Phenotypic Differences Between Natural and Selected Populations of Picea Abies. I. Frost Hardiness

The frost hardiness of non-juvenile Norway spruce [Picea abies (L.) Karst.] populations growing in northern Sweden (63°54' N) was monitored during 1996-1997. The investigated progenies originated from 12 natural populations and six seed orchards located between 58° N and 68° N in Sweden. Frost hardiness of needles was assessed by measuring chlorophyll fluorescence and electrolyte leakage after freezing. The loss of frost hardiness in 1-yr-old needles during spring occurred slightly earlier in populations originating north of 63°30' N than in those originating further south. Dehardening was slightly delayed in selected populations compared with natural populations of similar origin. The level of frost hardiness during autumn was higher in populations originating north of 63°30' N than in those originating south of this latitude, but there were no clear differences in frost hardiness between selected and natural populations of similar origin. The results are discussed in relation to climatic factors and inherent growth rhythms.     

Autumn frost hardiness of Pinus sylvestris offspring from seed orchard grafts of different ages

Autumn frost hardiness of offspring from clones (originating from 66.0–68.8° N) in six southern (62° N) and one northern (66.5° N) Pinus sylvestris seed orchards in Finland, was evaluated. One‐year‐old seedlings derived from open pollinated grafts of young, intermediate, and fairly mature age, and natural stand seedlings, were artificially freeze tested. The northerly located orchard produced the least injured offspring. There were no significant systematic differences between age classes. Neither were there any indications of decreased freezing injury with later age classes. The family variance was significant in four orchards. The estimated injury levels of orchard offspring were, in general, closer to those of natural stands in the neighbourhood of the orchard than natural stands at the place of clonal origin. It is suggested that not only differences in mother clone hardiness but also clonal differences in pollen contamination rate caused the observed family variation.     

Effects of nutritional factors on frost hardening in Larix leptolepis (Sieb & Zucc.) Gord.

A method for estimating frost hardiness in seedlings of Larix leptolepis in their first stage of acclimation is demonstrated. Cuttings of the shoot tip were frozen linearly from +2°C to ‐36°C. The viability was determined by TTC‐assay. Data were fitted to a logistic regression model, and hardiness was calculated from the fitting parameters. It was shown that hardiness is influenced by the duration of the fertilizer application period. No effect of the nutrient status of N, P or K in the shoots was found.     

Frost hardiness gradients in shoots and roots of picea mariana seedlings

Frost hardiness of tissues along the length of the stem and the root was investigated in first‐year black spruce (Picea mariana (Mill.) B.S.P.) seedlings. Frost hardiness of 1 cm long stem and root segments was evaluated based on Index of Injury, calculated from post‐freezing electrolyte leakage. Frost hardiness was tested approximately weekly beginning seven weeks after seedlings were transferred from an 18 to a 10 h photoperiod, both at day/night temperatures of 26°C/16°C. Trees were transferred to temperatures of 10°C day and 5°C night at a 10 h photoperiod after a further 18 days. Frost hardiness was greater at the terminal bud and least at the root tips. Although shoots were generally more frost hardy than roots, differences in hardiness along the stem and root axes were gradual, rather than abruptly differing at the shoot‐root interface. All tissues, including root tips, increased in frost hardiness after conditioning for 18 days under short photoperiods (10 h) and warm temperatures (26?C/16°C, day/night). Under cold temperatures (10°C/5°C, day/night) all tissues, excepting the root tips, tolerated — 16°C with little subsequent electrolyte leakage.     

The effect of photosynthetic photon flux density on development of frost hardiness in top and roots of Larix leptolepis seedlings

Seedlings of Larix leptolepis were grown in a growth chamber under short day conditions (8 h day) at 4 different photosynthetic photon flux densities (PPFD's). After a period of 8 weeks frost hardiness of the shoot tips and roots, dry matter content, dry weight (dw), content of glucose and starch were determined. The frost hardiness in the shoot tips increased from ‐15°C at a PPFD of 55μmol m^‐2s^‐1 to about ‐35°C at 440 μmol m^‐2s^‐1. No effect of PPFD was found on frost hardiness of the roots. A high PPFD results in a high dry matter content. The effect on dry matter content was most pronounced for the shoot tips and less pronounced for the roots. The total dry weight increased for both root and top with increasing PPFD. The height of the plants increased when the PPFD increased up to 220 μmol m^‐2s^‐1.     

