Establishment of Picea abies seedlings in a central European mountain grassland: an experimental study

Abstract. Secondary meadows in the Kru?né hory mountains, central Europe, are not colonized by woody species, although they have been abandoned for half a century and are surrounded by Picea abies (Norway spruce) forests. The causes of inhibition of establishment of Norway spruce seedlings in the meadows were tested experimentally. The experiment was started in a masting year to ensure sufficient seed input. Four treatments (vegetation cut; vegetation cut and litter removed; all above-ground biomass and topsoil removed; control) were combined in a factorial design. The effect of browsing, mainly by deer, was assessed by fencing half of the experimental plots. Seedling establishment differed among treatments (all pair-wise differences were significant) but was independent of fencing. In contrast, seedling survival was influenced both by the treatment and fencing. At the end of the second year, only the plots with all biomass and organic topsoil removed supported viable populations of spruce seedlings. Dense herb cover and a thick layer of slowly decaying litter are considered the main factors inhibiting the establishment of woody species.     

Effect of starvation on copulatory activity of Ercolania nigra (Lemche, 1935) (Mollusca,Opisthobranchia, Ascoglossa)

Feeding dominates copulatory behavior in the few opisthobranchs for which behavioral hierarchies have been established. However, the influence of starvation on the dominant role has not been investigated previously. The present study investigates copulatory activity in the ascoglossan opisthobranch Ercolania nigra (Lemche) subject to varying starvation regimes. Copulatory activity of E. nigra is significantly higher in unstarved animals than in animals starved for 2h, 6h, and 24h. The decrease in copulatory activity is independent of the duration of previous starvation within the range tested (2–24 h). Copulatory behavior dominates feeding behavior in 60–80% of the cases, regardless of the duration of previous starvation. Thus the behavioral hierarchy of E. nigra differs markedly from that of the opisthobranchs previously investigated.     

Structure of a pristine Picea abies forest in northeastern Europe

Abstract. The forest structure in a large, relatively homogeneous area of pristine Picea abies taiga in the southern boreal region west of the Ural mountains was studied along four 500-m long transects. P. abies dominated the forest in association with Abies sibirica and Betula spp. The mean volume of living trees was 216 m³/ha. This value varied among the four transects, from 138 - 252 m³/ha. Mean density of trees (DBH > 1 cm) (and variation over the transects) was 2 064/ha (1670 - 2710). Living trees classified as dying or seriously damaged made up 2.9 (2.5 - 3.5) % of the living tree volume. Betula was an important canopy component and made up 16% of the living tree volume. The number of dead standing trees varied from 195 - 325/ha, corresponding to a volume of 10.8 - 70.7 m³/ha. The density of trees with a broken stem was 90 - 170/ha and their estimated volume 7.6 - 41.3 m³/ha. Standing dead trees and trees with broken stems represented 10.4 and 8.9% of the total standing tree volume (living + dead), respectively. The mean volume of decaying logs on the forest floor was 117 (84.4–156.2) m³/ha, corresponding to 54 (35 - 113) % of the living tree volume. The canopy-forming trees were present in the understory as abundant saplings and suppressed individuals. The size distribution of the dominant tree species resembled the inverse J-shape. Generally, the forest was characterized by a high small-scale structural variation and a larger-scale relative homogeneity. This pattern is consistent with forest dynamics where the forest consists of a small-scale mosaic of patches in different stages of recovery following disturbance. Our results suggest that the ecology and dynamics of extensive areas of natural boreal forests can be driven by small-scale disturbance.     

Response of fine roots to an experimental gap in a boreal Picea abies forest

We examined the initial response of the quantity and distribution of fine roots to the creation of an experimental canopy gap with a diameter of 50 m in a mature managed Norway spruce forest. Under the canopy, the fine root length densities of trees, shrubs, and grasses and herbs were 3207, 707 and 2738 m m^–2, respectively. The fine root biomass of trees, shrubs, and grasses and herbs were 182, 47 and 52 g m^–2, respectively. Two growing seasons after gap creation hardly any fine tree roots were found in the middle part of the gap. The living tree roots in the gap edge zone were mainly located within a 5-m distance from the standing edge trees. The indices developed here to show the influence of trees on fine root lenght density clearly revealed the effect of the vicinity of living trees on fine root lenght density. The root densities of grasses, herbs and dwarf shrubs did not show a clear response to gap creation despite the increase of their foliage. Our results suggest that in boreal spruce forests a gap disturbance creates a distinct tree root gap and that the gap edge trees do not extend their root systems rapidly into the formed root gap.     

