Cumulative dressings of potassium metaphosphate and soluble PK-fertilizers for seedlings and transplants ofPicea sitchensis in two English nurseries

Summary Cumulative dressings of potassium metaphosphate tested on seedlings and transplants ofPicea sitchensis gave promising results from 1964 to 1966¹ at two nurseries — a sandy podsol and a sandy loam with better nutrient retention. The four experiments, which also included superphosphate alone, KC1/superphosphate, with and without topdressings of KNO₃, and potassium dihydrogen phosphate, were continued until 1969. On the podsol, potassium metaphosphate maintained its superiority for seedlings and also left larger residues in the surface soil. It had no special merit on the sandy loam or for transplants on either soil.     

Isobutylidene diurea and other nitrogen fertilizers for seedlings and transplants ofPicea sitchensis in two English forest nurseries

Summary Three slow-release nitrogen fertilizers — medium- and coarse-sized granules of IBDU (isobutylidene diurea) and formalized casein — were applied before sowing or transplanting to Sitka sprucePicea sitchensis, and their effects on growth compared with that on plots not given N or given four topdressings of ‘Nitro-Chalk’ (the source of N used in practice). The comparisons were made at two sites: a very sandy podsol and a sandy loam with better nutrient retention. The experiments with seedlings were continued for four years, those with transplants for three, and the same treatments were applied to the same plots each year. Responses to ‘Nitro-Chalk’ were large, especially during wet seasons. Formalized casein was always as effective as ‘Nitro-Chalk’ for transplants. IBDU (especially the coarse fraction) was almost as good as formalized casein for transplants at both sites and equalled it for seedlings on the sandy loam. For seedlings on the sandy podsol, both particle sizes of IBDU became steadily less effective in succeeding years, and no satisfactory reason can be offered for this decline.     

Fungal adhesion pad formation and penetration of root cuticle in early stage mycorrhizas ofPicea abies andLaccaria amethystea

Summary Primary events during the establishment of the fungus-root symbiosis in ectomycorrhizas are still little understood. No attention has been paid so far to the adhesion of hyphae to the root cuticle and penetration of this barrier, although the importance of the cuticle has been shown for pathogen-plant interactions. Early developmental stages of in vitro mycorrhization ofLaccaria amethystea onPicea abies after short periods of incubation in growth chambers under elevated CO₂ concentrations were studied by light and transmission electron microscopy. No structural changes in mycorrhization related to elevated CO₂ were found, but fine roots and mycorrhizas developed faster. Adhesion pad formation was observed at hyphal tips in contact with the root cuticle. The adhesion pad was connected to the outer cell wall layer of the hypha and reacted positively to the Swift reaction for cysteine rich proteins. Although the reaction cannot be considered as totally specific, findings are discussed in respect to hydrophobins, which have recently been found to be expressed during early steps in ectomycorrhizal development. The root cuticle was dissolved and penetrated by fungal tips of the fingerlike branching mycelium attached to the root surface. The findings are compared with well documented pathogenic fungus-plant interactions at the cuticle. The possibility of restriction of hyphal attack to that part of the cuticle covering cell junctions is discussed.     

Occurrence and types of ectomycorrhizae present in seedlings ofPicea glehnii in a natural forest in Hokkaido

Picea glehnii seedlings whose ages ranged from 2 to 5 years and which were growing in aP. glehnii-Abies sachalinensis natural forest where the surface had been scarified 6 years before, were assessed for the occurrence and types of ectomycorrhizae. Based on the macro- and microscopic characteristics, 34 types of ectomycorrhizae were classified. The basidiomycetes were clearly dominant and it was common to observe more than one type of mycorrhiza occurring in the same root tip. The diversity of mycorrhizal types and the presence of at least one type of ectomycorrhiza in a single seedling, observed in this work, indicate a very dynamic system of mycorrhizal formation inP. glehnii seedlings, which allows them to survive and grow in a forest surface subjected to soil scarification.     

