Phloem transport in Picea abies (L.) Karst. in mid-winter

Summary Translocation of ¹⁴C assimilates was studied on four different transport systems of Picea abies branches after induced activation in January. ¹⁴CO₂ assimilation of terminal shoots for 48 h at 25° C resulted in phloem loading and basipetal transport of ¹⁴C photosynthate into the following, older shoot generations. ¹⁴C import was enhanced, when these older shoot generations were kept in the dark. Microautoradiographs of the labelled terminal shoots showed that ¹⁴C assimilates were exported from needles via sieve elements of the leaf traces and loaded into the latest increment of the axial secondary phloem. No ¹⁴C label appeared in the obliterated sieve cells or in the tracheids. In addition, ¹⁴C photosynthate accumulated densely in the chlorophyllous cells of the cortex and in cells of the resin ducts, indicating certain sink activity. In the darkened 2-year-old shoot, imported ¹⁴C photosynthate was concentrated in the functional secondary phloem, while some ¹⁴C label was unloaded into the latest xylem increment. When 6-year-old shoots were exposed to ¹⁴CO₂ for 48 h in the light, ¹⁴C assimilates accumulated in the phloem of the leaf trace and in the latest increment of the axial secondary phloem. However, a substantial amount of radioactivity was unloaded into ray cells and phloem parenchyma cells. Thus, the presence of functioning phloem in needles and twigs of P. abies during winter allows long-distance translocation and radial distribution of assimilates according to existing source-sink relations.     

Fungal injury to seed tissues of Norway spruce,Picea abies (L.) Karst.

Norway spruce bore an abundance of cones in Finland in 2000, but these cones were often fungal-infected. The seeds had structural injuries that were revealed when seed samples were examined using light (LM) and a field emission scanning electron microscope (FESEM). Two main types of spores were found either in the tissues inside the seed coat or on the sarcotesta, the outermost layer of seed coat. The spores of Chrysomyxa pirolata appeared particularly in the nucellar tissue, where the cell walls were disintegrated at the middle lamellae and cytoplasm was disrupted. Degenerated remnants of fungal structures resembling aecial peridium were found close to aeciospores. The tissue of the megagametophyte differed also from that of a normal mature seed. Conidia of Thysanophora penicillioides were often encountered on the sarcotesta where the ordinary wax cover was missing. Fungal injury occurred in the nucellar layers that shelter the embryo and megagametophyte from desiccation and oxidation. Destruction of these structures together with rapid opening of the seed coat advance deterioration of seeds during storage and may cause unexpected economic losses in forest plant production.     

Performance of two Picea abies (L.) Karst. stands at different stages of decline

Summary A declining Picea abies (L.) Karst. stand produced as much foliage and branches as a healthy stand but less stemwood at a similar leaf area index and climate. Nutrient analyses revealed that most biomass components at the declining site had lower concentrations of calcium and magnesium, but similar nitrogen and potassium (except for lower potassium in younger needles) and higher phosphorus, manganese and aluminum than the respective components at the healthy site. Comparison of these data with the results from studies on the nutrition and growth of P. abies seedlings (Ingestad 1959) led to the conclusion that the healthy stand is in a balanced nutritional state, while trees at the declining stand have only 56% of the foliar magnesium concentration required to permit growth at a rate which could be achieved at their nitrogen status. It appears that acidic deposition, which involves an input of nitrogen and a leaching of cations from the soil, causes an imbalance in the availability of nitrogen and magnesium. Growth is eventually reduced as magnesium becomes limiting.     

Seasonal variation in the monoterpenes in needles of Picea abies (L.) Karst.

