influence on the drying time and okra samples were dried at 18 h. On the contrary in rehydration experiments maximum equilibrium rehydration values were achieved with the okras dried naturally. Experimental equilibrium rehydration (Req), theoretical equilibrium rehydration (Rmax) and diffusion coefficient (D) of okra dried naturally at 50 °C were calculated as 5.57 (g water/g dry matter), 5.96 (g water/g dry matter) and 2.17 × 10−10 (m2/s), respectively. Rehydration exponent (n) value, which is also important to identify the diffusion type of dried okra, was determined as between 0.332 and 0.383. Because of the exponent value n < 0.50, diffusion was defined as natural Fick type.