Effect of frost nights and day and night temperature during dormancy induction on frost hardiness, tolerance to cold storage and bud burst in seedlings of Norway spruce

For trees, the ability to obtain and maintain sufficient levels of frost hardiness in late autumn, winter and spring is crucial. We report that temperatures during dormancy induction influence bud set, frost hardiness, tolerance to cold storage, timing of bud burst and spring frost hardiness in seedlings of Norway spruce (Picea abies (L.) Karst.). Bud set occurred later in 12°C than in 21°C, and later in cool nights (7°C) than in constant temperature. One weekly frost night (−2.5°C) improved frost hardiness. Cool nights reduced frost hardiness early, but improved hardiness later during cold acclimation. Buds and stems were slightly hardier in 21°C than in 12°C, while needles were clearly hardier in 12°C. Cold daytime temperature, cool nights and one weekly frost night improved cold storability (0.7°C). Seedlings receiving high daytime temperatures burst buds later, and were less injured by light frost some days after bud burst.     

Warm temperature accelerates short photoperiod-induced growth cessation and dormancy induction in hybrid poplar (Populus × spp.)

There is increasing evidence that temperature, in addition to photoperiod, may be an important factor regulating bud dormancy. The impact of temperature during growth cessation, dormancy development, and subsequent cold acclimation was examined in four hybrid poplar clones with contrasting acclimation patterns: ‘Okanese’—EARLY, ‘Walker’—INT1, ‘Katepwa’—INT2, and ‘Prairie Sky’—LATE. Four day–night temperature treatments (13.5/8.5, 18.5/13.5, 23.5/8.5, and 18.5/3.5°C) were applied during a 60-day induction period to reflect current and predicted future annual variation in autumn temperature for Saskatoon, SK. Warm night temperature (18.5/13.5°C) strongly accelerated growth cessation, dormancy development, and cold acclimation in all four clones. Day temperature had the opposite effect of night temperature. Day and night temperatures appeared to act antagonistically against each other during growth cessation and subsequent dormancy development and cold acclimation. Growth cessation, dormancy development, and cold acclimation in EARLY and LATE were less affected by induction temperature than INT1 and INT2 suggesting that genotypic variations exist in response to temperature. Separating specific phenological stages and the impact by temperature on each clone revealed the complexity of fall phenological changes and their interaction with temperature. Most importantly, future changes in temperature may affect time to growth cessation, subsequently altering the depth of dormancy and cold hardiness in hybrid poplar.     

Chill-responsive dehydrins in blueberry: Are they associated with cold hardiness or dormancy transitions?

To survive winters, woody perennials of temperate zones must enter into endodormancy. Resumption of spring growth requires sufficient exposure to low temperature (chill units, CUs) in winter (chilling requirement), which also plays a role in the development of cold hardiness (cold acclimation). Physiological studies on dormancy breaking have focused on identifying markers, such as appearance or disappearance of proteins in response to varying degrees of chill unit accumulation. However, whether these changes are associated with dormancy transitions or cold acclimation is not clear. In the present study, greenhouse-grown blueberry (Vaccinium section Cyanococcus) plants were used to address this question. Three blueberry cultivars, Bluecrop, Tifblue, and Gulfcoast having chilling requirement of approximately 1 200, 900 and 600 CUs, respectively, were first exposed to 4°C for long enough to provide chill units equivalent to one-half of their respective chilling requirement. This treatment was expected to result in cold acclimation. A fraction of plants was then subjected to a 15/12°C (light/dark) regime for 2 weeks, a treatment expected to be “dormancy-neutral” but cause deacclimation. Before and after each treatment, cold hardiness and dormancy status of floral buds were determined; proteins were extracted from the buds collected on the same sampling date, and separated by one-dimensional SDS-PAGE. Dehydrin-like proteins were identified by immunoblotting, using anti-dehydrin antiserum. Results indicate that the chilling treatment resulted in cold acclimation as indicated by increased bud hardiness in all three cultivars. Data also indicate a distinct accumulation of three dehydrin-like proteins of 65, 60, and 14 kDa during cold acclimation. The cold hardiness and levels of dehydrin proteins decreased during the exposure to 15/12°C for 2 weeks. Results also confirmed that this treatment had no negative effect on chill unit accumulation. Densitometric scans of protein gels indicated a close association between the abundance of dehydrins and degree of cold hardiness in these cultivars. In addition, levels of the dehydrin proteins and cold hardiness remained about the same between 100% and >100% satisfaction of chilling requirement. These results suggest that changes in dehydrin expression are more closely associated with cold hardiness than with dormancy transitions.     

