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     

长白山北坡与西坡岳桦种群差异分析

本研究通过植被调查,从密度、胸径、树高、树龄、世代和种群分布格局等方面分析长白山北坡与西坡岳桦(Betula ermanii Cham.)种群的坡向差异及随海拔变化的坡向差异,探讨植被坡向差异的干扰因素.结果显示,整体上北坡岳桦种群胸径、二代木比重显著大于西坡,一代木比重则相反,说明尽管西坡生境优于北坡,但西坡植被演替...     

不同海拔长白山岳桦的生理变化

通过分析长白山国家级自然保护区内不同海拔(A1:1700 m, A2:1800 m, A3:1900 m, A4:2000 m, A5:2050 m)梯度岳桦叶片中各种生理指标含量的变化,探讨了林线树木适应高山环境的生理机制.结果表明:随着海拔的升高,比叶面积(SLA)显著减小,A5与A1相比下降了35.90%,差异达到显著水平;叶绿素含量随海拔梯度升高而降低,但叶绿素a/b比值(Chla/Chlb)和Car的相对含量(Car/Chl)随海拔梯度升高而增加;在海拔1900 m左右,MDA含量和MP均处于最低水平,各种酶的活性均为最低;当海拔超过2000 m,接近森林分布的界限时,MDA含量和MP升高,并达到最大值,各种酶的活性都出现了一定程度的下降.综合本次研究表明, 在海拔1900 m比较适合岳桦的生长;海拔超过2000 m,岳桦体内生理抗性下降,不利于岳桦的生长发育,因此高海拔限制了岳桦的分布.     

Intracellular resistance correlates with initial stage of frost hardening in willow (Salix viminalis)

Distributed model parameters of shoots of five clones of willow ( Salix viminalis ) were examined with electrical impedance analysis at the end of the growing season and with cold acclimation. The parameters were compared with regard to frost hardiness, linoleic (18:2) and linolenic (18:3) fatty acids, unsaturation/saturation ratio of fatty acids and dry weight content. The intracellular resistance (r_i) correlated best with changes in frost hardiness. The value of r_i rose from 1–2 Ω m in non-hardened to about 12 Ω m in hardened samples. In the initial stages of frost hardening, a linear relationship was found between r_i and frost hardiness and levels of 18:2 fatty acid, and an inverse relationship between r_i and levels of 18:3 fatty acid. The unsaturation/saturation ratio of fatty acids rose fairly rapidly in the initial stage of hardening. The dry weight content increased stepwise during the experimental period, and less steadily than r_i. In addition, equivalent circuit parameters changed in the prehardening phase, and were probably connected with cell differentiation and lignification. Frost hardiness by the visual method and by extracellular resistance, determined after controlled freezing tests, correlated well in the initial stages of hardening until about − 10°C but deviated upon further hardening.     

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.     

Effect of thawing time, cooling rate and boron nutrition on freezing point of the primordial shoot in norway spruce buds

BACKGROUND: Effects of cooling rates on bud frost hardiness have been studied but there is little information on bud responses to thawing. Since the cell wall pore size has been found to increase with boron (B) deficiency, B deficiency may affect the supercooling ability of buds in winter. METHODS: The effects of duration of thawing time and rate of cooling on bud frost hardiness of Norway spruce (Picea abies) were studied in a B fertilization trial in February 2003 and March 2005. Frost hardiness of apical buds was determined by differential thermal analysis (DTA) and visual scoring of damage. KEY RESULTS: In 2003, the freezing point of primordial shoots of buds (T(f)), i.e. the low-temperature exotherm (LTE), was, on average, -39 degrees C when buds were thawed for less than 3 h and the T(f) increased to -21 degrees C after 18 h of thawing. During the first 4 h of thawing, the rate of dehardening was 6 degrees C h(-1). In 2005, buds dehardened linearly from -39 degrees C to -35 degrees C at a rate of 0.7 degrees C h(-1). In 2003, different cooling rates of 1-5 degrees C h(-1) had a minor effect on T(f) but in 2005 with slow cooling rates T(f) decreased. In both samplings, at cooling rates of 2 and 1 degrees C h(-1), T(f) was slightly higher in B-fertilized than in non-fertilized trees. By contrast, at very short thawing times in 2003, T(f) was somewhat lower in B-fertilized trees. CONCLUSIONS: There was little evidence of reduced frost hardiness in trees with low B status. This study showed that buds deharden rapidly when exposed to above-freezing temperatures in winter, but if cooled again they reharden more slowly. According to this study, rapid dehardening of buds has to be taken into account in assessments of frost hardiness.     