Major and trace element concentrations in needles ofPicea abies: levels,distribution functions,correlations and environmental influences

Summary Eighteen major and trace elements (Al, As, Ba, Br, Ca, Cl, Co, Cu, Fe, K, Mg, Mn, Na, Sb, Sc, Sr, V, Zn) have been determined in needles ofPicea abies by neutron activation analysis. Trees from 31 sites with different characteristics were investigated. The sampled area was 140 km² and centered around the city of Winterthur, Switzerland.The effect of washing the needles before analysis was investigated. Washing was found to be essential for a meaningful determination of most elements. The values of the individual elements could be approximated by normal or by lognormal distributions. The width of these distributions varies greatly, being 14% for Cu and a factor of 5 for Mn. Many elements show highly significant positive or negative correlations.Whereas the levels of the major elements as determined here are in the generally accepted range, the values for most of the minor elements show very poor correspondence with published values. For some elements the present data seem to be the only available values. Distinct environmental influences were only manifest for Na, Cl, Br, and these elements show very high values at sites bordering highways.     

Differential energization of the tonoplast in suspension cells and seedlings from Picea abies

 Vacuolar ATPase (EC 3.6.1.3) and PPase (EC 3.6.1.1) were studied in suspension cells and seedlings from spruce [Picea abies (L.) Karst. Proton transport activity and uncoupler (1 μM nigericin) stimulated substrate hydrolysis were measured in tonoplast enriched membrane vesicles. In suspension cells the vacuolar PPase exhibited 1.8-fold activity of the ATPase. In roots and needles from 12-week-old spruce seedlings the vacuolar PPase was inactive, whereas the ATPase was active. Therefore, we investigated whether the preparation of spruce tonoplast vesicles from roots and needles inactivates the vacuolar PPase but not the ATPase. For this purpose, maize (Zea mays L.) tonoplast membranes exhibiting vacuolar PPase as well as ATPase activity were used as a probe and added to the homogenization medium prior to the preparation of spruce vesicles. The preparation of spruce vesicles was more inhibitory to the vacuolar ATPase than to the PPase. The comparison of vacuolar PPases from spruce suspension cells and maize roots revealed similar enzymatic properties. After isopycnic centrifugation on continuous sucrose gradients the vacuolar PPase from spruce suspension cells co-purified with the vacuolar ATPase. Together, these data show: (1) vacuolar PPases from spruce suspension cells and maize roots are similar, (2) the preparation of tonoplast vesicles from spruce roots and needles does not inactivate the vacuolar PPase, (3) tonoplasts of suspension cultured cells and seedlings from spruce are differentially energized by the vacuolar pyrophosphatase that may indicate a difference in pyrophosphate metabolism between embryogenic and differentiated spruce cells, and (4) tonoplast vesicles from spruce seedlings may allow investigations of the effect of pyrophosphate on the vacuolar ATPase in the absence of vacuolar PPase activity. Received: 2 July 1998 / Accepted: 14 September 1998     

Chlorophyll formation and the development of photosynthesis in dark grown seedlings of Picea abies

Abstract Seeds of Picea abies were germinated and grown in either darkness or constant light. The chlorophyll content and photosynthetic carbon dioxide uptake of developing seedlings of different ages was determined. Ten-day-old dark grown seedlings showed an instant ability for photosynthetic carbon dioxide uptake and also formed further chlorophyll most rapidly upon subsequent illumination. These activities progressively diminished when the dark growth period was extended. Light grown seedlings reached a maximum chlorophyll level after 15 days growth, and this preceded maximal photosynthetic development.     

Intraspecific variation in Heterobasidion annosum for mortality rate on Pinus sylvestris and Picea abies seedlings grown in pure culture

One-month-old Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) seedlings were inoculated in vitro with Heterobasidion annosum strains, four each of the P-, S- and F-intersterility groups. Variation among strains and between the IS groups in virulence, expressed in mortality rate, was detected during twelve months after inoculation. Most of the strains were more virulent on spruce than on pine, and mortality of spruce seedlings was significantly higher. The P strains displayed similar virulence on both hosts, while S strains caused higher mortality of spruce seedlings and significantly lower mortality of pine seedlings. Strains of the F group were less virulent, but killed significantly (P < 0.05) more spruce than pine seedlings. In the interspecific analyses with two hosts, the isolates and IS groups accounted for most of the explained variation in the host mortality.     

