Vegetative growth and frost hardiness of cloudberry (Rubus chamaemorus) as affected by temperature and photoperiod

The effect of photoperiod and temperature on growth and induction and development of frost hardiness in cloudberry ( Rubus chamaemorus L.) was examined in two experiments. The photoperiods were 8, 12 or 24 h and the temperatures were 18, 15, 12, 9, 4, 3, –3 or –4°C depending on the experiment. The level of hardiness was expressed as LT₆₆ or LT₅₀ (the lethal temperature for 66 or 50% of the plant material) for percentage of bud break and for the degree of coloring by triphenyltetrazolium chloride for rhizomes. The vegetative growth was clearly affected by daylength; petiole elongation, leaf growth, shoot dry weight and number of shoots per plant were all reduced under short days compared with long days. However, the photoperiod had no significant effect on hardening of buds or rhizomes. Hardening increased with successively decreasing temperatures. To get the maximum hardiness, plants had to be exposed to freezing temperatures.     

The long day leaf as a source of cold hardiness inhibitors

Short photoperiods followed by low temperatures induced cold hardiness in Acer negundo, Viburnum plicatum tomentosum, and Weigela florida. Hardiness was also obtained under long days and natural fall temperatures if the leaves were removed, either manually or by low temperature. Similarly, removal of leaves from plants exposed to long days at 5° brought about an accelerated rate of hardening. These observations suggested the presence of a hardiness inhibitor in the leaves which was counteracted by short days or removal of the leaves.     

Changing Environmental Effects on Frost Hardiness of Scots Pine During Dehardening

The effects of raised temperature and extended photoperiod onthe dehardening of quiescent and winter-hardy Scots pine saplingswere examined in an open-top-chamber experiment. The saplingswere exposed during winter to natural, square-curve fluctuating(between 1 and 11 °C with a 14 d interval), and constant(6 °C) temperatures with a natural and an extended (17 h)photoperiod. Frost hardiness of needles was determined by controlledfreezing tests and visual damage scoring. The constant 6 °Ctemperature treatment caused a gradual dehardening of needleswhereas under fluctuating temperatures the level of frost hardinessfluctuated. Trees exposed to extended photoperiods were lesshardy than under natural photoperiods after the initiation ofshoot elongation, but before this there were no clear differencesin frost hardiness between different photoperiodic treatments.The results indicate that the frost hardening competence ofScots pine changes during quiescence. Climate change; frost hardiness; hardening competence; photoperiod; Pinus sylvestris, Scots pine; temperature     

光周期对中华通草蛉自然越冬成虫及实验种群幼虫耐寒能力的影响

【目的】滞育诱导期进行短光照处理可影响昆虫耐寒性。为明确光周期对中华通草蛉Chrysoperla sinica (Tjeder)耐寒性的影响, 针对中华通草蛉滞育解除过程及非滞育虫态的耐寒性进行了一系列研究。【方法】测定了中华通草蛉自然越冬成虫的过冷却点（supercooling point, SCP）以及长光周期（15L∶9D）和短光周期（9L∶15D）条件下自然越冬成虫在滞育解除过程中在-12℃下的死亡率, 并测定了室内长、 短两种光周期下实验种群2龄和3龄幼虫的过冷却点（SCP）、 结冰点（freezing point, FP）以及-7℃下的死亡率。【结果】中华通草蛉12月份的自然越冬成虫SCP集中在-10~-14℃之间。SCP低于-12℃的个体占43.70%, 且-12℃处理1 d死亡率为62.00%。-12℃处理1 d条件下的长、 短光周期处理自然越冬成虫, 除处理0 d外, 长光周期处理死亡率均高于短光周期处理的, 且在处理15 （P=0.012）, 20 （P=0.01）和25 d （P=0.001）差异显著。中华通草蛉试验种群相同龄期幼虫在短光周期下的SCP和FP均高于长光周期下, 但差异不显著(P>0.05); 但在-7℃下, 2龄幼虫短光周期下的低温死亡率为67.00%±4.04%, 显著低于长光周期下的低温死亡率（78.00%±1.33%）（P=0.011）, 3龄幼虫短光周期条件下低温死亡率为24.33%±1.33%, 显著低于长光周期下的低温死亡率（53.00%±3.46%）（P=0.002）。【结论】中华通草蛉为结冰敏感型, 诱导滞育的短光照处理可提高其幼虫期及滞育解除过程中成虫的耐寒能力。     

Environmental control of cold hardiness in woody plants

The development of cold hardiness in 2 woody plant species (Acer negundo and Viburnum plicatum tomentosum) was shown to be independent of the induction of bud dormancy. Substantial hardiness levels were obtained under controlled conditions with long days and certain low temperatures—without dormancy development as a prerequisite.

