黄瓜的冷害及耐冷性

随着蔬菜反季节栽培面积的不断扩大,如何提高黄瓜(Cucumis sativus L.)耐冷性已成为选育新品种的研究重点。系统地综述近几年黄瓜耐冷性的鉴定、获得途径、冷害机理以及遗传和分子遗传学等方面的研究,以促进对黄瓜冷害机制的研究, 加速耐冷品种的培育。耐冷性鉴定时要从耐冷指数、低温发芽能力、MDA (丙二醛)含量和电解质渗漏率等几个方面综合鉴定。耐冷性的获得途径主要有冷驯化、激素处理、热激处理和培育耐低温品种,最重要的途径是耐冷品种选育。黄瓜冷害机理包括细胞膜的流动性降低及透性增加,光合作用被抑制,根系吸收减弱,可溶性糖含量减少,淀粉粒积累增加,微管的稳定性受到破坏等。黄瓜低温发芽能力由非加性基因决定,而幼苗时期主要由加性基因控制。黄瓜 耐冷的分子遗传学研究进展缓慢,目前已克隆出在低温锻炼中特异表达的功能未知的基因CCR18。今后还应研究黄瓜低温胁迫时的信号转导系统,以进一步揭示黄瓜的冷害机理;利用野生资源的抗逆性状,拓宽栽培黄瓜的遗传基础,选育适于保护地栽培的耐低温品种。     

Induction of chilling tolerance in cucumber (Cucumis sativus) seedlings by endogenous and applied ethanol

Five-day-old etiolated cucumber ( Cucumis sativus L.) seedlings cv. Marketmore held at 2°C for 72 h developed chilling injury, resulting in desiccation and collapse of the hypocotyl tissue and eventual plant death. Hypoxia-induced accumulation of ethanol and acetaldehyde led to tolerance of subsequent chilling, as evidenced by continued hypocotyl growth and freedom from injury. Attenuated accumulation of volatiles by applied bisulfite reduced the development of hypoxia-induced chilling tolerance in seedlings. In seedlings held in normoxia cold tolerance was induced by applied ethanol vapors, whereas acetaldehyde had a marginal effect, suggesting that hypoxia-induced cold tolerance may arise from the accumulation and activity of ethanol. Cold tolerance was also induced by exposure of seedlings to volatile anesthetics including n -propanol, n -butanol, chloroform and halothane, suggesting that ethanol activity may result from fluidization of membrane lipids. This view is consistent with results which showed that ethanol activity was not associated with lipid metabolism. However, development of cold tolerance in ethanol-enriched tissues was time dependent, indicating that ethanol activity probably also entails biosynthetic event(s).     

黄瓜的冷害及耐冷性

随着蔬菜反季节栽培面积的不断扩大,如何提高黄瓜(Cucumis sativus L.)耐冷性已成为选育新品种的研究重点.系统地综述近几年黄瓜耐冷性的鉴定、获得途径、冷害机理以及遗传和分子遗传学等方面的研究,以促进对黄瓜冷害机制的研究,加速耐冷品种的培育.耐冷性鉴定时要从耐冷指数、低温发芽能力、MDA(丙二醛)含量和电解质渗漏率等几个方面综合鉴定.耐冷性的获得途径主要有冷驯化、激素处理、热激处理和培育耐低温品种,最重要的途径是耐冷品种选育.黄瓜冷害机理包括细胞膜的流动性降低及透性增加,光合作用被抑制,根系吸收减弱,可溶性糖含量减少,淀粉粒积累增加,微管的稳定性受到破坏等.黄瓜低温发芽能力由非加性基因决定,而幼苗时期主要由加性基因控制.黄瓜耐冷的分子遗传学研究进展缓慢,目前已克隆出在低温锻炼中特异表达的功能未知的基因CCRl8.今后还应研究黄瓜低温胁迫时的信号转导系统,以进一步揭示黄瓜的冷害机理;利用野生资源的抗逆性状,拓宽栽培黄瓜的遗传基础,选育适于保护地栽培的耐低温品种.     

