First frost: Effects of single and repeated freezing events on acclimation in Picea abies and other boreal and temperate conifers

In experiments with needles of Picea abies, we tested the specific hypothesis that a single night of freezing acts as a signal that triggers a rapid increase in low temperature (LT) tolerance, and the more general hypothesis that repeated or prolonged freezing stimulates increased LT acclimation. In three growth chamber experiments involving acclimation under early- to mid-autumn light and temperature conditions followed by one or more freezing treatments, we found no significant effect of a single night of freezing on LT tolerance, and only limited and inconsistent effects of repeated and prolonged freezing. We also tested the effect of prolonged storage at −5 °C on LT tolerance on samples of three boreal and three temperate conifer species during acclimation under field conditions, and again found no consistent enhancement of LT tolerance attributable to freezing in either group. In agreement with our own and others’ anecdotal observations that some species can attain nearly maximal LT tolerance in the absence of freezing under field conditions, we conclude that freezing is neither required nor a major influence in LT acclimation, at least in well-studied boreal conifer species, while the effects of freezing on temperate conifers are not as well-documented. We conclude that freezing treatment of conifer seedlings to ensure sufficient hardiness for late planting seems to offer little practical advantage.     

Leaf respiratory acclimation to climate: comparisons among boreal and temperate tree species along a latitudinal transect

In common gardens along an ～900 km latitudinal transect through Wisconsin and Illinois, U.S.A., tree species typical of boreal and temperate forests were compared with respect to the nature and magnitude of leaf respiratory acclimation to contrasting climates. The boreal representatives were trembling aspen (Populus tremuloides Michx.) and paper birch (Betula papyrifera Marsh.), while the temperate species were eastern cottonwood (Populus deltoides Bartr ex. Marsh var. deltoides) and sweetgum (Liquidambar styraciflua L.). Assessments were conducted on seedlings grown from seed sources collected near southern and northern range boundaries, respectively. Nighttime rates of leaf dark respiration (R(d)) at common temperatures, as well as R(d)'s short-term temperature sensitivity (energy of activation, E(o)), were assessed for all species and gardens twice during a growing season. Little evidence of R(d) thermal acclimation was observed, despite a 12 °C range in average air temperature across gardens. Instead, R(d) variation at warm temperatures was linked most closely with prior leaf photosynthetic performance, while R(d) variation at cooler temperatures was most strongly related to leaf nitrogen concentration. Moreover, E(o) differences across species and gardens appeared to stem from the somewhat independent limitations on warm versus cool R(d). Based on this construct, an empirical model relying on R(d) estimates from leaf photosynthesis and nitrogen concentration explained 55% of the observed E(o) variation.     

Foliar temperature tolerance of temperate and tropical evergreen rain forest trees of Australia

Australian rain forests extend from tropical climates in the north to temperate climates in the south, providing an opportunity to investigate physiological responses to temperature of both temperate and tropical species within the same forest type. Eight, rain forest canopy tree species were selected to cover the 33 degrees latitudinal range of rain forests in eastern Australia. Temperature tolerance was measured in 6-year-old plants grown in a common environment, by exposing leaves to a series of high temperatures during late summer and a series of freezing temperatures during midwinter. Damage was evaluated based on chlorophyll fluorescence measurements made 2 h after exposure and by visual assessment of leaf damage made a week after exposure. Leaves of the tropical species were more heat tolerant and less frost tolerant than leaves of the temperate species, which is consistent with their climate distributions. In contrast, the temperature tolerance of the photosynthetic apparatus was unrelated to climate in a species' native habitat. However, the tropical species underwent significant photoinhibition during winter. All species maintained the integrity of the photosynthetic apparatus and avoided tissue damage over a similar span of temperatures (about 60 degrees C), reflecting the similar annual temperature ranges in Australia's temperate and tropical rain forests. Chlorophyll fluorescence measurements and visual assessment of leaf damage provided different estimates of the absolute and relative temperature tolerances of the species, thus emphasizing the importance of a direct assessment of tissue damage for determining a species' temperature tolerance.     

