Winter hardening and glutathione status in the bilberry (Vaccinium myrtillus) in response to trace gases (CO2, O3) and nitrogen fertilization

Bilberry plants (Vaccinium myrtillus L.) at a field site in northern Finland (65°N) were subjected to nitrogen fertilization [6.5 mmol m^?2 NH₄NO₃× Ca(OH)₂] at the beginning of 3 growing seasons in late May and to trace gas fumigation (CO₂ and O₃) for 5 months (May–September) in 1993–1995 in order to investigate frost resistance and glutathione concentrations during the winter hardening period, and to assess the correlation between these variables. Harvesting was performed twice in the autumn of both 1994 and 1995, and the two-year data for each harvest were pooled. The frost resistance of the bilberry stems increased by about 10°C during the hardening period between the two harvests. Nitrogen fertilization increased the frost resistance towards late autumn. The fumigation treatments had no marked effect on it. The combination of elevated CO₂ and nitrogen fertilization induced a decrease in frost resistance. Increases in total glutathione concentrations and the proportion of reduced glutathione (GSH) in the stems were evident during hardening. Nitrogen fertilization positively affected the total glutathione concentration and the proportion of GSH at the beginning of the hardening period but the effect disappeared during the hardening process. Trace gas fumigation as such had no marked effect on glutathione concentration. Increases in glutathione concentrations during hardening did not correlate with frost resistance, possibly due to different timing of the appearence of the response to fertilization treatment, i.e., glutathione responded in the beginning of hardening while frost resistance at the end. The lack of correlation with frost resistance, and especially the different responses to nitrogen fertilization, may reflect the indirect role of glutathione in the development of winter hardening, as a transport and storage form of reduced nitrogen and sulphur. In conclusion, winter hardening and glutathione status in the bilberry seems to be sensitive to nitrogen fertilization, and not affected by elevated CO₂ and O₃.     

Carbohydrate content of Eucalyptus gunnii leaves along an annual cycle in the field and during induced frost-hardening in controlled conditions

The annual changes in frost hardiness were studied for three Eucalyptus gunnii genotypes. Frost resistance evaluated on leaf discs by the electrolyte leakage method reached a maximum in the coldest period and a minimum in summer demonstrating winter frost hardening. Genotype 634 exhibited a higher intrinsic resistance than the other genotypes both in the hardened and in the non-hardened stages. Plants of this genotype were also frost acclimated in controlled conditions by a progressive decrease of culture temperature (25 to 0 °C) but the degree of hardening appeared to be lower in these conditions. The carbohydrate patterns in leaves varied with acclimation. In controlled conditions the leaves of genotype 634 exhibited a rise in sucrose, fructose and raffinose concentration up to a temperature of 10 to 7 °C which subsequently decreased. In natural conditions a comparison of the three genotypes allowed us to correlate the higher intrinsic resistance of genotype 634 to a higher soluble sugar content. During acclimation fructose and raffinose changes were also correlated to an increase in cold resistance even though the kinetics of these changes differed in controlled and natural conditions. The starch content was very low in the various genotypes in the different conditions but oligosaccharides such as stachyose and possibly verbascose were detected. The results point out the relationships occurring between increased frost resistance and changes in fructose and raffinose concentration in E. gunnii leaves.     

Plant resistance to cold stress: Mechanisms and environmental signals triggering frost hardening and dehardening

