Low-Temperature Adaptation of Winter Wheat Seedlings under Excessive Zinc Content in the Root Medium

Russian Journal of Plant Physiology - Cold adaptation of winter wheat (Triticum aestivum L.) seedlings was investigated under excessive zinc content (1000 μM) in the root medium. The ability...     

Storage of Porcine Articular Cartilage at High Subzero Temperatures

Objective: Transplantation of osteochondral allograft tissue can treat large joint defects but is limited by tissue availability, surgical timing, and infectious disease transmission. Fresh allografts perform the best but requirements for infectious disease testing delay the procedure with subsequent decrease in cell viability and function. Hypothermic storage at lower temperatures can extend tissue banking time without loss of cell viability and, therefore, increase the supply of allograft tissue. This study investigated the effects of different cryoprotectant solutions on intact AC at various subzero temperatures. Design: 10 mm porcine osteochondral dowels were immersed for 30 minutes in various combinations of solutions [(XVIVO, propylene glycol (51% w/w), sucrose (46% w/w)] cooled to various subzero temperatures (−10, −15, and −20 °C), and held for 30 min. After warming, 70 μm slices were stained with membrane integrity dyes, viewed under fluorescence microscopy and cell recovery calculated relative to fresh controls. Results: Results demonstrated excellent cell recovery (>75%) at −10°C provided ice did not form. Excellent cell recovery (>70%) occurred at −15°C in solutions containing 51% propylene glycol but formation of extra-matrix ice in other solutions resulted in significant cell loss. All groups had <6% cell recovery at −20°C and propylene glycol did not provide a protective effect even though extra-matrix ice did not form Conclusions: These results suggest that extra-matrix ice plays an important role in cell damage during cryopreservation. Excellent cell recovery can be obtained after storage at subzero temperatures if ice does not form. Hypothermic preservation at high subzero temperatures may extend AC storage time in tissue banks compared to current techniques.     

Toxicity of Anaerobic Metabolites Accumulating in Winter Wheat Seedlings during Ice Encasement

Ice encasement damages cold-hardened winter wheat without major disruption of cellular organelles. CO₂ accumulates during total ice encasement to higher levels in Kharkov than in less hardy Fredrick wheat. Partial ice encasement and exposure to a nitrogen atmosphere at -1 C allows greater CO₂ accumulation but neither treatment is as damaging as total ice encasement. Lactic acid accumulates to low levels only during the 1st day of encasement and thereafter remains constant. Exposure of plants to a combination of 50% CO₂ and 5% ethanol reduces survival, with a cultivar difference similar to that found in ice-encased plants. Plants in CO₂ and ethanol show a proliferation of membranes and nuclear condensation similar to that in cells of ice-encased plants. Permeability increases markedly in the presence of CO₂ and ethanol, to levels similar to or greater than those of iced plants. Ethanol alone does not increase permeability but in combination with CO₂ raises permeability of the less hardy Fredrick, although not of Kharkov, but reduces survival of both cultivars. A comparison of the endogenous levels of ethanol, CO₂, and lactic acid at the 50% kill point of plants due to ice encasement or due to externally supplied metabolite indicates that only CO₂ accumulates to independently toxic levels. Permeability and ultrastructural evidence suggest that CO₂ and ethanol in combination are the agents reducing plant viability during ice encasement.     

Motility of Colwellia psychrerythraea Strain 34H at Subzero Temperatures

We examined the Arctic bacterium Colwellia psychrerythraea strain 34H for motility at temperatures from −1 to −15°C by using transmitted-light microscopy in a temperature-controlled laboratory. The results, showing motility to −10°C, indicate much lower temperatures to be permissive of motility than previously reported (5°C), with implications for microbial activity in frozen environments.     

Differential Involvement of the Five RNA Helicases in Adaptation of Bacillus cereus ATCC 14579 to Low Growth Temperatures

Bacillus cereus ATCC 14579 possesses five RNA helicase-encoding genes overexpressed under cold growth conditions. Out of the five corresponding mutants, only the ΔcshA, ΔcshB, and ΔcshC strains were cold sensitive. Growth of the ΔcshA strain was also reduced at 30°C but not at 37°C. The cold phenotype was restored with the cshA gene for the ΔcshA strain and partially for the ΔcshB strain but not for the ΔcshC strain, suggesting different functions at low temperature.Bacillus cereus is a human pathogenic sporulated bacterium which is associated with emetic and diarrheal types of food-borne illnesses (4). B. cereus is widespread in the environment and in a wide range of foods. The growth domains of B. cereus strains range from psychrotrophic to nearly thermophilic and correlate with several phylogenetic clusters (15), which presumably permit B. cereus to colonize many different habitats with different thermal regimes. Many foods are stored refrigerated before consumption, and in such cases, B. cereus has to adapt to low-temperature conditions.B. cereus growth at low temperature takes place with a lag phase which may correspond to an adaptation phase (12). Cold is a stress which dramatically affects membrane fluidity, protein synthesis, and also the topology of nucleic acids (22). When exposed to low temperature, bacteria have to face a transient inhibition of protein synthesis mainly due to the presence of secondary structures in mRNA that are stabilized by cold conditions (16, 19). To overcome the translation interruption, cold-shocked cells synthesize cold-induced RNA helicases, which remove secondary structures from RNA duplexes in the presence of ATP, such as CsdA of Escherichia coli (19) or CshA of Bacillus subtilis (1). csdA and srmB deletion mutants of E. coli showed a cold-sensitive phenotype, and these RNA helicases have been described as involved in the biogenesis of the ribosomal 50S subunit at 20°C (10, 11). RNA helicases could also be involved in the degradation of mRNA by unwinding double-stranded mRNA, thereby allowing the action of RNase (8).We have recently shown that the deregulation of the expression of one RNA helicase gene of B. cereus ATCC 14579 increased the lag phase of B. cereus at a low temperature (7). In this context, our aim was to investigate the role of the five putative RNA helicases present in the genome of B. cereus ATCC 14579 in its adaptation at low temperature, close to the growth limit.     