Low temperatures given during the dark period with long days induced hardiness to a level not significantly different from that of short days. Giving plants continuous 10° temperatures under long days forced plants to harden as if they were under short days, even though they were not dormant.

Development of hardiness was shown to be a photoperiodic response. Increasing weeks of short days, followed by a low temperature hardening period in darkness, brought about a progressive increase in hardiness. The short day stimulus could be reversed by long days. Following 6 weeks of short days, the rate of hardening in darkness at 5° was over twice that of plants previously exposed to long days.

     

Low night temperature and inhibition of gibberellin biosynthesis override phytochrome action and induce bud set and cold acclimation, but not dormancy in PHYA overexpressors and wild-type of hybrid aspen

Juvenile trees of temperate and boreal regions cease growth and set buds in autumn in response to short day-lengths (SD) detected by phytochrome. Growth cessation and bud set are prerequisites for the development of winter dormancy and full cold hardiness. In this study we show that the SD-requirement for bud set and cold hardening can be overcome in hybrid aspen (Populus tremula L. × tremuloides Michx.) by low night temperature and inhibition of gibberellin (GA) biosynthesis. Bud set and increased cold hardiness were observed under normally non-inductive long day-length (LD) in wild-type plants, when exposed to low night temperature and paclobutrazol. In addition, the effect of PHYA overexpression could be overcome in transgenic plants, producing bud set and cold acclimation by treatment with: SD, low night temperature and paclobutrazol. After cold acclimation, the degree of bud dormancy was lower for wild-type plants prior treated with LD and transgenic plants (overexpressing PHYA), than SD-treated, wild-type plants. Thus, low night temperature in combination with reduced GA content induced bud set and promoted cold hardiness under normally non-inductive photoperiods in hybrid aspen, but was unable to affect development of dormancy. This might suggest separate signalling pathways from phytochrome regulating the induction of cold/cold hardiness and bud dormancy in hybrid aspen or alternatively, there might be one pathway that fails to complete its action in the transgenic and paclobutrazol treated plants.     

Photoperiod control of the initial phase of frost hardiness development in Pinus radiata

Abstract. The development of frost hardiness in Pinus radiata was investigated to establish whether there is quantitative relationship between photoperiod and the hardening process. Three controlled environment experiments were carried out. In the first, seedlings were exposed to a photoperiod that shortened from 13 h at a rate of 3 min d^?1 to 9.5 h. At intervals, the photoperiod was either held constant or lengthened. In the second experiment, seedlings were exposed to one of five constant photoperiods between 9 and 12 h for up to 90 d. In the third, seedlings were exposed to photoperiods shortening at rates of 1 or 5 min d^?1. Frost hardiness was also measured during the natural photoperiod-controlled stage of hardening in outdoor-grown seedlings. Frost hardiness developed at a constant rate in response to a shortening photoperiod once it had declined to about 12 h. This rate was consistent with the hardening rate that occurred in outdoor-grown seedlings. Hardening stopped when the photoperiod became constant, indicating a tight coupling between changes in photoperiod and hardiness development. When the photoperiod was held constant, the extent of frost hardiness was directly dependent on the photoperiod but the rate of hardening was apparently independent of the length of photoperiod. However, the rate of hardening was dependent on the rate at which the photoperiod shortened, increasing linearly with increases in the rate of change in photoperiod between 0 and 3 min d^?1. These results suggest shortening photoperiods control the first stage of the hardening process by regulating the rate of hardening. Frost hardening was inherently unstable once the maximum hardiness was reached since spontaneous dehardening occurred in spite of the controlled conditions. Dehardening also occurred when the photoperiod was lengthened suggesting that the cue for dehardening to commence was the shift from shortening to lengthening photoperiods.     