Ultrastructural and Extracellular Protein Changes in Cell Suspension Cultures of Populus euphratica Associated with Low Temperature-induced Cold Acclimation

Populus euphratica Olive is the only tree species that can grow in the saline land and also survive cold winters in northwest China, and it plays a very important role in stabilizing the vulnerable ecosystem there. A cell suspension culture was initiated from callus derived from plantlets of Populus euphratica. Cold acclimation was induced (LT50 of-17.5 ℃) in cell suspension at 4-5 ℃ in the dark for 30 days and the freezing tolerance increased from LT50 of-12.5 ℃ in nonacclimated cells to LT50 of-17.5 ℃ in cold-acclimated cells. Microvacuolation, cytoplasmic augmentation and accumulation of starch granules were observed in cells that were cold-acclimated by exposure to low temperatures. Several qualitative and quantitative changes in proteins were noted during cold acclimation. Antibodies to carrot extracellular (apoplastic) 36 kD antifreeze protein did not cross react on immunoelectroblots with extracellular proteins in cell suspension culture medium of Populus euphratica, indicating no common epitopes in the carrot 36 kD antifreeze protein and P euphratica extracellular proteins. The relationship of these changes to cold acclimation in Populus euphratica cell cultures was discussed.     

Low temperature responses of Nothofagus dombeyi and Nothofagus nitida, two evergreen species from south central Chile

Nothofagus dombeyi (Mirb.) Blume and Nothofagus nitida (Phil.) Krasser are closely related evergreen trees native to south central Chile. Nothofagus dombeyi is a pioneer in habitats subject to high daytime irradiances and nighttime freezing temperatures and has a wider altitudinal and latitudinal distribution than N. nitida, which is restricted to more oceanic climates. We postulated that N. dombeyi has a greater cold-acclimation capacity, expressed as a greater capacity to maintain a functional photosynthetic apparatus at low temperatures, than N. nitida. Because cold-acclimation may be related to the accumulation of cryoprotective substances, we investigated relationships between ice nucleation temperature (IN), freezing temperature (FT), and the temperature causing injury to 50% of the leaf tissues (LT(50)) on the one hand, and concentrations of total soluble carbohydrates (TSC), starch and proline on the other hand. Observations were made throughout a seasonal cycle in adults and seedlings in the field and in seedlings in the laboratory under cold-acclimation inductive and non-inductive conditions. In adults, LT(50) values were lower in N. dombeyi than in N. nitida, suggesting that N. dombeyi is the more frost tolerant species. Adults of both species tolerated freezing in autumn and winter but not in spring and summer. In the fall and winter, seedlings of N. dombeyi had a much lower LT(50) than those of N. nitida. Nothofagus nitida seedlings, in autumn and winter, exhibited freezing avoidance mechanisms. Although elevated TSC and proline concentrations may contribute to freezing tolerance in adults of both species, an increase in proline concentration is unlikely to be the dominant frost tolerance response in adults because proline concentrations were higher in N. nitida than in N. dombeyi. In seedlings, however, there were large differences in proline accumulation between species that may account for the difference between them in freezing tolerance. Starch concentration in both species decreased during winter. Chlorophyll fluorescence indicated that maximal photochemical efficiency (F(v)/F(m)) remained at optimal values (~0.8) throughout the year. The effective photochemical efficiency of PSII (PhiPSII) and relative electron transport rates (ETR(r)) decreased in winter in both species. In seedlings, fluorescence parameters were more affected in winter in N. nitida than in N. dombeyi. We concluded that adults and seedlings of N. dombeyi are hardier than adults and seedlings of N. nitida, which is consistent with its wider latitudinal and altitudinal distribution.     

