Quantitative assessment of the bacterial rhizosphere flora of Pinus contorta var. latifolia.

The bacterial flora associated with root systems of young and mature lodgepole pine was investigated by sampling forest-grown trees. Counts were performed and expressed on a surface-area basis to give a more realistic comparison of organism density or activity within the control soil, rhizosphere soil, and rhizoplane. On this basis, densities increased by an order of 10(4)- to 10(6)-fold from control soil to rhizoplane, with the degree of stimulation being inversely related to root radius.     

Nitrogen fixation associated with Suillus tomentosus tuberculate ectomycorrhizae on Pinus contorta var. latifolia

BACKGROUND AND AIMS: Tuberculate ectomycorrhizae are a unique form of ectomycorrhiza where densely packed clusters of mycorrhizal root tips are enveloped by a thick hyphal sheath to form a tubercle. The functional significance of such a unique structure has not previously been established. The purpose of the present study was to investigate and measure the potential nitrogenase activity associated with Suillus tomentosus/Pinus contorta tuberculate ectomycorrhizae in two stand ages, young and old, and across a range of nitrogen-poor soil conditions. METHODS: Short roots were compared with other mycorrhizae and non-mycorrhizal secondary roots using tuberculate ectomycorrhizae. Assessment of nitrogenase activity was determined and quantitative measurements were taken on tuberculate ectomycorrhizae in situ in a variety of different circumstances, by using an adaptation of the acetylene reduction assay. KEY RESULTS: Significant nitrogenase activity was measured associated with S. tomentosus/P. contorta tuberculate ectomycorrhizae whereas no nitrogenase activity was measured with non-tuberculate mycorrhizae or secondary roots without mycorrhizae. Average nitrogenase activity ranged from undetectable to 5696.7 nmol C2H4 g(-1) tubercle 24 h(-1). Maximum nitrogenase activity was 25,098.8 nmol C2H4 g(-1) tubercle 24 h(-1). Nitrogenase activity was significantly higher in young stands than in old stands of P. contorta. Season or some covariate also seemed to affect nitrogenase activity and there was suggestion of a site effect. CONCLUSIONS: Suillus tomentosus/P. contorta tuberculate ectomycorrhizae are sites of significant nitrogenase activity. The nitrogenase activity measured could be an important contribution to the nitrogen budget of P. contorta stands. Season and stand age affect levels of nitrogenase activity.     

Dihydroconiferyl alcohol in developing xylem of Pinus contorta

Dihydroconiferyl alcohol was identified by GC/MS as an endogenous constituent of developing xylem in Pinus contorta ssp. latifolia. Its role in regulating wood formation in P. contorta is discussed.     

A method for studying dead bole dynamics in Pinus contorta var. latifolia - Picea engelmannii forests

Rates of decomposition were determined for the boles of Pinus contorta var. latifolia Engelm. and Picea engelmannii Parry ex. Engelm. in five lower subalpine forest stands in the Southern Canadian Rocky Mountains. Stands ranged in age from 58 to 222 yr since last fire. The date of death of standing dead and fallen boles was determined by cross-dating their ring-width patterns to stand master ring- width chronologies. Boles could be dated which had been dead for up to 35 years in a 58-yr old stand and 100 yr in a 222-yr old stand. An empirical relationship between mass density and diameter of live trees was used to predict the mass density at death for trees already dead. The falling rate of dead standing boles was estimated from the dead bole's time of death and whether it was standing or on the ground at the time of observation. The falling rates for both Pinus contorta and Picea engelmannii ranged from 0.020 to 0.064/yr falling with most stands in the 0.050 range. No effect of size was found on the falling rate probably as a result of the small size of the boles (< 20 cm). Because of their dry condition standing dead boles do not decompose. An equation was developed which estimates the time a dead bole has been on the ground, given that it would have stood for some time before falling over and been subject to little decomposition during this time. Decomposition rates, using a negative exponential model, gave only marginally better r² than linear models. Pinus contorta had exponential decay rates from 0.0299 and 0.0171 mass density loss/yr for the most recent 15 and 25 yr in stands 58 yr old, to 0.0045 and 0.0035 mass loss/yr for the most recent 65 and 80 yr in stands 215 and 222 yr old. Picea engelmannii had exponential decay rates of 0.0054 and 0.0025 mass loss/yr for the most recent 20 and 65 yr in stands 99 and 215 yr old.     