Summary This investigation was conducted to obtain information about the fluctuations in composition and amount of needle monoterpenes during the development of spruce needles. Studies conducted with two Norway spruce clones clearly revealed the existence of fluctuations. In juvenile needles, the amounts of the oxygenated terpenes increase constantly with age during the first 2 months of needle growth. The hydrocarbon terpenes dominate within the first weeks, some of them even showing a very distinct first maximum within the first 3 weeks after bud burst. All terpenes, including the oxygenated ones, have a maximum in June/July, which favours the hypothesis of a substitution of the hydrocarbons later on. There are significant changes even in mature needles of Norway spruce. The terpene level of 1-year-old needles of the clonal trees increased from spring to early summer and then dropped again towards winter. In addition, fluctuations in mature needles were shown for a set of ten wild trees. Needles of the same age class, which emerged in 1986, were sampled 4 times from 1986 to 1988. The needle terpene concentrations of the 1 -year-old needles were considerably lower in spring at the time of bud burst than in autumn. The terpene level of older needles thus seems to be influenced by biosynthetic and catabolic activities.     

Performance of two Picea abies (L.) Karst. stands at different stages of decline

Summary The development of root tips and apparent ectomycorrhizas was compared in the Fichtelgebirge (FRG) over one growing season in two 30-year-old Picea abies stands, both on soils derived from phyllite but showing varying symptoms of decline. Visual symptoms of tree decline reflected a lower relative and absolute mycorrhizal frequency, a lower number of ectomycorrhizas per m² leaf area and an uneven vertical distribution of root tips and ectomycorrhizas. The number of apparent ectomycorrhizas per ground area was correlated with the amount of magnesium, calcium, and ammonium, and the pH in the free-drainage soil solution, and with the molar calcium to aluminium ratio in mineral soil extracts. The foliage concentrations of magnesium and calcium were correlated with the numbers of apparent ectomycorrhizas per m² leaf or ground area. These observations were used to formulate testable hypotheses concerning the role of the root system and the soil environment in forest decline.     

Naturally occurring monoterpenoids in needles of Picea abies (L.) Karst

Summary An analysis was made of the effects of different sampling and extraction techniques on the amounts and pattern of monoterpenoids isolated from needles of Norway spruce. The following isolation and analysis procedure was finally adopted: liquid nitrogen-cooled needles were pulverized by a microdismembrator, extracted with pentane overnight at 2°–3°C and concentrated to a volume not less than 3 ml/g fresh weight on a Vigreux column. The crude extract was injected splitless (with solvent split) onto a cold programmed temperature vaporized (PTV) precolumn of a gas chromatograph and the vaporizable compounds heated to a capillary column. This method was tested for production of artefacts and quantitative extraction and applied to needles of eleven 80-year-old spruce trees.     

Effect of auxins and ectomycorrhizal elicitors on wall-bound proteins and enzymes of spruce [Picea abies (L.) Karst.] cells

Summary Elicitors of the ectomycorrhizal fungus Hebeloma crustuliniforme and auxins (IAA, NAA and 2,4-D) were tested for their effects on apoplastic proteins and enzymes of suspension cultured cells of Picea abies (L.) Karst. The ectomycorrhizal elicitor increased the amount of some ionically wall-bound proteins (36, 28, 24, 21 kDa) and decreased the amount of others (61, 22 kDa). The elicitor triggered an H₂O₂ burst and enhanced the peroxidase (EC 1.11.1.7) activity of the Picea cells by increasing one of the two wall-bound peroxidase isoforms. Auxins significantly suppressed the elicitor induction of peroxidase but did not influence the elicitor-triggered H₂O₂ burst. The elicitors and auxin did not change the amount and the pattern of wall-bound invertase isoforms (EC 3.2.1.26) of spruce cells. However, auxin reduced the uptake of glucose by spruce cells and increased the acidification of the cell culture medium. Since Hebeloma lacks apoplastic invertase as well as a sucrose uptake system, utilization of plant-derived sucrose depends on the apoplastic plant invertase activity. Although the host invertase is constitutive, the fungus might be able to increase this invertase activity within a mycorrhiza by lowering the pH of the interface towards the pH optimum of the enzyme via the action of auxin. This fungus-released hormone could increase the H⁺ extrusion of plant cells by activation of the plant membrane H⁺-ATPases. Additionally, an auxin-dependent suppression of glucose uptake by cortical root cells could improve the glucose supply for the fungus. Furthermore, the fungal auxin might suppress the elicitor induced formation of defense enzymes, such as peroxidase.     