This introductory overview shows that cold, in particular frost, stresses a plant in manifold ways and that the plant’s response, being injurious or adaptive, must be considered a syndrome rather than a single reaction. In the course of the year perennial plants of the temperate climate zones undergo frost hardening in autumn and dehardening in spring. Using Scots pine (Pinus sylvestris L.) as a model plant the environmental signals inducing frost hardening and dehardening, respectively, were investigated. Over 2 years the changes in frost resistance of Scots pine needles were recorded together with the annual courses of day-length and ambient temperature. Both act as environmental signals for frost hardening and dehardening. Climate chamber experiments showed that short day-length as a signal triggering frost hardening could be replaced by irradiation with far red light, while red light inhibited hardening. The involvement of phytochrome as a signal receptor could be corroborated by respective night-break experiments. More rapid frost hardening than by short day or far red treatment was achieved by applying a short period (6 h) of mild frost which did not exceed the plant’s cold resistance. Both types of signals were independently effective but the rates of frost hardening were not additive. The maximal rate of hardening was − 0.93°C per day and frost tolerance of < − 72°C was achieved. For dehardening, temperature was an even more effective signal than day-length.     

Dormancy development during cold hardening of in vitro cultured Malus domestica Borkh. plants in relation to their frost resistance and cryotolerance

The development of dormancy, frost resistance and cryotolerance of in vitro apple plants (Malus domestica Borkh.), cv. Greensleeves during their exposure to cold hardening was studied. In vitro cultures were cold hardened at 4°C under a short photoperiod up to 25?weeks. The dormancy status, non-structural saccharides, proline, water content and frost resistance were evaluated for optimization of cryopreservation. According to regrowth tests, in vitro cultures exhibited endogenous dormancy after the maximal frost resistance was reached. The highest regeneration ability of shoot tips after cryopreservation by encapsulation–dehydration method coincided with the period of the plant’s dormant state and maximum of frost resistance. All studied saccharides and proline exhibited the maximal values at the beginning of cold hardening and/or the dormancy phase. Contrary to the accumulation of saccharides and proline, water content showed the inverse time behaviour. According to these results, the cold hardening-induced endodormancy, high frost resistance and accumulation of saccharides and proline are the important prerequisites for the successful cryopreservation of shoot tips of in vitro grown apple plants.     

Carbohydrate content and glycosidase activities following cold hardening in two grass species

The freezing resistance of the grass species Phleum pratense L. (timothy) and Phalaris arundinaces L. increases significantly after cold hardening. The content and composition of soluble carbohydrates were determined in the plants after short day treatment, cold hardening and dehardening. The amounts of mono-, di- and trisaccharides were reduced during the short day treatment, increased during cold hardening and decreased again during dehardening. The total amounts of soluble carbohydrates (mono-, di-, tri- and polysaccharides) were the same in hardened and dehardened plants, indicating that during hardening soluble polysaccharides (fructose polymers, fructans) were converted to mono- and oligosaccharides. Sucrose increased most after hardening conditions and, in P. arundinacea , a significant increase in 1-F-fructosylsucrose (isokestose) was also observed.
Invertase (β-fructofuranosidase. EC 3.2.1.26) activity increased following cold hardening and decreased following dehardening, while the α-galactosidase (EC 3.2.1.22) activity seemed to increase after dehardening. The glycosidases are probably involved in the mobilisation of polysaccharides during cold hardening.     

Frost hardiness in the juvenile and adult life phase of ivy (Hedera helix L.)

The common ivy (Hedera helix L.) remains juvenileat its northern, eastern and altitudinal distribution limits although juvenileparts are largely killed by severe frost spells. In order to explain thisdiscrepancy we investigated the seasonal course of frost resistance in variousorgans of juvenile and adult parts of the same H. helixplants. Maximum frost resistance of leaves (LT₅₀–25°C) and axis (xylem parenchyma:LT₅₀ –29°C; cambium: LT₅₀–35°C) was quite the same in juvenile and adultparts. Thus, H. helix is able to acquire full frostresistance in its juvenile phase. However, hardening of leaves was slower anddehardening of axis was faster in juvenile parts. Leaves of juvenile partsremain 2 to 4 K less resistant than those of adult parts untilattaining the maximum resistance. This explains why mainly leaves of thejuvenile parts were damaged following severe frost episodes with temperaturesbelow –20°C. The occurrence of H.helix in its juvenile phase at the frost-caused distribution limitsmay be explained as follows: Leaves of juvenile plants may occasionally bekilled by severe frosts, but regeneration from dormant eyes enables survival.Loss of leaves may impede the change to the adult phase, but even if the plantsbecame adult frost killing of rest buds (2 to 3 K less resistantthan leaves) would induce rejuvenation.     

