Micropropagation of western hemlock (Tsuga heterophylla [Raf.] Sarg.) from embryonic explants

An in vitro propagation protocol is described for western hemlock, an important forestry species in Canada. For shoot bud induction, embryonic explants were placed initially on one-third strength Schenk and Hildebrandt medium containing 5 M N⁶-benzyladenine (BA) or 5 M BA in combination with either 5 M kinetin or 5 M 2-isopentenyl adenine for 14 days. The explants were transferred to basal medium without cytokinins for 3 weeks, and then to basal medium containing 0.05% activated charcoal. Elongating shoots were subcultured every 4 weeks on charcoal medium. Shoots, 10 mm in stem height, were rooted either in agar or sterilized peat/perlite (1:1). Up to 70% of the shoots formed roots when they were transferred to the latter, moistened with 1/2 strength Gresshoff and Doy medium containing 5 M -naphthaleneacetic acid. About 90% of the plantlets survived transfer to greenhouse conditions.     

Five heavy metals in propagules of ten mangrove species of China

Five heavy metals in the propagules of 10 mangrove species in China have been investigated. The results revealed that the levels of five heavy metals in most of the propagules were lower than the background levels of the soil. The levels of copper, manganese, zinc, cadmium, and lead in the propagules varied at about 2.1–7.8 g/g, 3.9–28.0 g/g, 5.7–60.0 g/g, 0.014–0.057 g/g, and 0.018–0.038 g/g, respectively. On average, the levels of five heavy metals were in the order Zn > Mn > Cu > Cd > Pb. The biological absorption coefficients were 0.02–1.30 for copper, manganese, zinc, and cadmium and about 0.0007–0.0020 for lead in propagules, showing the following order: Zn > Cd > Cu > Mn > Pb. The above results indicate that the five heavy metals have low accumulation in most of the propagules of 10 mangrove species and are at safe levels for the effective utilization of mangrove resources.     

Adsorption capacities and related characteristics of wood charcoals carbonized using a one-step or two-step process

Sugi (Cryptomeria japonlca D. Don) wood powder was carbonized at varying temperatures by a onestep process up to 1000C and a two-step process using wood charcoal as the raw material up to 1600C. This study was conducted to evaluate the adsorptive properties of wood charcoal and discuss the mechanism of its adsorptive function in relation to the physical and anatomical characteristics of wood after carbonization. Anatomical characteristics of carbonized wood materials were directly observed under heating using an environmental scanning electron microscope (ESEM); the cell wall structures were analyzed by high-resolution transmission electron microscope (HRTEM). The largest weight losses were observed at the highest temperatures, in both the one-step and twostep processes but leveled off above 800C. Shrinkages in the tangential, radial, and longitudinal directions increased with carbonization temperature, peaking at 1000C. Direct observations by ESEM showed distinct shrinkage at around 400C. The first trial observations by HRTEM on the changes in the ultrastructure of cell walls of wood charcoals were done, and it was assumed to affect the formation of micropores. Adsorption was found to follow the Langmuir isotherm model. With the one-step carbonization process, the iodine adsorption capacities of the carbonized wood powders increased with increasing carbonization temperature, peaking at 800C, but decreased at higher temperatures. The wood powder carbonized at 1000C with the two-step process showed the highest capacity, but further heating up to 1400C drastically decreased the adsorption. The shrinkage of cells was related to the increases and decreases in its specific surface area. Specific surface area and total pore volume were evidently related to the adsorptive properties.Part of this paper was presented at the Second International Wood Science Seminar, Indonesia, November 6–7, 1998     

Maximum effective lumen and pit pore sizes of the earlywood and the latewood of never dried loblolly pine sapwood

