Chemical and physiological responses of four Turkish red pine (Pinus brutia Ten.) provenances to cold temperature treatments

Determining the adaptability to abiotic conditions and potential establishment success of tree species needs to be conducted before attempting to use a species in large-scale afforestation programs. In this study, the chemical and physiological performance of four Turkish red pine (Pinus brutia Ten.) provenances was investigated after exposure to artificial cold temperature treatments to determine their adaptability to cold environment for potential use in afforestation programs. Seeds were sown and raised for 24, 28, and 32 weeks and exposed to decreasing temperatures in an artificial freezer. Relative electrolyte leakage, chlorophyll fluorescence, and carbohydrate concentrations were measured to determine the variability between provenances. Results showed that diameter and height growth did not vary with origin for each of the three growth stages measured. Root electrolyte leakage values differed between provenances, confirming that cold stress was effectively causing physiological damages when plants were exposed to temperature at ?15 °C and below. The variability observed in the relationship between provenances and cold hardiness responses can be attributed to tree-to-tree variability within provenances and microsites conditions. There was generally no significant difference in chlorophyll fluorescence between provenances, also attributed to low genetic variation between provenances. Carbohydrate concentrations were also very variable and varied significantly among growth stages and provenances. High-altitude provenances had higher soluble carbohydrates concentrations in several cases, suggesting a relationship between altitude, soluble sugars, and cold hardiness. However, these trends were not consistent; therefore, we suggest that such hypotheses be confirmed through more comprehensive further studies.     

Seedling cold hardiness, bud set, and bud break in nine provenances of Pinus greggii Engelm.

Cold hardiness and timing of bud set and bud break are important processes that provide protection of nursery seedlings against low temperatures. Seedlings of 9 provenances of Pinus greggii from two different regions of Mexico were tested to determine cold hardiness, bud set, and bud break timing differences. Needle sections were exposed to freezing temperatures to determine an injury index of each provenance. In addition, bud set and bud break timing were recorded through the fall, winter and spring. There were significant differences in cold hardiness between seedlings from northern and southern provenances. At the maximum cold hardiness, the index of injury (LT₅₀) for northern provenances was LT₅₀ = −18 °C, compared to −12 °C for southern provenances. There was a considerable variation among the provenances in the proportion of seedlings that set terminal buds. Seedlings from northern provenances had greater proportions of seedlings that set a terminal bud than seedlings from southern provenances. There were also significant differences in the bud break timing in the following spring among the 9 provenances. Seedlings from northern provenances broke bud earlier than southern provenances. Cold hardiness, bud set, and bud break timing results may be useful to determine how far a specific seed source can be moved from its natural environment.     

A semi-physiological model of cold hardening and dehardening in walnut stem

It has been hypothesized that the increase in temperature in this century could lead to an increase in frost damage to plant tissues. Several models have been proposed to describe the development of cold hardiness, but never taking into account extreme climatic and/or physiological events. Our results on walnut tree (Juglans regia L.) show that cold hardiness was best correlated with average daily temperatures minimal temperatures over the last 15 days before sampling (T(min 15 days)), indicating that the freezing tolerance depended on the tree's climatic history. Moreover, this study also shows that the accumulation of sucrose and the water content (WC) decrease are an essential step towards cold hardiness. Thus, a simple linear model based on climatic (T(min 15 days)) and physiological (soluble sugars, WC) explanatory variables was developed to predict the cold hardiness level in walnut stem at any time during the leafless period. Each of the three input variables can be assigned a specific role contributing to the simulated function, cold hardiness. The extent and robustness of this relation was assessed on extreme physiological events on walnut trees bearing three main branches. On each tree, one branch was defoliated to limit the local carbohydrate and transpiration, one was girdled to increase local carbohydrate and prevent carbohydrate export and the third one was kept untreated as control. As expected, these treatments impacted both local carbon reserves and WC in the stems born by each main branch in comparison with the control on the same tree. The impact of these treatments on stem's freezing tolerance, as evaluated by an electrolyte leakage method (LT??), confirmed the direct impact of soluble sugar and WC on cold hardiness over a wide range of carbohydrate and WC. This is discussed in relation to the branch autonomy theory for carbon but also for water during summer growth and winter periods. The present study demonstrates the importance of physiological parameters in the prediction of cold hardiness and proposes a way to model cold hardiness with extreme climatic and/or physiological events.     