Assessment of seedling storability of Quercus robur and Pinus sylvestris

Pedunculate oak (Quercus robur L.) and Scots pine (Pinus sylvestris L.) seedlings were lifted on several occasions during autumn 1997 to determine the relationships between storability and frost hardiness. On each lifting date their physiological status was determined by assessment of shoot and root electrolyte leakage and frost hardiness, assessed as freeze-induced electrolyte leakage. Additional seedlings were simultaneously cold-stored for field planting and assessment of preplanting root growth potential in April 1998. First year field performance was determined the following winter. Storability and cold acclimation patterns differed between the two species. Both were negatively affected by early lifting, but oak was less sensitive with respect to survival, and pine attained tolerance to cold storage more rapidly and earlier with respect to growth increment. The correlations between shoot frost hardiness and performance suggest that freeze-induced shoot electrolyte leakage (SEL_diff?20) below a threshold of 5% is a good storability predictor for Scots pine in Denmark. A completely reliable criterion for pedunculate oak could not be established.     

Morphological and physiological differences in Scots pine seedlings of six seed origins

One-year-old seedlings of Scots pine (Pinus sylvestris L.) offour native seed origins (Loch Maree Islands, Glengarry/GlenMorriston, Glen Affric and Abernethy), a commercial Britishseedlot, and a seedlot from Hedesunda, in middle Sweden, werecompared at monthly intervals from October 1993 to April 1994.Seedling morphology, root condition, root frost hardiness andbud dry matter were determined at each date. There were clear morphological differences among seed origins.Seedlings raised from the commercial seedlot (A70) were largerbut had a poorer root:shoot ratio than the other seed origins.Of the native pines tested, the Loch Maree Islands origin allocateda larger proportion of its photosynthate to fine roots and needlesand smaller proportion to woody structures. Seedlings raised from the commercial British seedlot tendedto have poorer bud lignification than the other origins andalso, in autumn, higher electrolyte leakage rates from its fineroots. During winter, the Swedish origin had the lowest fineroot electrolyte leakage. Seedlings of all origins showed aprogressive increase in fine root hardiness towards mid-winterwith maximum hardiness (–7°C) in January. Dehardeningoccurred over subsequent months reaching –3°C in April.Differences among origins were evident. The Swedish seedlotdeveloped greater frost resistance than the other origins, hardeningbegan earlier in autumn and dehardening began later in spring.The commercial seedlot hardened later than the other originsbut reached a similar level of frost hardiness by January. Ofthe native pines, seedlings of the Loch Maree Islands originwere slowest to develop root hardiness.     

Variation in spring and autumn frost tolerance among provenances of Russian larches (Larix Mill.)

Abstract

Spring and autumn frost tolerance was measured using material from a range-wide (50–67° N, 38–158° E) provenance trial of four Russian larch species (Larix sukaczewii Dyl., L. sibirica Ledeb., L. gmelinii Rupr. and L. cajanderi Mayr.) growing in northern Sweden. Shoots were collected in early May and late September and frozen at ?8, ?12, ?16 and ?20°C. Cambial damage was assessed visually after development under ideal conditions for 2 weeks. Differences in frost damage among provenances were highly significant in both spring and autumn. Autumn frost damage was significantly correlated with provenance latitude and longitude and spring frost damage was significantly correlated with provenance longitude but not latitude. Frost damage was not correlated with provenance elevation. North-western provenances showed the least damage and far-eastern provenances the greatest damage in both spring and autumn. A possible explanation for less spring frost damage to north-western provenances is adaptation to maritime conditions in proximity to the Barents Sea, which is often ice free in late winter. This would counteract early loss of frost tolerance and bud flushing if warm spells occurred in late winter. North-eastern Siberian provenances did not show similar adaptation and may exhibit increased spring frost damage if global warming eventually results in the Arctic Ocean north of Siberia becoming ice free in late winter.     