Changes in endogenous abscisic acid and cold hardiness in Actinidia treated with triazole growth retardants

Soil drench of either paclobutrazol or uniconazole (0.4 mg/pot) was applied to plants of Actinidia arguta, Ananasnaja, to determine the effect on endogenous abscisic acid measured in November, January, and March, and the concomittant cold hardiness of the treated plants. Both paclobutrazol and uniconazole treatments significantly increased cold hardiness. Paclobutrazol was more effective in increasing ABA levels compared to uniconazole. Abscisic acid was found to be highest in January, corresponding to deep dormancy, and least in March when plants were undergoing vegetative bud break. Paclobutrazol delayed vegetative bud break by 6.3 days, while uniconazole delayed bud break by 2.9 days.     

Phytochrome control of short-day-induced bud set in black cottonwood

In trees and other woody perennial plants, short days (SDs) typically induce growth cessation, the initiation of cold acclimation, the formation of a terminal bud and bud dormancy. Phytochrome control of SD-induced bud set was investigated in two northern clones of black cottonwood (Populus trichocarpa Torr. & Gray) by using night breaks with red light (R) and far-red light (FR). For both clones (BC-1 and BC-2), SD-induced bud set was prevented when R night breaks as short as 2 min were given in the middle of the night. When night breaks with 2 min of R were immediately followed by 2 min of FR, substantial reversibility of bud set was observed for BC-1 but not for BC-2. By comparing the effects of the R night breaks on bud set and the length of specific internodes, we determined that the R night breaks influenced internode elongation in two opposing ways. First, the addition of a R night break to the SD treatment prevented the cessation of internode elongation that is associated with bud set. Those internodes that would not have elongated under SDs (and would have been found within the terminal bud) elongated in the R treatment. Second, the R night breaks decreased internode length relative to the long-day (LD) control. In contrast to the clonal differences in reversibility that we observed for bud set, the decrease in internode length (i.e. the second effect of R) was R/FR reversible in both clones. Based on these results, we conclude that internode elongation is influenced by two distinct types of phytochrome-mediated response. The first response is a typical response to photpperiod, whereas the second response is a typical “end-of-day” response to light quality. Our results demonstrate that SD-induced bud set in black cottonwood is controlled by phytochrome but that clonal differences have an important influence on the R/FR reversibility of this response. The availability of an experimental system in which SD-induced bud set is R/FR reversible will be valuable for studying the physiological genetics of photoperiodism in trees.     

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.     

Responses of transgenic <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> seedlings expressing a <Emphasis Type="Italic">Cucurbita pepo</Emphasis> antisense <Emphasis Type="Italic">PHYA</Emphasis> RNA to far-red radiation

The Nicotiana tabacum transgenic plants expressing a Cucurbita pepo antisense PHYA RNA were obtained. The seedlings of transgenic tobacco with reduced phytochrome A (PHYA) content displayed decreased sensitivity to continuous broad-band far-red radiation (λ > 680 nm). Under far-red irradiance transgenic seedlings showed less elongation of the hypocotyls, more rapid plastid development, more chlorophyll accumulation, less repression of lightdependent NADPH:protochlorophyllide oxidoreductase than wild-type plants that was in accordance with PHYA control of plant development. Dynamics of the far-red radiation dependent changes in low temperature chlorophyll fluorescence spectra for the transgenic and wild-type seedlings were consistent with the more rapid formation of photosynthetic apparatus in the seedlings with reduced PHYA.     

