Ethylene biosynthesis in a chilling-sensitive<Emphasis Type="Italic">Arabidopsis</Emphasis> mutant,<Emphasis Type="Italic">chs4-2</Emphasis>

We investigated chilling-induced changes in ethylene levels in Arabidopsis to find plants with distinct patterns of ethylene production in the cold-related biosynthetic pathway. The sensitive mutants identified here includedchs1-2,chs4-2, andchs6-2. Among these, plants of thechs4-2 mutant produced more ethylene than did the wild type after both were transferred from 4°C or 10°C to 22°C. This mutant also showed less freezing tolerance and more electrolyte leakage than the wild-type plants. Our results suggest a relationship between ethylene biosynthesis and chilling sensitivity in the mutant To determine which of the enzymes involved in ethylene biosynthesis were induced by chilling, we tested the activities of ACC synthase and ACC oxidase in both mutant and wild-type plants, and found greater activity by ACC synthase as well as a higher ACC content in the mutants after all the plants were transferred from 10°C to 22°C. However, ACC oxidase activity did not differ between mutant and wild-type plants in response to chilling treatment Therefore, we conclude thatchs4-2 mutants produce more ethylene than do other mutants or the wild type during their recovery from chilling conditions. Furthermore, we believe that ACC synthase is the key enzyme involved in this response.     

The effect of seed conditioning,short-term heat shock and salicylic,jasmonic acid or brasinolide on sunflower (Helianthus annuus L.) chilling resistance and polysome formation

The aim of this study was to develop the method for increasing resistance of sunflower seedlings ‘Wielkopolski’ to chilling. Seeds were conditioned at 25 °C for 2 days in water to 15, 20 and 25 % moisture content or in salicylic or jasmonic acid in concentration of 10^?2; 10^?3 and 10^?4 M or brassinolide in concentration of 10^?6; 10^?8 and 10^?10–15 % moisture content. After 2 days of incubation the conditioned seeds were heat shocked at 45 °C for 0, 30, 60, 120 and 240 min and 5 mm seedlings were exposed to chilling at 0 °C for 21 days. The effectiveness of the methods was assessed by evaluation of roots growth in Phytotoxkit Microbiotest, changes in the activity of dehydrogenases, the integrity of the cytoplasmic membrane and formation of polysomes after seedling were returned to 25 °C for 72 h. Seeds were conditioned at 25 °C for 2 days in water to 15 % moisture content and then heat shocked at 45 °C for 2 h decreased chilling injury of seedlings expressed by subsequent growth of the roots, electrolyte leakage, dehydrogenases activity and polysomes formation. Application of heat shock of 45 °C for 2 h during seed conditioning additionally provided seedling protection against subsequent chilling conditions. Brasinolide, salicylic acid or jasmonic acid applied during seeds conditioning exhibited further beneficial effect on seedling resistance to chilling. The most pronounced effect was obtained due to seed conditioning to 15 % moisture content in solutions of brassinolide in concentration of 10^?8 M. After 2 days of imbibition treated in this way seeds were exposed to heat shock at 45 °C for 2 h. The role of physiological events in improvement of sunflower chilling tolerance are discussed.     

Prior temperature exposure affects subsequent chilling sensitivity

The chilling sensitivity of small discs or segments of tissue excised from chillingsensitive species was significantly altered by prior temperature exposure subsequent to holding the tissue at chilling temperatures as measured by a number of physiological processes sensitive to chilling. This temperature conditioning was reversible by an additional temperature exposure before chilling, and mature-green and red-ripe tomato tissue exhibit similar chilling sensitivities. Exposing pericarp discs excised from tomato fruit (Lycopersicon esculentum Mill. cv. Castelmart), a chilling-sensitive species, to temperatures from 0 to 37°C for 6 h before chilling the discs at 2.5°C for 4 days significantly altered the rate of ion leakage from the discs, but had no effect on the rate of ion leakage before chilling and only a minimal effect on discs held at a non-chilling temperature of 12°C. Exposing chillingsensitive tissue to temperatures below that required to induce heat-shock proteins but above 20°C significantly increased chilling sensitivity as compared to tissue exposed to temperatures between 10 and 20°C. Rates of ion leakage after 4 days of chilling at 2.5°C were higher from fruit and vegetative tissue of chilling-sensitive species (Cucumis sativus L. cv. Poinsett 76, and Cucurbita pepo L. cv. Young Beauty) that were previously exposed for 6 h to 32°C than from similar tissue exposed to 12°C. Exposure to 32 and 12°C had no effect on the rate of ion leakage from fruit tissue of chilling tolerant species (Malus domestica Borkh. cv. Golden Delicious, Pyrus communis L. cv. Bartlett). Ethylene and CO₂ production were higher and lycopene synthesis was lower in chilled tomato pericarp discs that were previously exposed for 6 h to 32°C than the values from tissue exposed to 12°C for 6 h before chilling. Increased chilling sensitivity induced by a 6 h exposure to 32°C could be reversed by subsequent exposure to 12°C for 6 h.     