Response of Winter Wheat Seedlings Respiration to Long-Term Cold Exposure and Short-Term Daily Temperature Drops

Russian Journal of Plant Physiology - The effects of a long-term (6-day) exposure to low positive temperature (4°С, LT treatment) and short-term (3 h per a day for 6 days) daily...     

Adaptation to Water Stress in Wheat

Three experiments were designed to investigate to what extent adaptation to water stress take place. Wheat (Triticum aestivum L. cv. Kolibri) was grown in water culture at constant temperature, air humidity, and light intensity. When the plants were 16 days old, the potential of the root medium (ψ_r) was lowered by 1 bar every second day by means of polyethyleneglycol 1500 down to ?4 or ?7 bar and then remained at these levels. As a control one experiment was grown at ?0.7 bar. By regression it was found that when ψ_r was lowered by I bar, osmotic potential in leaf (ψ_π) decreased 1.46 bar, and leaf water potential (ψ_t) 0.68 bar, which mean an increase of turgor of 0.78 bar. At the same time the leaf water content did fall 0.30 g per g dry matter. Specific transpiration rate increased significantly after ψ_r was kept constant, but the increase in area of fresh leaves was strongly reduced due to wilting of old leaves. After an “adaptation” period during which ψ_r remained at ?0.7, ?4, and ?7 bar, respectively, for at least 1 week. ψ_r was altered so as to cover the range from 0 to ?14 bar and ψ_π, ψ_r, transpiration and diffusion resistance in stomata (r_s) were measured. The levels of ψ_π and ψ₁ were lower (more negative) and turgor potential higher in plants grown at low ψ_r. The transpiration in pre-stressed plants showed less sensitivity to the alteration of ψ_r than in the non-stressed plants. The values of ψ_r at which r_s increased greatly, were found to be about ?13, ?15, and ?18 bar for plants grown at ?0.7, ?4, and ?7 bar, respectively.     

Evidence for Involvement of at Least Six Proteins in Adaptation of Lactobacillus sakei to Cold Temperatures and Addition of NaCl

Lactobacillus sakei is a lactic acid bacterium widely represented in the natural flora of fresh meat. The aim of this study was to analyze the differences in protein expression during environmental changes encountered during technological processes in which L. sakei is involved in order to gain insight into the ability of this species to grow and survive in such environments. Using two-dimensional electrophoresis, we observed significant variation of a set of 21 proteins in cells grown at 4°C or in the presence of 4% NaCl. Six proteins could be identified by determination of their N-terminal sequences, and the corresponding gene clusters were studied. Two proteins belong to carbon metabolic pathways, and four can be clustered as general stress proteins. A phenotype was observed at low temperature for five of the six mutants constructed for these genes. The survival of four mutants during stationary phase at 4°C was affected, and surprisingly, one mutant showed enhanced survival during stationary phase at low temperatures.     

Maintenance of Membrane Fluidity during Development of Freezing Tolerance of Winter Wheat Seedlings

Fluidity of membrane lipids of shoot and root tissue and of chloroplasts from young wheat seedlings of contrasting freezing tolerance was investigated by measuring the motion and order parameters after spin labeling. A striking similarity was observed in membrane lipid fluidity of the five cultivars grown at 22 C. After cold hardening by growth at 2 C, a small change in membrane lipid fluidity was observed, but this was not correlated with the development of freezing tolerance, and there was no alteration in the transition temperature of membrane lipids. The results show that neither changes in membrane lipid fluidity nor transition temperature are a necessary feature of cold acclimation in wheat.     

Change in Lectin Specificity of Winter Wheat Seedlings in the Course of Infection with Mycoplasms

The activity of soluble lectins in leaves and roots of seedlings of winter wheat (Triticum aestivum L.) cultivar Mironovskaya 808 increased 1 day and 2 days, respectively, after infection with the mycoplasma Acholeplasma laidlawii 118. Analysis of acid-soluble proteins of wheat leaves by PAGE revealed the appearance of 22- and 20-kDa polypeptides, the disappearance of a 14-kDa polypeptide, and an increase in the content of polypeptides with molecular weights of 76, 48, 25, and 18 kDa. The 18-kDa polypeptide is a subunit of wheat germ agglutinin. A change in the activity of lectins may be a nonspecific response of plants to infection with the pathogen.     