Effects of Hormones on Morphogenesis and Cold Resistance in Berseem Clover (Trifolium alexandrinum L.)

Plants of berseem clover (Trifolium alexandrinum L.) cv. Taborwere raised under conditions inhibiting the acquisition of coldhardiness (non-hardened) or inducing cold hardiness (hardened).All non-hardened plants developed an elongated shoot and exhibitedconsiderable frost sensitivity, as measured by the extent ofthe reduction in yield of variable chlorophyll fluorescenceafter exposure to sub-zero temperature. Hardened plants developeda shorter shoot, with fewer leaves and a greater percentageof dry matter in the root system. These parameters were associatedwith a marked increase in frost resistance. Exogenous applicationof ABA to plants effected similar morphological modificationsin both hardening and non-hardening temperature regimes; plantsdeveloped a shorter primary shoot axis and leaves exhibiteda marked increase in frost hardiness. In berseem clover ABAcan thus substitute, at least partially, for the low temperaturetreatment required to induce cold hardiness. Spraying plantsraised under hardening conditions with gibberellic acid reversedthe effects of the hardening treatment, since they developedan elongated shoot and exhibited frost sensitivity comparableto non-treated plants grown under non-hardening conditions.It is concluded that these endogenous hormones are directlyinvolved in triggering changes in morphogenesis which accompanyphysiological and metabolic events associated with the inductionof plant cold hardiness. Key words: Frost resistance, morphogenesis, abscisic acid, giberellic acid, Trifolium alexandrinum     

Assessment of the frost sensitivity of Trifolium species by chlorophyll fluorescence analysis

The potential of the chlorophyll fluorescence technique in screening for frost sensitivity in a range of Trifolium species from different geographical origins was assessed by measuring the decrease in variable chlorophyll fluorescence (F_var) of leaves after freezing at - 5°C for 60 min. The method was rapid and the results obtained agreed well with a visual assessment of freezing injury carried out after leaves were returned to optimal growth conditions for 72 h. Trifolium alexandrinum (Berseem clover) cv. Tabor originating from Israel was shown to be the most frost sensitive species studied and Trifolium subterraneum (subterranean clover) cv. Mt. Barker, from temperate regions of Australia, the most frost resistant. On extended periods of freezing, frost damage increased and this was associated with a further reduction in variable chlorophyll fluorescence and in quenching capacity of the thylakoid membranes. These results thus indicate that substantial thylakoid membrane dysfunction is induced at freezing temperatures. Furthermore, it was found that frost hardening of the frost sensitive species T. alexandrinum for 21 days at 5°C reduced the extent of damage sustained by the thylakoid membranes as shown by higher fluorescence quenching capacity, smaller reduction in variable fluorescence (F_var) and higher initial fluorescence (F_o) when leaves of hardened plants were frozen at -5°C and -7°C.     

Application of Impedance Spectroscopy for Selecting Frost Hardy Varieties of English Ryegrass

The frost hardening and frost damage of 12 varieties of Englishryegrass (Lolium perenne) was studied by electrical impedancespectroscopy. For the measurement of the impedance spectrum(80 Hz to 1 MHz) a 10 mm length sample was cut from the stemabove the growing point, but the growing point was included.The impedance spectra were analysed by an asymmetric distributedcircuit model. The impedance spectra were measured at two phasesof hardiness and after freezing, i.e. (a) before hardening,(b) after hardening in controlled conditions, and (c) aftercontrolled frost exposure at -16 °C after hardening. Twomodel parameters, i.e. intra- and extracellular resistance,increased with hardening. The intracellular resistance and theskewness factor before hardening, and the ratio between thosetwo parameters before and after hardening, strongly correlatedwith hardening of different varieties of English ryegrass. Theextracellular resistance and the relaxation time decreased asa result of the frost exposure at -16 °C. Cold acclimation; electrical impedance; English ryegrass; frost hardiness; impedance spectroscopy; Lolium perenne     