Ecotype-dependent control of growth,dormancy and freezing tolerance under seasonal changes in <Emphasis Type="Italic">Betula pendula</Emphasis> Roth

Woody plants in the temperate and boreal zone undergo annual cycle of growth and dormancy under seasonal changes. Growth cessation and dormancy induction in autumn are prerequisites for the development of substantial cold hardiness in winter. During evolution, woody plants have developed different ecotypes that are closely adapted to the local climatic conditions. In this study, we employed distinct photoperiodic ecotypes of silver birch (Betula pendula Roth) to elucidate differences in these adaptive responses under seasonal changes. In all ecotypes, short day photoperiod (SD) initiated growth cessation and dormancy development, and induced cold acclimation. Subsequent low temperature (LT) exposure significantly enhanced freezing tolerance but removed bud dormancy. Our results suggested that dormancy and freezing tolerance might partially overlap under SD, but these two processes were regulated by different mechanisms and pathways under LT. Endogenous abscisic acid (ABA) levels were also altered under seasonal changes; the ABA level was low during the growing season, then increased in autumn, and decreased in winter. Compared with the southern ecotype, the northern ecotype was more responsive to seasonal changes, resulting in earlier growth cessation, cold acclimation and dormancy development in autumn, higher freezing tolerance and faster dormancy release in winter, and earlier bud flush and growth initiation in spring. In addition, although there was no significant ecotypic difference in ABA level during growing season, the rates and degrees of ABA alterations were different between the ecotypes in autumn and winter, and could be related to ecotypic differences in dormancy and freezing tolerance.     

Characteristics of Cold Acclimation and Deacclimation in Tuber-bearing Solanum Species

The effect of temperatures on cold acclimation and deacclimation in foliage tissues was studied in Solanum commersonii (Oka 4583), a tuber-bearing potato. The threshold temperature for cold acclimation was about 12 C. In a temperature range of 2 to 12 C, the increase in hardiness was dependent on the acclimating temperature; the lower the acclimating temperature, the more hardiness achieved. A day/night temperature of 2 C, regardless of photoperiod, appeared to the optimum acclimating temperature for the Solanum species studied. A subfreezing temperature hardened plants less effectively. The maximum level of hardiness could be reached after 15 days of cold acclimation. However, it took only 1 day to deacclimate the hardened plants to a preacclimation level when plants were subjected to a warm regime from cold. The degree of deacclimation was dependent on the temperature of the warm regime.     

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.     

A proposed role for the anaerobic pathway during low-temperature sweetening in tubers of Solanum tuberosum

Survival and growth of temperate zone woody plants under changing seasonal conditions is dependent on proper timing of cold acclimation and development of vegetative dormancy, shortening photoperiod being an important primary signal to induce these adaptive responses. To elucidate the physiological basis for climatic adaptation in trees, we have characterized photoperiodic responses in the latitudinal ecotypes of silver birch ( Betula pendula Roth) exposed to gradually shortening photoperiod under controlled conditions. In all ecotypes, shortening photoperiod triggered growth cessation, cold acclimation and dormancy development, that was accompanied by increases in endogenous abscisic acid (ABA) and decreases in indole-3-acetic acid (IAA). There were distinct differences between the ecotypes in the rates and degrees of these responses. The critical photoperiod and the photoperiodic sensitivity for growth cessation varied with latitudinal origin of the ecotype. The northern ecotype had a longer critical photoperiod and a greater photoperiodic sensitivity than the southern ecotype. Compared with the southern ecotypes, the northern ecotype was more responsive to shortening photoperiod, resulting in earlier cold acclimation, dormancy development, increase in ABA content and decrease in IAA content. However, at the termination of the experiment, all the ecotypes had reached approximately the same level of cold hardiness (−12 to −14°C), ABA content (2.1–2.3 µg g⁻¹ FW) and IAA content (17.2–20.3 ng g⁻¹ FW). In all ecotypes, increase in ABA levels preceded development of bud dormancy and maximum cold hardiness. IAA levels decreased more or less parallel with increasing cold hardiness and dormancy, suggesting a role of IAA in the photoperiodic control of growth, cold acclimation and dormancy development in birch.     