Activity of enzymatic antioxidant defense systems in chilled and heat shocked cucumber seedling radicles

Chilling whole cucumber seedlings that had 10‐mm long radicles for 4 days at 2.5°C significantly inhibited subsequent radicle growth both by increasing the time it took the seedlings to recover from chilling and attain a linear rate of radicle growth, and by decreasing the subsequent rate of linear growth. Exposing cucumber seedlings to 45°C for up to 20 min had no effect on subsequent radicle growth, while longer exposures produced reductions in growth. A heat shock at 45°C for 10 min induced the optimal protection to 4 days of chilling at 2.5°C by reducing chilling inhibition from 60 to 42%. Two hours after being chilled, heat shocked or heat shocked and then chilled, there was no difference in protein content of the apical 1 cm of the seedling radicle among these treatments and the non‐heat shocked, non‐chilled control. Two days after treatment, the protein content was still similar in tissue that had been heat shocked or heat shocked and chilled, while it was significantly reduced in tissue that had been chilled. In general, 2 h after treatment, the activity of the 5 antioxidant enzymes examined in this study [superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11), guaiacol peroxidase (GPX; EC 1.11.1.7) and glutathione reductase (GR; EC 1.6.4.2)] were reduced by chilling and unaffected or increased by heat shock. When heat shock was followed by chilling, there was a consistent effect of the heat shock treatment on preventing the loss of enzyme activity following chilling. This protective effect of the heat shock treatment was even more pronounced after 2 days of recovery at 25°C for SOD, CAT and APX. In contrast, the activity of GR and GPX was substantially higher in chilled tissue than in tissue that had been heat shocked before being chilled. Elevated levels of GR and GPX therefore appear to be correlated with the development of chilling injury, while elevated levels of SOD, CAT and APX appear to be correlated with the development of heat shock‐induced chilling tolerance.     

Application of phytohormones during seed hydropriming and heat shock treatment on sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis> L.) chilling resistance and changes in soluble carbohydrates

The objective of this study was to analyze the mechanism of some physiological processes accompanying acquisition of sunflower (Helianthus annuus L.) chilling resistance due to seeds hydropriming in the presence of salicylic acid, jasmonic acid, 24-epibrassinolide followed exposition of seeds to short-term heat shock treatment. The seeds were hydroprimed at 25 °C in limited amounts of water or solution of salicylic or jasmonic acid at 10^?2, 10^?3 and 10^?4 M concentration, 24-epibrassinolide at 10^?6, 10^?8 and 10^?10 M concentration. The seeds were incubated for 2 days, subjected to short-term heat shock (45 °C, 2 h) and chilled for 21 days at 0 °C. Sunflower chilling susceptibility and physiological responses were evaluated according to the inhibition of radicle growth, the inhibition of the number of lateral roots formation, the activity of catalase and changes in soluble carbohydrates in seedlings developing for 72 h at 25 °C. Hydropriming and short-term heat shock application explicitly reduced inhibition of roots as well as lateral roots development by allowing the germinating seeds to recover from the growth-inhibiting effects of chilling. Seeds hydropriming in solutions containing salicylic acid, jasmonic acid and 24-epibrassinolide followed heat shock treatment additionally promoted the activity of catalase and sugars metabolism, which stimulated seedlings development and alleviated the decrease of F _v/F _m caused by chilling conditions. These beneficial effects contributed to increased resistance of sunflower seedlings to chilling stress. The present study demonstrated that the most profitable effect on reducing negative effect of chilling may be achieved by short-term heat shock applied during hydropriming in water supplemented with 24-epiBL (10^?8 and 10^?10 M) or salicylic acid (10^?3 and 10^?4 M).     