On the mechanisms of frost injury and frost hardening of spruce chloroplasts

Hill reaction and noncyclic photophosphorylation of isolated class C chloroplasts of spruce (Picea abies (L.) Karst.), as well as ¹⁴CO₂ fixation by whole needles at constant laboratory conditions proceeded at high rates during spring and early summer, declined during late summer and autumn by about 60%, remained at this level during winter, and recovered quickly in early spring. During summer, the whole needles proved to be frost labile, since after exposure to-20°C and careful thawing, fast chlorophyll degradation occurred. In addition, only photosynthetically inactive chloroplasts could be isolated from those precooled needles. On the contrary, during winter the photochemical activities of plastids from freshly harvested needles did not differ from those of artificially frozen-thawed needles. When isolated spruce chloroplasts were exposed to the same subfreezing temperatures as the whole needles, no influence of freezing on the photochemical activities was observed, irrespective of whether the plastids were isolated from frost sensitive or frost hardened needles. It is concluded that frost damage to spruce chloroplasts is due to an attack of membrane toxic compounds or lytic enzymes which were liberated upon freezing from more labile compartments. Frost hardening of the chloroplasts, as determined by the stability of chlorophyll after exposure of the needles to low temperatures, as well as by the isolation of photosynthetically active chloroplasts from such precooled needles, appeared to depend at least on 2 processes: (i) an alteration of the composition of the photosynthetically active membranes and (ii) and additional stabilization of these membranes by protecting substances. The first process was indicated by a large increase (decrease) of the capability of isolated chloroplasts for PMS-mediated photophosphorylation which accompanied natural or artificial frost hardening (dehardening). Production of cryoprotecting compounds was suggested by a significant higher stability against NaCl observed with class C chloroplasts isolated from frost hardened needles as compared to that of plastids from frost labile material. The decrease of the capability for both, the ferricyanide dependent photoreactions of the plastids and the CO₂ fixation by whole needles, which was observed during the frost hardening phase, cannot be due to freezing injuries; it rather appears to be a consequence of the frost hardening process.     

Sodium hydrosulfide priming improves the response of photosynthesis to overnight frost and day high light in avocado (Persea americana Mill,cv. ‘Hass’)

Radiation frost events, which have become more common in the Mediterranean Basin in recent years, inflict extensive damage to tropical/subtropical fruit crops. During radiation frost, sub-zero temperatures are encountered in the dark, followed by high light during the subsequent clear-sky day. One of the key processes affected by these conditions is photosynthesis, which, when significantly inhibited, leads to the enhanced accumulation of reactive oxygen species (ROS) and damage. The use of ‘chemical priming’ treatments that induce plants' endogenous stress responses is a possible strategy to improve their coping with stress conditions. Herein, we studied the effects of priming with sodium hydrosulfide (NaHS), a donor of hydrogen sulfide (H₂S), on the response of photosynthesis to overnight frost and day high-light conditions in ‘Hass’ avocado (Persea americana Mill). We found that priming with a single foliar application of NaHS had positive effects on the response of grafted ‘Hass’ plants. Primed plants exhibited significantly reduced inhibition of CO₂ assimilation, a lower accumulation of hydrogen peroxide as well as lower photoinhibition, as compared to untreated plants. The ability to maintain a high CO₂ assimilation capacity after the frost was attained on the background of considerable inhibition in stomatal conductance. Thus, it was likely related to the lower accumulation of ROS and photodamage observed in primed ‘Hass’ plants. This work contributes toward the understanding of the response of photosynthesis in a subtropical crop species to frost conditions and provides a prospect for chemical priming as a potential practice in orchards during cold winters.     

