Variation of Microfibril Angle Between Four Provenances of Sitka Spruce (Picea sitchensis [Bong.] Carr.)

Abstract: The mean microfibril angle (MFA) of tracheids in Sitka spruce ( Picea sitchensis [Bong.] Carr.) from four provenances was determined by X-ray diffraction. Seeds were from four locations in North America: California, Oregon, Queen Charlotte Islands and Washington. Transplants were planted in 1976 and grown in Ireland (Shillelagh Forest). The mean MFA close to the pith was ca. 22° and decreased to ca. 11° in the outer rings. It was found that the MFA varies strongly not only as a function of the annual ring and the distance from pith but also as a function of the seed origin. Trees of California and Queen Charlotte Islands provenance had a larger mean MFA in general than trees of Washington and Oregon provenance. Also, the shapes of the cell cross sections were determined to some extent from the diffraction patterns. For samples of California and Washington provenance the average thickness of the cellulose crystallites was determined to be about 3.0 nm.     

Cytokinin pulse-mediated shoot organogenesis from cotyledons of Sitka spruce [Picea sitchensis (Bong.) Carr.] and high frequency in vitro rooting of shoots

Shoot formation from cotyledons of Sitka spruce was induced by a 2 h incubation in an aqueous solution of 400 μM benzyladenine, followed by culture on medium lacking plant growth regulators. Several basal media were assessed. Murashige and Skoog (1962) based medium was found to be optimal on which, after 25 weeks, 62.5% of cotyledons produced shoots with a mean of 10.7 shoots per cotyledon. Shoots, detached from cotyledons and propagated individually by axillary bud proliferation, rooted with a frequency of 84% following culture in half-strength MS-based medium lacking growth regulators under low illumination (1.5 μmol m⁻²s⁻¹). Rooted plants were successfully transferred to ex vitro conditions. This revised version was published online in June 2006 with corrections to the Cover Date.     

The influence of culture vessel head-space volatiles on somatic embryo maturation in Sitka spruce [Picea sitchensis (Bong.) Carr.]

The maturation of somatic embryos of Sitka spruce [Picea sitchensis (Bong.) Carr.] was found to be highly dependent on the method used to seal plastic Petri dishes. Large numbers of well-formed mature embryos developed if dishes were sealed with PVC cling-film (CF) whilst sealing with Parafilm M (PF) greatly reduced the numbers of embryos forming. Inclusion of potassium permanganate oxidation traps, normally used to deplete the atmospheric ethylene, greatly stimulated somatic embryo maturation under PF sealing. Similarly, traps of adsorption agents (Tenax, activated charcoal or soft white paraffin), capable of removing volatiles from the culture vessel head-space, stimulated somatic embryo maturation under PF sealing although to a lesser extent than the oxidation traps. Incorporation of silver nitrate or 2-chloroethylphosphonic acid (ethephon) in the culture medium indicated that ethylene was not the agent supressing somatic embryo maturation under PF sealing.Abbreviations ABA abscisic acid - CF PVC cling-film - PF Parafilm M     

Somatic embryogenesis in Sitka spruce (Picea sitchensis (Bong.) Carr.)

Embryonal-suspensor masses from immature embryos from cones of Sitka spruce (Picea sitchensis (Bong.) Carr.) proliferated on a modified Murashige & Skoog medium with N⁶-benzyl-aminopurine, kinetin, 2,4-dichlorophenoxyacetic acid and an organic nitrogen source. The slimy white embryonal-suspensor masses with proembryos were maintained on a solid proliferation medium with reduced amounts of growth regulators. Transfer of embryonal-suspensor masses to a non-woven polyester carrier with liquid maturation media containing ±2-cis-4-trans-abscisic acid and a reduced amount of inositol and organic nitrogen resulted in synchronized embryo formation. Further development was achieved on a medium without ±2-cis-4-trans-abscisic acid and organic nitrogen. Somatic embryos were successfully transferred ex vitrum.Abbreviations ABA ±2-cis-4-trans-abscisic acid - BAP N⁶-benzyl-aminopurine - ESM embryonal-suspensor masses - KIN kinetin - 2,4-D 2,4-dichlorophenoxyacetic acid     