Frost hardening and photosynthetic performance of Scots pine (Pinus sylvestris L.) needles. I. Seasonal changes in the photosynthetic apparatus and its function

Photosynthetic CO₂ uptake, the photochemical efficiency of photosystem II, the contents of chlorophyll and chlorophyll-binding proteins, and the degree of frost hardiness were determined in three-year-old Scots pine (Pinus sylvestris L.) trees growing in the open air but under controlled daylength. The following conditions were compared: 9-h light period (short day), 16-h light period (long day), and natural daylength. Irrespective of induction by short-day photoperiods or by subfreezing temperatures, frost hardening of the trees was accompanied by a long-lasting pronounced decrease in the photosynthetic rates of one-year-old needles. Under moderate winter conditions, trees adapted to a long-day photoperiod, assimilated CO₂ with higher rates than the short-day-treated trees. In the absence of strong frost, photochemical efficiency was lower under short-day conditions than under a long-day photoperiod. Under the impact of strong frost, photochemical efficiency was strongly inhibited in both sets of plants. The reduction in photosynthetic performance during winter was accompanied by a pronounced decrease in the content of chlorophyll and of several chlorophyll-binding proteins [light-harvesting complex (LHC)IIb, LHC Ib, and a chlorophyll-binding protein with MW 43 kDa (CP 43)]. This observed seasonal decrease in photosynthetic pigments and in pigment-binding proteins was irrespective of the degree of frost hardiness and was apparantly under the control of the length of the daily photoperiod. Under a constant 9-h daily photoperiod the chlorophyll content of the needles was considerably lower than under long-day conditions. Transfer of the trees from short-day to long-day conditions resulted in a significantly increased chlorophyll content, whereas the chlorophyll content decreased when trees were transferred from a long-day to a short-day photoperiod. The observed changes in photosynthetic pigments and pigment-binding proteins in Scots pine needles are interpreted as a reduction in the number of photosynthetic units induced by shortening of the daily light period during autumn. This results in a reduction in the absorbing capacity during the frost-hardened state. Received: 3 March 1997 / Accepted: 16 July 1997     

Effect of growth regulator and culture conditions on shoot multiplication and rhizome formation in ginger (Zingiber officinale Rosc.) in vitro

Summary Shoot multiplication of Zingiber officinale cv. V₃S₁₈ was achieved by meristem culture on a Murashige and Skoog (MS) basal medium supplemented with 26.6 μM 6-benzylaminopurine (BA), 8.57 μM indole-3-acetic acid (IAA), and 1111.1 μM adenine sulfate and 3% (w/v) sucrose. In vitro rhizome formation from in vitro-raised shoots was achieved on MS medium supplemented with 4.44 μM BA, 5.71 μM IAA, and 3–8% (w/v) sucrose after 8 wk of culture. Cultural variations such as photoperiod, carbohydrate, nutrient composition, and growth regulators were tested for the maximum yield of rhizomes. Among the different photoperiods used, a 24-h photoperiod helped in the formation of more rhizomes as compared with other photoperiods. Of the different carbohydrates used, sucrose helped to achieve rhizome formation as compared to other carbohydrates. The microrhizomes sprouted in a soil mixture within 2 wk of planting. The sprouted plantlets survived under field conditions with normal growth.     

Is tissue culture a viable system with which to examine environmental and hormonal regulation of cold acclimation in woody plants?

In vitro-grown saskatoon berry (Amelanchier alnifolia Nutt.) plantlets were exposed to various hormonal treatments, dormancy-inducing and cold acclimation conditions to determine if this in vitro system would be viable for dormancy/hardiness studies in woody plants. Low temperature induced significant hardiness levels in plantlets to ?27°C after 6 weeks at 4°C but did not approach liquid nitrogen levels of fully hardened, field-grown buds. Control plantlets were consistently killed at ?5°C throughout this period. Significant hardiness was attained under both short and long day/low temperature conditions; however, hardiness was reduced under continuous light or dark treatments. A pre-exposure to the typical short photoperiod regime of woody plants did not significantly increase the rate of acclimation in these plantlets. The presence/absence of phytohormones in the media have a pronounced influence on the ability of plantlets to cold acclimate. Hormone-free media increased hardiness to ?10.5°C after 2 weeks in treatment. Addition of abscisic acid (ABA) increased cold hardiness levels (?12°C) while addition of benzylaminopurine (BAP) to this hormone-free media decreased hardiness to ?5.3°C. A combination of BAP and ABA treatments produced LT₅₀ values intermediate between individual applications of either hormone. Conversely, α-naphthaleneacetic acid (NAA) could not counteract the ABA-induced hardiness. ABA treatments alone were not able to harden plantlets to the extent attained under low temperature acclimation conditions. Further, ABA could not maintain the hardiness levels of cold-acclimating treatments and plantlets de-acclimated to ?9°C in BAP + ABA media. Subculturing in itself significantly elevated cold hardiness in plantlets to ?9°C on BAP + NAA media within 3 days after subculture and thereafter plantlets dehardened to ?5°C. While tissue culture has value in specific cases, caution should be taken when using tissue-cultured plantlets as a system to evaluate environmental regulation of cold acclimation in woody plants, in part, due to the influence of phytohormones in the media.     