Performance of two Picea abies (L.) Karst. stands at different stages of decline

Summary The water relations of Picea abies in a healthy stand with green trees only and a declining stand with trees showing different stages of needle yellowing were investigated in northern Bavaria. The present study is based on observations of trees differing in their nutritional status but apparently green on both sites in order to identify changes in the response pattern which might be caused by atmospheric concentrations of air pollutants and could lead to the phenomenon of decline. Transpiration was measured as water flow through the hydroactive xylem using an equilibrium mass-flow measurement system. Total tree transpiration was monitored diurnally, from July 1985 until October 1985 at both sites. The relationship between transpiration and meteorological measurements indicated that transpiration was a linear function of the vapor pressure deficit. No differences in transpiration of green trees were observed between the two sites. Canopy transpiration was 57%–68% of total throughfall and 41%–54% of total rainfall. Due to this positive water balance, soil water potential at 10 and 20 cm depths remained close to-0.02 MPa (max.-0.09 MPa) for most of the summer. Soil water potential was correlated with the difference between the weekly precipitation and transpiration. No differences in the water relations of apparently healthy trees in the two P. abies stands were observed. It is concluded that differences between green trees at the two sites in terms of nutrient relations or growth rate cannot be explained by changes in whole-tree transpiration or soil water status.     

The significance of structure for imbibition in seeds of the Norway spruce,Picea abies (L.) Karst.

Since the observations of those regularly handling Norway spruce [Picea abies (L.) Karst.] seeds with regard to their imbibition frequently disagree with earlier opinions that this process is markedly inhibited by the seed coat, we decided to examine the morphological factors influencing imbibition in seeds of different colour and different provenances. The seed coat, consisting of the sarcotesta, sclerotesta and endotesta, was found to have little influence on the passage of water, despite the presence of sclereids full of wax lamellae. No differences in seed coat structure were observed between provenances or colours of seeds. The cells of the endotesta were lignified in the area of the micropyle, however, and stood out lip-like on the outer surface of the micropyle after imbibition. An opening in the sclerotesta filled with parenchyma cells was also seen at the chalazal end of the seed. Neither of these openings, which were covered by accumulations of wax, served as the main route for the passage of water, though the micropyle opened up slightly after only 24 h incubation, when the lignified cells bordering it swelled differently from the rest of the endotesta. The progress of water into the seed soon discontinued, however, as the tip of the nucellar cap, covered with wax and crystals, effectively plugged the micropyle. This opening of the micropyle may be the reason why the IDS method does not always succeed in separating viable from non-viable spruce seeds sufficiently well by their density. Imbibition was mostly regulated by the lipophilic layers surrounding the endosperm, which are mainly of nucellar origin, and particularly the megaspore membranes, the outer and inner exine. Imbibition was further hampered by the impermeable nucellar cap, which covered about 3/4 of the length of the endosperm and had merged with the outer exine at its edges. Deposits of wax were observed both between the exines and between the endotesta and the nucellar layers at the edges of the nucellar cap. Waxes may serve as a defence against diseases at the sites of water penetration, while simultaneously increasing the significance of the nucellar endosperm covers as regulators of imbibition.     