Effects of Temperature on Cold Acclimation and Deacclimation in Flower Buds of Evergreen Azaleas

The effects of various storage temperature/duration combinations(5, 10 and 17°/4, 8, 12 and 16 weeks) on cold acclimationand deacclimation of flower buds were studied in four speciesof evergreen azaleas having different natural distribution andcold hardiness. The freezing process and the exotherm temperaturedistribution of florets in excised whole buds determined bydifferential thermal analysis were used as the diagnostics todetermine the degree of bud acclimation and deacclimation. Theacclimation in buds lasted for as long as 12 to 16 weeks at5°C storage, and from 8 to 12 weeks at 10°C, and itappeared to be maintained after the chilling requirement forbreaking bud dormancy had been satisfied. Therefore, bud acclimationseems to be maintained independently from bud dormancy. Thedehardening effect on acclimated buds occurred as a result ofshort exposures to higher temperatures or long exposures tolower temperatures, and there was no relation between the rateof deacclimation and the degree of hardiness in each species.Among three storage temperatures examined, 5°C was the mosteffective for the maintenance of cold acclimation in flowerbuds and the small difference of floret water contents at 5and 10°C storage is not significant. (Received August 28, 1982; Accepted February 4, 1983)     

Freezing exposure releases bud dormancy in Betula pubescens and B. pendula

Bud dormancy in woody plants is released by long-term exposure to non-freezing chilling temperatures, whereas freezing temperatures have been considered to have little or no effect. However, the present results demonstrate that short-term exposure to freezing can release bud dormancy in Betula pubescens (Ehrh.) and B. pendula (Roth). Short-term freezing during the dormancy induction phase improved the release of bud dormancy only if an adequate level of dormancy had been reached. In fully dormant or chilled plants both the percentage and the speed of bud-burst increased, the more so the lower the temperature. Our results rule out the possibility that endogenous abscisic acid could be directly involved in the physiological control of bud dormancy release. The fast, easily applicable method presented here for bud dormancy release could further investigations into the biochemical and biophysical background to the process. The mechanisms of bud dormancy release and its relationship to cold acclimation are discussed in the light of these results, as also are the implications of the findings for modelling of bud dormancy.     

Independent activation of cold acclimation by low temperature and short photoperiod in hybrid aspen

Temperate zone woody plants cold acclimate in response to both short daylength (SD) and low temperature (LT). We were able to show that these two environmental cues induce cold acclimation independently by comparing the wild type (WT) and the transgenic hybrid aspen (Populus tremula x Populus tremuloides Michx.) line 22 overexpressing the oat (Avena sativa) PHYTOCHROME A gene. Line 22 was not able to detect the SD and, consequently, did not stop growing in SD conditions. This resulted in an impaired freezing tolerance development under SD. In contrast, exposure to LT resulted in cold acclimation of line 22 to a degree comparable with the WT. In contrast to the WT, line 22 could not dehydrate the overwintering tissues or induce the production of dehydrins (DHN) under SD conditions. Furthermore, abscisic acid (ABA) content of the buds of line 22 were the same under SD and long daylength, whereas prolonged SD exposure decreased the ABA level in the WT. LT exposure resulted in a rapid accumulation of DHN in both the WT and line 22. Similarly, ABA content increased transiently in both the WT and line 22. Our results indicate that phytochrome A is involved in photoperiodic regulation of ABA and DHN levels, but at LT they are regulated by a different mechanism. Although SD and LT induce cold acclimation independently, ABA and DHN may play important roles in both modes of acclimation.     