Ethylene effects on cambial activity and cell wall formation in hypocotyls of Picea abies seedlings

Ethylene regulation of cell division in the vascular cambium and cell wall formation was studied in hypocotyls of Norway spruce ( Picea abies [L.] Karst.) seedlings. Cuttings from 6-week-old seedlings were placed in water culture to which compounds affecting the synthesis and action of ethylene were added. After a 3-week treatment period, growth, ethylene production, morphology and cell wall composition of the hypocotyls were determined. Addition of high concentrations of the potent ethylene releasing agent 2-chloroethylphosphonic acid (ethrel), which increased ethylene emission by more than twice compared to control plants, inhibited the expansion of xylem cells while stimulating the incorporation of cell wall material, especially cellulose. Addition of small amounts of ethrel, which slightly stimulated ethylene emission, led to increases in the size of xylem cells, the amount of phloem tissue and the number of intercellular spaces in the cortex, and thus to increased hypocotyl diameter. However, no significant change in cell wall composition was detected. When ethylene production was decreased by adding Co²⁺ to the nutrient solution, differentiation of new xylem was disturbed, but the rate of cell division was not affected. Although the incorporation of cell wall material was inhibited, the proportions of lignin and cellulose in the wall appeared to remain unchanged. Silver ions stimulated the expansion of both xylem and cortex cells, but had no significant effect on cell wall formation. We conclude that ethylene has a role in regulating the incorporation of wall carbohydrates.     

Evidence for two photoreceptors controlling growth in de-etiolated seedlings

The effect of blue light on hypocotyl extension in de-etiolated seedlings of lettuce, cucumber and tomato was investigated under conditions which precluded the involvement of phytochrome. Small but highly inhibitory amounts of blue light were added to a high intensity background illumination from low pressure sodium lamps. A log-linear response for inhibition of hypocotyl extension against the blue light fluence rate was obtained for lettuce and cucumber, and inhibition in tomato was also related to the blue light fluence rate. The added blue light did not alter phytochrome photostationary state and its effect was independent of the total fluence rate. Growth inhibition by Pfr could be demonstrated in tomato and cucumber but not in lettuce. The results indicate that two photoreceptors may normally be involved in the control of seedling growth but their relative importance varies greatly between species.Abbreviations HIR high irradiance reaction - Pfr far red absorbing form of phytochrome - Pr red absorbing form of phytochrome     

Chromosomal and pTi genotypes of agrobacterium strains isolated from Populus tumors in two nurseries

Abstract Agrobacterium tumefaciens strains isolated from aspen tumors have been characterized in order to study Ti plasmid stability in different chromosomal backgrounds under natural conditions. Chromosomal and pTi genotypes were characterized by DNA-DNA hybridization. Single tumors contained one to four strains of A. tumefaciens as characterized by their chromosomal genotypes and one or two different nopaline-type pTi genotypes. Genotypes of pTi were associated with identical chromosomal backgrounds in different tree nurseries, suggesting that pTi plasmids are stably maintained in the small set of strains causing the present aspen crown gall outbreaks.     