Effect of heat shock on the chilling sensitivity of trichomes and petioles of African violet (Saintpaulia ionantha)

Chilling at 6°C caused an immediate cessation of protoplasmic streaming in trichomes from African violets ( Saintpaulia ionantha ), and a slower aggregation of chloroplasts in the cells. Streaming slowly recovered upon warming to 20°C, reaching fairly stable rates after 4, 15, 25 and 35 min for tissue chilled for 2 min and for 2, 14 and 24 h, respectively. The rate of ion leakage from excised petioles into an isotonic 0.2  M mannitol solution increased after 12 h of chilling and reached a maximum after 3 days of chilling. A heat shock at 45°C for 6 min reduced chilling-induced rates of ion leakage from excised 1-cm petiole segments by over 50%, namely to levels near that from non-chilled control tissue. Heat-shock treatments themselves had no effect on the rate of ion leakage from non-chilled petiole segments. Protoplasmic streaming was stopped by 1 min of heat shock at 45°C, but slowly recovered to normal levels after about 30 min Chloroplasts aggregation was prevented by a 1 or 2 min 45°C heat-shock treatment administered 1.5 h before chilling, but heat-shock treatments up to 6 min only slightly delayed the reduction in protoplasmic streaming caused by chilling. Tradescantia virginiana did not exhibit symptoms associated with chilling injury in sensitive species (i.e. cessation of protoplasmic streaming in stamen hairs and increased ion leakage from leaf tissue).     

Effect of temperature conditioning on chilling injury of cucumber cotyledons: possible role of abscisic Acid and heat shock proteins

Endogenous abscisic acid levels and induced heat shock proteins were measured in tissue exposed for 6 hours to temperatures that reduced their subsequent chilling sensitivity. One-centimeter discs excised from fully expanded cotyledons of 11-day-old seedlings of cucumber (Cucumis sativus L., cv Poinsett 76) were exposed to 12.5 or 37°C for 6 hours followed by 4 days at 2.5 or 12.5°C. Ion leakage, a qualitative indicator of chilling injury, increased after 2 to 3 day exposure to 2.5°C, but not to 12.5°C, a nonchilling temperature. Exposure to 37°C before chilling significantly reduced the rate of ion leakage by about 60% compared to tissue exposed to 12.5°C before chilling, but slightly increased leakage compared to tissue exposed to 12.5 or 37°C and held at the nonchilling temperature of 12.5°C. There was no relationship between abscisic acid content following exposure to 12.5 or 37°C and chilling tolerance. Five heat shock proteins, with apparent molecular mass of 25, 38, 50, 70, and 80 kilodaltons, were induced by exposure to 37 or 42°C for 6 hours, and their appearance coincided with increased chilling resistance. Heat shock treatments reduced the synthesis of three proteins with apparent molecular mass of 14, 17, and 43 kilodaltons. Induction of heat shock proteins could be a possible cause of reduced chilling injury in tissue exposed to 37 or 42°C.     