Performance of two Picea abies (L.) Karst. stands at different stages of decline

Summary Photosynthetic rates and nutrient contents of spruce needles were measured in a region with high levels of air pollution in NE Bavaria, Germany (FRG), and compared to spruce grown under clean air conditions at Craigieburn, in the South Island of New Zealand (NZ). The absolute rates of CO₂ uptake, the slope of the CO₂ response curve at 240 l l^–1 internal CO₂ concentration, and the change of photosynthetic rates with needle age at ambient and saturated CO₂ concentrations were virtually identical at both measuring sites. These results confirm an earlier conclusion, that there is no long-term effect of atmospheric pollutants directly on photosynthetic CO₂ uptake rates with persistent exposure at the FRG site to high levels of anthropogenic air pollution. Photosynthetic capacity at saturating CO₂ concentration was three times higher in the NZ spruce. Needles with high photosynthetic capacity in NZ had lower nitrogen and higher calcium concentrations per unit dry weight but higher concentrations of nitrogen, phosphorus, potassium, magnesium and calcium per unit leaf area, and twice the specific leaf weight.     

Performance of two Picea abies (L.) Karst. stands at different stages of decline

Summary This is the first in a series of papers on the growth, photosynthetic rate, water and nutrient relations, root distribution and mycorrhizal frequency of two Norway spruce forests at different stages of decline. One of the stands was composed of green trees only while the other included trees ranging in appearance from full green crowns to thin crowns with yellow needles. In this paper we compare the growth and carbohydrate relations of the two stands and examine relationships among growth variables in ten plots. The declining stand produced 65 percent of the wood per ground area compared with the stand in which all trees were green because its foliage produced less wood at any level of leaf area index. The difference in foliage efficiency between the sites could not be explained by differeneces in climate, competition or stand structure. The declining stand appeared to have lower carbon gain as indicated by a smaller increase in reserve carbohydrates before bud break, and weaker sinks for carbohydrates as indicated by less use of the stored carbohydrates than the healthy stand. Thus, growth reduction was probably related to factors which affect both photosynthesis and, even more, the sinks for carbohydrate.     

Proportion of fungal mantle,cortex and stele of ectomycorrhizas in Picea abies (L.) Karst. in different soils and site conditions

Anatomical variability of ectomycorrhizal short roots in Norway spruce (Picea abies (L.) Karst) stands was investigated in five stands differing in site quality class (I^a–V) and soil type. Ten root samples per stand were randomly collected from the forest floor and the subsequent 20 cm soil layer. Thin transverse or axial sections (5 m) of randomly taken short roots were examined by light microscopy (AXIOPHOT; magnification 200–800×). All analyzed short root tips of Norway spruce in different stands were colonized by ectomycorrhizal fungi. Thickness of the mantle (T_mantle) and cortex (T_cortex) and diameter of the root (D_root) were measured in four crossing radial directions for transverse sections and in two radial directions for axial sections. The proportions of the mantle (PS_mantle), cortex (PS_cortex) and stele (PS_stele) of the root cross-sectional area (CSA) were calculated. Mean T_mantle and T_cortex varied from 16.5 ± 0.6 to 29 ± 1.3 m and 83.9 ± 1.7 to 108.4 ± 2.4 m, respectively; significant differences between stands were found. The number of cell rows in the cortex (4–6) did not vary between different stands, thus thickness of cortex depended on cell size. Mean PS_mantle, PS_cortex and PS_stele of the root CSA varied from 17.7 to 28.1%, from 58.9 to 66.9%, and from 13.4 to 15.8%, respectively. No differences between stands were revealed in mean CSA ratio of cortex and stele. It can be concluded that irrespective of big differences in soil and site conditions, including the influence of fungal symbionts, a 4:1 relation on a CSA basis between the cortex and stele is inherent to Norway spruce.     

Within-population variation in susceptibility to Agrobacterium tumefaciens A281 in Picea abies (L.) Karst

Summary The purpose of this study was to investigate the genetic variation of susceptibility to Agrobacterium within a Picea abies population. Tumor formation was studied in 16 open-pollinated families belonging to a central Swedish population of Picea abies. Strain A281 of Agrobacterium tumefaciens was used to infect 3-monthold seedlings in a five-block greenhouse experiment. The analysis of variance showed strong significance for the between-family variation of tumor-formation percentages, varying from 28% to 73%. The most susceptible material will be suitable for experiments on the production of transgenic plants in vitro using disarmed Agrobacterium strains.     