Interaction between atmospheric and pedospheric nitrogen nutrition in spruce (Picea abies L. Karst) seedlings

The effect of NO₂ fumigation on root N uptake and metabolism was investigated in 3-month-old spruce (Picea abics L. Karst) seedlings. In a first experiment, the contribution of NO₂ to the plant N budget was measured during a 48 h fumigation with 100mm³m^?3 NO₂. Plants were pre-treated with various nutrient solutions containing NO₂ and NH₄⁺, NO₃^? only or no nitrogen source for 1 week prior to the beginning of fumigation. Absence of NH₄⁺ in the solution for 6d led to an increased capacity for NO₃^? uptake, whereas the absence of both ions caused a decrease in the plant N concentration, with no change in NO₃^? uptake. In fumigated plants, NO₂ uptake accounted for 20–40% of NO₃^? uptake. Root NO₃^? uptake in plants supplied with NH₄⁺plus NO₃^? solutions was decreased by NO₂ fumigation, whereas it was not significantly altered in the other treatments. In a second experiment, spruce seedlings were grown on a solution containing both NO₂ and NH₄⁺ and were fumigated or not with 100mm³m^?3 NO₂ for 7 weeks. Fumigated plants accumulated less dry matter, especially in the roots. Fluxes of the two N species were estimated from their accumulations in shoots and roots, xylem exudate analysis and ¹⁵N labelling. Root NH₄⁺ uptake was approximately three times higher than NO₃^? uptake. Nitrogen dioxide uptake represented 10–15% of the total N budget of the plants. In control plants, N assimilation occurred mainly in the roots and organic nitrogen was the main form of N transported to the shoot. Phloem transport of organic nitrogen accounted for 17% of its xylem transport. In fumigated plants, neither NO₃^? nor NH₄⁺ accumulated in the shoot, showing that all the absorbed NO₂ was assimilated. Root NO₃^? reduction was reduced whereas organic nitrogen transport in the phloem increased by a factor of 3 in NO₂-fimugated as compared with control plants. The significance of the results for the regulation of whole-plant N utilization is discussed.     

Nitrogen limitation in mycorrhizal Norway spruce (Picea abies) seedlings induced mycelial foraging for ammonium: implications for Ca and Mg uptake

Although many studies support the importance of the external mycelium for nutrient acquisition of ectomycorrhizal plants, direct evidence for a significant contribution to host nitrogen nutrition is still scarce. We grew nonmycorrhizal seedlings and seedlings mycorrhizal with Paxillus involutus (Batsch) Fr. in a sand culture system with two compartments separated by a 45-m Nylon mesh. Hyphae, but not roots, can penetrate this net. Nutrient solutions were designed to limit seedling growth by nitrogen. Hyphal density in the hyphal compartment, host N status and shoot growth of mycorrhizal seedlings significantly increased in response to NH₄ ⁺ addition to the hyphal compartment. Labeling the compartment only accessible to hyphae with ¹⁵NH₄ ⁺ showed that the increase in N uptake in the mycorrhizal seedlings was a result of hyphal N acquisition from the hyphal compartment. These results indicate that hyphae of P. involutus may actively forage into N-rich patches and improve host N status and growth. In the mycorrhizal seedlings, which received additional NH₄ ⁺ via their external mycelium, the increase in NH₄ ⁺ supply less negatively affected Ca and Mg uptake than in nonmycorrhizal seedlings, where the additional NH₄ ⁺ was directly supplied to the roots. This was most likely due to the close link of NH₄ ⁺ uptake and H⁺ extrusion, which, in the nonmycorrhizal seedlings, lead to a strong acidification in the root compartment, and subsequently reduced Ca and Mg uptake, whereas in the mycorrhizal seedlings the site of intensive NH₄ ⁺ uptake and acidification was in the hyphal and not in the root compartment. Our data support the idea that the ectomycorrhizal mycelium connected to an N-deficient host may actively forage for N. The mycelium may also be important as a biological buffer system ameliorating negative influence of high NH₄ ⁺ supply on cation uptake.     