Heat shock proteins and chilling sensitivity of mung bean hypocotyls

Excised mung bean (Vigna radiata L.) hypocotyl sections wereexposed to 40 C for up to 4 h in the presence or absence of50 µM cycloheximide (CHX) before being held at a non-chilling(20 C) or chilling (2.5 C) temperature. Mung bean hypocotyltissue is chilling sensitive, and the rate of solute leakageis highly correlated with the extent of chilling injury. A 3h heat shock at 40 C reduced chilling-induced solute leakageby up to 40%, but leakage was similar to non-heat-shocked hypocotylswhen CHX was present. Specific proteins were labelled when hypocotylswere exposed to [³⁵S] methionine during the last hour of heatshock. The nine most intense bands on the autoradiographs ofSDS-PAGE gels of extracted protein corresponded to molecularweights of 114, 79, 73, 70, 60, 56, 51, 46, and 18 kDa. The18 kDa band reached a maximum after 1 h at 40 C and then rapidlydecreased in intensity as the heat shock continued, becomingundetectable at 4 h. The four most intense bands after 3 h at40 C corresponded to molecular weights of 79, 70, 51, and 46kDa. The synthesis of these four hsps was markedly reduced whenthe hypocotyl sections were exposed to CHX during heat shock.During chilling for 6 d, the levels of hsps 79 and 70 remainedsignificantly higher in tissue that was heat shocked prior tochilling than in tissue that was not heat shocked. In contrast,the levels of hsps 51 and 46 were similar in bothheat-shockedand control tissues. Heat-shock-induced chilling tolerance waslost between 6 and 9 d ofstorage at 2.5 C; this loss coincidedwith the decay of hsps 79 and 70 to control levels. These resultssuggest that heat shock induces an increase in both chillingtolerance and the de novo synthesis of specific heat shock proteins;namely hsps 79 and 70. This is the first report showing a relationshipbetween heat-shock-induced chilling tolerance and specific heat-shock-inducedproteins. Key words: Ion leakage, protein synthesis, Vigna radiata     

Arabidopsis: Sensitivity of growth to high temperature

Arabidopsis thaliana seedlings as measured by an electrolyte leakage assay, have been found to be extremely sensitive to high temperature stress as compared to a high temperature tolerant variety (Tracy) of soybean. Over 50% ion leakage occurred in Arabidopsis leaves during a 15-minute exposure to 50°C, indicating a heat killing time of less than 15 minutes. In contrast, the heat killing time for soybean at 50°C was over five times longer. When soybean or Arabidopsis seedlings in culture plates were exposed to 37°C for 2 hours and then returned to 23°C, they suffered no apparent short-term or long-term damage. Soybean seedlings given a 42°C, treatment for 2 hours also showed no damage. Arabidopsis seedlings after a 42°C treatment for 2 hours showed no apparent immediate damage, but 48 hours after return to 23°C severe damage symptoms were visible and after 96 hours all the seedlings were dead. Both soybean and Arabidopsis seedlings synthesize heat shock proteins (hsps) when exposed to 42°C for 2 hours. The hsps synthesized are of similar molecular weights, although the relative abundances of the different size classes are very different in the two plants. Even though hsps are produced in Arabidopsis seedlings after a 2 hour exposure to 42°C their presence is not sufficient for the seedlings to recover from the effects of rhe heat shock when returned to 23°C. Our results show that Arabidopsis has a heat sensitive genotype. This along with its other characteristics should make it a good model system in which to assay in transgenic plants, the functions of homologous and heterologous genes that might be candidates for determining heat tolerance in plants.     

Exposure to alcohol vapours reduces chilling-induced injury of excised cucumber cotyledons, but not of seedlings or excised hypocotyl segments

Cucumber (Cucumis sativus L. cv. Poinsett 76) seedlings withfully expanded cotyledons, and excised cotyledons, first trueleaves, hypocotyl segments and fruit mesocarp discs were exposedto vapours from a series of aqueous alcohol solutions of 0 to320 mM methanol, ethanol, n-propanol, n-butanol, and n-pentanolduring chilling at 2.5C for 5 d. Certain concentrations ofeach alcohol reduced subsequent chilling-induced ion leakagefrom the cotyledons and leaves. Exposure of cotyledons to certainmethanol or ethanol solutions also reduced chilling-inducedethylene production, but not carbon dioxide production. In contrast,exposing cucumber seedlings with fully expanded cotyledons tothe same series of alcohol concentrations that resulted in reducedchilling-induced ion leakage and ethylene production of excisedcotyledons actually increased chilling injury of the seedlings.The hypocotyl region directly below the cotyledons was the siteof chilling-induced injury and contained the most chilling-sensitivehypocotyl tissue. Exposing hypocotyl segments excised from thissensitive region to alcohol solutions did not significantlyreduce chilling-inducedion leakage. Exposing excised cucumbercotyledons or hypocotyl segments to equivalent osmotic nonvolatilesolutions of mannitol and glycerol at 2.5C or to alcohol solutionsat 12.5C had no significant effect on the rate of ion leakage.For the series of alcohols used, the relationship between thelog of the alcohol concentration that minimized chilling-inducedion leakage from cucumber cotyledon discs held at 2.5C for5 d and the log of the partition coefficient of the alcoholinto olive oil or the log of the molecular weight of the alcoholswas highly significant. The same concentrations of alcoholsthat reduced chilling-inducedion leakage also reduced stomatalaperture as measured as decreased porosity of excised cotyledons.The correlation between reduced chilling injury and stomatalconductance of cotyledons exposed to a series of ethanol solutionswas highly significant. It appears that alcohols may reducechilling injury of cucumber cotyledons by inducing stomata closure.Sufficient endogenously synthesized ethanol accumulated in discsheld in N2 at 10C for 1 d to confer tolerance to chilling at2.5C for 5 d. Key words: Anaerobic, Cucumis sativus, ethanol, ion leakage, stomatal conductance     