Growth Cessation and Autumn-frost Hardiness in One-year-old Picea abies Progenies from Seed Orchards and Natural Stands

One-year-old seedlings of Norway spruce [Picea abies (L.) Karst.] from seed orchards and natural stands were compared with respect to growth cessation and autumn-frost hardiness. The correlation among traits related to growth, growth cessation and frost hardiness on the same plants in two sets of environmental conditions was assessed. Total height, the degrees of shoot lignification and frost hardiness, and the timing of height growth cessation and budset were recorded at two nurseries in central and northern Sweden. Nine seed-orchard seed-lots were compared with seed from 26 natural stands originating from 56° N to 66° N in Sweden. Latitude explained 55-87% of the statistical variation among stands in the analysed traits at the central nursery and 49-84% at the northern nursery. On average, the seed-orchard progeny performed similarly to progeny from natural stands located 1-2° south of the origin of the seed-orchard clones. Reference material representing a geographical gradient was found to be a valuable aid when interpreting the results of growth cessation and frost-hardiness evaluations.     

Genetic variation in the natural pollen cloud of pinus sylvestris: a study based on progeny testing

Seeds from open pollination of individual female stroboli on different days in June 1990 in a coastal clone archive of Pinus sylvestris (L.) in northern Sweden (latitude 63°39’ N) were used for a short‐term progeny experiment to study differences over time in the genetic composition of the background pollen. Analysis of progeny growth and cold acclimatization in artificial whole‐plant freezing tests suggested no genetic differences in the fertilizing pollen between pollination days. A clinal variation with the latitude of pollen origin was observed for reference progenies from fertilization with pollen of known geographical origin. Comparisons between open‐pollinated progenies and reference progenies indicate that the pollen cloud originated from approximately 1° latitude (based on height growth) or 1.5° latitude (based on freezing injury) north of the pollination site.     

Freeze testing and field mortality of Pinus sylvestris (L.) in northern Sweden

Six short‐term freezing experiments were carried out on 24 one‐year‐old single‐tree open pollinated families of Pinus sylvestris (L.) representing four populations in northerly Sweden (latitude 62°10'N to 66°50'N). Two experiments were conducted in a climate chamber, two in a greenhouse and two outdoors. For freeze testing the plants were exposed to ‐10°C for three hours in a freezing chamber. A freezing experiment in a climate chamber of 28 one‐year‐old full‐sib families (half diallel with two parents from each population) was also carried out. The results were related to the mortality of the same open pollinated families over the first 18 years in the field. The ranking of the populations was the same in all six freezing experiments as in the field. Significant differences in frost resistance between one‐year‐old open pollinated families were found within all populations except the most northern one. The correlations within populations between frost resistance and field mortality varied considerably between populations and freezing experiments. The results from the two greenhouse experiments showed the best correlations with the cumulative field mortality after 18 years. The freezing test of the 28 full‐sib families resulted in the same mutual ranking, according to the general combining ability of parents within populations, as was obtained from the freezing tests of single tree families.     

Timing of cold temperature exposure affects root and shoot frost hardiness of Picea Mariana container seedlings

Seventeen‐week‐old black spruce seedlings were hardened under short daylengths and one of three short day length environments, which were either warm (24/16°C, day/night) throughout a 10 week hardening period (WW), cool (10/5°C) throughout hardening (CC), or warm for three weeks followed by seven weeks of cool temperatures (WC). Greatest root and shoot frost hardiness resulted from the exposure of seedlings to three weeks of warm followed by seven weeks of cool temperatures. Seedlings receiving warm temperatures throughout hardening increased in root and shoot frost hardiness, but to a lesser extent than seedlings exposed to cool temperatures. The frost hardiness of woody roots was generally greater than that of fine roots, but the extent of the difference in frost hardiness depended on the time since bud initiation and on the hardening treatment.     