Will loss of snow cover during climatic warming expose New Zealand alpine plants to increased frost damage?

If snow cover in alpine environments were reduced through climatic warming, plants that are normally protected by snow-lie in winter would become exposed to greater extremes of temperature and solar radiation. We examined the annual course of frost resistance of species of native alpine plants from southern New Zealand that are normally buried in snowbanks over winter (Celmisia haastii and Celmisia prorepens) or in sheltered areas that may accumulate snow (Hebe odora) and other species, typical of more exposed areas, that are relatively snow-free (Celmisia viscosa, Poa colensoi, Dracophyllum muscoides). The frost resistance of these principal species was in accord with habitat: those from snowbanks or sheltered areas showed the least frost resistance, whereas species from exposed areas had greater frost resistance throughout the year. P. colensoi had the greatest frost resistance (−32.5°C). All the principal species showed a rapid increase in frost resistance from summer to early winter (February–June) and maximum frost resistance in winter (July–August). The loss of resistance in late winter to early summer (August–December) was most rapid in P. colensoi and D. muscoides. Seasonal frost resistance of the principal species was more strongly related to daylength than to temperature, although all species except C. viscosa were significantly related to temperature when the influence of daylength was accounted for. Measurements of chlorophyll fluorescence indicated that photosynthetic efficiency of the principal species declined with increasing daylength. Levels of frost resistance of the six principal alpine plant species, and others measured during the growing season, were similar to those measured in tropical alpine areas and somewhat more resistant than those recorded in alpine areas of Europe. The potential for frost damage was greatest in spring. The current relationship of frost resistance with daylength is sufficient to prevent damage at any time of year. While warmer temperatures might lower frost resistance, they would also reduce the incidence of frosts, and the incidence of frost damage is unlikely to be altered. The relationship of frost resistance with daylength and temperature potentially provides a means of predicting the responses of alpine plants in response to global warming.     

Oryzalin-Induced Changes in Water Status and Cytoskeleton Proteins of Winter Wheat Seedlings upon Cold Acclimation and ABA Treatment

The effect of oryzalin (a specific inhibitor of tubulin polymerization in plant cells) on water retention by the leaves and roots of winter wheat (Triticum aestivum L.) seedlings was studied. The cultivars differing in their frost resistance were compared after their acclimation to low temperature (3°C for 3 or 7 days) and after treatment with ABA. In control untreated plants, oryzalin reduced the water-retaining capacity (WRC) of leaves and roots. Both hardening and ABA lowered the effect of the inhibitor on WRC in leaves, whereas their effects on water retention by roots were opposite, i.e., hardening weakened and ABA intensified the effect of oryzalin. Oryzalin-induced reduction of WRC decreased in the following sequence of cultivars: weakly frost resistant moderately frost resistant highly frost resistant. It was more pronounced in the leaves than in the roots, the latter being characterized by the lower WRC and lower frost resistance. After three-day-long hardening of plants, an additive effect of hypothermia and ABA on oryzalin-induced decrease in WRC of leaves and the lack of such effect in the roots were observed. The immunochemical analysis of the composition and content of cytoskeletal proteins with Western blotting showed that in the leaves the actin/tubulin ratio was higher than in the roots. The treatment of nonacclimated plants with ABA lowered the content of - and -tubulins and actin in roots but did not affect the level of actin in leaves. Hardening negated the effects of ABA on cytoskeletal proteins. Oryzalin produced the greatest inhibitory effect on WRC and an increase in frost resistance in ABA-treated plants in the experiments with leaves of the weakly frost resistant cultivar before and after hardening. Organ- and cultivar-specific and ABA-mediated dependence of WRC on cytoskeletal proteins and microtubules and microfilaments formed by them is supposed to result from their effect on the state of intracellular water and water permeability of the plasma membrane. In the course of cold acclimation of plants and upon their treatment with ABA, this dependence was more distinctly expressed in leaves than in roots, and especially in the plants of the weakly frost resistant cultivar.     