Cold-acclimation induced changes in freezing tolerance and translatable RNA content in Citrus grandis and Poncirus trifoliata

Cold-acclimation-induced changes in freezing tolerance and translatable RNA content were compared in seedlings of a relatively cold sensitive citrus species, Citrus grandis L. Osb. cv. Thong Dee (pummelo), and the cold-hardy citrus relative, Poncirus trifoliata L. Raf. cv. Pomeroy (trifoliate orange). Cold acclimation of pummelo (10 days at 15°C followed by 4 weeks at 10°/5°C, day/night) resulted in a decrease in LT₅₀ from −6 to −8°C, while in trifoliate orange (acclimated for 7 weeks at 5°C), the LT₅₀ decreased from −9 to −18°C. Qualitative changes in the in vitro translation profile, revealed by two-dimensional gel electrophoresis, were observed following cold acclimation in both species. An mRNA for a large polypeptide (ca 160 kDa) was detected following cold acclimation of trifoliate orange. A similar change was not observed in pummelo following cold acclimation.     

Relationship of cold acclimation,total phenolic content and antioxidant capacity with chilling tolerance in petunia (Petunia × hybrida)

Extracts from Petunia × hybrida plants, which had been subjected to cold pretreatment to induce chilling tolerance, were analyzed for specific phenolic acids, such as gentisic acid, and assessed for their antioxidant capacity by their ability to reduce (decolorize) the 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt radical (ABTS^*). Gentisic acid was induced in significant quantities by the third week of cold acclimation and levels remained constant up to the fourth week. Cold acclimation induced accumulation of total phenolics, which was positively related to antioxidant capacity. Petunia plants recovered from chilling injury following 3 weeks of cold pretreatment with an increase in total phenolics, which suggested some form of antioxidant protection. However, antioxidant capacity was only moderately related to chilling tolerance, which indicated that factors other than total phenolics may play a role in the chilling tolerance in petunia. These data suggest that the 5 °C cold pretreatment may have initially caused injury that impeded acclimation at the outset, and that subsequent phenolic metabolism was related to protective functions in petunia.     

Cold acclimation of wheat (Triticum aestivum): Properties of enzymes involved in proline metabolism

Two cultivars of wheat ( Triticum aestivum L.), a winter wheat, Kharkov, and a spring wheat, Glenlea, were acclimated under controlled conditions at 2 temperatures, 5°C and 25°C with a 12-h photoperiod. Water content, protein and proline concentrations were determined. Enzymatic properties (activity and apparent energy of activation) were investigated for enzymatic systems involved in 2 pathways of proline metabolism, the glutamic acid and ornithine pathways. Four enzymes were studied, proline dehydrogenase (PDH, EC 1.5.1.2), glutamate dehydrogenase (GDH, EC 1.4.1.2-4), glutamine synthetase (GS, EC 6.3.1.2) and ornithine transaminase (OT, EC 2.6.1.13). Cold acclimation led to an accumulation of proline, a decrease in water content and an increase in soluble protein, especially in winter wheat. For both cultivars, cold acclimation modulated enzyme properties of PDH and GDH. Increased activities of GS and OT were observed as a result of cold acclimation in both cultivars, with the greatest increase in Kharkov. The apparent energy of activation of these 2 enzymes decreased, particularly for Kharkov, which accumulated proline in cold conditions.     

The role of the liver in lipid metabolism during cold acclimation in non-hibernator rodents

Cold exposure increases the demand for energy substrates. Cold acclimation of rats led to a 3-fold increase in fatty acid (FA) beta-oxidation (P<0.01) for ex vivo livers perfused at 37 degrees C. This increase was preserved following perfusion at 25 degrees C (P<0.001). In vitro measurement of absolute rates of hepatic beta-oxidation revealed no significant difference following cold acclimation, implying changes in fatty acid flux through beta-oxidation rather than increased oxidation capacity. Total FA uptake was increased one-third following perfusion at 25 degrees C (P<0.001) and cold acclimation (P<0.05) and cold acclimation led to diversion of tissue FA from storage to beta-oxidation (P<0.01). In separate experiments, in vivo hepatic lipogenesis rates for saponifiable lipids doubled (P<0.01) and cholesterol synthesis increased one-third (P<0.001). Taken together these data suggest the oxidation and synthesis of lipids occur simultaneously in hepatic tissue possibly to increase prevailing tissue FA concentrations and to generate heat through increased metabolic flux rates.     

