Frost damage in Pinus sylvestris L. stems assessed by chlorophyll fluorescence in cortical bark chlorenchyma

? Several techniques were used to investigate the threshold minimum temperature inducing severe frost damage during winter in Scots pine (Pinus sylvestris L.) needles and stems from southern provenances in the Iberian Peninsula.

? Chlorophyll (Chl) fluorescence, electrolyte leakage (EL), visual scoring (VS), and the normalized difference vegetation index (NDVI) were compared.

? Chl fluorescence, and in particular the maximum potential photosystem II (PSII) efficiency — estimated through the dark-acclimated F_V/F_M ratio-, was found to be a simple, non-destructive indicator of freezing tolerance in needles, as expected.

? Moreover, the existence of a Chl-containing tissue, the cortical bark chlorenchyma, allowed us the use of Chl fluorescence as a new non-destructive indicator of frost damage in live tissues of Pinus sylvestris L. stems.

? Freezing tolerance values of southern provenances are compared with those found by other authors in the northern distribution limit of the species.

     

Frost hardiness of nutrient-loaded two-year-old <Emphasis Type="Italic">Picea abies</Emphasis> seedlings in autumn and at the end of freezer storage

The effects of nutrient loading (NLOAD) on the frost hardening and dehardening of Picea abies (L.) Karst. seedlings were investigated under nursery conditions. Before NLOAD, second-year container seedlings were either short-day (SD) treated for 3 weeks in July or left for the natural photoperiod (CO). By mid-September, after 5 weeks of NLOAD, the fertilization of three foliar nutrient concentration levels (low = L-SD, medium = M-SD, and high = H-SD) for the SD-treated seedlings and one (medium = M-CO) for the CO-seedlings was completed. The NLOAD resulted in foliar nitrogen concentration 10.6, 16.1, 22.3, and 17.5 g kg⁻¹ for L-SD, M-SD, H-SD and M-CO seedlings, respectively. The NLOAD had no effects on the morphology or dry mass variables of the seedlings, while SD-treatment reduced the dry mass of shoots, but not that of roots. The frost hardiness (FH) of different batches of the seedlings was assessed by the visual scoring of damage in their needles, stems and buds after their controlled exposure to freezing during frost hardening and dehardening. The low nutrient concentration in the SD-treated seedlings (L-SD seedlings) resulted in poor FH, to an even lower extent than that of the M-CO seedlings. The NLOAD did not affect the dehardening of the seedlings at the end of the freezer storage in the following spring.     

The use of field and artificial freezing studies to assess frost tolerance in natural populations of Pinus oocarpa

The susceptibility of Pinus oocarpa to freezing temperatures limits the commercial deployment of the highly productive Pinus patula × P. oocarpa hybrid in South Africa. Identifying P. oocarpa germplasm with increased frost tolerance is important. Twenty-three P. oocarpa provenances, originating from Mexico, Honduras, Guatemala and Nicaragua, were therefore assessed for their tolerance to freezing conditions by analysing field survival after frost events, subjecting needles to freezing temperatures and assessing damage using the electrolyte leakage test, and exposing young plants to freezing temperatures in a semi-controlled environment and scoring tissue damage based on a visual assessment. The performance of many of the provenances represented in the field and artificial freezing studies were similar and there was a strong correlation between provenance ranking in the whole plant freezing and electrolyte leakage test. We therefore support the use of these techniques as a means to assess cold tolerance in P. oocarpa at the provenance level. Provenances from north-western Mexico demonstrated more frost tolerance than those from southern Mexico. Provenances representing Honduras and Guatemala appear to be highly susceptible to frost.     

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.     