Short-day treatment during the growing period limits shoot growth and increases frost hardiness of hybrid aspen plants in the nursery

In Finland, under nursery conditions hybrid aspen may continue their shoot growth until early September. Thus, frost hardening is usually delayed. To solve this problem, we used a three-week period of short-day (SD) treatment between late July and mid-August. During autumn after frost exposure, frost hardiness (FH) was assessed three times with a stem-browning test. The re-sults showed that after SD treatment shoot growth ceased and FH increased when compared to untreated hybrid aspen. Furthermore, the height of SD-treated hybrid aspen varied much less than that of the control plants. We conclude that SD treatment in the nursery during the growing period can be used as a supplementary method for producing well-hardened and uniform hybrid aspen plants.     

Growth and hardening of four provenances of containerized white spruce (Picea glauca (Moench) Voss) seedlings in response to the duration of 16 h long-night treatments

Long-night treatments of 16 h were applied to four provenances of 10-week-old Picea glauca (Moench) Voss seedlings for durations of 0 (control), 4, 8, 12 and 16 d. Their effects on growth and on the development of frost hardiness were determined during the fall. Height growth cessation was hastened by long-night treatments, with 4, 8, 12 and 16 d treatment seedlings ceasing growth approximately 5, 10, 13 and 15 d earlier than control seedlings. The rate of hardening was affected by the duration of the long-night treatment while the level of hardening was affected by both treatment duration and the latitude of origin of the seedlings. Specifically, seedlings receiving the 8, 12 and 16 d treatments had achieved approximately the same level of hardiness by mid-October, which was greater than the 0 and 4 d treatments. However, hardiness levels of the 12 and 16 d treatments were higher in late September than with the 8 d treatment. The level of frost hardiness in late November increased with both increasing duration of long-night treatments (0 to 8 d) and increasing latitude of origin.     

Relationships among cold hardiness, root growth potential and bud dormancy in three conifers

Greenhouse-cultured, container-grown ponderosa pine (Pinus ponderosa var. scopulorum Engelm.), interior Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) and Engelmann spruce (Picea engelmannii (Parry) Engelm.) were cold acclimated and deacclimated in growth chambers over 19 weeks. Stem cold hardiness, total new root length at 14 days and days to bud break were measured weekly. Relationships among cold hardiness, root growth potential (RGP) and bud dormancy suggest that cold hardiness, which can be measured quickly, could provide a useful basis for estimating the two other parameters. During cold acclimation, there was a lag period in which stem cold hardiness remained at -15 degrees C and RGP was at a minimum, in all three species. Douglas-fir and Engelmann spruce buds remained fully dormant during this lag period. Ponderosa pine buds had no chilling requirement for the loss of dormancy, and reached quiescence during the lag period. Immediately following the lag period, as stem cold hardiness progressed to -22 degrees C, RGP increased to a high plateau in all three species, and Douglas-fir and Engelmann spruce buds approached quiescence. Cold deacclimation and bud development began immediately on exposure to warm, long days, but RGP remained high until stem cold hardiness returned to approximately -15 degrees C. At bud break, cold hardiness and RGP were at the minimum.     

Effect of stock type characteristics and time of planting on field performance of aspen (Populus tremuloides Michx.) seedlings on boreal reclamation sites

Aspen (Populus tremuloides Michx) has great potential as a reclamation species for mining sites in the boreal forest, but planting stock has shown poor field performance after outplanting. In this study we tested how different aspen seedling characteristics and planting times affect field outplanting performance on reclamation sites. We produced three different types of aspen planting stock, which varied significantly in seedling size, root-to-shoot ratio (RSR), and total non-structural carbohydrate (TNC) reserves in roots, by artificially manipulating shoot growth during seedling production. All three stock types were then field-planted either in late summer, late fall, or early spring after frozen storage. Seedlings were outplanted onto two reclaimed open-pit mining areas in the boreal forest region of central and east-central Alberta, Canada, which varied significantly in latitude, reclamation history, and soil conditions. Overall, height growth was better in aspen stock types with high RSR and TNC reserves. Differences in field performance among aspen stock types appeared to be more strongly expressed when seedlings were exposed to more stressful environmental site conditions, such as low soil nutrients and moisture. Generally, aspen seedlings planted with leaves in the summer showed the poorest performance, and summer- or fall-planted seedlings with no shoot growth manipulation had much greater stem dieback after the first winter. This indicates that the dormancy and hardening of the stem, as a result of premature bud set treatments, could improve the outplanting performance of aspen seedlings, particularly those planted during summer and fall.     