Effect of chilling and subsequent storage at 20°C on electrolyte leakage and phospholipid fatty acid composition of tomato pericarp

Mature green tomato fruit ( Lycopersicon esculentum cv. Caruso) were stored at 1°C or 20°C and analyzed on day 0, 18 and 22 for electrolyte leakage, ripening-associated changes in pigmentation and phospholipid fatty acid composition. Chilled fruit were also analyzed 4 days after they were returned to 20°C. Fruit did not ripen significantly during chilling and subsequent storage at 20°C, and showed visible chilling injury symptoms only at 20°C. Electrolyte leakage increased in control and chilled fruit, indicating enhanced membrane permeability during both ripening and chilling. Returning the fruit to ambient temperature gave an apparent decrease in electrolyte leakage. Phospholipid and linolenic acid content and double bond index decreased during ripening at 20°C. The small changes in phospholipid fatty acid composition during chilling cannot account for the enhanced membrane permeability. The significant decrease in percentage of linolenic acid and in double bond index in the total lipids, but not in the phospholipids, upon returning the fruit to 20°C suggests loss of galactolipid polyunsaturated fatty acids     

Responses of poplar to chilling temperatures: proteomic and physiological aspects

Abstract: The effects of cold acclimation on primary metabolism in actively growing poplar ( Populus tremula L. × P. tremuloides Michaux) were studied. Three-month-old poplar plants were exposed to chilling stress (4 °C) and compared to plant material kept at a control temperature (23 °C). This treatment did not affect the survival of the plants but growth was almost stopped. The freezing tolerance of the adult leaves increased from - 5.7 °C for the control plants to - 9.8 °C after 14 days of exposure to 4 °C. During acclimation, the evolution of soluble carbohydrate contents was followed in the leaves. Sucrose, glucose, fructose and trehalose accumulated rapidly under chilling conditions, while raffinose content increased after one week at 4 °C. Proteomic analyses, by bidimensional electrophoresis, performed during this stage revealed that a large number of proteins had higher expression, while much less proteins disappeared or had a lower abundance. MALDI-TOF-MS analyses enabled ca. 30 spots to be proposed for candidate proteins. Among the accumulating or appearing proteins proposed, about a third presented similarities with chaperone-like proteins (heat shock proteins, chaperonins). In addition, dehydrins and other late embryogenesis abundant proteins, i.e., stress-responsive proteins, detoxifying enzymes, proteins involved in stress signalling and transduction pathways were also activated or newly synthesised. Finally, cold exposure induced a decrease in the candidate proteins involved in cell wall or energy production.     

Effect of chilling, heat shock, and vigor on the growth of cucumber (Cucumis sativus) radicles

Chilling at 2.5°C reduced the subsequent growth of cucumber ( Cucumis sativus L.) radicles at 25°C. The reduction in radicle growth was linear for 1–3 days of chilling at ≈10% per day of treatment, but then it increased in a non-linear pattern until subsequent radicle growth was all but eliminated by 6 days of chilling. A heat shock of 40°C for 4–12 min increased chilling tolerance such that 4 days of chilling caused only a 36% decrease in radicle growth, compared to 66% for seedlings not heat shocked. Heat shocks were only able to protect that part of radicle growth that was in excess of the linear decrease in radicle growth projected from 0–3 days. There appear to be two effects of chilling on radicle growth. The first inhibition of subsequent growth was linear and was not affected by heat shocks. The second inhibition was much more severe; it appeared after 3 days of chilling and could be prevented by heat shock. Seeds classified with different levels of vigor (i.e., different initial rates of growth) did not respond significantly different to chilling stresses following heat-shock treatments.     