Nitrogen retranslocation, allocation, and utilization in bare root Larix olgensis seedlings

We quantified biomass accumulation and nitrogen (N) retranslocation, allocation, and utilization of Changbai larch (Larix olgensis) seedlings subjected to four fertilization treatments (24, 59, 81, 117 kg·ha-1 N) with an unfertilized control during summer and autumn 2009. Ammonium phosphate (18-46-0) was the fertilizer used in all treatments. On both sampling dates, the needles had greater biomass and N content than new (2009) stems and old (2008) stems, and coarse, medium and fine roots (diameters of >5, 2-5 mm, and 0-2 mm, respectively). Higher N concentration was observed in old stems and coarse roots than that in new stems and medium roots. In mid-summer, fine roots had higher N concentration than coarse roots. The treatment with 24 kg·ha-1 N had the greatest biomass and N content in needles and old stems, and highest net N retranslocation (NRA) and amount of N derived from soil. On September 21, no N translocation was observed, while the treatment with 24 kg·ha-1 N had the highest N utilization efficiency and fertilizer efficiency. Vector analysis revealed that all four fertilization treatments induced N excess relative to the control. The treatments with 59, 81, 117 kg·ha-1 N induce N excess compared with treatments at 24 kg·ha-1 N. We conclude that the traditional local fertilizer application rates exceeded N requirements and N uptake ability for Changbai larch seedlings. The application rate of 24 kg·ha-1 N is recommended.     

Ice nucleation and frost resistance of Pinus canariensis seedlings bearing needles in three different developmental states

Frost resistance and ice formation in different developmental states of needles of P. canariensis seedlings were assessed. Regrowth after frost damage was used to determine the overall frost survival capacity. Two distinct freezing exotherms (E1, E2) were registered. E1 was between ?1.7 and ?2.0 °C. Initial frost damage (LT₁₀) was 1.5–2.7 °C below E1. E2 was between ?5.6 and ?6.0 °C, and either corresponded with LT₅₀ or occurred in between LT₁₀ and LT₅₀. Current year needles were less frost resistant than 1-year-old needles. The overall recuperation capacity of seedlings revealed that frost survival may be underestimated when only needle damage is assessed. Freezing of seedlings with or without roots had no effect on the frost resistance of needles but recuperation capacity was significantly affected. Seedlings survived ?10 °C during summer indicating that they withstand the lowest naturally occurring frosts in Tenerife.     

Tephrosia species and provenances for improved fallows in southern Africa

Seeking an alternative to Sesbania spp. tree fallows, a Tephrosia species and provenance trial was conducted at Msekera Research Station, Chipata (Zambia) to evaluate eleven Tephrosia vogelii and three Tephrosia candida provenances. They were tested for biomass production, quality of biomass, resistance to root-knot nematodes, nitrogen release, and for their effects on soil nitrogen dynamics. At the end of 1.5 years, the T. candida provenances 02970, 02971 and 02972 from Madagascar produced two times greater amount of aboveground biomass than the T. vogelii provenances. There was little variability among the T. vogelii provenances in terms of litter and biomass production. Weed growth was significantly greater under T. vogelii than T. candida provenances. While Tephrosia vogelii provenance 98/02 from Zambia and T. candida 02972 were highly tolerant to the Meloidogyne incognita nematodes, T. vogelii provenances 02977, 98/03, 02973 from Kenya, Zambia and Malawi, respectively, were highly susceptible to the nematodes. The Tephrosia species and provenances showed a wide variability in terms of N, lignin and polyphenol concentration in their foliage. Mineralization of N in the foliage of T. candida provenances 02970 and 02971 and T. vogelii provenances 98/04 and 02974 from Malawi occurred rapidly within 14 weeks of incubation. At the end of the 2-year growth period, there was significantly greater total inorganic N under T. candida provenance 02972 (12.5 mg kg⁻¹) than T. vogelii (5 mg kg⁻¹) provenance Mungwi 98/02. Maize (Zea mays L.) yields after T. candida provenances were greater than those after T. vogelii provenances. Further testing of the most promising provenances is needed for their effects on subsequent maize yields under a range of farm conditions. This revised version was published online in June 2006 with corrections to the Cover Date.     