Carbohydrate Metabolism and Frost Hardiness in Pine and Spruce Seedlings Grown at Different Photoperiods and Thermoperiods

Large changes occur in carbohydrate contents of pine (Pinus silvestris L.) and spruce (Picea abies (L.) Karst.) seedlings cold-hardened by photoperiod or by combined photo- and thermo-period. The largest change is in sucrose content, which is almost doubled after six weeks short-day (6/18 h) treatment; and more than doubled (spruce) or more than tripled (pine), when also temperature is lowered (10/5°C). Development of frost hardiness is strongly correlated with the change in carbohydrate contents. At dehardening, the carbohydrate content decreases rapidly, especially in pine, and the raffinose formed during the rest period disappears within 2–4 weeks. Frost hardiness decreases in parallel. The content of soluble carbohydrates may thus play a role in frost hardiness, although it is not the only factor. Bud formation at cold acclimation is not directly correlated with the changes in carbohydrate content and hardiness.     

Migration, invasion and decline: changes in recruitment and forest structure in a warming-linked shift of European beech forest in Catalonia (NE Spain)

Altitudinal upward shifts of species' ranges have occurred across a wide range of taxonomic groups and geographical locations during the twentieth century in response to current climate warming. However, actual data of plant species' altitudinal shifts are still scarce and not always clear. Here we provide a more detailed investigation of a previously reported European beech Fagus sylvatica forest altitudinal shift in the Montseny Mountains (Catalonia, NE Spain) now based on field photographic survey and on the population age structure and the recruitment patterns in the high Fagus limit (HFL), the central forest area (CFA) and the low Fagus limit (LFL). Monitoring of the lowest altitudinal range shows that beech forest is being progressively replaced by Mediterranean holm oak forest. Holm oaks are characterized by recruitment rates more than three times higher than those of beech in the LFL in the last decades. The percentage of young individuals in the LFL is only half that in the HFL and CFA. In the highest altitudinal range, present day and early 20th century photographs show that the HFL has gained density and has shifted altitudinally upwards, advancing with establishment of new, vigorous outpost trees (13 individuals per each 100 m of tree-line). They are mostly (89%) younger than 35 yr old and mostly (97%) located up to 70  m (with a few up to 105 m) ground surface distance above the current tree line (36–51 m altitude) at the highest altitudes (1600–1700 m). The beech forest upward shift is a likely consequence of warming, but land-use practice changes (cessation of burning by shepherds) have made it possible. These changes in vegetation distribution and population structure constitute a new indication of the complex global change effects on life in mountain ecosystems.     

Assessing frost hardiness of Pinus bungeana shoots and needles by electrical impedance spectroscopy with and without freezing tests

Aims Nursery and forest operations require that frost hardiness results be produced faster than can be provided by controlled freezing tests. There is a great challenge to develop a rapid method for predicting frost hardiness that might not necessitate controlled freezing tests. The aim of this study was to examine the assessment of the frost hardiness of shoots and needles of Pinus bungeana by electrical impedance spectroscopy (EIS) with and without controlled exposure to freezing.Methods The frost hardiness of current-year shoots and needles of P. bungeana in an 8-year-old provenance field trial was measured at Shisanlin Nursery in Beijing, China, from September 2006 to January 2007 by means of EIS and conventional electrolyte leakage (EL). In the same plants, but without controlled freezing test, were monitored the EIS parameters in current-year shoots and needles.Important findings The results showed that (i) after controlled freezing tests, the frost hardiness estimated by EIS parameters (extracellular resistance, r e, and membrane time constant, τ m) was significantly correlated with the frost hardiness assessed by EL method (r = 0.95) and (ii) for the samples not exposed to controlled freezing treatment, the relaxation time τ 1 for shoots and β for needles had greater correlations with the frost hardiness estimated by EL after controlled freezing tests relative to the other parameters (r = ?0.90 for shoots and r = 0.84 for needles, respectively). The parameters r e of shoots and needles and τ m of needles might be applied for measuring frost hardiness of samples after exposed to controlled freezing tests. The frost hardiness results can be obtained within 48 h. The parameters τ 1 of shoots and β of needles could be used for estimating the frost hardiness of samples without using a controlled freezing test. The frost hardiness results can be obtained within 24 h.     