Random amplified polymorphic DNA diversity of marginal and central populations in Pinus contorta subsp. latifolia.

Fifteen populations of lodgepole pine (Pinus contorta subsp. latifolia) were surveyed for diversity across 52 random amplified polymorphic DNAs (RAPDs). The objective was to compare single-locus and multilocus structures in four marginal, three intermediate, and eight central populations. Single-locus estimates indicated average observed and expected heterozygosity to be 0.19 and 0.17, respectively. When these estimates were split into population categories, a clear trend of increasing diversity was detected in the direction of marginal to central populations. F-statistics indicated an excess of heterozygotes, with F(IS) ranging from -0.08 for marginal populations to -0.15 for central populations and averaging -0.12 over 15 populations. The estimates of F(ST) decreased towards the margins of the species range, indicating increased population differentiation. Forty-nine of 52 RAPDs tested neutral in the Ewens-Watterson analysis. Multilocus analysis showed significant two-locus and high-order gametic disequilibria in all 15 populations. The most prominent components of the two-locus analysis were the variance of disequilibrium (VD, 46.2%) and the multilocus Wahlund effect (31.9%). This high value for VD indicated that founder effects could explain much of the observed multilocus associations. When analyzed by population categories, the VD showed a decreasing trend indicating that variation due to founder effects was more prominent in marginal populations. The two-locus Wahlund effect (WC) that is characteristic of strong population subdivision was highest in the central populations. This indicated significant levels of gene flow between populations with different allelic combinations.     

Construction of a framework map in Pinus contorta subsp. latifolia using random amplified polymorphic DNA markers.

We report on the construction of the first random amplified polymorphic DNA (RAPD) framework map in Pinus contorta subsp. latifolia. Genomic DNA of haploid megagametophytes from 90 open-pollinated seeds originating from a single tree were amplified with 840 random decamer oligonucleotide primers by the polymerase chain reaction. Three-hundred twenty-eight RAPD markers with fragment sizes that ranged between 260 and 3080 base pairs were found segregating at 110 random decamer oligonucleotide primers. Of these 328 RAPD markers, 148 were mapped to 16 framework linkage groups and 77 were mapped as accessory markers onto the framework linkage groups, on a support interval of minimal LOD score of 3. The 16 framework maps cover a distance of 2287 cM. The estimate of genome size was 2407 cM with a 95% confidence interval of 2304-2459 cM.     

Surface colonization of lodgepole pine (Pinus contorta var. latifolia [Dougl. Engelm.]) roots by Pseudomonas fluorescens and Paenibacillus polymyxa under gnotobiotic conditions

Indirect immunofluorescence techniques and confocal scanning laser microscopy were used to identify rhizobacterial strains on the root surfaces of pine seedlings, which were grown from seeds under gnotobiotic conditions. Conifer plant growth promoting rhizobacterial strains Paenibacillus polymyxa L6 and Pw-2, and the forest soil isolate Pseudomonas fluorescens M20, were inoculated onto surface-disinfested pine seeds, singly, or in dual combinations: strains L6 + M20, or strains Pw-2 + M20. Segments containing particular root microsites (root tip, root hair zone, or areas of lateral root emergence) were sampled randomly from roots 7 or 13 weeks after inoculation, and the colonization of roots by each bacterium was observed. Root segments were also sampled from individual roots at six different points along the length of the root, and the qualitative colonization of younger areas, closer to the root tip, contrasted with that of older areas, closer to the root base. The ability of strain M20 to colonize root areas adjacent to sites of lateral root emergence improves in the presence of either P. polymyxa strain, while the ability of the P. polymyxa strains to colonize these areas was not affected. More rhizobacteria were also generally observed on younger root tissues than on areas closer to the root base.     