Genetic diversity within and among southern African provenances of <Emphasis Type="Italic">Uapaca kirkiana</Emphasis> Müell. Årg using morphological and AFLP markers

Domestication of Uapaca kirkiana Müell. Arg is a high priority for improving rural livelihoods of smallholder farmers in southern Africa. Domestication efforts require knowledge of adaptive traits and intra-specific variation. Morphological traits and amplified fragment length polymorphic (AFLP) markers were used to assess genetic variation in twelve provenances of U. kirkiana collected from southern Africa. Assessment of morphological traits showed significant differences (P < 0.05) between provenances. Provenances from Zimbabwe and Zambia showed faster growth than those from Malawi (except Phalombe) and Tanzania. Morphological traits exhibited strong genetic differentiation between provenances and within provenances. The degree of provenance differentiation of traits (Q _ST = 0.03–0.139, Q _ST mean = 0.092) was in the same range with that of AFLP loci (F _ST mean = 0.089) detected among provenances and within provenances (0.002 < F _ST < 0.259). The differentiation for some morphological traits could be attributed to local adaptation and human selection of the U. kirkiana trees at the site of origin. Mean Nei’s (H) genetic diversity of AFLP showed high diversity within the provenances (H = 0.181–0.321, H mean = 0.256). An analysis of molecular variance (AMOVA) revealed higher genetic variation (90.8%) within provenances than among provenances (9.2%). There was no geographical pattern of variation in growth and morphological traits among the seed sources. Chipata provenance from Zambia was the most diverse while Mapanzure from Zimbabwe was the least diverse, though it was superior in height and earliest in fruiting. The pattern of genetic diversity indicates low selection in some areas and high gene flow which would be counteracting it. Therefore regional and country collections and conservation strategies should consider differences by focussing on the main range of the species by paying particular attention to unique populations within countries.     

Late frost sensitivity of juvenile <Emphasis Type="Italic">Fagus sylvatica</Emphasis> L. differs between southern Germany and Bulgaria and depends on preceding air temperature

Fagus sylvatica, the dominant native forest tree species of Central Europe, is sensitive to late frost events. Advanced leaf flushing due to climate warming may lead to more frequent frost damage in the future. Here, we explore local adaptation to late frost events at both continental and regional scales and test how moderate climate warming (+1.5°C) affects late frost sensitivity. Short-term leaf injury and height growth after a late frost event were quantified in a common garden experiment with 2-year-old F. sylvatica seedlings. The fully crossed three-factorial design consisted of a late frost manipulation, a continuous warming manipulation and selected provenances (three provenances from western Bulgaria and three from southern Germany). Late frost led to leaf injury and reduced height growth (−7%). Provenances differed in their late frost sensitivity at the regional scale, and local adaptation was detected. At the larger scale, the Bulgarian provenances showed reduced height growth (−17%), while the German provenances did not exhibit growth reduction. The warming treatment prevented late frost damage, while height growth declined by 19% in the reference temperature treatment. This surprising finding was attributed to advanced leaf maturity in the warming treatment. The impact of late frost events on F. sylvatica in a warmer world will depend on timing. An event that damages leaves immediately after leaf flushing appears negligible a few days earlier or later, thereby complicating projections. Local adaptation to late frost is evident at a regional scale. Management strategies should aim at maximizing genetic diversity to adapt to climate change.     

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.     

