Photoactivation of the latent water-oxidizing complex in photosystem II membranes isolated from dark-grown spruce seedlings

Photosystem II membranes (D-PSII) were isolated from dark-grown spruce seedlings. All major PSII proteins except the 17- and 23-kDa extrinsic proteins were present in D-PSII. O₂ evolution and Mn content in D-PSII were negligible, while PSII-donor activity showed a value comparable to that of NH₂OH-treated PSII membranes (NH₂OH-L-PSII) from light-grown seedlings. Light incubation of D-PSII with 1 m M MnCl₂, 50 m M CaCl₂ and 100 μ M DCIP at pH 5.3 resulted in activation of the latent water-oxidizing complex. Accomplishment of photoactivation of PSII membranes from dark-grown spruce seedlings clearly indicates that only ligation of Mn²⁺ to the apo-water oxidizing complex is required for expression of O₂ evolution, and that protein synthesis is not involved in the photoactivation process. There was no essential difference between 'photoactivation' of naturally Mn-free PSII membranes and 'photoreactivation' of artificially Mn-depleted PSII membranes on kinetics, pH dependence, Mn²⁺-concentration dependence. However, kinetics and pH dependence of photoactivation were appreciably different in spruce PSII membranes and in PSII membranes of angiosperms such as wheat and spinach.     

Cell structural changes in the needles of Norway spruce exposed to long-term ozone and drought

Effects of ozone and/or drought on Norway spruce needles werestudied using light microscopy and electron microscopy. Saplingswere exposed to ozone in open-top chambers during 1992–1995and also to drought in the late summers of 1993–1995.Samples from current and previous year needles were collectedfive times during 1995. Ozone increased the numbers of peroxisomesand mitochondria, which suggests that defence mechanisms againstoxidative stress were active. The results from peroxisomes suggestthat the oxidative stress was more pronounced in the upper sideof the needles, and those from mitochondria that defence wasmore active in the younger needle generation. Possibly due tothe good nitrogen status and the active defence, no ozone-specificchloroplast alterations were seen. At the end of the season,older needles from ozone treatments had smaller central vacuolescompared with other needles. Cytoplasmic vacuoles around thenucleus were increased by ozone in the beginning of the experiment,and did not increase towards the end of the season as in thecontrols. These results from vacuoles may indicate that ozoneaffected the osmotic properties of the cells. Decreased numberand underdevelopment of sclerenchyma cells and proliferationof tonoplast were related to nutrient imbalance, which was enhancedby drought. Larger vascular cylinders and more effective starchaccumulation before and after the drought periods compensatedfor the reduced water status. Numbers of peroxisomes and mitochondriawere increased in the drought-exposed needles before the onsetof drought treatments of the study year, i.e. these changeswere memory effects. Interactions between ozone and droughtwere few.     

Nutrient content and photosynthesis of young yellowing Norway spruce trees (Picea abies L. Karst.) following calcium and magnesium fertilisation

Severely yellowed ten-year-old spruce trees growing in the Vosges Mountains on an acidic soil were fertilised with Magnesium lime during the spring of 1990. The effects of this treatment were assessed 18 months later. A very significant improvement of the mineral status of the trees was detected, with increasing Mg contents in the needles, and as a consequence, reduced yellowing and improved chlorophyll content. Only slight differences with control trees were observed for height increase. Effects of this improved nutrition on photosynthesis were tested measuring net CO₂ assimilation rates and chlorophyll a fluorescence. Light-saturated net assimilation rates of current-year needles were high, reaching 5.3 mol m^–2 s^–1 on a total needle area basis. The improvement in chlorophyll and Mg content had no significant effect on net assimilation rates or on any parameter describing photochemical functions of both current-and previous-year needles. Despite the strong inter-individual variability in needle chlorophyll and Mg contents (ranging from 0.2 to 0.8 mg g^–1 fresh weight, and 0.05 to 0.5 mg g^-1 dry weight respectively), photochemical efficiency of PS II under limiting irradiance only decreased significantly on older needles displaying Mg contents below 0.1 mg g^–1. It is concluded from these results that spruce trees exhibit a high degree of plasticity with regard to Mg deficiency on acidic soils, and that improved Mg nutrition and increased chlorophyll content do not necessarily improve photosynthesis and height growth.Abbreviations A light-saturated net CO₂ assimilation rate (mol m^–2 s^–1) - g_w light-saturated needle conductance to water vapour (mmol m^–2 s^–1) - _wp and _wm pre-dawn and mid-day needle water potential (MPa) - osmotic potential of sap expressed from needles (MPa) - PFD photosynthetic photon flux density (mol m^–2 s^–1) - F_v/F_m photochemical efficiency of PS II after 20 min dark adaptation - F/F_m ^' photochemical efficiency of PS II reaction centres after 10 min at a PFD of 220 mol m^–2 s^–1     