The effect of cultivation technique on root and shoot biomass production by young Sitka spruce (Picea sitchensis (Bong.) Carr.) trees on surface water gley soils

In Ireland much of the land available for plantation establishment requires some degree of soil cultivation and drainage to improve its suitability for tree growth. The method of cultivation and drainage normally varies depending upon the soil type and its drainage characteristics. Little research has been carried out on the impact of practices such as mounding and mole drainage upon rooting and biomass production in young crops. The research reported in this paper was carried out on four sites where the general soil type was surface water gley. Three of the sites studied were afforestations on old farmland, while the fourth site was a reforestation of a windblown stand. The cultivation methods investigated included mounding, mole drainage with mounds, mole drainage only and ripping. Sitka spruce trees ranging in age from five to thirty years were excavated from these experimental sites to describe the impact of different soil cultivation techniques upon root architecture and above and below ground biomass production. The results showed that while the effect of mound drainage upon water table level varied from site to site, the establishment of trees on mounds led to an increase in above and below ground biomass production. Mole drainage of these sites without mounds, did not lead to any increase in root or shoot development when compared with trees growing on uncultivated/undrained ground. This revised version was published online in June 2006 with corrections to the Cover Date.     

The mycorrhizal community in a forest chronosequence of Sitka spruce [Picea sitchensis (Bong.) Carr.] in Northern England

Demography and fungal diversity of the belowground ectomycorrhizal community in a chronosequence of Sitka spruce [Picea sitchensis (Bong.) Carr.] in Northumberland, Northern England, were analysed; mycorrhizal root samples were taken from 6-, 12-, 30- and 40-year-old stands, and fungal fruiting bodies were collected in autumn to complement the survey. Naturally germinated seedlings less than 1 year of age (taken from the 30-year-old stand) were also examined. A total of 118,000 mycorrhizal root tips were extracted from 40 soil cores (ten per age class) and from the complete root systems of 25 seedlings and separated into active and senescent root tips according to their morphology and anatomy. Active tips were distinguished according to their mycobionts which were characterised and identified microscopically. Although almost 100% of all fine roots were mycorrhizal, EM fungal diversity throughout the chronosequence was low, consisting of a total of 16 species of which three were only found as fruiting bodies. Of the six mycobionts found most regularly below ground, Tylospora fibrillosa was the most common, colonising about 70% of all root tips and more than 90% of those of seedlings and young trees. Root density and mycorrhizal diversity increased, but percentage of vital root tips decreased with increasing tree age, levelling off in the 30- and 40-year-old stand. Among the five subdominant fungal species, Dermocybe crocea was found to have its peak of distribution in the 12-year-old stand and Russula emetica, Lactarius rufus, Hymenoscyphus ericae agg. and the unidentified Piceirhiza sulfo-incrustata in the 30- and 40-year-old stands. The possible correlations between the mycorrhizal community structure and biotic and abiotic factors are discussed.     

Diurnal water storage in the stems of Picea sitchensis (Bong.) Carr.

Abstract. Two models of the relationship between diurnal variation in shoot water potential and transpiration in 14-year-old Picea sitchensis (Bong.) Carr. were compared. The first model was a physiologically based resistance-capacitance (R-C) analogue with its associated differential equations. The second was a non-physiological discrete-difference (D-D) or stochastic transfer function model. The RC model included only the effect of water storage in the phloem and bark while the D-D model implicity included all storage mechanisms. The R-C and D-D models explained similar fractions (62% and 68% respectively) of the variation in shoot water potential due to diurnal changes in transpiration rate. However, the D-D model had fewer parameters than the R-C model. The results from the D-D model showed that the resistance to flow from soil to shoots along the trunk, (RT), was 5 × 10³ MPa kg^-1s and the capacitance of the phloem and bark treated as a single store, (C_s), was 1.6 kg MPa^-1. It is suggested that the resistance to flow into storage (R_s) is much greater than R_T and can be disregarded. A non-linear version of the D-D model suggested [hat resistance to flow in the trunk increases with increasing transpiration rate.     