Combined action of antioxidant defense system and osmolytes in chilling shock-induced chilling tolerance in Jatropha curcas seedlings

Low non-freezing temperature is one of the major environmental factors affecting growth, development and geographical distribution of chilling-sensitive plants, Jatropha curcas is considered as a sustainable energy plants with great potential for biodiesel production. In this study, chilling shock at 5 °C followed by recovery at 26 °C for 4 h significantly improved survival percentage of J. curcas seedlings under chilling stress at 1 °C. In addition, chilling shock could obviously enhance the activities of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and glutathione reductase (GR), and the levels of antioxidants ascorbic acid (AsA) and glutathione (GSH), as well as the contents of osmolytes proline and betaine in leaves of seedlings of J. curcas compared with the control without chilling shock. During the process of recovery, GR activity, AsA, GSH, proline and betaine contents sequentially increased, whereas SOD, APX and CAT activities gradually decreased, but they markedly maintained higher activities than those of control. Under chilling stress, activities of SOD, APX, CAT, GR and GPX, and contents of AsA, GSH, proline and betaine, as well as the ratio of the reduced antioxidants to total antioxidants [AsA/(AsA + DHA) and GSH/(GSH + GSSG)] in the shocked and non-shock seedlings all dropped, but shocked seedlings sustained significantly higher antioxidant enzyme activity, antioxidant and osmolyte contents, as well as ratio of reduced antioxidants to total antioxidants from beginning to end compared with control. These results indicated that the chilling shock followed by recovery could improve chilling tolerance of seedlings in J. curcas, and antioxidant enzymes and osmolytes play important role in the acquisition of chilling tolerance.     

Expression of heat shock proteins in response to high and low temperature extremes in diapausing pharate larvae of the gypsy moth,Lymantria dispar

Diapausing pharate first instars of the gypsy moth, Lymantria dispar, respond to high temperature (37–41°C) by suppressing normal protein synthesis and synthesizing a set of seven heat shock proteins with M_rs of 90,000, 75,000, 73,000, 60,000, 42,000, 29,000, and 22,000 as determined by SDS-PAGE. During recovery at 25°C from heat shock, synthesis of the heat shock proteins gradually decreases over a period of 6 h, while normal protein synthesis is restored. A subset of these same heat shock proteins is also expressed during recovery at 4°C or 25°C from brief exposures to low temperature (-10 to 20°C), and its expression is more intense with increased severity of cold exposure. During recovery at 4°C after 24 h at ?20°C, both 90,000 and 75,000 M_r heat shock proteins are expressed for more than 96 h. While normal protein synthesis is suppressed during heat shock and recovery from heat shock, normal protein synthesis coincides with synthesis of the heat shock proteins during recovery from low temperatures, thus implying that expression of the heat shock proteins is not invariably linked to suppression of normal protein synthesis. Western transfer, using a monoclonal antibody that recognizes the inducible form of the human 70,000 M_r heat shock protein, demonstrates that immunologically related proteins in the gypsy moth are expressed at 4°C and during recovery from cold and heat shock.     