Growth responses of Typha orientalis presl to controlled temperatures and photoperiods

Experiments are described in which seedlings of Typha orientalis Presls were grown for up to 6 months under precise conditions of temperature and photoperiod; photosynthesis was by natural daylight and did not vary between treatments. Variable treatments were imposed either from the seedling stage or on large plants raised under constant conditions.In general, total dry matter production increased as photoperiod increased from 8 to 16 h and also as day or night temperature increased, maximum production occurring when there was a warm day (30 or 27°C) and a small temperature drop (to 22°C) at night. The distribution of dry matter was also markedly affected by the imposed variables, leaf growth being favoured by high temperatures (to 30°C) and long photoperiods, and production of roots and rhizomes by low temperatures (to 10°C) and short photoperiods. None of the treatments resulted in floral initiation. The results are considered in relation to growth in the natural habitat.     

Regulation of the loss of frost hardiness in Pinus radiata by photoperiod and temperature

Abstract. In controlled environments, the interactive effects of warm (16: 8°C, day: night) and cool (12: 4°C, day: night) temperatures and long (13.5 h) and short (10 h) photoperiods on the dehardening of seedlings of Pinus radiata D. Don were investigated. In another experiment, the effect of four photoperiods from 9 to 14 h was examined. In a third, dehardening at constant temperatures from 5 to 17°C was followed. There was no evidence for an interaction between photoperiod and temperature. Dehardening was temporarily delayed by photoperiods below about 10 h, but there was no other quantitative effect of photoperiod. At constant temperatures, the rate of dehardening was initially constant but declined as the minimum summer frost hardiness was reached. In the initial phase the rate of dehardening was a linear function of temperature, increasing from 0.05°C day⁻¹ at 8°C to 0.30 °C day⁻¹ at 17°C. Temperature controlled the loss of frost hardiness by regulating the rate of dehardening.     

The Role of Light in Cold Acclimation of Hedera helix L. var. Thorndale

The role of light in cold acclimation of Hedera helix L. var. Thorndale appears to differ from that reported for winter annuals. Although light greatly enhances the degree of hardiness attained, cold acclimation is not obligatorily linked to a light requirement. Photoperiods, varying from 8 to 24 hours, received during the cold acclimation period were equally effective in promoting maximum hardiness. Relatively low light intensities and short photoperiods stimulated maximum hardiness, and proportional increases in hardiness in response to increased photoperiods were demonstrated only in stems of prestarved plants. Exclusion of CO₂ and high concentrations of photosynthetic inhibitors decreased hardiness, but in no instance was hardiness reduced to the level of the dark control. The data are only compatible with a photosynthetic role of light if it is assumed that only a small portion of the total photosynthates are required to elicit maximum hardiness. Alternatively, the light stimulation which was elicited by low light intensities, short photoperiods, in the absence of CO₂, and in the presence of photosynthetic inhibitors, may be a light signal similar to a phytochrome response.     

Control of summer and winter diapause in the leaf-mining fly Pegomyia bicolor Wiedemann (Dipt., Anthomyiidae)

Effects of photoperiod and temperature on diapause induction and termination were investigated in both aestival and hibernal pupae of Pegomyia bicolor Wiedemann under field and laboratory conditions. In the field, summer diapause had occurred already in part of the first pupal population; the proportion of diapause gradually rose as the day length and temperature increased. This fly is a short-day species with a pupal summer and winter diapause. Summer diapause was induced by both long day-lengths and mild temperatures. The whole larval life is sensitive to photoperiod. Winter diapause was induced mainly by low temperatures, especially in the first 10 days after pupation. High temperatures strongly enhanced summer diapause induction regardless of photoperiod. The diapause-averting influence of short photoperiods was fully expressed only at moderately low temperatures. High temperatures delayed diapause development, resulting in a rather long summer diapause; whereas low temperatures hastened it, leading to a short winter diapause and showing a low thermal threshold for diapause development. In the field, the post-diapause development started in January, the coldest month, suggesting that the thermal requirements for post-diapause development is also low.     