Pinus contorta var. yukonensis var. nov. (Pinaceae) from south–central Yukon,Canada

Pinus contorta Douglas ex Loudon var. yukonensis W. L Strong (Yukon pine), a new variety, is described from south–central Yukon, Canada. It is a stout tree with long lower branches, often with paired lower and forked upper stems, and weakly platy to smooth gray bark on the lower stem, when < 70 years old; with bark on the lower stem becoming furrowed and platy on older trees. This contrasts with the narrow columnar habit, simple stem, and scaly bark associated with the more common inland var. latifolia. In several respects, var. yukonensis is more similar to var. contorta (a coastal variant), although occupying a different ecological regime and spatially disjunct (i.e., a high latitude and high elevation variant). Variety yukonensis may represent the taxon alluded to in previous leaf oil terpene and genetic studies, and a possible Beringian survivor.     

Multilocus structure in Pinus contorta Dougl.

We studied isozyme variation at 21 loci in 66 populations from three subspecies of Pinus contorta Dougl.; 35 in spp. latifolia, 20 in spp contorta and 11 in spp. murrayana. The objectives were to assess gametic disequilibria and multilocus structure. There was considerable differentiation of allele frequencies at 19 polymorphic loci across the 66 populations and within the subspecies. Allele frequencies at many loci correlated with geographic variables. Genetic variability varied considerably among populations within subspecies but the subspecies means were similar. The mean number of polymorphic loci and the mean heterozygosity over 19 polymorphic loci were, respectively, 13 and 0.194 in latifolia, 12 and 0.196 in murrayana, and 12 and 0.180 in contorta. The mean heterozygosity correlated with longitude and altitude across the 66 populations and with latitude in latifolia. Gametic disequilibria were evident in 40 populations; 29 in latifolia, eight in murrayana and three in contorta. Gametic disequilibria correlated with latitude across the 66 populations and with longitude in latifolia. The single-locus F _ST averaged 0.0339 in latifolia, 0.0567 in murrayana, and 0.0764 in contorta. The multilocus F _STM was 0.1227 in latifolia, 0.2926 in murrayana, and 0.3328 in contorta. Multilocus Wahlund and founder effects, migration patterns, and natural selection, probably played significant roles in generating and maintaining the multilocus genetic structure in P. contorta in general and the subspecies latifolia in particular.     

Northernmost North American Pinus contorta var. latifolia (lodgepole pine) sociations and vegetation diversity relative to its central range east of the Rocky Mountains

Lodgepole pine (Pinus contorta var. latifolia) stands were sampled in central Yukon, Canada (61.5–64°N latitude), which represented the northernmost 9% of the tree's North American range. Within this area, lodgepole pine occupied only ? 2% of the landscape. This study determined: 1) what forest sociations occurred (i.e. structural dominance‐types); 2) how plant growth form composition and richness differed from the central portion of the species’ geographical range; and 3) if stands were biased towards occurring on more thermally favorable south‐facing slopes. Five lodgepole pine sociations were recognized among 100 relevés: Rhododendron groenlandicum (Labrador tea); Cladonia arbuscula (green reindeer lichen); Calamagrostis purpurascens (purple reedgrass); Hylocomium splendens (stairstep moss) and Alnus viridis (green alder, n = 4 relevés). Rhododendron stands were proportionally more common on low gradient sites and had more total plant cover than the other sociations. Cladonia and Calamagrostis stands were typically associated with dry coarse‐textured soils and warm dry sites, respectively; whereas the composition of the Hylocomium sociation reflected the detrimental influences of atypically dense forest canopies on understory vascular plants. Only the Calamagrostis sociation was unique to the study region. Species richness among common northern lodgepole pine sociations averaged 16–19 taxa per relevé (p > 0.05). Northern compared to central range (n = 1394) relevés were compositionally different based on little overlap of their datasets in the ordination space. Northern vegetation had less (p < 0.001) total plant (129% vs 184%), deciduous shrub (9% vs 26%), broad‐leaved herb (5% vs 25%), and bryophyte (27% vs 54%) cover; had greater macro‐lichen cover (13% vs 5%) and lower floristic richness (11 vs 24 taxa) and was less than half as phytosociological diverse. Lodgepole pine stands in the northernmost portion of their range were not biased towards occurring on south‐facing slopes, which suggested an ecological potential for range expansion.     