Winter water relations of timberline larch (Larix leptolepis Gord.) on Mt. Fuji

 A krummholz mat of larch, Larix leptolepis, is the predominant growth form near the upper limit (near 2500 m) of the timberline ecotone on the south slope of Mt. Fuji, Japan. On the south-eastern slope, the tree line is lowered to 1600 m because of the last volcanic eruption. The extent and causes of winter desiccation were compared in timberline larch between 2500 m and 1600 m elevation over two winters. Bark abrasion due to wind-blown fine volcanic gravels caused a decrease in bark resistance to water loss and resulted in severe desiccation damage to current-year shoots of krummholz larches in the winter of 1986 – 87 at 2500 m, whereas abraded shoots at 1600 m maintained high water content during both winters. In the winter of 1985 – 86, shoots of krummholz larches at 2500 m did not experience bark abrasion and high water contents were maintained. Experimental abrasion of shoot surfaces resulted in similar results at each elevation. Thus, in timberline larch at 2500 m, abrasion by wind-blown fine volcanic gravels is the primary factor causing winter desiccation damage and krummholz formation. Based on field experiments, the estimated amount of water movement to non-abraded shoots was the same for the two elevations. At 2500 m, water movement to abraded shoots was less than to non-abraded shoots, but the reverse situation was noted at 1600 m. Water supply to abraded shoots at 2500 m was limited and insufficient to compensate for water loss. A cause of limited water supply at 2500 m may be xylem embolism. Received: 13 January 1995 / Accepted: 31 January 1996     

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 importance of hardening and winter temperature for growth in mountain birch populations

Seedlings of five mountain birch populations (Betula pubescens Ehrh. ssp. czerepanovii) from Fennoscandia and Iceland were raised and grown at natural daylengths at Tromsø, Norway (69°N) and different temperatures during late summer and fall season, followed by winter temperature treatment at ambient and +4 °C above ambient temperatures at Bergen, Norway (60°N). The experiment took place during two seasons (2000/01 and 2001/02). The following summer shoot and biomass growth were reduced as a result of winter warming and subsequent premature dehardening in early flushing provenances and treatments. Biomass increased in plants grown at low hardening temperature when compared with high temperature treatment. As a conclusion, increased winter temperatures would tend to increase the risk of spring frost damage and reduce growth in birch seedlings, because the differences between the frost hardening and ambient temperatures are decreasing, and because the time from budbreak to dehardening is shortened. The results are discussed in relation to simultaneous experiments with frost hardiness in the same populations and treatments.     

Ecology of the pteridophytes on the southern slopes of Mt. Kilimanjaro – I. Altitudinal distribution

140 taxa of 61 genera in 24 families of pteridophytes were recorded on the southern slopes of Mt. Kilimanjaro. These represent about one third of the entire pteridophyte flora of Tanzania. The families richest in species are the Aspleniaceae, the Adiantaceae, the Dryopteridaceae, the Thelypteridaceae and the Hymenophyllaceae. Due to its luxuriant montane rain forest, which receives a precipitation of up to over 3000 mm, Mt. Kilimanjaro is distinctly richer in pteridophyte species than other volcanoes in East Africa. However, compared with the mountains of the Eastern Arc, the number of pteridophytes on Mt. Kilimanjaro is smaller. This can be explained by the widely destroyed submontane (intermediate) forest rather than by the higher age of the Eastern Arc Mts.The altitudinal distribution of all ferns was investigated in 24 transects. On the southern slopes of Mt. Kilimanjaro they were found in an altitudinal range of 3640 m. Cyclosorus quadrangularis, Azolla nilotica, Azolla africana andMarsilea minuta are restricted to the foothills, while Polystichum wilsonii, Cystopteris nivalis and Asplenium adiantum-nigrum are species found in the highest altitudes.Based on unidimensionally constrained clustering and on the analysis of the lowermost and uppermost occurrence of species, floristic discontinuities within the transects were determined. From these data and from an evaluation of the distribution of ecological groups and life forms, 11 altitudinal zones could be distinguished: a colline zone (–900 m asl), a submontane zone (900–1600 m asl) with lower and upper subzones, a montane zone (1600-2800 m asl) divided into 4 subzones, a subalpine zone (2800–3900 m asl) with lower, middle and upper subzones, and finally a (lower) alpine zone above 3900 m. The highest species numbers were observed in the lower montane forest belt between 1600 and 2000 m altitude. The zonation of ferns found at Mt. Kilimanjaro corresponds well with the vegetational zonation described by other authors using bryophytes as indicators in different parts of the humid tropics.     