The Effects of Cold Acclimation on Several Enzyme Activities in Cucumber Seedling

The changes in activities of SOD, peroxidase, catalase and ATPase in chilling sensitive cucumber (Cucumis sativus L.) seedlings using biochemical and cytochemical methods were studied. The results indicated that the activities of SOD, peroxidase and catalase enhanced dramatically in cold acclimated cucumber seedlings and the three enzymes remained stable under chilling stress. Consequently, the ability of cleaning up free radical of oxygen and peroxidates increased. The cold-tolerant character of plasmalemma ATPase activity was developed after low-temperature acclimation. All these changes provided the possibility for protecting the stability of membrane structure and metabolism from chilling injury, and for the enhancement of cold tolerance by low temperature acclimation.     

Genotype-specific differences in chilling tolerance of maize in relation to chilling-induced changes in water status and abscisic acid accumulation

Four inbred maize lines differing in chilling tolerance were used to study changes in water status and abscisic acid (ABA) levels before, during and after a chilling period. Seedlings were raised in fertilized soil at 24/22°C (day/night), 70% relative humidity. and a 12-h photoperiod with 200 μmol m⁻² s⁻¹ from fluorescent tubes. At an age of 2 weeks the plants were conditioned at 14/12°C for 4 days and then chilled for 5 days at 5/3°C. The other conditions (relative humidity, quantum flux, photoperiod) were unchanged. After the chilling period the plants were transferred to the original conditions for recovery. The third leaves were used to study changes in leaf necrosis, ion efflux, transpiration, water status and ABA accumulation. Pronounced differences in chilling tolerance between the 4 lines as estimated by necrotic leaf areas, ion efflux and whole plant survival were observed. Conditioning significantly increased tolerance against chilling at 5/3°C in all genotypes. The genotypes with low chilling tolerance had lower water and osmotic potentials than the more tolerant genotypes during a chilling period at 5/3°C. These differences were related to higher transpiration rates and lower diffusive resistance values of the more susceptible lines. During chilling stress at 5/3°C ABA levels were quadrupled. Only a small rise was measurable during conditioning at 14/12°C. However, conditioning enhanced the rise of ABA during subsequent chilling. ABA accumulation in the two lines with a higher chilling tolerance was triggered at a higher leaf water potential and reached higher levels than in the less tolerant lines. We conclude that chilling tolerance in maize is related to the ability for fast and pronounced formation of ABA as a protective agent against chilling injury.     

Chilling sensitivity of the frost-tolerant potato Solanum commersonii

Frost tolerance has been reported in the shoots of wild, tuberiferous potato species such as Solanum commersonii when the plants are grown in either field or controlled conditions. However, these plants can survive as underground tubers and avoid unfavorable environmental conditions altogether. As such, leaf growth and photosynthesis at low temperature may not be required for survival of the plants. In order to determine the temperature sensitivity of S. commersonii shoots, we examined leaf growth, development and photosynthesis in plants raised at 20/16°C (day/night). 12/9°C and 5/2°C. S. commersonii leaves grown at 5°C exhibited a marked decrease in leaf area and in total chlorophyll (Chl) content per leaf area when compared with leaves grown at 20°C. Furthermore, leaves grown at 5°C did not exhibit the expected decrease in either water content or susceptibility to low-temperature-induced photoinhibition that normally characterizes cold acclimation in frost-tolerant plants. Measurements of CO₂-saturated O₂ evolution showed that the photosynthetic apparatus of 5°C plants was functional, even though the efficiency of photosystem II photochemistry was reduced by growth at 5°C. A decrease in the resolution of the M-peak in the slow transients for Chl a fluorescence in leaves grown at 12 and 5°C and in all leaves exposed to high light at 5°C indicated that low temperature significantly affected processes on the reducing side of Q_A, the primary quinone electron acceptor in photosystem II. Thus S. commarsonii exhibits the characteristics of a plant that is limited by chilling temperatures. Although S. commersonii can tolerate light frosts, its sensitivity to chilling temperatures may result in shoot dieback in winter in its native habitat. The plants may avoid both chilling and freezing temperatures by overwintering as underground tubers.     