Mycorrhizas and C and N transformations in the rhizospheres of Pinus sylvestris,Picea abies and Betula pendula seedlings

Scots pine (Pinus sylvestris L.), Norway spruce (Picea abies L.) and silver birch (Betula pendula L.) seedlings were grown in a greenhouse for four months in three different soils. The soils were from a field afforestation site on former agricultural land: soil from a pine site, soil from a spruce site and soil from a birch site. Pots without seedlings were included. The aim was to discover, independent of the effects of the different quality of aboveground litter and microclimate under the tree species, whether the roots change the microbial activities and chemical characteristics of the soil, whether the changes are dependent on the tree species, and whether the changes vary in different soils. Pine, spruce and birch had, on average, five, one and six meters of roots, respectively. Birch had by far the highest number of short root tips, on average 11 450 per seedling, compared to 1900 and 450 in pine and spruce seedlings, respectively. The majority of the short roots of pine and spruce were brown sheathed mycorrhizas, and those of birch were mycorrhizas in an early stage of development. The seedlings caused no major changes in either the soil pH or the concentrations of nutrients in the soils, but did affect the microbial characteristics of the soils. The effect of the tree species did not differ in different soils. Microbial biomass C and N, C mineralization rate and the concentration of ergosterol were all higher under birch and pine than under spruce and in plantless soils. Nitrate concentrations were lowest under pine and birch, but rates of net N mineralization, nitrification and denitrification did not differ under different seedlings. The stimulative effect of pine and especially birch on soil microbes was possibly due to them having more roots and releasing more root exudates to soil. There were, however, indications that not only the length/mass of roots determined the changes in microbial activities, but also differences in root activities per unit of root or in the quality of root exudates.     

Soluble and microsomal glutatione S-transferase activities in pea seedlings (Pisum sativum L.)

Epicotyl and primary leaves of pea seedlings (Pisum sativum L., var. Alaska) were found to contain soluble and microsomal enzymes catalyzing the addition of glutathione to the olefinic double bond of cinnamic acid. Glutathione S-cinnamoyl transfer was also obtained with enzyme preparations from potato slices and cell suspension cultures of parsley and soybean.The pea transferases had pH-optima between pH 7.4 and 7.8 K_m-values were 0.1–0.4 mM and 1–4 mM for cinnamic acid and glutathione, respectively. V-values were between 2–15 nmol mg^-1 protein x min.Chromatography on Sephacryl S-200 indicated that the soluble pea glutathione S-cinnamoyl transferase activity existed in molecular weight forms of 37,000, 75,000, and 150,000. The glutathione-dependent cleavage of the herbicide fluorodifen was catalyzed by a different soluble enzyme activity which eluted in molecular weight positions of 47,000 and/or 82,000.The microsomal fraction from pea primary leaves also catalyzed the conjugation of the carcinogen benzo[]pyrene with glutathione.Abbreviations GSH glutathione - DDE 1,1-Dichloro-2,2-bis-(4-chlorophenyl)-ethylene - DDMU l-Chloro-2,2-bis-(4-chlorophenyl)-ethylene     

Soluble proteins and dehydrins in nitrogen-fertilized Scots pine seedlings during deacclimation and the onset of growth

Deacclimation of Scots pine ( Pinus sylvestris ) in northern Finland (65°N) was studied from the perspective of protein metabolism, which was related to nitrogen fertilization. Two-year-old seedlings were fertilized with calcium ammonium nitrate (0, 442 and 884 kg N ha⁻¹) in the summer prior to the study. Needles were harvested for analyses at 3-week intervals during the natural deacclimation period from mid-March to the beginning of June 1998. Deacclimation was followed by monitoring various physiological variables: freezing resistance decreased and osmotic potential increased during the experiment and needle dry weight increased from bud flushing onwards. The concentration of soluble proteins in needles was higher in fertilized seedlings but decreased transiently in the 884 kg N ha⁻¹ fertilized seedlings before budbreak. The abundance of several small polypeptides (17–32 kDa) decreased in the spring. A 60-kDa protein, identified by immunoblotting as a dehydrin, was detected in all treatments. The quantity of this dehydrin decreased with resumption of growth, along with the appearance of 50- and 56-kDa dehydrins. The concentration of these dehydrins decreased during dehardening more rapidly in fertilized seedlings than in the control plants. The fertilized seedlings started to grow earlier than the unfertilized plants. In conclusion, nitrogen fertilization provided good reserves for new growth but did not affect deacclimation of pine needles.