Genetic diversity in a seed production population vs. natural populations of Sitka Spruce

Isozyme analysis was applied to estimate the level of variation and the genetic structure of a seed-production population (i.e., seed orchard) and 10 range-wide natural populations of Sitka spruce (Picea sitchensis (Bong.) Carr.). Gene diversity and heterozygosity estimates were comparatively high in both the seed orchard and the natural populations studied. The seed orchard population showed a significantly higher number of alleles per locus and percentage of polymorphic loci. Though not significant, mean heterozygosity of the seed orchard was higher than that observed for all natural populations. Genetic distance analysis indicated that the seed-orchard population was genetically similar to three natural populations from which the parent trees were selected. Parent trees sampling breadth has been identified as the major cause for the observed increased level. The impact of recurrent selection and seed orchard biology and management on maintaining the genetic diversity is discussed.     

Early needle senescence and thinning of the crown structure of Picea abies as induced by chronic SO2 pollution. II. Field data basis, model results and tolerance limits

Field data on the sulphur and cation budget of growing Norway spruce canopies (Picea abies [L.] Karst.) are summarized. They are used to test a spruce decline model capable of quantifying effects of chronic SO₂ pollution on spruce forests. At ambient SO₂ concentrations, acute SO₂ damage is rare, but exposure to polluted air produces reversible thinning of the canopy structure with a half-time of a few years. Canopy thinning in the spruce decline model is highest (i) at elevated SO₂ pollution, (ii) in the mountains, (iii) at unfertilized sites with poor K⁺, Mg²⁺ or Zn²⁺ supply, (iv) at low spruce litter decomposition rates, and (v) acidic, shallow soils at high annual precipitation rates in the field and vice versa. Model application using field data from Würzburg (moderate SO₂ pollution, alkaline soils, no spruce decline) and from the Erzgebirge (extreme SO₂ pollution, acidic soils in the mountains, massive spruce decline) predicts canopy thinning by 2–11% in Würzburg and by 45–70% in the Erzgebirge. The model also predicts different SO₂-tolerance limits for Norway spruce depending on the site elevation and on the nutritional status of the needles. If needle loss of more than 25% (damage class 2) is taken to indicate ‘real damage’ exceeding natural variances, then for optimum soil conditions SO₂ tolerance limits range from (27.3 ± 7.4) μg m^?3 to (62.6 ± 16.5) μg m^?3. For shallow and acidic soils, SO₂ tolerance limits range from (22.0 ± 5.5) μg m^?3 to (37.4 ± 7.5) μ m^?3. These tolerance limits, which are calculated on an ecophysiological data basis for Norway spruce are close to epidemiological SO₂-toIerance limits as recommended by the IUFRO, UN-ECE and WHO. The observed statistical regression slope of the plot (damaged spruce trees vs. SO₂-pollution) in west Germany is confirmed by modelling (6% error). Model application to other forest trees allows deduction of the observed sequence of SO₂-sensitivity: Abies > Picea > Pinus > Fagus > Quercus. Thus, acute phytotoxicity of SO₂ seems not to be involved in ‘forest decline’. Chronic SO₂-pollution induces massive canopy thinning of Abies alba and Picea abies only at unfavourable sites, where natural stress factors and secondary effects of SO₂pollution act together to produce tree decline.     

A long-term record of carbon exchange in a boreal black spruce forest: means, responses to interannual variability, and decadal trends