Different responses of northern and southern ecotypes of Betula pendula to exogenous ABA application

We investigated responses of northern and southern ecotypes of silver birch (Betula pendula Roth) to exogenous abscisic acid (ABA) under controlled environmental conditions to determine the role of ABA in cold acclimation and dormancy development. Abscisic acid was sprayed on the leaves and changes in freezing tolerance, determined by the electrolyte leakage test, and bud dormancy were monitored. Applied ABA induced cold acclimation but had no effect on growth cessation in seedlings grown in long day conditions (LD, 24-h photoperiod at 18 degrees C). It enhanced freezing tolerance and accelerated growth cessation in seedlings grown in short day conditions (SD, 12-h photoperiod at 18 degrees C), and slightly enhanced freezing tolerance in seedlings grown at low temperature (LT, 24-h photoperiod at 4 degrees C) in both ecotypes. There were distinct ecotypic differences in ABA-induced cold acclimation and dormancy development. The northern ecotype was more responsive to applied ABA than the southern ecotype, resulting in more rapid development of freezing tolerance in all treatments, and earlier dormancy development in SD. When plants were grown in a photoperiod just above the critical photoperiod for the ecotype (defined as the longest photoperiod that induces growth cessation), applied ABA caused growth cessation and dormancy development. Compared with ABA-treated seedlings grown in SD, dormancy development was delayed in ABA-treated seedlings exposed to a near-critical photoperiod, but even in this treatment dormancy developed faster in the northern ecotype than in the southern ecotype.     

毛白杨抗冻性低温诱导过程中糖的效应(英文)

在低温锻炼和结合蔗糖处理的低温锻炼下 ,对毛白杨 (Populustomentosa)幼苗枝条和叶片的可溶性糖含量 ,以及幼苗存活率和抗冻性分别进行了测定 .结果表明 ,低温锻炼明显提高了幼苗枝条和叶片的可溶性糖含量 ,同时也提高了幼苗的存活率和抗冻性 .2 0 %蔗糖处理对上述低温锻炼的作用有加强的效果 .进一步分析发现 ,无论是单纯的低温锻炼还是结合蔗糖预处理的低温锻炼 ,幼苗枝条中可溶性糖含量的提高程度均较叶片明显 ,而且它们的含量增加与幼苗抗冻性的提高密切相关 .这表明低温锻炼所引起的可溶性糖积累可能参与了幼苗抗冻性的诱导     

Relationship between freezing tolerance and shoot water relations of western red cedar

Freezing tolerance and shoot water relations parameters of western red cedar (Thuja plicata Donn) seedlings were measured every 2 weeks from October 1989 to April 1990. Freezing tolerance, measured by freeze-induced electrolyte leakage, showed seasonal shifts in the temperature causing 50% foliage electrolyte leakage (LT(50)). The LT(50) value was -4 degrees C in October, it decreased to -20 degrees C in February and then increased to -6 degrees C in April. The foliage index of injury at -10 degrees C (II(-10)) also showed seasonal shifts from a high of 98% in October to a low of 18% in February followed by an increase to 82% in April. Osmotic potentials at saturation (Psi(s(sat))) and turgor loss point (Psi(s(tlp))) were, respectively, -1.07 and -1.26 MPa in October, -1.57 and -2.43 MPa in January, and -1.04 and -1.86 MPa in April. Dry weight fraction (DWF) increased and symplastic volume at full turgor (V(o)) decreased during the fall-winter acclimation phase, whereas DWF decreased and V(o) increased during the late winter-spring deacclimation phase. Relationships between seasonal patterns of freezing tolerance and shoot water relations parameters showed that LT(50) and II(-10) decreased linearly as Psi(s(tlp)) and V(o) decreased and DWF increased. There was no discernible difference in the relationship during fall acclimation or spring deacclimation. The freezing dehydration index at -10 degrees C (FDI(-10)) declined from 0.69 in November to 0.41 in February and increased to 0.56 in April. The value of II(-10) decreased linearly as FDI(-10) decreased, although a measurement made on actively growing spring foliage did not fit this relationship. The results indicate that seasonal changes in freezing tolerance of western red cedar are partially due to changes in tissue water content, symplastic volume, passive osmotic adjustment and FDI(-10).     