海拔梯度对长白山北坡岳桦水分利用效率的影响

以分布于长白山北坡海拔1800～2050 m的岳桦林为对象,以叶片碳稳定同位素(δ13C)值作为岳桦长期水分利用效率指示值,探讨海拔梯度对岳桦林水分利用效率的影响.结果表明:随海拔升高,岳桦林土壤体积含水量、比叶质量极显著增加,而叶片含水量和土壤温度显著降低;岳桦叶片δ13C值与海拔呈极显著正相关,增幅为1.013‰·(100 m)-1,与土壤体积含水量、比叶质量呈显著正相关,与生长季土壤平均温度、叶片含水量呈显著负相关.温度不是长白山林线的唯一限制性因子,海拔梯度上水热条件的差异及其交互作用可能造成岳桦生长期间的生理干旱.     

Differences in frost hardiness of two Norway spruce morphotypes growing at Mt. Brocken, Germany

Norway spruce (Picea abies (L.) Karst.) exhibits strong ecotypic variation along altitudinal gradients in morphological traits, e.g. slenderness of crowns or arrangement of second-order branches. We were interested whether montane and lowland morphotypes differ in a key trait for the survival in cold environments, i.e. frost hardiness, and asked: (i) are montane morphotypes more resistant to frost damage and (ii) do they have a lower risk of frost damage by late frosts in spring than lowland morphotypes?We used the electrolyte leakage-method to measure frost hardiness on a monthly basis from October 2006 to May 2007 in stands of the montane and lowland morphotypes at Mt. Brocken in the Harz Mountains, Germany.LT₅₀ (i.e. the temperature that results in 50% of maximum electrolyte leakage) was assessed by freezing treatments in a frost chamber and was significantly influenced by morphotype, month and minimum ambient temperatures. LT₅₀ was significantly lower in the montane than in the lowland morphotype, with −107 °C and −49 °C, respectively. However, the interactions between morphotype with minimum ambient temperature or month were not significant. Thus, as frost hardiness of the two morphotypes responded to temperature in the same way, both morphotypes can be supposed to be exposed to the same risk of frost damage during hardening in autumn and dehardening in spring.     

Leaf anatomy as a constraint for photosynthetic acclimation: differential responses in leaf anatomy to increasing growth irradiance among three deciduous trees

Interspecific variation in the response to transfer from low to high growth irradiance with respect to anatomical and photosynthetic characteristics was studied in mature leaves of three tree species, Betula ermanii Cham., Acer rufinerve Sieb. et Zucc. and Fagus crenata Blume, which occur in different successional stages in temperate deciduous forests. Transfer from low to high irradiance increased the light-saturated rate of photosynthesis per unit leaf area ( P _max) significantly in B. ermanii and A. rufinerve , but not in F. crenata . Leaves of B. ermanii grown at low irradiance were relatively thick and had vacant spaces along the mesophyll cell surfaces which was not occupied by chloroplasts or other organelles. After transfer to high irradiance, chloroplasts enlarged to fill the space along with P _max without an increase in leaf thickness. Leaves of A. rufinerve were plastic in mesophyll cell surface area and in leaf thickness, both of which increased after the transfer to high irradiance, along with an increase in the amount of chloroplasts and in P _max. On the other hand, F. crenata had little mesophyll cell surface unoccupied by chloroplasts and leaf anatomy was not changed after the transfer. In all species, P _max was strongly correlated with chloroplast surface area adjacent to the exposed mesophyll surface across different growth irradiances. An increase in P _max was observed only when chloroplast volume also increased. We conclude that light acclimation potential is primarily determined by the availability of unoccupied cell surface into which chloroplasts expand, as well as by the plasticity of the mesophyll that allows an increase in its surface area.     

Altitudinal variations in germination and growth responses ofReynoutria japonica populations on Mt Fuji to a controlled thermal environment

The authors examined altitudinal variations in the thermal responses of seed germination and seedling growth inReynoutria japonica (=Polygonum cuspidatum) under controlled environmental conditions. Seed populations were collected from different altitudes on Mt Fuji in Japan. The mean seed weight of the upland populations (above 1500 m) was significantly (1.5-fold) heavier than that of the lowland populations (below 1400 m). Under the lowest temperature regime of 15/10°C (day/night) the upland populations showed a significantly higher percentage and speed of germination than the lowland populations; this was not significant under higher temperature regimes. These results indicate that the germination traits of the upland populations on Mt Fuji are favorable for colonization in their cold habitats (low temperature and short growing season). Growth and shoot development were compared between the seedlings grown from seeds collected at altitudes of 700 and 2420 m. The upland seedlings showed a significantly larger biomass and leaf area than the lowland seedlings at 15°C, but there was no difference at 25°C. The difference in biomass at 15°C was attributed to the difference in seed weight. The upland seedlings produced a significantly larger number of branches with smaller and more numerous leaves at both 15°C and 25°C. these developmental traits of the upland seedlings were considered to represent the adaptation of the life form to upland environments. It was concluded that theR. japonica populations along an altitudinal gradient on Mt Fuji can be classified into two ecotypes, whose distribution border lies at an altitude of about 1400–1500m. In this study, the seed weight and germination traits of twoR. japonica seed populations collected in Chiba Prefecture were briefly compared with those of the lowland populations on Mt Fuji.     