We present a decadal (1994–2004) record of carbon dioxide flux in a 160‐year‐old black spruce forest/veneer bog complex in central Manitoba, Canada. The ecosystem shifted from a source (+41 g C m⁻², 1995) to a sink (−21 g C m⁻², 2004) of CO₂ over the decade, with an average net carbon balance near zero. Annual mean temperatures increased 1–2° during the period, consistent with the decadal trend across the North American boreal biome. We found that ecosystem carbon exchange responded strongly to air temperature, moisture status, potential evapotranspiration, and summertime solar radiation. The seasonal cycle of ecosystem respiration significantly lagged that of photosynthesis, limited by the rate of soil thaw and the slow drainage of the soil column. Factors acting over long time scales, especially water table depth, strongly influenced the carbon budget on annual time scales. Net uptake was enhanced and respiration inhibited by multiple years of rainfall in excess of evaporative demand. Contrary to expectations, we observed no correlation between longer growing seasons and net uptake, possibly because of offsetting increases in ecosystem respiration. The results indicate that the interactions between soil thaw and water table depth provide critical controls on carbon exchange in boreal forests underlain by peat, on seasonal to decadal time scales, and these factors must be simulated in terrestrial biosphere models to predict response of these regions to future climate.     

An attempt to predict long-term effects of atmospheric nitrogen deposition on the yield of Norway spruce stands (Picea abies (L.) Karst.) in southwestern Sweden

Long-term field experiments in Norway spruce stands on fertile sites (site indices 27–35 m) in southwestern Sweden were analysed with respect to volume increment. Three treatments were included (0=No fertilization, N = Fertilization with N, NP = Fertilization with N and P).Volume growth was monitored for 18 years in 10 blocks. No significant differences in annual volume increment between the treatments were detected. Volume increments in the N treatment were 97%, 99% and 107% as high as those in the 0 treatment for the periods 1–5, 6–10 and 11–15 years after the first fertilization. Corresponding values for the NP treatment were 104%, 108% and 110%, indicating that P has a small positive effect.The amount of N-fertilization would correspond to an annual N deposition of 20 kg ha^-1 during the next 30 years in southwestern Sweden. For this period, the results imply that this N deposition would not affect the growth of Norway spruce stands on fertile sites.     

Distinct sources of gene flow produce contrasting population genetic dynamics at different range boundaries of a Choristoneura budworm

Populations are often exposed to multiple sources of gene flow, but accounts are lacking of the population genetic dynamics that result from these interactions or their effects on local evolution. Using a genomic clines framework applied to 1,195 single nucleotide polymorphisms, we documented genomewide, locus‐specific patterns of introgression between Choristoneura occidentalis biennis spruce budworms and two ecologically divergent relatives, C. o. occidentalis and Choristoneura fumiferana, that it interacts with at alternate boundaries of its range. We observe contrasting hybrid indexes between the two hybrid zones, no overlap in “gene‐flow outliers” (clines showing relatively extreme extents or rates of locus‐specific introgression) and variable linkage disequilibrium among those outliers. At the same time, correlated genomewide rates of introgression between zones suggest the presence of processes common to both boundaries. These findings highlight the contrasting population genetic dynamics that can occur at separate frontiers of a single population, while also suggesting that shared patterns may frequently accompany cases of divergence‐with‐gene‐flow that involve a lineage in common. Our results point to potentially complex evolutionary outcomes for populations experiencing multiple sources of gene flow.     

Latitudinal cline of requirement for far‐red light for the photoperiodic control of budset and extension growth in Picea abies (Norway spruce)

To test for the effects of far‐red light on preventing budset in Picea abies , seedlings of six populations originating from latitudes between 67°N and 47°N were grown for 4–8 weeks in continuous incandescent (metal halogen) light at 300 µmol m⁻² s⁻¹ and 20°C and then transferred, at the same temperature, to a daily regime of 8 h incandescent light (300 µmol m⁻² s⁻¹) followed by 16 h cool white fluorescent light (40 µmol m⁻² s⁻¹). (Cool white lamps are deficient in far‐red light, with a R/FR ratio of 7.5 compared with 2.0 for the incandescent lamps.) All the seedlings from 67° and 80% of those from 64° stopped extension growth and set terminal buds within 28 days of the change of regime. The seedlings from 61° and further south continued growing, as did control seedlings from 67° grown as above but with incandescent light at 20 µmol m⁻² s⁻¹ replacing cool white illumination. To distinguish between a clinal and ecotypic pattern of variation, the interval between 64° and 59° was investigated by growing populations originating from that area in the same regimes as before. After 28 days in the cool white day‐extension regime, the percentage budset was 86 for the population from 64°, 0 for the population from 59° and 25–50 for the intermediate populations; i.e. the populations showed a clinal variation in requirement for far‐red light according to latitude. Thus northern populations of Picea abies appear to behave as 'light‐dominant' plants for the photoperiodic control of extension growth and budset, whereas the more southern populations behave as 'dark‐dominant' plants.