Effect of seed size on seedling growth response to elevated CO2 in Picea abies and Picea rubens

Several previous studies have observed that species and individuals with large seeds respond more positively to elevated CO (2) than those with small seeds. We explored the reasons for this pattern by examining the relationship between seed size and CO (2) response in Picea abies and P. rubens using growth analysis. The large seeded species (P. abies) responded more positively to elevated CO (2) than the small seeded species (P. rubens). At the intraspecific level, P. abies individuals from large seeds responded more positively to elevated CO (2) than individuals from small seeds, however, there was no significant intraspecific variation in CO (2) response in P. rubens. The greater CO (2) response of plants from large seeds was not simply the result of a larger starting capital compounded at the same rate as in plants from small seeds. Elevated CO (2) increased relative growth rate to a greater extent in individuals from large seeds. This effect appears to be related to differences in time of establishment, source to sink ratio and nutrient availability with seed size. These results are significant not only in understanding the potential effect of rising atmospheric CO (2) concentrations on plant populations, but also in understanding the factors affecting plant success at current atmospheric CO (2) levels due to the elevation of CO (2) within the litter layer that occurs at many germination sites.     

Genetic transformation of Norway spruce (Picea abies (L.) Karst) using somatic embryo explants by microprojectile bombardment

Stable transformation of Norway spruce tissue has been obtained following bombardment of mature somatic embryos with pRT99gus, a plasmid that contains neo coding for NPTII, and gusA, coding for -glucuronidase, both fused to the CaMV 35S promoter. At least 8 lines have been stably transformed (over 15 months in culture) following bombardment and selection on kanamycin. Polymerase chain reaction analyses showed a high frequency of cotransformation of the gusA and neo genes. The frequency of coexpression of the selected and unselected markers was 100%. DNA/DNA hybridization of one transformed line provided conclusive evidence of stable integration and showed copy numbers of over 10 plasmid sequences per genome. None of the transformed lines has remained embryogenic.     

温度对副珠蜡蚧阔柄跳小蜂发育和繁殖的影响

通过观察副珠蜡蚧阔柄跳小蜂Metaphycus parasaissetiae Zhang and Huang在18,21,24,27,30,33和36℃恒温条件下的生长发育情况,求得其世代发育起点温度和世代有效积温分别为13.10℃和215.00日.度,建立世代发育历期预测式为N=[215.00±44.32]/[T-(13.10±3.24)],世代发育速率与温度之间的关系拟合为S形曲线V=-e(1.33+37.73/T),推算出该蜂在海南1年可发生18代。所设试验温度范围内,该蜂对橡副珠蜡蚧Parasaissetia nigra(Nietner)的寄生率在13.33%～20.00%范围内波动,30℃时寄生率最高;雌性比率在27℃时最大为70.03%。随着试验温度的升高,成蜂羽化相对集中,羽化持续时间逐渐缩短,羽化高峰提前。30℃条件下雌蜂终生产卵量最多,平均25.77粒。雌蜂寿命随试验温度的升高而逐渐缩短,同一恒温条件下补充20%蜂蜜水可显著延长雌蜂寿命。27～30℃可视为有利于副珠蜡蚧阔柄跳小蜂发育繁殖的适温范围,最适温度为30℃。     

Biodegradation of polyethylene by the thermophilic bacterium Brevibacillus borstelensis

AIM: To select a polyethylene-degrading micro-organism and to study the factors affecting its biodegrading activity. METHODS AND RESULTS: A thermophilic bacterium Brevibaccillus borstelensis strain 707 (isolated from soil) utilized branched low-density polyethylene as the sole carbon source and degraded it. Incubation of polyethylene with B. borstelensis (30 days, 50 degrees C) reduced its gravimetric and molecular weights by 11 and 30% respectively. Brevibaccillus borstelensis also degraded polyethylene in the presence of mannitol. Biodegradation of u.v. photo-oxidized polyethylene increased with increasing irradiation time. Fourier Transform Infra-Red (FTIR) analysis of photo-oxidized polyethylene revealed a reduction in carbonyl groups after incubation with the bacteria. CONCLUSIONS: This study demonstrates that polyethylene--considered to be inert--can be biodegraded if the right microbial strain is isolated. Enrichment culture methods were effective for isolating a thermophilic bacterium capable of utilizing polyethylene as the sole carbon and energy source. Maximal biodegradation was obtained in combination with photo-oxidation, which showed that carbonyl residues formed by photo-oxidation play a role in biodegradation. Brevibaccillus borstelensis also degraded the CH2 backbone of nonirradiated polyethylene. SIGNIFICANCE AND IMPACT OF THE STUDY: Biodegradation of polyethylene by a single bacterial strain contributes to our understanding of the process and the factors affecting polyethylene biodegradation.     