Comparison of G6PDH Activity and LT50 Between P.tomentosa and P.suaveolens During Freezing Acclimation

The differences of glucose-6P dehydrogenase(G6PDH) activity and freezing resistance induced by freezing acclima-tion between cuttings of freezing-sensitive P.tomentosa and freezing-resistant P.suaveolens were compared for exploring the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation.After 5d of freezing acclimation at -3℃ ,the LT50 of P.tomentosa has deereased from -6.2℃ incontrol cuttings to -14.3℃ in freezing acclimated ones,and the increase of LT50 of P.tomentosa has decreased from -6.2℃ in control cutting to -14.3℃ in freezing acclimated ones,and the increase of G6PDH activity was observed in freezing acclimated cuttings as compared with control ones.Whereas,when P.suaveolens was freezing acclimated at -20℃ for 5d,the LT50 has decreased from -27.1℃ in control cuttings to -43.5℃ in freezing acclimated ones,and the activity of G6PDH increased considerably.In addition,the increase of LT50 and the decrease of G6PDH activity resulting from 2d of deacclimation at 25℃ were found in two kinds of freezing acclimated cuttings.It is concluded that the increase in the activity of G6PDH may associate with the inherited freezing resistance of species and the enhancemen of freezing resistance of cuttings,and may play an important role in the antifreeze process under freezing temperature,which would provide the basis for the study on the molecular mechanism of freezing resistance in P.suaveolens and the cloning of gene associated with freezing resistance.     

Frost hardening and dehardening in Abies procera and other conifers under differing temperature regimes and warm-spell treatments

Frequent bud frost damage in cultivation of Abies procera Rehderand pending climate changes are the background for this studyof cold hardiness under varying acclimation regime (in closed-topchambers) and experimental warm spells during the cold season.LT₅₀ values were established by freezing tests at differenttimes of year. Damage and deaths were assessed on leader buds,subapical lateral buds, needles and cambium. Minor parallelexperiments involved Abies nordmanniana, Picea abies and Piceasitchensis. Lower acclimation temperatures resulted in deeperfrost hardiness during late autumn but less during spring, comparedwith ambient temperature controls. Elevated temperatures resultedin less deep frost resistance. Apical buds generally developeddeeper frost hardiness than lateral buds but less deep thanthe cambium, varying with species, however. Frost damage inbuds ranged from death over partially destroyed bud contentsresulting in distorted shoots to buds seemingly remaining dormant.Responses to warm spells differed with duration, timing andspecies, ranging from dramatic decrease in frost hardiness withor without subsequent recovery to no reaction. Furthermore,the reactions did not show any clear relation to dormancy level.For A. procera, exposure to fluctuating temperatures appearedto be particularly problematic. This explains why this speciesdevelops best in coastal climates, and in sites sheltered fromtemperature extremes either by hedging, a winter snow cover,or topography. The Christmas tree production will suffer severelyon sites with harsh temperatures due to losses of lateral andterminal buds, which destroy the crown symmetry. Clipping ofgreenery is less influenced by frost damages, although the developmentof normal branch whorls is often disturbed.     

Differential responses of silver birch (Betula pendula) ecotypes to short-day photoperiod and low temperature

We investigated interrelations of dormancy and freezing tolerance and the role of endogenous abscisic acid (ABA) in the development of silver birch (Betula pendula Roth) ecotypes in controlled environments. Short-day treatment induced growth cessation, bud set and dormancy development, as well as initiation of cold acclimation and an increase in freezing tolerance. Subsequent low temperature and short days (12-h photoperiod) resulted in a significant increase in freezing tolerance, whereas bud dormancy was gradually released. The concentration of ABA increased in response to short days and then remained high, but ABA concentrations fluctuated irregularly when the dormant plants were subsequently exposed to low temperature during short days. Although there was a parallel development of freezing tolerance and bud dormancy in response to short days, subsequent exposure to low temperature had opposite effects on these processes, enhancing freezing tolerance and releasing dormancy. Compared with the southern ecotype, the northern ecotype was more responsive to short days and low temperature, exhibiting earlier initiation of cold acclimation, growth cessation and an increase in ABA concentrations in short days, and higher freezing tolerance, faster dormancy release and greater alteration in ABA concentrations when subsequently exposed to low temperature during short days. The rates and extent of the increases in ABA concentration may be related to increases in freezing tolerance and dormancy development during short days, whereas the extent of the fluctuations in ABA concentration may play an important role in enhancing freezing tolerance and releasing dormancy during a subsequent exposure to low temperature during short days.     