Summary Measurements of the air pressure required to initially displace a saturating liquid and allow a slow continuous stream of air bubbles to pass through wood cross sections of different thickness, together with the equilibrium surface tension of the saturating liquid, make it possible to calculate the maximum effective opening radii. Previous measurements were made for wood as a whole over complete annual rings. Measurements reported here were made separately for earlywood and latewood. Extrapolating plots of the maximum effective opening radius-cross section thickness, for thicknesses below the maximum fiber length, to zero thickness gave maximum lumen radii of 16 m for the earlywood and 10.3 m for the latewood. The values are only slightly greater than the calculated average values. Extrapolating the plots in the opposite direction to zero opening radius gave approximate maximum lumen or fiber lengths for the earlywood of 6 mm and for the latewood of 5 mm. The maximum effective opening radii for cross sections thicker than the maximum fiber length give maximum effective communicating pore radii. These values continue to decrease, with increasing thickness of the cross sections due to the decreasing probability of the largest openings falling in any one series path through the structures. The maximum effective pit pore radius for passage through fifty pits in series was 0.8 m for the earlywood and 0.28 m for the latewood.Paper No. 3787 of the Journal Series of North Carolina State University Agricultural Experiment Station, Releigh, N. C.     

Ultrastructure of pits inPinus banksiana lamb

Summary The cross-sectional view of pitting between various cell types inPinus banksiana Lamb. was studied at the ultrastructural level. Cell types inPinus banksiana include longitudinal tracheids, ray tracheids, ray parenchyma cells, buffer cells and epithelial cells. Two common characteristic features of bordered pit-pairs between longitudinal tracheids are an initial pit border and a thickened torus at the center of the pit membrane. The shape and size of the pit border and torus of bordered pit-pairs between two compression wood cells, and between the last-formed latewood longitudinal tracheid and first-formed earlywood longitudinal tracheid were different from those in the earlywood and latewood longitudinal tracheids. The pit border on the ray tracheid side varied in size and shape due to wall dentation. No initial pit border was found on the pit border of the ray tracheid side. The shape of bordered pit-pairs between two ray tracheids varied considerably due to irregularity of the dentate cell wall. The size of bordered pit-pairs in longitudinal tracheids was between 16 m to 20 m, which was twice the diameter of bordered pit-pairs in ray tracheids. Bordered pitpairs at the end wall of two ray tracheids appeared to be the smallest at 5 m, Pit aspiration occurred in the bordered pit-pairs with or without a torus. In the heartwood zone, some half-borders pit-pairs between tracheary and ray parenchyma cells showed an additional secondary wall on the ray parenchyma cell side. Plasmodesmata were found in the half-bordered pit-pairs as well in the simple pit-pairs. Blind pits were observed between a ray tracheid and a longitudinal tracheid. Bordered pit-pairs between two buffer cells were also observed. The possible functions of buffer cells were discussed.Use of transmission electron microscope provided by the Science Instrumentation Lab, Lakehead University and the technical assistance provided by Mr. A. MacKenzie, Director of Science Instrumentation Lab are gratefully appreciated     

Tissue culture micropropagation of Douglas-fir

A procedure was developed for plantlet production from embryos of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco]. The Medium for Conifer Morphogenesis, used at full strength, and supplemented with 10 M benzyladenine for 17 days produced an average of 6.8 shoots on more than 90% of the embryos. The percentage of shoot-forming embryos as well as the average number of shoots per embryo varied significantly among eight seedlots. For secondary multiplication, 89% of the adventitious shoots produced axillary buds on MCM with 5 M benzyladenine. However, 0.5 M BA was more suitable for the elongation of axillaries. Rooting ranged from 0–87% depending upon the treatment. The highest percentage was obtained with a 7-week incubation in peat:perlite containing 1/5-strength medium, 1% sucrose and 2.7 M naphthaleneacetic acid, followed by 5 weeks on peat:perlite with 1/5-strength major and minor salts, full iron and organics, 1% sucrose and 0.1% charcoal.     