Effect of damaged pine needles on growth and development of pine caterpillar larvae

Chinese pine caterpillar (Dendrolimus tabulaeformis) larvae were fed with pine needles of different degrees of damage to evaluate the effects of pine needles on the growth and development of larvae. The results showed that the nutritional index of the larvae declines after feeding on the damaged pine needlings. The lowest amount of food ingested and voided feces, the lowest nutritional index, slowest development, lightest pupae and most mortality were found in those pine caterpillar larvae fed with pine needles which were 50% damaged. The damaged pine needles significantly affected the population dynamics of Chinese pine caterpillars. The nutritional indices of larvae fed with 25% and 75% damaged pine needles were similar. The nutritional index of the dark morphs was higher than that of the tinted morphs, however, their mortality was lower than that of the tinted morphs. This phenomenon was reversed at the later stage of development when the larvae were fed on 50% damaged pine needles. __________ Translated from Journal of Beijing Forestry University, 2005, 27(6): 83–88 [译自: 北京林业大学学报, 2005, 27(6): 83–88]     

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.     

Shading Reduces both Visible and Invisible Frost Damage to Norway Spruce Seedlings in the Field

Two-year-old cuttings of Norway spruce were subjected to nightfrosts in spring on an exposed site in southern Sweden. Shadingwas used to assess the influence of sunlight on the extent ofdamage resulting from night frost. Chlorophyll fluorescencewas measured in needles in flushing shoots, and in shoots atthe stage of bud burst. The F_v:F_m ratio was significantly lowerfor plants exposed to light, compared with shaded plants onthe days following the night frost (minimum temperature –6°C).The effect was similar both in 1-year-old and current year needles.The low F_v:F_m ratios indicate damage to photosystem H, causedby an interaction between sub freezing temperatures and highlight intensity. Shading also increased the survival of flushingshoots. It is suggested that regeneration of Norway spruce onsites exposed to frost should be carried out in partial shade,for example under a shelterwood.     

Winter Frost Hardiness of two Chilean Provenances of Nothofagus procera in Scotland

The frost hardiness of the shoots of individual trees withintwo Chilean provenances of Nothofagus procera (Poepp & Endl.)Oerst. was measured once in each of the months January, February,November and December 1989 and January and February 1990. Therewere significant (P<0.05) differences of frost hardinessbetween provenances but only one tree could be shown to be significantlymore frost hardy than the others within the same provenance.During the winter of 1989/90 both provenances were hardy toabout –14°C (temperature killing 50 per cent of shoots)in December, but the shoots dehardened to about –9°Cin January before hardening again in February. This patternof alternate hardening and dehardening seemed to mirror changesin air temperature and could render N. procera liable to frostdamage where (as happened in 1988/9 in the UK) mild spells occurin winter followed by severe frosts.     

Effect of Ca on Al-induced activation of antioxidant enzymes in the needles of hinoki cypress (Chamaecyparis obtusa)

The effect of CA(NO₃)₂ on the active oxygen scavenging system in hinoki cypress (Chamaecyparis obtusa) seedlings cultured in a nutrient solution containing aluminum was examined. The hinoki cypress seedlings were transferred to nutrient solutions containing 5 mM AlCl₃ together with various concentrations of Ca(NO₃)₂ in pots containing glass beads and Teflon tips. The growth in height and dry matter allocation to each organ was little influenced over a period of 12 weeks by either Al or the concentration of Ca(NO₃)₂. The activity of superoxide dismutase (SOD) in the needles was stimulated by Al, and the effect of Al was lowered significantly by simultaneous application of 25 mM Ca(NO₃)₂. At week 1, the activity of catalase (CAT) in the needles was increased by Al, but the effect was no longer observed at week 12. The Al concentration in the roots was increased by treatment with Al, whereas the Al concentration in needles was not. These results indicate that rhizospheric Al stress stimulates antioxidative enzyme activities in hinoki cypress needles and the activation of the enzymes is suppressed by addition of Ca. The transmission of Al stress to the needles, which induced a change in the enzyme activity, is not caused by the transfer of the Al ion itself from roots to needles. This work was supported in part by funding from the Japan Science and Technology Corporation, the CREST program 1996–2001, and the Center for Forest Decline Studies.     