Temperature step response of dehardening in Pinus sylvestris seedlings

The time constant of the dehardening of one‐year‐old Scots pine (Pinus sylvestris L.) seedlings was studied in three experiments started at different times of the year. The seedlings were transferred from outside to the different controlled conditions in January (Exp. 1), March (Exp. 2) and April (Exp. 3). Changes in the frost resistance of the seedlings were followed on the basis of the temperature response of the specific impedance difference in the shoot. The time constant for dehardening at a temperature of 7.5°C was found to be about 12 days in Exp. 1, about 8 days in Exp. 2, and about 5 days in Exp. 3. the terminal bud burst when the frost resistance of the shoot had risen to about ‐10°C.     

Effect of maternal soil treatment on first year growth and autumn frost hardiness of Pinus sylvestris L. full‐sib families

After‐effects of maternal soil treatment, controlling for photoperiod and weather conditions, were tested in a nursery experiment with Pinus sylvestris L. seedlings. Three full‐sib crosses were replicated in 4 different soil treatment plots of a grafted experimental seed orchard (63"54'N). Weight of filled seeds, percentage of empty seeds, first year hypocotyl and epicotyl length, and autumn frost hardiness (from artificial freeze testing) were evaluated. Systematic effects from maternal soil treatment were small compared to family differences for all characters studied. However, epicotyl length of the progenies showed a positive relationship with N needle content of the mother grafts. The results also indicated an important interaction between maternal soil treatment and autumn frost hardiness of the families. It is suggested that the altered mother tree treatment could lead to a change in progeny performance, corresponding to a 1.5 degree difference in latitudinal origin of natural stand seedlings. The results demonstrate the advantages of providing the mother tree with uniform conditions in early progeny testing.     

Coffee shade with Mimosa scabrella Benth. for frost protection in southern Brazil

Damage by radiative frosts is a major limiting factor for coffee cultivation in southern Brazil (south of 20° S latitude). The use of Mimosa scabrella (bracatinga) as a shade tree, to modify the local energy balance and thus prevent damage to the coffee plants, has been evaluated from 1986 to 1994. The study was carried out near Londrina, Parana State (23°23' S, 51°11′ W). During the experimental period, several radiative frosts with intensity ranging from moderate to very severe occurred at the site. Minimum coffee leaf temperatures during these events were 2 to 4 °C higher in the shaded plots. Due to frost protection, coffee bean yields on the average of 7 harvests were higher on the shaded plots. The potential of this system for frost protection in southern Brazil is discussed.     

Effects of nitrogen fertilization and temperature on frost hardiness of Aleppo pine (Pinus halepensis Mill.) seedlings assessed by chlorophyll fluorescence

In a 14-week study, 1-year-old Aleppo pine seedlings were grownin two growth chambers. Seedlings were artificially hardenedby decreasing photoperiod and temperature. In each chamber halfof the seedlings were fertilized with nitrogen (8.4 mg seedling^–1).In order to determine the relative importance of the hardeningenvironment versus fertilization, each chamber was programmedto decrease night temperatures down to a low of 8 or 4°C.Chlorophyll fluorescence and frost hardiness was measured fivetimes during the experiment. A sample of seedlings from eachtreatment was exposed to an artificial frost at –5°Cand the freezing effects were assessed by measurements of chlorophyllfluorescence and visual evaluation of needle damage. Seedlingsincreased their frost hardiness during the experiment in allthe treatments but the ratio of variable to maximal chlorophyllfluorescence (F_v/F_m) measured before freezing did not vary duringthe experiment. This indicates that Aleppo pine maintains itsphotosynthetic ability during hardening in contrast to otherconiferous species from colder climates. The effect of nitrogenfertilization on frost hardiness was small in comparison withchamber effect. Nitrogen fertilization slightly delayed theacquisition of hardening in the coldest chamber. Seedlings inthe warmest chamber did not become fully resistant to –5°C,but in the coldest chamber, where night temperature reached4°C, all the seedlings were resistant to the frost. Severedamage caused by frost could be related to a rapid rise of minimalfluorescence (F₀) but the best index of damage was the dropof F_v/F_m after freezing.     