热激对水稻幼苗耐冷性及热激蛋白合成的诱导

萌发的水稻种子经42℃热激处理后其幼苗的耐冷性明显增强,膜伤害程度降低,脯氨酸含量增加,超氧物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)活性和抗氧化物质抗坏血酸含量增加,而膜脂过氧化的关键酶脂氧合酶(LOX)活性及其产物丙二醛(MDA)含量下降.并且热激诱导萌发的水稻胚合成78、70、64、60、46、38、24、17、16kD的热激蛋白(HSP),其中属于HSP70的内质网结合蛋白(BiP)的合成与水稻幼苗耐寒性的提高有关.     

Involvement of antioxidant defense system in chill hardening-induced chilling tolerance in Jatropha curcas seedlings

Jatropha curcas L. is a sustainable energy plant with great potential for biodiesel production, and low temperature is an important limiting factor for its distribution and production. In this present work, chill hardening-induced chilling tolerance and involvement of antioxidant defense system were investigated in J. curcas seedlings. The results showed that chill hardening at 10 or 12 °C for 1 and 2 days greatly lowered death rate and alleviated electrolyte leakage as well as accumulation of the lipid peroxidation product malondialdehyde (MDA) of J. curcas seedlings under severe chilling stress at 1 °C for 1–7 days, indicating that the chill hardening significantly improved chilling tolerance of J. curcas seedlings. Measurement of activities of the antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), and glutathione reductase (GR), and the levels of the antioxidants ascorbic acid (AsA) and glutathione (GSH) showed the chill hardening at 12 °C for 2 days could obviously increase the activities of these antioxidant enzymes and AsA and GSH contents in the hardened seedlings. When the hardened and non-hardening (control) seedlings were subjected to severe chilling stress at 1 °C for 1–7 days, the chill-hardened seedlings generally maintained significantly higher activities of the antioxidant enzymes SOD, APX, CAT, POD, and GR, and content of the antioxidants AsA and GSH as well as ratio of the reduced antioxidants to total antioxidants [AsA/(AsA + DHA) and GSH/(GSH + GSSG)], when compared with the control without chill hardening. All above-mentioned results indicated that the chill hardening could enhance the chilling tolerance, and the antioxidant defense system plays an important role in the chill hardening-induced chilling tolerance in J. curcas seedlings.     

Heat shock tolerance and antioxidant activity in moth bean seedlings treated with tetcyclacis

Moth bean (Vigna aconitifolia (Jacq.) Marechal cv. Jaadia) seeds were germinated in the presence of 0, 18, or 36 M solutions of the gibberellin biosynthesis inhibitor, tetcyclacis. After 72 h, seedlings were exposed to 22 or 48°C for 90 min. The 48°C temperature dramatically increased total electrolyte and sugar leakage from the seedlings, particularly in the controls. Tetcyclacis reduced electrolyte and sugar leakage at 48°C by 15–35% compared to the 48°C controls. High temperature increased malondialdehyde concentration in control seedlings but not in treated seedlings indicating that tetcyclacis inhibited high temperature-induced lipid peroxidation. Relative to the control, tetcyclacis tended to increase the total activities of catalase and peroxidase in the seedlings. In contrast, tetcyclacis tended to decrease ascorbic acid oxidase activity, particularly at 48°C. These results suggest that tetcyclacis conferred at least some heat shock tolerance to moth bean seedlings. This increased tolerance was correlated with increased activities of some antioxidant systems.     

Chilling-induced changes of vacuolar proton pumps in hypocotyls of <Emphasis Type="Italic">Vigna unguiculata</Emphasis>

Vigna unguiculata (cowpea) is a legume adapted to high temperatures and is sensitive to low temperatures. Temperature is one of the limiting factors of growth and yield for many crops but its effect on cowpea metabolism is not known. We investigated the effect of chilling on activity of vacuolar proton pumps (V-ATPase and V-PPase) and their protein content in tonoplast vesicles of cowpea hypocotyls. Seedlings grown for 7 days at 10 or 4°C were used for experiments. Chilling treatment at 10 or 4°C markedly suppressed growth of cowpea seedlings. Following chilling at 10 and 4°C, activity of both proton pumps and the relative amount of V-PPase and subunit A of V-ATPase were significantly increased. Both substrate hydrolysis and H⁺ transport activities of V-PPase remained at relatively high levels during chilling treatment. For V-ATPase, treatment at 10°C for 6 days increased the ATP hydrolysis activity. However, the H⁺ transport activity of the enzyme was increased when treated for 4 days but was markedly decreased when treated for 6 days. Our results provide evidence for different regulation for these vacuolar proton pumps, indicating that V-PPase is the more stable proton pump throughout chilling stress.     