Foliar freezing resistance of Australian alpine plants over the growing season

We assessed the freezing resistance of leaves ex situ of 25 Australian alpine plant species. We compared the freezing resistance of forb, graminoid and shrub species from three alpine summits of different altitudes; from a low altitude site just above treeline, to a fully alpine tundra site. Foliar freezing resistance (LT₅₀) in spring varied from ?5.9°C to ?18.7°C and standardized LT₅₀ values within species were significantly related to site altitude. Additionally, when comparing all the species in the study, freezing resistance was significantly related to site; the LT₅₀ of samples from a low‐altitude summit (1696 m) were significantly lower than those of samples from mid‐ (1805 m) and high‐altitude (1860 m) summits. The LT₅₀ of juvenile foliage did not differ significantly from that of adult foliage. Shrubs were highly resistant to freezing. At the highest summit, we examined the course of seasonal freezing resistance from early summer to early autumn across three alpine plant communities that differed in the time of natural snowmelt; from sheltered (snowpatch) to exposed (open heath). No differences in freezing resistance over the growing season were detected for exposed or sheltered communities and there were no consistent trends indicating frost hardening over the growing season. Overall, the common Australian alpine species we investigated appear well adapted to freezing conditions throughout the snow‐free growing season. We have no evidence to suggest that freezing temperatures soon after snowmelt in spring are especially damaging to the alpine plants at these summits.     

The effects of long-term elevation of air temperature and CO on the frost hardiness of Scots pine

The frost hardiness of 20 to 25-year-old Scots pine (Pinus sylvestris L.) saplings was followed for 2 years in an experiment that attempted to simulate the predicted climatic conditions of the future, i.e. increased atmospheric CO₂ concentration and/or elevated air temperature. Frost hardiness was determined by an electrolyte leakage method and visual damage scoring on needles. Elevated temperatures caused needles to harden later and deharden earlier than the controls. In the first year, elevated CO₂ enhanced hardening at elevated temperatures, but this effect disappeared the next year. Dehardening was hastened by elevating CO₂ in both springs. The frost hardiness was high (相似文献   

Glycosidases and B group vitamins produced by six yeast strains from the digestive tract of Phoracantha semipunctata larvae and their role in the insect development

Six strains of yeasts were isolated from P. semipunctata larval gut in order to determine their role in insects nutrition. P. semipunctata larvae possess a rich glycosidasic equipment acting on oligosaccharides, heterosides and polysaccharides which is particularly characterized by an activity on hemicellulose. The yeast strains hydrolyze oligosaccharides, heterosides and some polysaccharides, such as starch and pectin, but no activity was detected on cellulose. -Glucosidase was secreted in the medium by all strains except one. B group vitamins were also secreted by the yeasts.

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Résumé Six souches de levures ont été isolées du tube digestif de larves de P. semipunctata afin de déterminer leur rôle dans la nutrition de l'insecte. Les larves de P. semipunctata possèdent un riche équipment enzymatique agissant sur les oligosaccharides, les hétérosides et les polysaccharides (en particulier sur les hémicelluloses). Les souches de levures hydrolysent les oligosaccharides, les hétérosides et quelques polysaccharides, tels que l'amidon et la pectine, mais on n'a détecté aucune activité sur la cellulose. Toutes les souches, sauf une, excrètent dans le milieu de la -glucosidase. Enfin, ces levures excrètent également des vitamines du groupe B.

     

Anthocyanins and glutathione S-transferase activities in response to low temperature and frost hardening in Vaccinium myrtillus (L.)