Role of phytochrome B in the development of cold tolerance in cucumber plants under light and in darkness

Cold tolerance of cucumber (Cucumis sativus L.) seedlings was investigated using wild-type plants and the phytochrome B-deficient mutant (lh-mutant). Plants were subjected for 6 days to intermittent short-term cooling (12°C for 2 h per day) and to continuous chilling under conditions of 16-h photoperiod (day/night = 16/8 h) and permanent illumination. “Dehardening” process was initiated by the transfer of plants to either light or dark conditions at 23°C. It was concluded that phytochrome B participates in the development of cold tolerance in cucumber plants under stress conditions, i.e., under short-term intermittent chilling at nights and during dehardening in continuous darkness.     

Chilling tolerance of maize,cucumber and rice seedling leaves and roots are differentially affected by salicylic acid

Salicylic acid (SA) is one component of a complex signalling pathway that is induced by a number of biotic and abiotic stresses. Exposing seedling radicles to aqueous solutions of 0.5 m M salicylic acid for 24 h before chilling at 2.5°C for 1–4 days reduced the chilling-induced increase in electrolyte leakage from maize and rice leaves, and cucumber hypocotyls, but not from their radicles. The SA treatments that induced chilling tolerance in the aerial portion of the seedlings did not induce chilling tolerance in the radicles, even though the SA treatments were applied to the radicles. A comparison of activity among five antioxidant enzymes showed that SA did not alter enzyme activities in the radicles, but that chilling tolerance induced by SA in the aerial portions of maize and cucumber plants was associated with an increase in the activity of glutathione reductase and guaiacol peroxidase.     

Characterization of cold-induced changes in the fatty acids profile of rice seedlings

Rice, a staple food for more than one half of the world’s population, is one of the most cold-sensitive cereals. Breeding programs aimed at increasing rice production are expected to reduce cold-imposed grain losses. Several reports have demonstrated that cold induce differential effects on the fatty acids profile of membranes in chilling-sensitive and chilling-tolerant plants. In this work, we evaluated changes in fatty acid (FA) composition as a potential screening tool to evaluate chilling sensitivity of rice accessions. Cold exposure led to the preferential accumulation of the polyunsaturated linolenic and linoleic FAs and reduction of palmitic and stearic FAs, besides showing increased lignoceric acid content in roots of the variety. Similarly, roots of cold-exposed line Quila 66304 also presented preferential accumulation of linolenic and linoleic FAs and reduction of palmitic and stearic FAs. Cold exposure also led to enhanced levels of palmitic acid in shoots of Amaroo and, in a smaller extent, in shoots of Quila 66304. Linolenic acid was reduced in the shoots of both Amaroo and Quila 66304, while oleic acid content was reduced in shoots of Amaroo and slightly increased in shoots of Quila 66304. Double-bond index analysis indicated that 18 carbons FAs DBI for roots might be a good screening tool for cold response in rice. Results in this report demonstrate that cold-induced changes in FA profile represent a useful screening tool for early identification of differences in cold acclimation potential among rice accessions.     

Metabolic changes associated with cold-acclimation in contrasting cultivars of barley

Cereal plants become more resistant to freezing when first exposed to a period of cold-acclimation. Many physiological and molecular changes have been shown to occur at low temperatures, but the role and the contribution of each to frost resistance is still poorly understood. Two cultivars of barley ( Hordeum vulgare L.), the winter barley Onice and the spring barley Gitane, were acclimated under controlled conditions under an 8-h photoperiod at 4°C (light) and 2°C (dark) for 21 days. Changes in free proline, ABA, water-soluble carbohydrates and free fatty acids were measured to assess their involvement in cold-acclimation and to explain the different frost-resistant capacities of the two cultivars. Exposure of barley plants to low temperature resulted in an equal increase in proline in both cultivars. During the first days of cold acclimation, ABA levels showed a peak in the frost-resistant cultivar, lasting about 24 h, followed by a decrease. The water soluble carbohydrates reached their highest content after 3 days of hardening, although after 14 to 21 days of acclimation the carbohydrate content was similar to that of unhardened plants. The frost-resistant Onice had a much higher free fatty acid content than the frost-sensitive Gitane. Furthermore in Onice 86% of free farty acids was represented by unsaturated molecular species. Inolenic acid alone being 71%. In contrast, in the frost-sensitive cultivar only 31% of free fatty acids was unsaturated and a large amount of 9-oxo-nonanoic acid, a product present in the linolenic acid cascade, was also detected.
The ABA content after 2 days of hardening and the free fatty acid composition were clearly different between the two cultivars and may explain, at least in part, the different frost-resistant capacities of Onice and Gitane.     