Stomatal patchiness in conifers: experiments with Picea abies (L.) Karst. and Abies alba Mill.

Summary Strong evidence for the occurrence of pronounced stomatal patchiness in needles of Picea abies (L.) Karst. and Abies alba Mill. was found using various indirect methods. Anatomical investigations revealed a septate leaf anatomy for both species, a phenomenon expected if a patchy distribution of stomatal aperture is present. Calculation of some photosynthetic characteristics (e.g. carboxylation efficiency) from gas exchange measurements is shown to be markedly affected by the patchy distribution of stomatal apertures on the needles. The importance of stomatal patchiness in connection with air pollution related forest decline symptoms as well as an hypothesis suggesting a possible role of the phenomenon as a protective mechanism against photoinhibition are discussed.     

Dendrochronology of Norway spruce (Picea abies (L.) Karst.) from two range centres in lowland Poland

In Europe, spruce grows in two main regions which meet in Poland, one to the north and east, the other to the south and west. The northeastern area ranges from the northern treeline extending from Norway to Siberia, to southern Sweden, north-eastern Poland and the southern Ural mountains. The southwestern spruce region reaches the mountainous areas of the Alps and the Balkans, the mountains and uplands of the Sudety Mountains, the Carpathians and neighbouring lowlands. Opinions about the distribution of Norway spruce have changed over the years, and its scarcity in the centre of Poland has been strongly debated. The favoured current theory is that Norway spruce once had a continuous distribution in Poland. It is assumed that the rare occurrence in the central Polish lowland is due to a combination of unfavourable soil conditions and previous management activity. The main aim of this work was to analyse climate–growth relationships of Norway spruce in eastern Poland and distinguish regions with similar increment patterns with regards to spruce range. Spruce growth in northern Polish sites is positively correlated with rainfall from May to July. Tree-ring widths in southern sites are more correlated with March temperature. Selected homogenous regions are the same as range types. Trees from the so-called “spruceless area” seem to have similar climate–growth relationships to trees from the southern region. This finding does not settle the question of the origin of the trees, but it does indicate that similar environmental conditions exist in these two areas and proves that the climate was not a limiting factor there.

CO2 fixation in stem slices of Picea abies (L.) Karst: microautoradiographic studies

Summary Microautoradiography was used to show that chlorophyllous cells of young Picea abies stem slices are able to fix ¹⁴CO₂, in the dark as well as in the light. The amount of ¹⁴CO₂ fixed in the dark is much lower than that in the light. In the dark the concentration of radioactive label is equally high in all chlorophyllous cells of the stem. In the light, however, a gradient of radioactive assimilates extends from the stem surface to its centre, with the highest concentration being located in the phelloderm and the outer one-third of the cortex. This is in spite of even illumination and CO₂ supply across the whole stem slice. In the dark, stem slices with and without bark show the same amount of radioactive label in the chlorophyllous cells of xylem, perimedullary region and pith. In the light, however, the concentration of radioactive assimilates in these cells is much higher in stem slices with bark than in stem slices without bark. It is assumed therefore that light fixation products of phelloderm and cortex are transported radially into the tissue inside the cambium.     

Uptake of water and solutes through twigs of Picea abies (L.) Karst

Summary Uptake of water and magnesium chloride solution was investigated through the outer surface of twigs of Picea abies (L.) Karst. Water uptake was determined by using pressure/volume (P/V) curves of the twigs as a basis for calculation to avoid problems of superficial extraneous water. When water was sprayed on bark and needles of 3- to 7-year-old twigs at a xylem water potential of -1.00 MPa, they absorbed as much as 80 mm³ water in 200 min/g twig dry weight as the twig water potential recovered to -0.15 MPa. With fluorescent dyes, pathways for absorption of water and solutes through the twig bark were found, particularly through the radially orientated ray tissue. In addition to uptake by mass flow, magnesium could also diffuse along a concentration gradient from the twig surface into the xylem. In the field, the magnitude of these uptake processes would depend on the concentration of elements deposited by atmospheric precipitation, the concentration gradient between the plant surface and the xylem sap, the xylem water potential and the intensity and duration of each precipitation event.     