Localised impact of Sitka spruce (Picea sitchensis (Bong.) Carr.) on soil permeability

A typical upland soil catena afforested with Sitka spruce (Picea sitchensis (Bong.) Carr.) was chosen to examine the localised effect of trees on soil permeability. A borehole permeameter was used to measure soil permeability at 0.2 m and approximately 2 m distance from the stem of 20 trees at a fixed measurement depth of 0.25 to 0.45 m. In the case of the near-tree measurements, this corresponded to soil beneath the main root plate of each conifer. Two principal elements of the soil catena: the ferric podzol of the mid-slope and histosol soil of the foot-slope were investigated.The preliminary data set shows that within the ferric podzol element, the permeability of the soil beneath individual conifers was a factor of 5.4 less than that of the adjacent soil. In contrast, within the histosol sub-tree permeabilities could not be distinguished from those of soil 2 m away from each tree. The decrease in sub-tree permeability within the podzol may be caused by sensitivity of the Bsl horizon to consolidation by tree weight or by enhanced illuviation resulting from changes in local soil chemistry. The histosol may be less sensitive to such processes. The results of a consolidation test applied to the Rawls and Brakensiek model of soil permeability supported the possible role of consolidation in the reduction of soil permeability beneath conifers in podzolic soil. Additional data on soil bulk density, porosity and texture are required to corroborate either the consolidation or illuviation hypotheses. As the Bsl horizon of ferric podzol soil is typically slowly permeable, a further decline may (i) restrict root development and thus, increase windthrow hazard, and (ii) increase the lateral flow of water within podzolic Eag horizons and thus affect stream acidification. Deep ploughing of a site prior to afforestation may mitigate such impacts.     

Development of a set of highly polymorphic genomic microsatellites (gSSRs) in Sitka spruce (Picea sitchensis (Bong.) Carr.)

Robust, polymorphic microsatellite DNA markers (simple sequence repeats—SSRs) are valuable tools for a range of tree conservation and breeding applications. SSRs are routinely used in the study of population genetic structure and diversity, pedigree reconstruction and genetic linkage mapping. Their abundance in the genome, co-dominant inheritance and potential for cross-species amplification make microsatellites highly prized markers. This paper characterises 22 novel genomic polymorphic microsatellite loci for Sitka spruce (Picea sitchensis (Bong.) Carr.). Amplification of DNA from Sitka spruce material was carried out both with a set of unrelated trees to obtain diversity statistics for each locus, and with the progeny of a full-sib family to test simple Mendelian inheritance. Observed heterozygosity ranged from 0.38 to 0.91 and allele number per locus ranged from 6 to 21, with a mean of 12.2. In addition, the primer pairs were tested with DNA from Norway spruce (P. abies) and white spruce (P. glauca) to investigate their potential for cross-species amplification and ten loci amplified in all three species. The results from these genomic microsatellites are compared to data generated from microsatellites derived from Picea EST libraries. In summary, this novel, highly polymorphic markers represent a significant addition to the rapidly expanding Picea genomics tool-box. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Ectomycorrhizal colonisation of Sitka spruce [Picea sitchensis (Bong.) Carr] seedlings in a Scottish plantation forest

 The ectomycorrhizal (ECM) colonisation of seedling Sitka spruce (Picea sitchensis) was examined in an uneven-aged plantation forest in southern Scotland. The extent of ECM colonisation of individual seedlings was 43.8–97.2%, with an overall mean of 80.3 ± 1.1%. A total of 13 ECM morphotypes were differentiated, with 1–4 ECM types colonising an individual seedling. ECM colonisation was dominated by a single species, Tylospora fibrillosa, which accounted for 72.4–97.7% of the ECM colonisation recorded, on a plot mean basis. Other ECM types appeared to be distributed very patchily, only two types (Lactarius sp. and Mycelium radicis atrovirens Melin) exceeding a mean of 10% colonisation in any one plot. No significant correlations were recorded between ECM colonisation and seedling growth, or between ECM colonisation and soil pH, loss-on-ignition, or water content. Accepted: 16 October 1997     

The Effect of Explant Type on the Establishment of Sitka Spruce [Picea sitchensis (Bong.) Carr.] in Culture