Temporal resolution of cold acclimation and de-acclimation in the Antarctic collembolan, Cryptopygus antarcticus

Abstract.  The Antarctic collembolan, Cryptopygus antarcticus (Willem), can switch its supercooling point (SCP) between 'winter' and 'summer' modes of cold hardiness over a matter of hours. High resolution temporal scaling of the acquisition and loss of cold hardiness is undertaken by assaying changes in the proportion of animals freezing below −15 °C in response to cooling rate, acclimation temperature, and access to food and moisture. Rapid de-acclimation to the 'summer' modal state is readily achieved after 1–6 h in response to warming and access to food; however, rapid acclimation to the 'winter' modal state is only evident in response to slow cooling and narrow ranges of temperature (0–5 °C). The rapid loss of cold tolerance at higher temperatures with access to food, in particular, emphasizes this species' opportunistic responses to resource availability in the short polar summers. Cold hardiness is apparently more readily traded off against nutrient acquisition than vice versa in this maritime Antarctic species.     

The influence of thermal parameters on the acclimation responses of pinfish Lagodon rhomboides exposed to static and decreasing low temperatures

Pinfish Lagodon rhomboides acclimation rates were determined by modelling changes in critical thermal minimum ( T _{crit min}, ° C) estimates at set intervals following a temperature decrease of 3–4° C. The results showed that pinfish gained a total of 3·7° C of cold tolerance over a range of acclimation temperatures ( T _acc, ° C) from (23–12° C), that cold tolerance increased with exposure time to the reduced temperature at all T _acc, but that the rate of cold tolerance accruement (mean 0·14° C day⁻¹) was independent of T _acc. A highly significant ( P < 0·001) multivariate predictive model was generated that described the acclimation rates and thermal tolerance of pinfish exposed to reduction in water temperature: log₁₀ T _{crit min}= 0·41597 − 0·01704 T _acc+ 0·04320 T _plunge− 0·08376[log₁₀ ( t + 1)], where T _plunge is plunge temperature (° C) and t is the time (days). A comparison of the present data, with acclimation rate data for other species, suggests that factors such as latitude or geographic range may play a more important role than ambient temperature in determining cold acclimation rates in fishes.     

Effects of acclimation temperature on thermal tolerance and membrane phospholipid composition in the fruit fly Drosophila melanogaster

Adaptative responses of ectothermic organisms to thermal variation typically involve the reorganization of membrane glycerophospholipids (GPLs) to maintain membrane function. We investigated how acclimation at 15, 20 and 25 degrees C during preimaginal development influences the thermal tolerance and the composition of membrane GPLs in adult Drosophila melanogaster. Long-term cold survival was significantly improved by low acclimation temperature. After 60 h at 0 degrees C, more than 80% of the 15 degrees C-acclimated flies survived while none of the 25 degrees C-acclimated flies survived. Cold shock tolerance (1h at subzero temperatures) was also slightly better in the cold acclimated flies. LT50 shifted down by ca 1.5 degrees C in 15 degrees C-acclimated flies in comparison to those acclimated at 25 degrees C. In contrast, heat tolerance was not influenced by acclimation temperature. Low temperature acclimation was associated with the increase in proportion of ethanolamine (from 52.7% to 58.5% in 25 degrees C-acclimated versus 15 degrees C-acclimated flies, respectively) at the expense of choline in GPLs. Relatively small, but statistically significant changes in lipid molecular composition were observed with decreasing acclimation temperature. In particular, the proportions of glycerophosphoethanolamines with linoleic acid (18:2) at the sn-2 position increased. No overall change in the degree of fatty acid unsaturation was observed. Thus, cold tolerance but not heat tolerance was influenced by preimaginal acclimation temperature and correlated with the changes in GPL composition in membranes of adult D. melanogaster.