The distribution of lignin in sprucewood as determined by ultraviolet microscopy

The distribution of lignin in black spruce has been determined quantitatively by the study of 0.5 m transverse sections in a UV microscope. The average lignin concentration in the compound middle lamella was about twice that in the secondary wall. The lignin concentration of the middle lamella at the cell corners of adjacent tracheids was nearly four times that in the secondary wall but the volume of the secondary wall was much greater than the volume of the middle lamella. Thus, for earlywood, 72% of the total lignin was in the secondary wall leaving only 28% in the compound middle lamella and cell corner middle lamella regions. The corresponding values for latewood were 82% and 18% respectively. Use of oblique longitudinal sections of 0.1 m thick permitted the resolution of the compound middle lamella. The lignin concentration in the true middle lamella was found to be equal to that in the cell corner middle lamella and the primary wall lignin content to be about twice that in the secondary wall.

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Zusammenfassung Die Verteilung des Lignins in Fichtenholz wurde quantitativ durch Untersuchung von 0,5 m dicken Querschnitten unter dem UV-Mikroskop bestimmt. Die mittlere Ligninkonzentration war in der Mittelschicht etwa doppelt so hoch wie in der Sekundärwand. Die Ligninkonzentration der Mittelschicht war in den an die Tracheiden anstoßenden Zellecken annähernd viermal höher als in der Sekundärwand, wogegen das Volumen der Sekundärwand wesentlich größer war als das der Mittelschicht. Dagegen befand sich beim Frühholz 72% des gesamten Lignins in der Sekundärwand und nur 28% fanden sich in der Mittelschicht selbst und in ihren Zelleckbereichen. Die entsprechenden Werte für Spätholz betragen 82% bzw. 18%. Die Anwendung von schrägen Längsschnitten von 0,1 m Dicke erlaubte die Auflösung der Mittelschicht. Die Ligninkonzentration in der Mittllamelle war gleich groß wie in der in den Zellzwickeln befindlichen Mittellamelle und der Ligningehalt der Primärwand war etwa doppelt so groß wie derjenige in der Sekundärwand.

     

Water circulation, groundwater outflow and nutrient dynamics in Mida creek, Kenya

The relationship between physical hydrodynamic processes and nutrients dynamics was investigated in Mida creek, a groundwater influenced mangrovefringed creek in Kenya between March 1996 and May 1997. The research involved spot and timeseries measurement of nitrate–nitrite, ammonia, silicates, phosphates, salinity, temperature, sealevel as well as tidal currents at seven stations located in the front, middle and backwater zones of the creek. Groundwater level as well as total dissolved solids' concentration, salinity, temperature and nutrients' concentration were also measured once every month in shallow wells (watertable<5m) located in the upper region of the creek. Results of the study show that nutrient concentrations vary with the tide and that, though there is no river drainage, they are of the same magnitude as in mangrove creeks with substantial river runoff. The peak concentrations of NH ₄ ⁺ –N (5.45M), NO ₂ –NO ₃ (5.63M), PO ₄ ^3– –P (0.58M) and SiO ₃ ^2– –Si (81.36M) in the creek occurred during flood tide, 2–3h before high waters. The (NO ₂ ^– + NO ₃ ^– )–N concentrations declined rapidly during ebb tide, reaching the minimum levels during low water. Contribution of groundwater seepage to the net nutrients flux (particularly on nitrite–nitrates) is largest in dry seasons. The study shows that groundwater outflow sustains the mangroves during periods of severe salinity stress and nutrients deficiency in dry seasons. This is essentially by limiting salinity increase and by boosting nutrient supply in dry seasons.     

Cultural factors influencing adventitious shoot and plantlet formation from slash pine cotyledons

Cultural factors affecting in vitro shoot and subsequent plantlet formation of slash pine (Pinus elliotti Engelm.) cotyledons were investigated. Basal media composition, N⁶-benzylaminopurine (BAP) concentration and exposure time significantly influenced bud induction in cotyledons cultured under a continuous photoperiod of 35–40 mol m^–2 s^–1 at 24 ± 1 °C. The largest number of adventitious shoots was obtained after 28 days exposure to 66 M BAP-supplemented modified Gresshoff and Doy 1 (GD1) medium. Relatively high frequencies of large shoots were obtained after a 14-day exposure to 22 M BAP-supplemented Brown and Lawrence (BL) or 66 M BAP-supplemented GD1. Adventitious shoots derived from 21- or 28-day exposures to BAP developed more slowly and were smaller in size than those derived from a 14-day exposure to the cytokinin. Shoot differentiation and subsequent growth were also influenced by basal media, media concentration, and presence of activated charcoal in the medium. The percentage of cotyledons forming shoots was highest on half-strength GD1 medium containing activated charcoal. Rooting was achieved in vitro under a continuous photoperiod of 60–70 mol M^–2 S^–1. Roots were formed when excised shoots were planted on GD 1/2 medium supplemented with 2.68 M 1^–1 a-napthaleneacetic acid (NAA) with or without BAP for 14 days. The proposed technique of slash pine propagation using cotyledon explants can produce up to 100 seedlings per embryo.     