Freezing injury in primary and secondary needles of Mediterranean pine species of contrasting ecological niches

? Pine seedlings show a marked ontogenetic difference between primary and secondary needles, the latter prevailing with a different timing among species.

? Using the electrolyte leakage method following an artificial freezing test, we aimed at (1) determining the differences in freezing tolerance between primary and secondary needles in eight pines of contrasting thermal habitats: P. canariensis, P. pinea, P. halepensis, P. brutia, P. pinaster, P. nigra, P. sylvestris and P. radiata, (2) evaluating the relation between freezing tolerance and sclerophylly and (3) relating freezing tolerance with the climate of origin.

? Primary needles were significantly more sensitive to freezing than secondary needles in Pinus halepensis, P. brutia, P. pinaster and P. nigra, whereas no differences were found in P. canariensis, P. pinea and P. radiata. LT50 was uncorrelated with needle sclerophylly but very highly correlated with the mean temperature of the coldest month at the seed source.

? Results support an adaptive role of secondary needles in the mountain Mediterranean pines P. nigra, P. pinaster and P. brutia, while the more complex responses in coastal Mediterranean pines can be interpreted in the light of seedling ontogeny and species’ ecological niches.

     

Freezing tolerance of conifer seeds and germinants

Survival after freezing was measured for seeds and germinants of four seedlots each of interior spruce (Picea glauca x engelmannii complex), lodgepole pine (Pinus contorta Dougl. ex Loud.), Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and western red cedar (Thuja plicata Donn ex D. Donn). Effects of eight seed treatments on post-freezing survival of seeds and germinants were tested: dry, imbibed and stratified seed, and seed placed in a growth chamber for 2, 5, 10, 15, 20 or 30 days in a 16-h photoperiod and a 22/17 degrees C thermoperiod. Survival was related to the water content of seeds and germinants, germination rate and seedlot origin. After freezing for 3 h at -196 degrees C, dry seed of most seedlots of interior spruce, Douglas-fir and western red cedar had 84-96% germination, whereas lodgepole pine seedlots had 53-82% germination. Freezing tolerance declined significantly after imbibition in lodgepole pine, Douglas-fir and interior spruce seed (western red cedar was not tested), and mean LT50 of imbibed seed of these species was -30, -24.5 and -20 degrees C, respectively. Freezing tolerance continued to decline to a minimum LT50 of -4 to -7 degrees C after 10 days in a growth chamber for interior spruce, Douglas-fir and lodgepole pine, or after 15 days for western red cedar. Minimum freezing tolerance was reached at the stage of rapid hypocotyl elongation. In all species, a slight increase in freezing tolerance of germinants was observed once cotyledons emerged from the seed coat. The decrease in freezing tolerance during the transition from dry to germinating seed correlated with increases in seed water content. Changes in freezing tolerance between 10 and 30 days in the growth chamber were not correlated with seedling water content. Within a species, seedlots differed significantly in freezing tolerance after 2 or 5 days in the growth chamber. Because all seedlots of interior spruce and lodgepole pine germinated quickly, there was no correlation between seedlot hardiness and rate of germination. Germination rate and freezing tolerance of Douglas-fir and western red cedar seedlots was negatively correlated. There was a significant correlation between LT50 after 10 days in the growth chamber and minimum spring temperature at the location of seedlot origin for interior spruce and three seedlots of western red cedar, but no relationship was apparent for lodgepole pine and Douglas-fir.     