Genotype and environment: two factors related to autumn cold hardiness on Persian walnut (Juglans regia L.)

Context

J. regia timber is appreciated for high-value wood products. In new plantations, biotic or abiotic events which could affect wood quality should be monitored. Autumn frosts could affect annual shoot development, with consequent loss of timber value or even tree death. In southern Europe, climate change forecasts include erratic and severe autumn frost events.

Aims

The relationship between genotype and environment regarding susceptibility to autumn frost damage was examined in four provenances of Juglans regia L., planted at two ecologically different sites, one subject to Mediterranean and the other Atlantic weather conditions.

Methods

Annual budsticks from eight trees per provenance were collected in November 2010 from each site. The samples were then submitted to freeze–thaw cycles down to ?8, ?13 and ?18 °C, plus a control treatment (keeping samples at 5 °C). Damage to the stem was assessed using the index of freezing injury calculated from relative electrolyte leakage at each temperature considered. Frost damage to stem, apical and lateral buds was recorded by visual scoring.

Results

Differences in cold acclimation between sites were detected, with provenances exhibiting differences on senescence. A ‘provenance?×?site’ interaction was found in some of the analyses, but the same ranking of susceptibility was detected for all provenances at both sites and in all the tissues analysed. The differences between provenances could be related to their geographical origins, where an altitude gradient was observed.

Conclusions

The genetic component was important in the expression of autumn cold hardiness and, together with productive traits, should be considered in new afforestation projects.     

Seasonal variation in photochemical activity and hardiness in clones of Norway spruce (Picea abies)

We investigated changes in photochemical activity and cold hardiness of detached needles of three clones of Picea abies (L.) Karst. by measuring variable chlorophyll fluorescence (F(v)/F(m)), before and after artificial freezing, from September to June. Photochemical activity varied considerably during the study, but only minor differences in photochemical activity among the clones were observed before freezing. Photochemical activity was high during early fall and then declined from November until April. Photochemical activity was at a minimum in April and then increased quickly to high values in May. During the period from late September to October, and also during the winter, differences in F(v)/F(m) ratios after artificial freezing to below -10 degrees C were observed among clones, indicating clonal differences in cold hardiness and hardiness development. The clone having an average height of 2.3 m after 11 years showed consistently lower cold hardiness than clones that had reached average heights of 4.0 and 5.0 m. There were also differences in the temperature requirement for bud flushing among clones.     

Cold acclimation of Pinus contorta and Pinus sylvestris assessed by chlorophyll fluorescence

Needle samples of six provenances each of lodgepole pine (Pinus contorta Dougl. var. latifolia) and Scots pine (Pinus sylvestris L.), originating from latitudes 55 to 68 degrees N in western Canada and northern Sweden, were collected during the autumn and subjected to freezing temperatures in the range of -8 to -29 degrees C on three occasions in September and October. Needle injury was assessed by two different methods: visual assessment and chlorophyll a fluorescence. Chlorophyll a fluorescence data showed a highly significant correlation with the visual assessments of injury, indicating that the technique can be used as a simple, non-destructive and objective measure for rapid detection of freezing injury and for ranking of needle materials with respect to development of cold acclimation. The analyses showed that, during the autumn, lodgepole pine needles were more hardy and acclimated to low temperatures earlier than Scots pine needles.     

Frost hardiness of 16 European provenances of sessile oak growing in Scotland

The frost hardiness of 16 European provenances of sessile oakQuercus petraea (Matt.)Liebl. originating from six Europeancountries was examined from autumn until budburst in springusing the method of relative conductivity. There were significant differences of frost hardiness betweenprovenances and a strong relationship between phenology andfrost hardiness. In spring, provenances that burst bud earlydehardened earlier than provenances that burst bud later. Inautumn, provenances that stopped growing early were more frosthardy than provenances that continued to grow. The interprovenance ranges of frost hardiness were greatestin spring and autumn and least in midwinter. Consequently, itwas not possible to demonstrate significant differences of frosthardiness between provenances in December and January. Overall, German, Polish and Danish provenances were more frosthardy than French, Austrian and British provenances. Frenchprovenances were least hardy at all times.     