The measurement of tensile creep in thin wood strips

Summary Tensile creep of 100 m thick wood strips cut parallel to the grain was measured using a high resolution high accuracy creep apparatus. Rigorous attempts were made in both design and use of the apparatus to reduce errors to a minimum: the standard deviation of the random errors about a smoothed curve was assessed as 0.34 m. By comparing elongation of strips under load with control strips at zero load viscous and visco-elastic elongation may be accurately measured.     

Agrobacterium-mediated transformation of cork oak (<Emphasis Type="Italic">Quercus suber</Emphasis> L.) somatic embryos

A simple method is presented for the unreported genetic transformation of cork oak (Quercus suber L.). Pro-embryo masses (PEMs) were induced on immature zygotic embryos applied to medium supplemented with 2.3 M 2,4-dichlorophenoxyacetic acid. The established PEMs were inoculated with Agrobacterium tumefaciens LBA4404/p35S GUS INT/pCAMBIA 1301 strain. Transformants were selected on hygromycin (HYG) 94 M-supplemented medium. Viable embryos constituted 13% of those selected on HYG during 4 months. Expression of -glucuronidase at 4 months following co-cultivation confirmed transformation in 5.8% embryos selected on HYG. This method forms a basis for genetic transformation of cork oak somatic embryos.     

Microdistribution of metal elements in wood impregnated with a copper-chrome-arsenic preservative as determined by analytical electron microscopy

Summary The distribution of Cu, Cr and As in the walls of tracheids of Scots pine impregnated with Tanalith C at a retention of 2.5 lb/ft³ has been examined at the submicroscopic level in the electron microscope microanalyser, EMMA-4, using the absolute method, as well as in the conventional electron microscope. All three elements are present in all regions examined (probe area 0.2 m diameter). The concentration of Cu ranges from 0.4–0.9% (w/w), that of Cr is about 0.8%, while As ranges from 2%–2.6%. The morphology of this fine deposit is clearly dictated by the run of the cellulose microfibrils. There is in the wall an occasional coarse deposit, almost entirely copper, and the inner face of the tracheid wall is covered by a thin layer 20–30 m thick which contains all these elements at high concentration.     

Field measurements of gas exchange in Avicennia marina and Bruguiera gymnorrhiza

Diurnal gas exchange characteristics were measured simultaneously in two mangrove species, Avicennia marina and Bruguiera gymnorrhiza, over 7 d in summer (February–March), to compare their productivity. The study was undertaken in the Beachwood Mangroves Nature Reserve, Durban, South Africa, using fully expanded leaves of young and mature trees at the top of the canopy. Gas exchange was strongly influenced by photosynthetic photon flux density (PPFD), leaf temperature and the accompanying leaf to air vapour pressure deficit ( w). Carbon dioxide exchange was saturated at a PPFD of about 600 mol m^-2s^-1 in B. gymnorrhiza compared to 800 mol m^-2s^-1 in A. marina. Maximal CO₂ exchange occurred between 12h00 and 14h00 and was consistently greater in A. marina (8.8 mol m^-2s^-1) than in B. gymnorrhiza (5.3 mu;mol m^-2s^-1). Mean internal CO₂ concentrations ( c_i) were 260 l l^-1 in A. marina and 252 l l^-1 in B. gymnorrhiza. Photorespiratory activity was 32% in A. marina and 30% in B. gymnorrhiza. Mean water use efficiency (WUE) was 8.0 mol mmol^-1 in A. marina and 10.6 mol mmol^-1 in B. gymnorrhiza. Diurnal leaf water potentials ranged from –0.8 to –3.5 MPa and were generally lower in A. marina.     