Variation in total and specific RNA during inwintering of two contrasting populations of Picea abies

Seedlings of a southern (Romanian) and a northern (Swedish) population of Picea abies (L.) Karst. were cultivated in a phytotron under continuous lighl and 20°C for 10 weeks. Subsequently they were exposed to a regime of long nights (16 h). initially at 20°C but with stepwise lowering to 10°C over 12 weeks. Samples of needles were taken for total RNA extraction and studies of gene expression at intervals from just before the change of regime and onwards. At the same time samples were taken for abseisic acid (ABA) determination. Extension growth, degree of budrest and frosl tolerance were also recorded. The main results were as follows. (1) Extractable total RNA per g fresh weight of needles increased by 150–250% through the period of long nights. (2) The capacity of the total RNA to direct protein synthesis in a cell-free system declined in response to long nights to a minimum value of 5% (northern population) and 23% (southern population) of the reference values under continuous light. (3) In vitro translatability was highly correlated with the size of the polyadenylated fraction of the total RNA, which declined also when normalized to needle fresh weight rather than total RNA. (4) A peak of abscisic acid content in the southern population at 3 days into the long night treatment coincided with a trough in the in vitro translatability of the total RNA. (5) The steady state levels of rbc S-mRNA as a fraction of total RNA declined rapidly to a final value of about 15% of the reference value in both populations, the decline being more rapid for the northern population. (6) In contrast. ubiquitin-mRNA showed an initial increase, particularly in the northern population, and at the end of the long night treatment was only 40% under the reference value. (7) Rapid changes in the molecular characters occurred during the first 3–4 weeks when the morphological and physiological changes were most rapid.     

Influence of liming and acidification on the activity of the mycorrhizal communities in a Picea abies (L.) Karst. stand

A study of mycorrhizal activity was conducted in a mature Norway spruce ((Picea abies) [L.] Karst.) stand subjected to soil treatments of liming and acidification for six years (Höglwald research project). Samples were collected five times during one growing season using a soil corer. All the turgescent, not shriveled mycorrhizal tips were sorted out and identified on the fungal species level as far as yet possible. The proportion of each mycorrhizal type on the plots was calculated. The results revealed a shift in the mycorrhizal communities caused by both acid rain and liming. Data are in agreement with the findings of the more comprehensive study on the mycorrhizal communities carried out by another research group on the same plots (Taylor et al., submitted)The activity of the predominant types of mycorrhizas, Piceirhiza gelatinosa, Piceirhiza nigra, Russula ochroleuca-P. abies, Tuber puberulum-P. abies, Tylospora sp.-P. abies, Xerocomus badius-P. abies, was investigated by staining hand sectioned tips with FDA and their fluorescence. Different FDA-hydrolysing activities of the mycorrihizal types had been found in a previous-year study on the same plots and were confirmed during the second year. The proportion of the different stages of activity of the mycorrhizal tissues was calculated on the type level and in connection to the soil treatments. X. badius-P. abies and R. ochroleuca-P. abies displayed the most active fungal tissues and proportion increased on the acidic plots while Tuber puberulum-P. abies and Piceirhiza nigra were the most active types and occurred in higher proportion after liming. Thus, the overall activities of the mycorrhizas were only slightly changed by the treatments. In addition to the mycorrhizal effect acidification reduced while liming enhanced the meristematic activity of the short root tips. The same tendency was found by studying root production on the same plots (Hahn and Marschner, 1998). Although nearly 3000 mycorrhizal tips were studied, the data are still limited, allowing no statistical validation. This is, however, the first investigation connecting overall activity of the mycorrihizal tissues with the proportion of the mycorrhizal types as influenced both by alterations of the forest soil caused by acid rain and liming. The results are interesting and reasonable but further investments are necessary to validate the general conclusions.     