太白山南坡土壤种子库的物种组成与优势成分的垂直分布格局

本文通过野外植被调查和室内试验,采用物理方法从1220份土样中挑选种子,进行分类统计,研究了秦岭太白山南坡土壤种子库物种组成与优势成分的垂直格局。主要研究结果有：(1)从种子数量上看,在不同的海拔段,优势科不同;从物种数来看,海拔1600m和海拔2600m处的优势科数相对较多,有4个科;蔷薇科是种数最多的优势科。(2)桦木科的糙皮桦(Betula utilis)在16个海拔样地(海拔样地共有21个)中都是优势成分之一。糙皮桦和巴山冷杉(Abies fargesii)的种子储量在中等海拔呈现峰值;较高海拔处,糙皮桦作为优势成分之一所占的比例较大;而在低海拔则相对较小。(3)从种子库中种类组成上来看,草本种类占优势;从各样地土壤种子库的生活型组成并结合优势种类来看,乔木不仅在个体组成上占优势,大部分单优种类也是来自于乔木种类。灌木种类在土壤种子库中无论是种类组成,还是在个体组成上都没有表现出优势。本文为研究太白山南坡植物群落演替动态及其沿海拔梯度的变化提供了基础。     

Intergenerational acclimation in aphid overwintering

Abstract.  1. When first instar nymphs and adults of the grain aphid Sitobion avenae (Fabricius) (Hemiptera: Aphidiae) were maintained in long-term cultures (>6 months) at 20 °C and 10 °C, the LT₅₀ decreased from −8 and −8.8 °C to −16.0 and −13.5 °C, respectively.
2. When aphids from the 20 °C culture were transferred to 10 °C, there was a progressive increase in cold tolerance through three successive generations. Transfer of newly moulted pre-reproductive adults reared at 10 °C for three generations back to 20 °C resulted in a rapid loss of cold hardiness in their nymphal offspring.
3. In all generations reared at 10 °C, first born nymphs were more cold hardy than those born later in the birth sequence. The LT₅₀ of nymphs produced on the first day of reproduction in the first, second and third generations maintained at 10 °C were −14.8, −17.0 and −16.6 °C, respectively. Thereafter, nymphal cold hardiness decreased over the subsequent 14 days of reproduction in each generation at 10 °C with mean LT₅₀ values of −10.3, −12.6 and −14.8 °C, respectively. By contrast, the cold tolerance of first born nymphs of aphids reared continuously at 20 °C did not differ in comparison with later born siblings. The LT₅₀ of adult aphids was also unaffected by ageing.
4. The ecological relevance of these findings is discussed in relation to the overwintering survival of aphids such as S. avenae .     

Geographical variation in egg cold hardiness: a study on the adaptation strategies of the migratory locust Locusta migratoria L.

Abstract. 1. For many species of insect, cold hardiness is an important trait that enables a population to develop in the next season and to extend its range. To elucidate the role of cold hardiness of the migratory locust Locusta migratoria L. in its outbreak and distribution areas, egg cold hardiness was examined in locusts derived from four locations from latitude 18°23'N to latitude 41°10'N in eastern China.
2. The supercooling points of eggs from different geographic populations did not differ significantly for the first development stage, with an average ± SE of −24.5 ± 0.51 °C, or for the second stage, −22.06 ± 0.68 °C, however there was a significant difference for the embryonic development phase among the four geographical populations. The egg supercooling point increased gradually from neonatal egg to old egg; eggs prior to hatching always had a much higher supercooling point.
3. Comparisons of the cold hardiness of four populations were carried out by validating the close correlation between latitude and the effects of cold on hatching, low lethal temperature (Ltemp₅₀), and low lethal time (Ltime₅₀). There were significant differences among the four populations; the northern population was more cold hardy than the southern population, and the two mid-latitude populations were intermediately cold hardy.
4. The cold hardiness of all populations was enhanced to various degrees by short-term cold acclimation at 0 °C and 5 °C. For most populations, a 2-day acclimation period seemed to be optimal.