Lateral fluxes of organic carbon in a mangrove forest partly contaminated with sewage wastes

Lateral fluxes of macrodetritus, particulate matter and dissolved organic carbon were determined. Samples were collected monthly at the mangrove forest–inshore water boundary and within the forest. Floating macrodetritus (mainly mangrove leaves) was collected by a net (2mm mesh size). Dissolved and particulate matter were separated by filtration (0.45m filter) and centrifugation. Dissolved and particulate carbon were determined by a total organic carbon analyser (SHIMADZU model TOC5000). There was a significant export of mangrove litter to the adjacent marine environment during spring tides. Rates of import and export of particulate matter in the forest were not statistically different. Translocation of material within the forest was clearly demonstrated. A relatively high export of macrodetritus was recorded at the marine fringe (mainly colonised by Sonneratia alba). There was low export of litter in the terrestrial fringe zone, a monospecific stand occupied by Avicennia marina (Forsk) Vierh. Net organic carbon export from the entire forest was 79×10⁶g C y^–1, dissolved organic carbon accounted for about 78% of the total export.     

Studies on opposite wood in conifers part II: Histology and ultrastructure

Summary A study has been made of the histology and ultrastructure of opposite wood in Larix laricina, Picea rubens, and Pinus resinosa. The width of the growth rings varied considerably, in one case from 0.1–1.0 mm, with the wide rings containing a much higher proportion of latewood than the narrow ones. The earlywood tracheids were square in outline and more regularly arranged than in normal wood. In the latewood they were sometimes irregular and distorted. The S₃ layer in the tracheids was 0.2 m thick in the earlywood and 0.4–0.8 m in the latewood, as compared to a thickness in normal wood of 0.1–0.2 m in both zones. The S₃ was often buckled in the latewood and was terminated towards the lumen by a spiral thickening. The cell wall structure of the tracheid pit border was described. Normal coniferous wood might be regarded as an intermediate between opposite wood and compression wood.This paper is dedicated to Dean Edwin C. Jahn in honor of his 70th birthday.     

Moisture content-water potential relationship of red pine sapwood above the fiber saturation point and determination of the effective pore size distribution

Summary The pressure membrane and pressure plate techniques were used to establish the moisture content-water potential (M-) relationship of red pine (Pinus resinosa Ait.) sapwood in desorption above the fiber saturation point. The moisture content-water potential relationship is required for the development of a model of drying considering the gradient of water potential as the driving force of moisture in wood. This relationship was established at 18, 56 and 85 °C for radial desorption. The results obtained demonstrate that water potential increases with temperature T at a given moisture content M. There is no significant variation of /T with temperature. Also, there is no plateau at intermediate moisture contents as was the case for the M- relationship of aspen sapwood established in a previous work. The effective integral and differential pore size distributions inferred from the M- relationship are also presented. The largest proportion of effective pore openings was found for a radius of 0.2 m. This value can be related to the pit membrane openings of red pine.This research project is currently supported by the Natural Sciences and Engineering Research Council of Canada under grant no. OGP0121954     

Lignin in woody tissues after chemical pretreatment and bacterial attack

Summary Tangential latewood sections (60 m) of Scots pine sapwood were differently treated with chlorite. The subsequently incubated two strains of Bacillus polymyxa caused a weight loss up to about 25%, including a loss of lignin of about 42%.UV-microspectrophotometry of 1 m-cross-sections prepared from the cultured woody tissues demonstrated that bacteria caused neither quantitative nor qualitative changes of the remaining lignin.The lignin, which was dissociated from the pretreated woody cell wall by bacteria, could not be respired, suggesting that the lignin is a ballast to these bacteria that inhibits the dissimilation of the carbohydrates in the wood.We appreciate the financial support by Deutsche Forschungsgemeinschaft. Thanks are due to Mrs. R. Endeward for her assistance during the microspectrophotometric evaluations and to Mr. U. Engel and Dr. O. Faix for carrying out the IR-measurements     