Ectomycorrhizal syntheses with Picea abies and three fungal species: a case study on the use of an in vitro technique to identify naturally occurring ectomycorrhizae

Ectomycorrhizal syntheses between Picea abies and the fungal associates Scleroderma citrinum, Boletus luridus, and Tricholoma vaccinum were carried out using Melin's Erlenmeyer flask technique. The symbioses of S. citrinum were characterized by a mantle composed of an outer prosenchymatous and an inner synenchymatous layer. The mantles of B. luridus and T. vaccinum were solely prosenchymatous. Rhizomorphs were produced in all treatments, but only in association with S. citrinum were they differentiated with additional, enlarged hyphae. All synthesized ectomycorrhizae were white or whitish to light orange and greyishorange. On large-scale root sampling in two differing Picea abies forests in Switzerland, nine out of a total of 22 morphological types of ectomycorrhizae were white or yellow in colour and were, therefore, comparable with the synthesized ectomycorrhizae. These nine natural types generally had distinct mantle features (irregular synenchyma, gelatinous matrix, cystides, thick-walled hyphae), but mostly lacked clamp connections. Synthesized ectomycorrhizae, on the other hand, lacked distinct mantle characteristics and always had clamp connections. Natural and synthesized white or yellow ectomycorrhizae did not coincide morphologically and thus identification of the fungal partners of natural symbioses by means of in vitro-synthesis with potential ectomycorrhizal fungi was not possible in the present study.     

Comparative studies on the mycorrhization of Larix decidua and Picea abies by Suillus grevillei

Summary Mycorrhization of Picea abies has been achieved, for the first time, with six strains of Suillus grevillei by a new culture method, using activated charcoal paper and liquid medium as a substrate. Mycorrhization of P. abies and Larix decidua was compared, and the process was found to be significantly different in the two tree species. S. grevillei is not incompatible with P. abies, but it forms mycorrhizae more readily with L. decidua. Hyphal growth was clearly stimulated on the surface of roots of Larix but retarded on Picea. A well organized Hartig net was formed with both tree species, but wall protuberances were frequently observed on the outer cell walls of Picea cortex cells when the Hartig net was not fully developed. No conspicuous cell wall reactions occurred in Larix roots. Cell wall protuberances may be comparable to those in transfer cells and are interpreted as an alternative to Hartig net development. Anatomical differences between roots of Larix and Picea, and physiologically active substances such as recognition factors on the root surfaces, are discussed with respect to their responsibility for the different reactions of S. grevillei.     

Changes in root growth patterns of (Picea abies) spruce roots by inoculation with an ectomycorrhizal fungusPisolithus tinctorius and jasmonic acid treatment

Symbiosis between fungi and plant roots forming a mycorrhiza involves extensive interactions at the molecular level between both partners. The role of plant hormones in the regulation of mycorrhizal infection is not known to involve jasmonates. Their endogenous levels increase during pathogen attack; however, little has been done on their involvement in mycorrhizae. In our recent work, root growth patterns of 2-month-old spruce seedlings after inoculation withPisolithus tinctorius and/or jasmonic acid (JA) treatment were studied using a paper-sandwich technique. Changes in root length, the degree of branching, presence and length of root hairs, and infection parameters were followed using a stereomicroscope. The first mycorrhizal contact of hyphae with roots was significantly accelerated upon treatment with 0.5 M JA. Interactions between root hairs and fungal hyphae were seen by scanning electron microscopy. The multiplication of root hairs of non-mycorrhized seedlings treated with 5.0 M JA and changes of the root surface were observed by the same technique.     

Abiesin,a biflavonoid of abies webbiana

Abiesin, a new biflavonoid, has been isolated from the leaves of Abies webbiana and identified as 5,3″,7″- trihydroxy-7,4′,4?-trimethoxy-(3′,6″)-biflavone.     

Biological activity, identification and quantification of gibberellins in seedlings of Norway spruce (Picea abies) grown under different photoperiods

Short photoperiod induces growth cessation in seedlings of Norway spruce ( Picea abies (L.] Karst.). Application of different gibberellins (GAS) to seedlings growing under a short photoperiod show that GA₉ and GA₂₀ can not induce growth. In contrast application of GA, and GA₄ induced shoot elongation. The results indicate that 3β-hydroxylation of GA₉ to GA₄ and of GA₂₀ to GA₁ is under photoperiodic control. To confirm that conclusion, both qualitative and quantitative analyses of endogenous GAs were performed. GA₁, GA₃, GA₄, GA₇, GA₉, GA₁₂, GA₁₅, GA₁₅, GA₂₀, GA₂₉, GA₃₄ and GA₅₁ were identified by combined gas chromatography-mass spectrometry in shoots of Norway spruce seedlings. The effect of photoperiod on GA levels was determined by using deuterated and ¹⁴C-labelled GAs as intermal standards. In short days, the amounts of GA₉, GA₄ and GA₁ are less than in plants grown in continuous light. There is no significant difference in the amounts of GA₃, GA₁₂, and GA₂₀ between the different photoperiods. The lack of accumulation of GA₉ and GA₂₀ under short days is discussed.