Relationships between foliar phosphorus fractions of Pinus sylvestris var. mongolica and soil available phosphorus

In order to find out the best foliar diagnostic index of phosphorus (P) nutrition in Mongolian pine (Pinus sylvestris var. mongolica) in the southeastern Keerqin Sandy Lands, the concentrations of total nitrogen (N), inorganic P, organic P and total P in needles of different ages and soil available P were examined. The results show that in the study area, soil available P was rather low (0.12–0.63 mg/kg) and was significantly correlated with inorganic P (cPi) and total P (cPt) concentrations in current year needles of Mongolian pine. The significant correlation between soil available P and needle cPt derived from the significant correlation between cPi and cPt. Compared with cPt, cPi did reflect the level of soil P supply more accurately and more directly. __________ Translated from Chinese Journal of Applied Ecology, 2008, 19(3): 494–498 [译自: 应用生态学报]     

冷湿层积法对印度冷杉(Abies pindrow)和长叶云杉(Picea smithiana）种子萌发的影响

研究赤霉素(GA3),冷湿和温度 对五个种源的印度冷杉(Abies pindrow)和长叶云杉(Picea smithiana)种子萌发的影响.种子被浸泡在GA3 (10 mg(L-1)中24小时,然后在3(5(C温度的条件下冷藏15天.设计4个温度(10(C, 15(C, 20(C 和 25(C)条件来促进种子的萌发.结果表明,浸泡和冷湿处理明显增加种子的萌发率.在10 (C时种子的萌发率最高.总体结果表明,种子被浸泡在GA3 (10 mg(L-1)中24小时,冷湿藏15天,可以有效地促进印度冷杉和长叶云杉的种子萌发.统计数据表明,浸泡处理、温度和种源以及与温度的相互作用都对种子的萌发有明显的影响.     

Effects of ozone on gas exchange,frost resistance,flushing and growth of different provenances of European silver fir (Abies alba Mill.)

Photosynthesis, respiration, and transpiration were studied in different Abies alba provenances during and alter ozone exposure at three different levels (0/control, 125, and 250 ppb O₃, respectively). The after-effects of ozone on frost resistance and growth rate were also analyzed. Pronounced reductions of photosynthesis and transpiration rates and an increased respiration were registered as a response to O₃. Five months after exposure the winter frost resistance was significantly reduced, and growth depressions were registered during the following year. Differences in ozone sensibility and regeneration ability between provenances could be demonstrated.     

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.     

Comparison of foliar nutrient concentrations between natural and artificial forests of Pinus sylvestris var. mongolica on sandy land, China

In order to examine the causes of degradation of Pinus sylvestris var. mongolica plantations on sandy land, the foliar concentrations of N, P, K and C were analyzed and compared between the field grown P. sylvestris var. mongolica trees from two provenances （natural forests and plantations）. The results indicated that natural tree needles had lower N, P and C concentrations, and higher K concentrations than those of plantation tree needles. For plantation tree needles, ratios of N： P, P. K and N： K increased with tree age before 45 years old; but they were not clear for the natural tree needles. Compared with the conclusions reported on Pinus spp., we found that the foliar N and P concentrations were in the optimal range for both natural and plantation tree needles. This result suggested that N or P might not be the absolute limit factors in plant nutrient for P sylvestris var. mongolica on sandy land. However, foliar K concentrations in both natural and plantation tree needles were much lower than those reported on Pinus spp. （〉4.80 g kg-1）.The N： P ratio of natural needles was in the adequate ranges, but N： P ratio of plantation needles was out of the adequate ranges. These results indicated that there was a better balanced nutrition status in the natural forest than in the plantations. If only considering the foliar nutrient concentrations of P sylvestris var. mongolica from different provenances, it might be concluded that the degradation phenomenon of P. sylvestris var. mongolica plantations was not induced by nutrition deficiency of absolute nutrients of N and P, but might be induced by other mineral nutrients or by the effectiveness of N and P nutrients. The unbalanced nutrition status and relatively quick decomposition of needles in the plantations might also contribute to the degradation.