Soil solution chemistry in lodgepole pine (Pinus contorta ssp. latifolia) ecosystems,southeastern Wyoming,USA

Concentrations of the principal inorganic and organic solutes in the soil root-zone were measured in six contrasting lodgepole pine (Pinus contorta ssp.latifolia) forest ecosystems for five years (1979–1983). Consistent temporal changes in the principal inorganic solutes (Ca, Mg, K, Na, SO₄, Cl) were observed in all the forest stands and years of study: high concentrations at the initiation of snowmelt in the spring were followed by rapid declines to rather constant values in the mid to late stages of snowmelt. Except for K, concentrations of these solutes differed significantly between sites and between years. Sulfate was the principal mobile anion in the root-zone soil solutions, but contributions of bicarbonate and organic anions also were important.The pH of root-zone solutions was relatively high (6.0), did not change significantly as snowmelt proceeded, and was significantly lower in high-clay soils. No consistent trends in bicarbonate alkalinity were observed and soil atmosphere CO₂ concentrations were only about 10 to 20 times above atmospheric levels, peaking at the end of the snowmelt interval. Concurrent changes in the concentrations of dissolved organic carbon, non-volatile acid-neutralizing capacity, and total Al and Fe indicated that these soil-forming metals were transported vertically in the soil as organic complexes. Precipitation of these complexes was more rapid and more complete in the soils with high clay content than in the coarser soils. Moreover, organic anions comprised up to 30% of the total anionic charge in the coarse-textured soils but less than 10% in the fine soils.Little seasonal or spatial variation of inorganic N and P concentrations was observed in root-zone solutions, probably as a result of high biotic demand for these limiting nutrients. Flux of N and P in these ecosystems was predominately via organic forms so that losses of these nutrients was strongly linked to the mobility of dissolved organic carbon. However, a two-fold increase in the organic N:P and C:P ratios was observed during passage of melt water from the forest floor to mineral soil, evidence of more rapid mineralization of organic P.     

Role of cytokinin in differentiation of secondary xylem fibers

The differentiation of secondary xylem fibers was studied in cultured hypocotyl segments of Helianthus annuus L. It is shown that cytokinin is both a limiting and controlling factor in the early stages of fiber differentiation. In the absence of kinetin or zeatin, there was no fiber differentiation. However, cytokinin could induce fiber differentiation only in the presence of indoleacetic and gibberellic acids. First fibers were observed in the tissue after 12 days in culture, and their number increased linearly during the following 2 weeks. At low cytokinin levels, there was a positive correlation between cytokinin concentration in the medium and the number of fibers formed in the explants. A similar correlation was also found at low gibberellic acid concentrations. At high concentration, zeatin was more effective than kinetin. It seems that later stages of fiber differentiation can occur in the absence of cytokinin. It is proposed that the mechanism which controls and determines the early stages of fiber differentiation is based on an interaction of three major hormonal signals: indoleacetic acid plus gibberellic acid from the leaves with zeatin from the root apices.     

Soil and xylem water potential and soil water content in contrasting Pinus contorta ecosystems,Southeastern Wyoming,USA

Summary The relationships between volumetric soil water content (), in situ soil water potential (_soil) and predawn xylem pressure potential (_predawn) were quantified in four contrasting lodgepole pine ecosystems in Wyoming, USA. On three of the sites, changes in _soil correlated closely with _predawn, but on a porous soil derived from coarse granitic parent material, _predawn declines occurred much sooner than corresponding declines in _soil, possibly because of local depletion of rhizosphere moisture and low molecular diffusivity of water in that soil. Exptrapolation of laboratory-derived characteristic curves for soil moisture to field conditions yielded different relationships between and _soil than curves derived from in situ measurements, probably because of disruption of soil structure and porosity during sample collection and handling in laboratory studies. Although a close correlation between and _predawn was observed, future efforts at modelling the soil-plant-atmosphere continuum should be directed towards a more detailed understanding of the complex relationships between _soil at varying depths and plant water stress.     