Effects of cold-hardening on compatible solutes and antioxidant enzyme activities related to freezing tolerance in Ammopiptanthus mongolicus seedlings

Cold acclimation is associated with many metabolic changes that lead to an increase of freezing tolerance. In order to investigate the biochemical process of cold acclimation in Ammopiptanthus mongolicus, seedlings were acclimated at 2℃ under 16-h photoperiod （150 μmol·m^-2·s^-1 photosynthetically active radiation） for 14 d. Freezing tolerance in seedlings increased after 14 d of cold-hardening. Contents of protein, proline and solute carbohydrate in cotyledon increased after cold acclimation. Patterns of isozymes of superoxide dismutase （SOD）, peroxidase, catalase and polyphenol oxidase （PPO） were investigated. The activities of SOD, peroxidase and PPO in cold acclimated plants were increased during cold-hardening. We deduced that compatible solutes and antioxidant enzymes play important roles in development of freezing tolerance during cold acclimation in this evergreen woody plant.     

Characterization and Role of Glucose-6-phosphate Dehydrogenase of Populus suaveolens in Induction of Freezing Resistance

Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) was purified from the leaves of 8-week-old Populus suaveolens cuttings. The enzyme activity in the absence and presence of reduced dithiothreitol (DTTred) was determined. The results show that the G6PDH activity is not inactivated by pre-incubation with DTTred, indicating that the purified enzyme probably presented in cytosol of P suaveolens. The catalytic characteristics and kinetic parameters of cytosolic G6PDH purified from P.suaveolens cuttings were also studied. The results show that G6PDH is characterized by Km value of 360/amol.L^-1 for G6P and 16μmol.L^-1 for NADP, a pH range of 7.3-8.9, and the maximum activity around pH 8.2. The enzyme activity is inhibited by various metabolites such as NADPH, NADH, GTP, UTP, ATP, AMP, ADP, CoA, acetyl CoA, fructose-6-phosphate (F6P), erythrose-4-phosphate (E4P), ribose-5-phosphate (R5P) and 3-phosphoglycerate (3-PG) (all at 1 mmol‘L i except for NADPH and NADH)to different extents. NADPH is the most effective inhibitor of enzyme activity, with an inhibition of 72.0%. The addition of metal ions such as MgC12, CaC12 and KC1 (all 1.0 mmol.L^-1) to the standard reaction mixture has no remarkable influence on the cytosolic G6PDH activity. However, CdCl2 (1.0 mmol.L-1) causes high inhibitory effect on the enzyme activity. To explore the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation, the changes in the cytosolic G6PDH activity and freezing resistance (expressed as LTs0) of P. suaveolens cuttings during freezing acclimation at -20℃ were investigated. The results reveal that freezing acclimation decreases LTs0 of cuttings, and increases the activity of cytosolic G6PDH compared with control ones,while 2 d of de-acclimation at 25 ~C result in a decrease in cytosolic G6PDH activity, and caused an increase in LT50. Furthermore,the change in cytosolic G6PDH activity is found to be closely correlated to the degree of freezing resistance of cuttings during freezing acclimation. It is suggested that cytosolic G6PDH may be involved in the induction of freezing resistance of cuttings.     

Verification of the resistance of a LEA gene from Tamarix expression in Saccharomyces cerevisiae to abiotic stresses

The role of late embryogenesis abundant （LEA） proteins in stress tolerance was examined by using a yeast expression system. LEA protein tolerance to the abotic stresses in plants involved in salt, drought and freezing stresses and additional tolerance to heat, NaHCO3 （salt-alkali） and ultraviolet radiation was also investigated. The transgenic yeast harboring the Tamarix LEA gene （DQ663481） was generated under the control of inducible GAL promoter （pYES2 vector）, yeast cells transformed with pYES2 empty vector were also generated as a control. Stress tolerance tests showed that LEA yeast transformants exhibited a higher survival rates than the control transformants under high temperature, NaHCO3, ultraviolet radiation, salt （NaCl）, drought and freezing, indicating that the LEA gene is tolerant to these abiotic stresses. These results suggest that the LEA gene is resistant to a wider repertoire of stresses and may play a common role in plant acclimation to the examined stress conditions.     