Environmental regulation and physiological basis of freezing tolerance in woody plants

Cold acclimation of plants is a complex process involving a number of biochemical and physiological changes. The ability to cold acclimate is under genetic control. The development of freezing tolerance in woody plants is generally triggered by non-freezing low temperatures but can also be induced by mild drought or exogenous abscisic acid, as well as by short photoperiod. In nature, the extreme freezing tolerance of woody plants is achieved during sequential stages of cold acclimation the first of which is initiated by short photoperiods and non-freezing low temperatures, and the second by freezing temperatures. Although recent breakthroughs have increased our knowledge on the physiological molecular basis of freezing tolerance in herbaceous species, which acclimate primarily in response to non-freezing low temperatures, very little is known about cold acclimation of woody plants. This article attempts to review our current understanding of the physiological aspects that underline cold acclimation in woody plants.     

Temperature-driven plasticity in growth cessation and dormancy development in deciduous woody plants: a working hypothesis suggesting how molecular and cellular function is affected by temperature during dormancy induction

The role of temperature during dormancy development is being reconsidered as more research emerges demonstrating that temperature can significantly influence growth cessation and dormancy development in woody plants. However, there are seemingly contradictory responses to warm and low temperature in the literature. This research/review paper aims to address this contradiction. The impact of temperature was examined in four poplar clones and two dogwood ecotypes with contrasting dormancy induction patterns. Under short day (SD) conditions, warm night temperature (WT) strongly accelerated timing of growth cessation leading to greater dormancy development and cold hardiness in poplar hybrids. In contrast, under long day (LD) conditions, low night temperature (LT) can completely bypass the short photoperiod requirement in northern but not southern dogwood ecotypes. These findings are in fact consistent with the literature in which both coniferous and deciduous woody plant species’ growth cessation, bud set or dormancy induction are accelerated by temperature. The contradictions are addressed when photoperiod and ecotypes are taken into account in which the combination of either SD/WT (northern and southern ecotypes) or LD/LT (northern ecotypes only) are separated. Photoperiod insensitive types are driven to growth cessation by LT. Also consistent is the importance of night temperature in regulating these warm and cool temperature responses. However, the physiological basis for these temperature effects remain unclear. Changes in water content, binding and mobility are factors known to be associated with dormancy induction in woody plants. These were measured using non-destructive magnetic resonance micro-imaging (MRMI) in specific regions within lateral buds of poplar under SD/WT dormancing inducing conditions. Under SD/WT, dormancy was associated with restrictions in inter- or intracellular water movement between plant cells that reduces water mobility during dormancy development. Northern ecotypes of dogwood may be more tolerant to photoinhibition under the dormancy inducing LD/LT conditions compared to southern ecotypes. In this paper, we propose the existence of two separate, but temporally connected processes that contribute to dormancy development in some deciduous woody plant: one driven by photoperiod and influenced by moderate temperatures; the other driven by abiotic stresses, such as low temperature in combination with long photoperiods. The molecular changes corresponding to these two related but distinct responses to temperature during dormancy development in woody plants remains an investigative challenge.     

Inheritance of the photoperiodically induced cold acclimation response in Cornus sericea L., red-osier dogwood

Summary Cold acclimation responses of latitudinal ecotypes of Cornus sericea L. (C. stolonifera Michx.) and F₁, F₂ and BC₁ hybrid progenies were measured under natural photoperiod conditions in St. Paul, MN and artificially shortened photoperiods in the glass-house. The 65 °N and 62 °N ecotypes (Alaska and Northwest Territories, respectively) were characterized by a short night length for hardiness induction, the 42 °N ecotype (Utah A and B) by a long night length for hardiness induction, while the F₁ was intermediate to the parents. Results from reciprocal crosses indicated there was no significant unilateral maternal influence on cold acclimation. Acclimation responses of the F₂ were highly variable but generally ranged between the parental extremes. However, three individuals from the 42 ° × 62 °N crosses exhibited greater cold resistance than the northern parent on two successive freezing test dates. F₂ plants were also found with less freezing resistance than the southern parent. Backcrosses to the southern parent produced progeny with acclimation patterns resembling that of the southern parent and were significantly less hardy than the F₂ in early freezing tests.Scientific Journal Series Paper No. 12,075 of the Minnesota Agricultural Experiment Station     

Rhizome transition to storage organ is under phytochrome control in lotus <Emphasis Type="Italic">(Nelumbo nucifera)</Emphasis>