Water Status Changes in Shoots of a Seaweed Ascophyllum nodosum at Subzero Temperatures

The gradient freezing and NMR spectroscopy were used to study the physical state of water in apices of the intertidal seaweed Ascophyllum nodosum at freezing temperatures. In the apices exposed to temperatures below –10°C, two fractions of bound water were revealed. The slow (T₂ 50 ms) fraction of bound water was completely frozen at –25°C, and its freezing rate was temperature-sensitive. This fraction was apparently associated with protoplasmic water and cell-wall polysaccharides. The fast fraction (T₂ < 10 ms) of bound water was presumably due to water-soluble globular proteins. The freezing rate for this fraction depended on neither the temperature nor the amount of water. The presence of unfrozen water in apical cells at –40°C was demonstrated. The role of this water fraction in maintaining the native structure of biomacromolecules and apex survival is discussed.     

Adaptive Changes in ATPase Activity in the Cells of Winter Wheat Seedlings during Cold Hardening

A cytochemical study of ATPase activity in the cells of cold hardened and nonhardened winter wheat (Triticum aestivum L. cv. Nongke No. 1) seedlings was carried out by electron microscopic observation of lead phosphate precipitation. ATPase activity associated with various cellular organelles was altered during cold hardening. (a) At 22°C, high plasmalemma ATPase activity was observed in both cold hardened and nonhardened tissues; at 5°C, high activity of plasmalemma ATPase was observed in hardened tissues, but not in unhardened tissues. (b) In nonhardened tissues, tonoplast and vacuoles did not exhibit high ATPase activity at either 22 or 5°C, while in hardened tissues high activity was observed at both temperatures. (c) At 5°C, ATPase activity of nucleoli and chromatin was decreased in hardened tissues, but not in nonhardened tissues. It is suggested that adaptive changes in ATPase activity associated with a particular cellular organelle or membrane may be associated with the development of frost resistance of winter wheat seedlings.     

Effects of Self-sown Wheat on Levels of the Take-all Disease on Seedlings of Winter Wheat Grown in a Model System

Abstract The mechanisms of build-up of inoculum of the take-all fungus, Gaeumannomyces graminis var. tritici. and infection of crop plants from self-sown (volunteer) wheat were analysed in a factorial experiment in a glasshouse. Treatments comprised sowing-date, inoculum density, soil aeration and texture, volunteer density and control of volunteers. Timing of treatments was related to field practice by the use of cumulative day-degrees and by consideration of sowing date and the geometry of seed placement. Prior sowing of volunteers, which were exposed to soil inoculum. resulted in significant increases in the incidence and severity of disease on subsequently sown seedlings. Increasing the density of volunteer seedlings increased the levels of subsequent infection. This effect, however, was significantly influenced by sowing date and density of the initial inoculum. The use of glyphosate to kill volunteers did not markedly affect the carry-over of disease.     

Changes in Hormonal Status and Expression of Lectin Genes in Wheat Seedlings Treated with Stevioside and Exposed to Low Above-Zero Temperatures

Russian Journal of Plant Physiology - Effect of presowing treatment with stevioside (10–8 M) on activity, content, and expression of the wheat-germ agglutinin (WGA) gene, as well as the...     

Ultrastructural and Enzymic Studies of Cell Membranes from Ice-encased and Noniced Winter Wheat Seedlings

A marked increase in the amount of cisternal-like cytoplasmic membranes was observed after ice encasement of winter wheat (Triticum aestivum L.) seedlings. Linear sucrose gradients were employed to separate the various membrane components of the microsomal membrane fraction. NADH- and NADPH-cytochrome c reductase, two specific enzyme markers for plant endoplasmic reticulum (ER) were used to locate the ER in the linear gradients. The identity of the ER fraction was confirmed by determining the effect of EDTA and Mg²⁺ in the preparative media on the distribution of NADH- and NADPH-cytochrome c reductase activity within the gradient. In the presence of EDTA which dissociates ribosomes from ER, peaks of activity for the two enzymes were observed at a density corresponding to that for “smooth” ER. When the media also contained an appropriate concentration of Mg²⁺ to maintain the attachment of ribosomes to the ER, the peaks of activity for the enzymes shifted to a density corresponding to that for “rough” ER. NADH-cytochrome c reductase activity was similar for 24 C-grown and 2 C-grown iced seedlings, but significantly lower for 2 C noniced seedlings. No preferential increase in uptake of radioactive leucine or choline in the ER was observed during ice encasement. The accumulation of electron microscopically visible membrane arrays was not inhibited by the presence of protein synthesis inhibitors at concentrations which severely inhibited incorporation of [1-¹⁴C]leucine into membrane protein, but did not affect survival and growth of the seedlings. These observations indicate that the apparent proliferation of ER during ice encasement does not result from net membrane synthesis, but rather from reorganization of existing membrane elements within the cell.