Anthocyanin (Acy) contents and GST activities of bilberry (Vaccinium myrtillus L.) were investigated in two experiments conducted in June (Exp. I: active growth) and in August September (Exp. II: beginning of frost hardening) in Northern Finland (65 degrees N). Bilberry plants were subjected to +2 degrees C and +18 degrees C in Exp. I or +5/0 degrees C (day/night) and +18/+13 degrees C (day/night) in Exp. II. GST activities were assessed using either 1-chloro-2,4-dinitrobenzene (CDNB) or trans-cinnamic acid (tCA) as substrates. We found temperature to have no effect on Acy during either active growth or frost hardening. Acy increased several-fold from active growth to the beginning of frost hardening, but no increment was observed during the development of frost hardening. This suggests a role of Acy in photoprotection at low temperatures rather than their direct involvement in the development of freezing tolerance. The lack of response of GST activity to frost hardening and to temperature in autumn may indicate an indirect role of GSTs in frost hardening as protective enzymes. GST activity was the same with the two substrates studied (CDNB, tCA), supporting the assumption that GSTs could catalyze reactions with endogenous phenylpropanoids.     

Phospholipid Involvement in Frost Tolerance

Changes in frost tolerance and in phospholipid content were studied in the leaves of winter rape plants (Brassica napus L. var. oleifera L. cv. Górczański) grown under natural or artificially controlled conditions. Frost hardening was found to be a three-stage process. During the first stage, occurring at low but above freezing environmental temperatures, phospholipid changes do not seem to be directly related to the leaf frost tolerance. This stage of hardening is possibly related to a metabolic shift caused by the cessation of growth. The achievement of the second level of frost tolerance in the fully turgid leaves depends on the occurrence of sub-freezing temperature and is related to increase in phospholipid level. It was shown that freezing brought about phospholipid degradation which was reversible only in slightly injured leaves with a relatively high phospholipid content. The third stage of hardening is related to frost-induced dehydration of the cells and may overlap the second one.     

Modifications of abscisic acid level in winter oilseed rape leaves during acclimation of plants to freezing temperatures

An almost twofold increase in abscisic acid (ABA) content was observed in the leaves of winter oilseed rape plants (Brassica napus L., var. oleifera L., cv. Jantar) grown in the cold (>0°C). This ABA increase took place during the first three days of cold treatment. After 6 days of plant growth in the cold, the level of ABA started to decline or remained constant, depending on the calculation basis: dry weight or disc area units, respectively. The exposure of cold-acclimated plants to night frost (–5°C for 18 h) induced a further increase (65%) in the ABA level, which begun during the first few hours after thawing. The comparison of time courses of frost resistance increments and ABA content changes showed that modifications of ABA level in the cold-treated leaves preceded those of frost resistance, whereas in the frost-pretreated tissues the ABA increase occurred later than that of frost tolerance. Possible interrelations between ABA content, frost tolerance and tissue water potential modifications in the low temperature-affected tissues are discussed.     

Effects of artificial frost hardening and winter stress on net photosynthesis, photosynthetic electron transport and RuBP carboxylase activity in seedlings of Pinus silvestris

Net photosynthesis of seedlings of Pinus silvestris has been measured and compared with the activities of photosynthetic electron transport and extracted RuBP carboxylase. The effects of prolonged frost hardening (photoperiod 8 h, + 3°C) followed by winter stress at subzero temperatures were analysed. There was a parallel effect of frost hardening and winter stress on the photosynthetic properties of both intact seedlings and isolated chloroplast thylakoids. The activity of extracted RuBP carboxylase was less affected by the treatments. In relation to earlier works we conclude that the decay of net photosynthesis in winter climate is determined by the electron transport properties of the chloroplast thylakoids, i.e. by the pool sizes of photosynthetically active plastoquinone. The results of this work justify the definition of two phases in the response of conifers towards autumn and winter climates: I. Frost hardening occurs at temperatures slightly above zero and it does not affect the efficiency of photosynthesis as defined by the quantum yield at rate limiting light absorption. II. Winter stress occurs at subzero temperatures and it is characterized by a suppression of the photosynthetic efficiency as a result of damage within the photosynthetic apparatus.