We examined photoperiodic response of lotus (Nelumbo nucifera) rhizome morphogenesis (its transition to a storage organ) by using seed-derived plants. Rhizome enlargement (increase in girth) was brought about under 8, 10 and 12 h photoperiods, whereas the rhizomes elongated under 13 and 14 h photoperiods. Rhizomes elongated under 14 h light regimes supplied as 8 h of natural light plus 6 h supplemental hours of white, yellow or red light, but similar treatments with supplemental blue, green or far red light, caused enlargement in girth of the rhizomes. A 2 h interruption of the night with white, yellow or red light, in plants entrained to 8 h photoperiod brought rhizome elongation, whereas 2 h-blue, green or far red light night breaks still resulted in rhizome increase in girth. The inhibitory effect of a red (R) light night break on rhizome increase in girth was reversed by a far-red (FR) light given immediately afterwards. Irradiation with R/FR/R inhibited the rhizome increase in girth. FR light irradiation following R/FR/R irradiation cancelled the effect of the last R light irradiation. It was demonstrated that the critical photoperiod for rhizome transition to storage organ is between 12 and 13 h photoperiod. It was also evident that the optimal light quality range for interruption of dark period (night break) is between yellow and red light and that a R/FR reversible reaction is observed. From these results, we propose that phytochrome plays an important role in photoperiodic response of rhizome increase in girth in lotus. This is the first report on phytochrome-dependent morphogenesis of storage organs in rhizomous plants.     

Habitat temperature and the temporal scaling of cold hardening in the high Arctic collembolan, Hypogastrura tullbergi (Schäffer)

Abstract.  1. Cold tolerance is a fundamental adaptation of insects to high latitudes. Flexibility in the cold hardening process, in turn, provides a useful indicator of the extent to which polar insects can respond to spatial and temporal variability in habitat temperature.
2. A scaling approach was adopted to investigate flexibility in the cold tolerance of the high Arctic collembolan, Hypogastrura tullbergi , over different time-scales. The cold hardiness of animals was compared from diurnal warming and cooling phases in the field, and controlled acclimation and cooling treatments in the laboratory. Plasticity in acclimation responses was examined using three parameters: low temperature survival, cold shock survival, and supercooling points (SCPs).
3. Over time-scales of 24–48 h, both field animals from warm diurnal phases and laboratory cultures from a 'warm' acclimation regime (18 °C) consistently showed greater or equivalent cold hardiness to animals from cool diurnal phases and acclimation regimes (3 °C).
4. No significant evidence was found of low temperature acclimation after either hours or days of low temperature exposure. The cold hardiness of H. tullbergi remained 'seasonal' in character and mortality throughout was indicative of the summer state of acclimatization.
5. These data suggest that H. tullbergi employs an 'all or nothing' cryoprotective strategy, cold hardening at seasonal but not diel-temporal scales.
6. It is hypothesised that rapid cold hardening offers little advantage to these high Arctic arthropods because sub-zero habitat temperatures during the summer on West Spitsbergen are rare and behavioural migration into soil profiles offers sufficient buffering against low summer temperatures.     

Causes of variation in cold hardiness among fast-growing willows (Salix spp.) with particular reference to their inherent rates of cold hardening

Causes of variation in cold hardiness in the autumn were assessed among closely related, fast‐growing clones of willow of northern/continental and southern/maritime origins, under controlled regimes and natural conditions. Cold hardiness was assessed by controlled freezing followed by injury analysis, based on measurements of chlorophyll fluorescence (stems) and electrolyte leakage (leaves). During growth at a given temperature, the cold hardiness of the clones' stems was negatively correlated with their rate of growth. This apparently phenotypic variation was independent of temperature and, hence, the absolute growth rate. At later stages, cold hardiness of stems varied mainly with respect to genetic differences in the timing and rate of cold hardening. Cold hardening began up to 7 weeks earlier in northern/continental clones, and their rates of hardening in cool temperature regimes were up to three times higher than in southern/maritime clones. Ranking of clones with respect to rates was essentially the same whether natural or abrupt reductions of day length were used to trigger cold hardening. Results closely agreed with those of a previous field trial. Comparisons of rates at cool and warm temperatures suggest that cold hardening became increasingly dependent on cool temperatures with time. Increasing sucrose‐to‐glucose ratios, and especially dry‐to‐fresh weight ratios, paralleled early cold hardening. Before leaves were shed in the autumn they underwent cold hardening in parallel with stems, eventually allowing them to tolerate temperatures down to ?10 °C.