Five different types of shoot tip explant from three-year-oldand 13-year-old Sitka spruce trees were established on a rangeof media and shoot growth was recorded. While Schenk and Hildebrandtmedium gave the greatest shoot elongation, Webb and Street mediumproduced the healthiest-looking shoots. Poorest growth was recordedfor the explants cultured without the crown or any vasculartissues from the previous year's growth. [¹⁴C]sucrose tracerstudies indicated that poor uptake was not responsible for thereduced growth and it was suggested that damage due to inadequatewater uptake may be responsible. Results demonstrated that peelingthe protective scales from resting buds and trimming the shoottissues below the crown to a wedge shape and excluding the barkwas a reliable method of explant preparation for establishingmature and juvenile Sitka spruce in culture. Picea sitchensis, Sitka spruce, mature, juvenile, crown, [¹⁴C]sucrose, micropropagation     

Effect of carbenicillin and cefotaxime on somatic embryogenesis of Sitka spruce (Picea sitchensis (Bong.) Carr.)

Carbenicillin prevented the development of mature somatic embryos,reduced early stage embryos by >90% and tissue growth by50%. Cefotaxime had no effect on overall tissue growth, butreduced the development of early and mature embryos by 20% and66–80%, respectively. Carbenicillin should not be usedfor transformation of Sitka spruce. Key words: Sitka spruce, somatic embryogenesis, antibiotic sensitivity     

A set of microsatellite markers for use in Sitka spruce (Picea sitchensis) developed from Picea glauca ESTs

Few microsatellite markers have been specifically developed for Picea sitchensis. In January 2004 the appearance of over 10 000 sequences of expressed regions of DNA from P. glauca in GenBank presented an opportunity for the development of additional microsatellite markers in Sitka spruce. Mono‐ and dinucleotide repeat sequences were located in these sequences and primers were designed around these regions. Amplification was attempted in Sitka material from a broad geographical range and the level of polymorphism was assessed. Primers were also tested in progeny of a controlled cross. Nine polymorphic loci that demonstrated Mendelian inheritance in Sitka were discovered in this study.     

Effect of stand age on greenhouse gas fluxes from a Sitka spruce [Picea sitchensis (Bong.) Carr.] chronosequence on a peaty gley soil

The influence of forest stand age in a Picea sitchensis plantation on (1) soil fluxes of three greenhouse gases (GHGs – CO₂, CH₄ and N₂O) and (2) overall net ecosystem global warming potential (GWP), was investigated in a 2‐year study. The objective was to isolate the effect of forest stand age on soil edaphic characteristics (temperature, water table and volumetric moisture) and the consequent influence of these characteristics on the GHG fluxes. Fluxes were measured in a chronosequence in Harwood, England, with sites comprising 30‐ and 20‐year‐old second rotation forest and a site clearfelled (CF) some 18 months before measurement. Adjoining unforested grassland (UN) acted as a control. Comparisons were made between flux data, soil temperature and moisture data and, at the 30‐year‐old and CF sites, eddy covariance data for net ecosystem carbon (C) exchange (NEE). The main findings were: firstly, integrated CO₂ efflux was the dominant influence on the GHG budget, contributing 93–94% of the total GHG flux across the chronosequence compared with 6–7% from CH₄ and N₂O combined. Secondly, there were clear links between the trends in edaphic factors as the forest matured, or after clearfelling, and the emission of GHGs. In the chronosequence sites, annual fluxes of CO₂ were lower at the 20‐year‐old (20y) site than at the 30‐year‐old (30y) and CF sites, with soil temperature the dominant control. CH₄ efflux was highest at the CF site, with peak flux 491±54.5 μg m⁻² h⁻¹ and maximum annual flux 18.0±1.1 kg CH₄ ha⁻¹ yr⁻¹. No consistent uptake of CH₄ was noted at any site. A linear relationship was found between log CH₄ flux and the closeness of the water table to the soil surface across all sites. N₂O efflux was highest in the 30y site, reaching 108±38.3 μg N₂O‐N m⁻² h⁻¹ (171 μg N₂O m⁻² h⁻¹) in midsummer and a maximum annual flux of 4.7±1.2 kg N₂O ha⁻¹ yr⁻¹ in 2001. Automatic chamber data showed a positive exponential relationship between N₂O flux and soil temperature at this site. The relationship between N₂O emission and soil volumetric moisture indicated an optimum moisture content for N₂O flux of 40–50% by volume. The relationship between C : N ratio data and integrated N₂O flux was consistent with a pattern previously noted across temperate and boreal forest soils.