In vitro and ex vitro rooting of micropropagated shoots using three green ash (Fraxinus pennsylvanica) clones

Shoot multiplication using seedling materials was achieved by subculture on Murashige and Skoog salts with Gamborg's B5 vitamins (MSB5) medium containing a combination of 5 M 6-benzyladenine (BA), 5 M thidiazuron (TDZ), and 1 M indole-3-butyric acid (IBA) with three green ash (Fraxinus pennsylvanica) clones, SD1009 (South Dakota origin), SD2002 (South Dakota origin), and KA2018 (Kansas origin). Shoots were rooted using in vitro and ex vitro methods. For in vitro rooting studies, elongated shoots were transferred to rooting plugs supplied with liquid MSB5 medium containing a 3×3 factorial arrangement of two different auxins, -naphthaleneacetic acid (NAA) and IBA, at three concentrations (0, 5, and 10 M). The most effective treatment for in vitro root number, root length, and shoot height was 5 M IBA. The three clones also were tested for ex vitro rooting using a quick dip in 1 mM NAA, 1 mM IBA, or control (no auxin). The maximal ex vitro rooting response occurred when shoot explants of the three clones were dipped in 1 mM IBA. Significant clonal differences were noted in response to in vitro and ex vitro rooting treatments. Rooted plantlets were acclimated to the greenhouse.     

The validity of the use of SEM/EDAX as a tool for the detection of UF resin penetration into wood cell walls in particleboard

Summary The interpretation of data from scanning electron microscopy (SEM) combined with energy dispersive analysis of X-rays (EDAX) is dependent on the size of the excitation volume, the magnitude of which for organic materials such as wood has been little studied. From the analysis of a model system comprised of a thin layer of gold sandwiched between two layers of an epoxy plastic, it was shown that about 90% of all excitations arise from a volume about 6 m wide and 4 m deep. However, these dimensions vary with the orientation of the gold relative to the specimen surface, and the specimen tilt. Theoretical predictions based on these data support, but not conclusively, the previously published SEM/EDAX evidence of the penetration of urea formaldehyde resins into the wood cell wall in particleboard manufacture.This paper was written while A. J. Bolton was Heritage Visiting Scientist at the Forest Products Laboratory, U.S. Department of Agriculture, and Visiting Professor at the Department of Forestry, University of Wisconsin, both at Madison, Wisconsin, U.S.A. This support is gratefully acknowledged. The authors would also like to acknowledge the assistance of E. Edwards in machining the SEM specimens     

Electromagnetic shielding efficiency of the electric field of charcoal from six wood species

Six wood species were carbonized under various carbonization temperatures and nonoxygen conditions to obtained charcoal. The effects of wood species, rate of temperature rise, and carbonization temperature on the electromagnetic shielding efficiency (ESE) of the electric field were investigated. The wood species used in this study were Japanese cedar, China fir, western hemlock, red oak, fortune paulownia, and Taiwan acacia. Tested materials were carbonized in a high-temperature oven under the following conditions: rate of temperature rise 1°–5°C/min; carbonization temperature 500°–1100°C, with temperature intervals of 100°C; maximum temperature maintained for 1h; and flow rate of nitrogen 300ml/min. The electromagnetic insulation strength system was used to detect the ESE of the electric field of charcoal. It was found that western hemlock and fortune paulownia charcoal showed maximum ESE values of of 36 and 61dB generated at a carbonization temperature of 1000°C. The charcoals derived from four other wood species showed maximum ESE values of 28dB for Japanese cedar, 23dB for China fir, 32dB for red oak, and 38dB for Taiwan acacia, respectively, at a carbonization temperature of 1100°C. The ESE value for fortune paulownia charcoal was similar to those of metal nets. The relations between ESE and logarithmic values of resistivity (log) could be represented by a negatively exponential formula.Part of this report was presented at the 50th Annual Meeting of the Japan Wood Research Society, Kyoto, April 2000