Seasonal alteration in amount of Ca(2+) in apical bud cells of mulberry (Morus bombciz Koidz): an electron microscopy-cytochemical study

Subcellular localization of calcium in apical bud cells of mulberry (Morus bombciz Koidz) was investigated by a combination of calcium antimonate precipitation and electron microscopy (EM), over a 10-month period extending from July to May. Calcium antimonate deposits, an indication of Ca(2+) localization, were found mainly in the intercellular spaces and vacuoles of tissues collected in summer (e.g., July 10). Few deposits were seen in the cytosol and the nucleus, indicating that when plants were actively growing, the cytoplasm had a low concentration of Ca(2+). As the day length became shorter (e.g., August 8), Ca(2+) deposits increased in the cytosol and the nucleus. When plant dormancy and cold hardiness were rapidly developing in September and October, a large number of Ca(2+) deposits were seen in the cytoplasm and the nucleus, where they remained for about 60 days. Tissue samples collected during midwinter (e.g., January 20) displayed few Ca(2+) deposits in the cytoplasm and the nucleus compared with the sample collected on November 1. The number of Ca(2+) deposits remained low even when day length increased in the spring (e.g., May 5). We conclude that the seasonal dynamics of intracellular Ca(2+) concentration bear a close relationship with growth cessation and the development of dormancy and cold hardiness in temperate woody perennials.     

Seasonal Changes in the Frost Hardiness of Provenances of Picea sitchensis in Scotland

Changes in the natural level of frost hardiness of shoots offour provenances of Picea sitchensis were monitored over twogrowing seasons by detaching shoots from 7 to 10-year-old treesgrowing in a nursery in Scotland, and subjecting them to freezingtemperatures under conditions which simulated night frosts. Six seasonal phases of frost hardiness were identified (Fig.3).

1. During each autumn, killing temperatures (the level of hardiness)decreased from –5°C to below –20°C, beginningseveral weeks after shoot elongation ceased. Alaskan provenanceshardened in September, apparently in response to shorteningday lengths alone, whereas an Oregon provenance did not hardenuntil November, after repeated frosts. Queen Charlotte Islandsprovenances were intermediate.
2. From November to March allprovenances were hardy to below –20°C,which is adequateto prevent direct freezing injury at mostplantation sites.
3. In March-April, several weeks before bud-burst, old shootsdehardenedto killing temperatures of about –10°Cin responseto warm temperatures, and southerly provenancesdid so beforenortherly ones.
4. During bud-burst the newly-emergingshoots were hardy to only–3°C to –5°C untilthey were about 3.5 cmlong. All provenances burst bud at thesame time and were equallyfrost susceptible at this time.
5. DuringMay-July the elongating shoots fluctuated in hardinessbetween–5°C and –10°C apparently in responsetofluctuating ambient temperatures.
6. In August 1980 there wasa period of late summer dehardeningto killing temperaturesof about –3°C.

Seasonal changes in hardiness are discussed in relation to changesin shoot growth and environmental factors. The main opportunitiesfor selecting frost hardy genotypes seem to be in the rate ofautumn hardening, the time of pre-bud burst dehardening, andthe time of bud-burst.     

杉木苗期抗寒性的遗传变异和家系选择

近20年来,我国杉木遗传改良工作进展很快,但杉木育种目标多局限于速生丰产,过于单一。目前杉木的栽培区域不断从中心产区向边缘产区扩展延伸,出现了对不良气候、土壤条件的适应性问题。例如,1987年冬浙江各地苗圃遭受早霜冻害十分严重,使上百万株杉苗     

Chlorophyll fluorescence of stem cambial tissue reflects dormancy development in Juglans nigra seedlings