Effects of seed hydropriming in presence of exogenous proline on chilling injury limitation in <Emphasis Type="Italic">Vigna radiata</Emphasis> L. seedlings

Three-day-old seedlings (t ₀ stage) of Vigna radiata (L.) Wilczek obtained from seeds hydroprimed (H) and hydroprimed with proline (HPro) were examined. H and HPro slightly improved mung bean seed germination and seedlings growth at 5°C. The best growth was observed in the seedlings obtain from HPro5 (5 mM) seeds in comparison with the seedlings obtained from the control-non-primed seeds and H seeds. Exposure of mung bean seedlings grown from non-primed seeds to chilling for 4 days induced chilling injury: membrane lipid peroxidation, decrease in endogenous proline level and inhibition of growth of roots and hypocotyls. The seedlings obtain from HPro seeds grew better during the time of chilling and after rewarming at 25°C. The possible role of HPro in chilling injury limitation is discussed.     

Heat shock and the protection against metal toxicity in wheat leaves1

Abstract. Wheat (Triticum aestivum L. var. TAM-W101) leaf segments exhibited an acquired protection against metal toxicity following exposure of the seedlings to heat shock temperatures in the dark. The acquired protection of the leaf segments to cadmium was 400-fold greater than leaf segments from seedlings kept at 25°C and exhibited the greatest change in protection of the five metals tested. Increased protection against aluminium and iron toxicity was also detected in the leaf segments from heat shocked seedlings. The concentration of aluminium at which a 50% loss of 2,3,5-triphenyltetrazolium chloride reduction occurred was 5.5 mol m^?3 in control leaf segments and 20 mol m^?3 in the leaf segments from heat shocked seedlings. The 50% loss of 2,3,5-triphenyltetrazolium chloride reduction in the leaf segments from heal shocked seedlings was four-fold higher for iron. A small, yet reproducible change in the 2,3,5-triphenyltetrazolium chloride reduction was observed for copper and no change in reduction was observed for zinc treatments in the leaf segments from heat shocked seedlings. These data indicate that exposure of wheat seedlings to heat shock temperatures results in the acquisition of protection against metal toxicity to otherwise lethal concentrations of aluminium, cadmium, and iron.     

Lipid response to short-term chilling shock and long-term chill hardening in Jatropha curcas L. seedlings

Low non-freezing temperature is one of the major environmental factors that affect metabolism, growth, development and geographical distribution of chilling-sensitive plants, Jatropha curcas, a chilling-sensitive plant, which is considered as a sustainable energy plant with great potential for biodiesel production. Our previous studies showed that short-term chilling shock at 5 °C for 4 h and long-term chill hardening at 12 °C 1 or 2 days could improve chilling tolerance of J. curcas seedlings, but lipidomic response to chilling shock and chill hardening has not been elucidated. In this study, membrane lipid composition change in J. curcas seedlings during chilling shock and chill hardening was investigated by liquid chromatography-electrospray ionization-mass spectrometry (LC–ESI–MS) approach. The results indicated that the relative abundances of nine classes and 72 species of membrane lipids, such as phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylinositol (PI), lysophosphatidylcholine (lysoPC) and lysophosphatidylglycerol (lysoPG), two glycolipids digalactosyldiacylglycerol (DGDG) and monogalactosyldiacylglycerol (MGDG) and a sulfoquinovosyldiacylglycerol (SQDG), were significantly changed, and the degree of unsaturation of above-mentioned cellular membrane lipids with fatty acid differing in chain lengths and the number of double bonds also increased in varying degrees during chilling shock and chill hardening. These results suggested that remodeling and increase in the degree of unsaturation of membranes lipids may be a common physiological basis for short-term chilling shock- and long-term chill hardening-induced chilling tolerance of J. curcas seedlings.