Dependence of dormancy release on temperature in different origins of Pinus sylvestris and Betula pendula seedlings

The chilling requirement of rest completion and the high temperature requirement of growth initiation were determined in three origins of silver birch (Betula pendula Roth.) seedlings and five origins of Scots pine (Pinus sylvestris L.) seedlings. In both the pine and birch the chilling requirement was highest in maritime Scottish origins and lowest in the most continental Finnish and Russian origins. The requirement for southern mountainous Spanish and Bulgarian pine origins was in between. In terms of the high temperature requirement, there were no clear differences between origins. These results suggest that owing to their high chilling requirement, which prevents the beginning of growth and the loss of frost hardiness during the frost‐exposed season, origins from a maritime climate could be the most tolerant under climatic warming.     

Frost Hardiness of Red Alder (Alnus rubra) Provenances in Britain

The phenology and frost hardiness of shoots of 15 provenancesof Alnus rubra growing in Scotland were measured over one autumn,winter and spring. Dates of budset (in September) and the onsetof rapid frost hardening (in October-November) occurred about2 days earlier for each degree latitude of origin northwards,except for an Idaho provenance. However, all provenances dehardenedat about the same time in March and burst their buds between8 and 14 April. Assuming that rapid frost hardening in the autumnwas triggered primarily by shortening daylengths, Alaskan provenancesof A. rubra seemed better adapted to British conditions thansouthern British Columbian provenances, which have been mostcommonly planted. However, even Alaskan provenances are proneto spring frost damage. Scottish A. glutinosa and Alaskan A.sinuata set buds and frost hardened 1–2 weeks before eventhe Alaskan A. rubra, and burst their buds 2–3 weeks laterin April-May. All three species were hardy to below –30°Cfrom December to mid-March.     

Marker trait association for autumn cold acclimation and growth rhythm in pinus sylvestris

The main objective of this study was to identify quantitative trait loci (QTLs) involved in the genetic control of growth rhythm and autumn cold acclimation in Pinus sylvestris L. Eighteen full‐sib families of P. sylvestris with parental origins between latitudes 62 and 67° N in Sweden were assessed for autumn cold acclimation (frost injury measured from artificial freeze testing) and growth rhythm during the first two growing seasons. One family north × south with 108 individuals was used for the construction of one linkage map for the male and one for the female parent using 286 markers from random amplification of polymorphic DNA. Marker trait associations (QTLs) for shoot elongation, growth cessation and cold acclimation were found on both maps. Some QTLs were verified in two additional north × south families. Proportions of the additive genetic variance explained by the markers (R ²/_G) indicated that genetic factors of large effect were involved in all investigated traits. Single or multiple markers accounted for between 16 and 47% of the additive variance in annual shoot elongation, between 42 and 79% of the additive variance in growth termination, and between 7 and 31% of the additive variation in autumn cold acclimation. The study confirms that there are major QTL regions on different linkage groups controlling a large part of the variation for growth rhythm and autumn cold acclimation in P. sylvestris.     

Within‐population variation in autumn frost hardiness and its relationship to bud‐set and height growth in Picea abies

Artificial freezing tests were performed with two sets of 12 full‐sib families of Picea abies. Each set was selected from a complete diallel cross performed within a natural population. Significant differences in autumn frost hardiness were observed between the two populations, which originate from the same altitude and longitude approximately 60 km apart. Substantial variation in frost hardiness was observed within both populations. Significant pheno‐typic correlations at the individual level were found between freezing injury and the two traits terminal bud‐set and height growth year one. However, no significant relationships were present between freezing injury and bud‐set at the family level, indicating that bud‐set cannot be used to predict autumn frost hardiness for families.