Assessment of chlorophyll fluorescence (CF) of forest tree seedlings can provide important insight into physiological function, dormancy status, and stress resistance. This evaluative tool has been measured routinely using foliage on conifer seedlings to assess seedling physiological status during winter dormancy. Absence of foliage during dormancy has thus far precluded the potential application of CF to seedling quality assessment of temperate deciduous hardwood seedlings. Because stems contain chlorophyll, however, assessment of CF using stem tissue may serve as an effective alternative tissue type to facilitate CF measurements. We collected Juglans nigra L. (black walnut) seed from two provenances (Alabama and Indiana, USA) and subjected 1-year-old container seedlings from these provenances to a simulated hardening regime (i.e., progressively decreasing temperatures and photoperiods) in a growth chamber environment over an 18-week period; CF of stem tissue (evaluated as ΦPSII, efficiency of photosystem II) was sampled periodically at seven time intervals. Though both provenance and measurement period significantly affected ΦPSII, measurement period had a much more pronounced effect. Values for ΦPSII fluctuated (ranging from 0.45 to 0.72) during the simulated growth and hardening regimes, generally decreasing over time for both provenances. Our results suggest that physiological status of temperate deciduous seedlings may be effectively evaluated during dormancy by assessing CF of stem tissue.     

不同种源非洲桃花心木苗期抗寒能力和生长节律研究

2008年1月上旬至2009年7月下旬,对非洲引进的3个桃花心木种源的幼树进行每周1次的观测.结果显示:种源间抗寒能力、树高生长和地径生长以及速生期都存在明显差异.种源3-1324与4-1325在苗期表现出一定的抗寒性,种源1-1320苗期对低温敏感.总体上,非洲桃花心木幼树1年抽梢4～7次,1～2月为树高生长休眠期,...     

Fast, nondestructive measurement of frost hardiness in conifer seedlings by VIS+NIR spectroscopy

Frost hardiness development from mid-August to mid-November was evaluated in seedlings of three provenances of Norway spruce (Picea abies (L.) Karst.) and three provenances of Scots pine (Pinus sylvestris L.) raised at nurseries in north, central and south Sweden. Measurements of the visible + near infrared (VIS+NIR) spectra of shoots were made simultaneously with estimates of frost hardiness based on electrolyte leakage following artificial freezing. Nine physiological variables known to influence frost hardiness were measured throughout the experiment. Multivariate analysis showed that VIS+NIR spectra explained 69% and 72% of the variation in frost hardiness in Scots pine and Norway spruce, respectively. Stem lignification, dry weight fraction, and starch, glucose, fructose, galactose, sucrose, raffinose and stachyose concentrations together explained 80% and 85% of the variation in frost hardiness in Scots pine and Norway spruce, respectively when used as independent X variables in a partial least squares model. These physiological variables could be related to varying degrees with variation in the VIS+NIR spectra. We conclude that VIS+NIR spectroscopy provides a rapid nondestructive technique for measuring frost hardiness in conifer seedlings based on causal relationships between the spectra and the physiology of seedling frost hardiness.     

Overwinter Storability of Conifer Planting Stock: Operational Testing of Fall Frost Hardiness

Operational stock-testing facilities that estimate overwinter storability of seedlings (ability to survive and grow after storage) need a reliable method that provides fast results to forest nurseries. We compared three methods using container-grown seedlings of Douglas-fir, interior spruce, lodgepole pine, and western larch from forest nurseries in British Columbia. On three to nine dates in autumn, frost hardiness at −18°C was estimated using visible injury of foliage or stems (VI), electrolyte leakage from needles or stems (EL), and chlorophyll fluorescence of shoots (CF). Seedlings were placed into overwinter cold storage (−2°C). In the spring, stored seedlings were planted in nursery beds; survival and growth were assessed after one growing season. There were close correlations (r ≥ 0.93) between the assessment methods. Seedlings lifted after they reached thresholds of 69% or higher for CF and 25% or lower for EL and VI had over 90% survival at harvest and doubled shoot dry weight compared with seedlings lifted earlier. Measuring CF was the fastest and most easily replicated method to estimate successful storability, and reduced testing time by 6 days relative to VI tests.     

华山松抗寒性的地理变异

本文报道了1980－1988年间26个产地的华山松实生苗和幼树的冻害情况，看到原产云贵高原亚热带山区的华山松，引种到暖温带气候区种植极不抗寒，基本不能越冬，云贵高原区域内种源的抗寒性也有某种程度的差异，尤以西部（云南保山地区）最弱，东部和北部（云南北部、四川西南部和贵州西部）较强，因而不经试验，不宜把云贵高原华山松种子调入暖温带使用。