Patterns and mechanisms of transpiration in a large subalpine Norway spruce (Picea abies (L.) Karst.)

In situ water relations of a large subalpine Norway spruce (Picea abies) were analyzed by simultaneous measurements of sap flow at different crown positions. In the diurnal scale, transpiration varied greatly, both spatially and temporally. Over longer periods, however, different parts of the crown transpired in fairly constant proportions. The average estimated transpiration was about 3.5 times greater in the upper than in the lower half and decreased 1.6-fold from south to north. Water intercepted from rain, fog and dew buffered and significantly decreased the transpiration. The effect was strongest in those parts which were least coupled to the free atmosphere. The top of the crown seemed to experience a regular shortage of water shortly after starting transpiration, when it was forced to switch from internal reserves to sources in the soil. Further, lower branches then started transpiring, which may have led them to compete for the water. An enhanced nocturnal sap flow during warm and dry winds (Foehn) indicated that the tree also transpired at night. Shaded twigs had more capacity to intercept water externally than twigs in the sun. The significance of the crown structure for interaction with water in both liquid and vapour phases is discussed.     

Respiration and photosynthesis in cones of Norway spruce (Picea abies (L.) Karst.)

Summary Dark respiration and photosynthetic carbon dioxide refixation in purple and green Picea abies cones were investigated from budbreak to cone maturity. The rate of dark respiration per unit dry weight and CO₂ refixation capacity decreased during cone maturation. At the beginning of the growing season, photosynthetic CO₂ refixation could reduce the amount of CO₂ released by respiration in green and purple cones by 50% and 40%, respectively. The seasonal performance of the components of the cone carbon balance was calculated using information on the seasonal course of respiration, refixation capacity and the light response curves of cone photosynthesis, as well as the actual light and temperature regime in the field. The daily gain of CO₂ refixation reached 28%–34% of respiration in green and 22%–26% in purple cones during the first month of their growth, but decreased later in the season. Over the entire growth period refixation reduced carbon costs of cone production in both cone colour polymorphs by 16%–17%.     

Isolation of two cytokinin metabolites from the rhizosphere of Norway spruce seedlings (Picea abies L. Karst.)

Roots of young Norway spruce seedlings were incubated under hydroculture conditions in a synthetic nutrient medium containing either ³H-isopentenyladenosine, isopentenyladenosine or zeatin riboside. When feeding with ³H-isopentenyladenosine a new radiaolabelled metabolite was found in the feeding solution as well as in root extracts. Isopentenyladenosine and zeatin riboside were metabolised and for both compounds an unknown metabolite was detected in the feeding solution. The metabolites were purified by solid phase extraction, HPLC and partially characterised. A major characteristic of the metabolites is their reactivity in the presence of NH₄OH, which results in the formation of the cytokinin bases isopentenyladenine or zeatin, respectively. UV-spectra and the chemical characteristics indicate that the new metabolites are closely related. The GC-MS analysis revealed, that the metabolites are true derivatives of isopentenyladenine and zeatin. The biogenesis of the new metabolites is discussed with regard to plant microbial interactions.Abbreviations Ck(s) = cytokinin(s) - GC-MS = gas chromatography-mass spectrometry - iP = isopentenyladenine - [9R]iP = isopentenyladenosine - [9G]iP = isopentenyladenine-9-glucoside - [9R-MP]iP = isopentenyladenosine-5-monophosphate - Z = trans-zeatin - [9R]Z = trans-zeatin riboside