The function of phytochrome A

Knowledge of the photoperceptive function of phytochrome A has improved substantially thanks to the availability of mutants lacking phytochrome A and transgenic plants transformed with the PHYA gene in sense or anti-sense orientation. In imbibed seeds, phytochrome A mediates very-low-fluence responses. In etiolated seedlings, phytochrome A mediates very-low-fluence responses, high-irradiance responses under continuous far-red light, responsivity amplification to phytochrome B and red-light enhancement of the phototropic response to blue light. In light-grown seedings, phytochrome A modulates the extent of response to reductions in red/far-red ratio perceived by phytochrome B, perceives daylength extensions and night interruptions affecting flowering, and perceives light treatments resetting endogenous rhythms. Under natural radiation these abilities are manifested during seed germination and seedling de-etiolation under dense canopies or extremely low light fluences, and during early neighbour detection, but other processes await experimental evaluation. Phytochrome A affects growth and development throughout the whole life cycle of angiosperms.     

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.     

Cold-acclimation of Hibiscus rosa-sinensis L. and Hibiscus syriacus L. in natural and controlled environments

Abstract Seasonal cold-acclimation patterns and the effects of photoperiod and temperature on cold-hardiness of Hibiscus rosa-sinensis L. and Hibiscus syriacus L. were determined. Field-grown H. rosasinensis consistently failed to survive freezing at - 2°C. Two genotypes of field- and container-grown H. syriacus initiated cold-acclimation in mid September, in response to decreasing daylength, and continued to an ultimate midwinter hardiness level of - 27°C in early February. Controlled environment experiments using combinations of short days (SD) and cool day/night temperatures were unable to induce even minimal cold acclimation of H. rosasinensis. In controlled environments, H. syriacus attained a moderate amount of cold tolerance at warm temperatures and long days (LD). Low night temperature combined with LD, warm day produced the same degree of cold-acclimation as the SD treatments. While not essential, SD enhanced H. syriacus cold-acclimation in controlled environments. A - 5°C frost treatment of intact plants did not enhance cold-hardiness of H. syriacus.     

Alfalfa water status and cold hardiness as influenced by cold acclimation and water stress

Abstract Water stress at a nonacclimating temperature (18–20°C) increased the cold hardiness of Medicagosativa L. (alfalfa) plants. This increased cold hardiness was retained when the previously water-stressed plants were cold acclimated (2–9°C) in the absence of water stress. Water stress during cold acclimation also increased cold hardiness. Alfalfa was demonstrated to suffer injury, measured as decreased growth following freezing, at sub-lethal temperatures. During cold acclimation the turgor potential (ψ) of watered plants increased, whereas the solute potential and the water content per unit dry weight decreased. The large positive psgrdap of acclimated plants indicates that the decreased water content per unit dry weight is related to an increased proportion of tissue dry matter rather than to tissue dehydration.     

Ectopic expression of oat phytochrome A in hybrid aspen changes critical daylength for growth and prevents cold acclimatization

Survival of temperate-zone tree species under the normal summer-winter cycle is dependent on proper timing of apical growth cessation and cold acclimatization. This timing is primarily based on the perception of daylength, and through evolution many tree species have developed photoperiodic ecotypes which are closely adapted to the local light conditions. The longest photoperiod inducing growth cessation, the critical photoperiod, is inherited as a quantitative character. The phytochrome pigment family is the probable receptor of daylength, but the exact role of phytochrome and the physiological basis for the different responses between photoperiodic ecotypes are not known. This report shows for the first time that over-expression of the oat phytochrome A gene ( PHYA ) in a tree significantly changes the critical daylength and effectively prevents cold acclimatization. While the critical daylength for elongation growth in the wild-type of hybrid aspen ( Populus tremula × tremuloides ) was approximately 15 h, transgenic lines with a strong expression of the oat PHYA gene did not stop growing even under a photoperiod of 6 h. Quantitative analysis of gibberellins (GA) as well as indole-3-acetic acid (IAA) revealed that levels of these were not down-regulated under short days in the transgenic plants expressing high levels of oat PHYA , as in the wild-type. These results indicate that photoperiodic responses in trees might be regulated by the amount of PHYA gene expressed in the plants, and that the amount of phytochrome A (phyA) affects the metabolism of GAs and IAA.