Changes in freezing tolerance, plasma membrane H+-ATPase activity and fatty acid composition in Pinus resinosa needles during cold acclimation and de-acclimation

It has previously been suggested that plasma membrane ATPase (PM H+-ATPase, EC 3.6.1.3.) is a site of incipient freezing injury because activity increases following cold acclimation and there are published data indicating that activity of PM H+-ATPase is modulated by changes in lipids associated with the enzyme. To test and extend these findings in a tree species, we analyzed PM H+-ATPase activity and the fatty acid (FA) composition of glycerolipids in purified plasma membranes (PMs) prepared by the two-phase partition method from current-year needles of adult red pine (Pinus resinosa Ait.) trees. Freezing tolerance of the needles decreased from -56 degrees C in March to -9 degrees C in May, and increased from -15 degrees C in September to -148 degrees C in January. Specific activity of vanadate-sensitive PM H+-ATPase increased more than two-fold following cold acclimation, despite a concurrent increase in protein concentration. During de-acclimation, decreases in PM H+-ATPase activity and freezing tolerance were accompanied by decreases in the proportions of oleic (18:1) and linoleic (18:2) acids and increases in the proportions of palmitic (16:0) and linolenic (18:3) acids in total glycerolipids extracted from the plasma membrane fraction. This pattern of changes in PM H+-ATPase activity and the 18:1, 18:2 and 18:3 fatty acids was reversed during cold acclimation. In the PM fractions, changes in FA unsaturation, expressed as the double bond index (1 x 18:1 + 2 x 18:2 + 3 x 18:3), were closely correlated with changes in H+-ATPase specific activity (r2 = 0.995). Changes in freezing tolerance were well correlated with DBI (r2 = 0.877) and ATPase specific activity (r2 = 0.833) in the PM fraction. Total ATPase activity in microsomal fractions also closely followed changes in freezing tolerance (r2 = 0.969). We conclude that, as in herbaceous plants, simultaneous seasonal changes in PM H+-ATPase activity and fatty acid composition occur during cold acclimation and de-acclimation in an extremely winter hardy tree species under natural conditions, lending support to the hypothesis that FA-regulated PM H+-ATPase activity is involved in the cellular response underlying cold acclimation and de-acclimation.     

Cold acclimation induced accumulation of phenolic compounds and freezing tolerance in Ammopiptanthus mongolicus

Ammopiptanthus mongolicus, the only freezing tolerant evergreen broad-leaved shrub, local species of the Alashan desert, northwest sand area of China, can survive -30°C or even lower temperature in winter. In the present study, the secondary products phenolics in A. mongolicus cotyledons were determined to study the effects of phenolics on cold tolerance. Cytochemical localization of phenolics in cotyledon cells was observed by electron microscopy and the content of phenolic compounds was assayed by spec-trophotometric measurement. The results showed that the freezing tolerance of A. mongolicus seedlings increased after acclimation at 2-6°C for 14 days, which accompanied the increase of the content of phenolic compounds in cotyledons. Cytochemical observation showed that phenolic deposits were mainly localized in vacuoles and in close proximity to tonoplast, and also in the cytoplasm. The amount and the size of phenolics droplets increased obviously in cytoplasm and vacuoles after cold acclimation, predominantly ag-gregated along membranes of vacuoles and tonoplast. No phenolic deposits were found in cell walls. As hydrogen-or elec-tron-donating agents, phenolics may protect plant cells against reactive oxygen species formed during chilling or freezing stress and improve the freezing tolerance of cold-acclimated A. mongolicus seedlings.     

南京地区引种的6种常绿阔叶树种抗寒性测试

选取南京地区已引种成功的6种常绿阔叶树种,运用越冬适应性观察与电导法作为对比测试手段,分析这些树种的抗寒能力。结果表明,采用低温循环仪自动降温程序模拟冰冻处理来测试叶片的相对电导率,计算出各树种的半致死温度(LT50)能定量地反映这些树种本身的抗寒能力高低,并且与它们在南京地区越冬适应性的观察基本相符。这些研究结果对预测常绿阔叶树种抗寒能力以及在较高纬度地区园林中的应用具有指导意义。