扁担木叶片和次生木质部解剖和水分生理特征的可塑性

基于叶片和次生木质部解剖特征及水分生理指标观测,研究了淮北相山不同群落中扁担木的表型可塑性.结果表明,扁担木叶片结构表现出中生特点：叶为异面叶,较薄,角质层不甚发达,气孔密度较小.次生木质部表现出旱生特点：导管频率和复孔率较高,导管分子短而窄,纤维很短,射线很低.扁担木叶片、次生木质部的解剖和水分生理特征均表现出一定的可塑性,其可塑性指数高低顺序为次生木质部解剖特征（0.24）＞水分生理特征（0.19）＞叶片解剖特征（0.18）.与侧柏林和混交林相比,灌丛中扁担木个体对干旱生境有一定的适应能力,表现在次生木质部导管分子短,导管频率高,单孔率低,木纤维短,射线矮小,具有较大的相对输导率和较小的脆性指数;叶片水势、组织含水量、自由水含量较低,叶面积、比叶面积较小,而束缚水含量、束缚水自由水比值较高.扁担木的解剖和生理可塑性,使之能忍受群落演替早期的干旱生境,更好地适应演替后期的中生环境,从而成为广布种和混交林中的优势种.     

In vitro differentiation of plantlets from embryonic explants of lodgepole pine (Pinus contorta Dougl. ex Loud.)

A protocol is described for plantlet formation in juvenile tissues of Pinus contorta. Shoots were induced on embryonic, cotyledonary and hypocotyl explants cultured on a defined medium supplemented with cytokinin. The concentration of salts, vitamins and cytokinin (benzylamino purine) in the medium, as well as different temperature regimes, strongly influenced the frequency of bud formation. Differentiation of shoot primordia and their subsequent development was also markedly affected by cytokinin exposure times. Bud development and elongation were enhanced by elimination of the phytohormone, reducing the strength of mineral salts, vitamins and sucrose in the medium, as well as by the inclusion of charcoal. Rooting was induced by treating the shoots with a sterilized rooting powder containing indole-butyric acid and culturing them in agar-solidified medium containing reduced mineral salts, vitamins, sucrose and charcoal. The number of chromosomes and their structure were found to be normal in the regenerated plantlets.     

The refilling of embolized xylem in Pinus sylvestris L.

Abstract. The hypothesis that ray parenchyma cells are actively involved in the refilling of embolized xylem of Pinus sylvestris L. was tested by killing the ray parenchyma and comparing rehydration of killed stems with that of control material. Killing of ray parenchyma was achieved using hot water or sodium azide. In most experiments, the available water for refilling was at negative water potential. Experiments were done on three kinds of plant material: small branch segments, potted seedlings and small potted trees. In all experiments, there was no indication that the azide-killed xylem was slower to refill than the control material and it was concluded that the parenchyma has no role in the refilling process, which therefore must be purely physical or physicochemical. Stems treated with hot water did not refill; we suggest that this may be caused by high temperatures decreasing the water permeability of the tracheid wall. The refilling of small branch segments may be explained by surface tension forces (capillary action), which inside the tracheid lumen may lower the water potential down to -9.7kPa; this may be enough to draw in water from the available water that in experiment one was at -2kPa. In the case of seedlings or saplings, capillary action cannot explain refilling, because the xylem water potentials were always lower than those estimated from tracheid radii. Condensation of water during diurnal cycles of warming and cooling is also unlikely to contribute to refilling significantly. To account for refilling in these cases, it is supposed that the tracheid wall may be chemically active and able to lower the water potential below the value expected by capillarity.     

The role of plant hormones in higher plant cellular differentiation. II. Experiments with the vascular cambium, and sclereid and tracheid differentiation in the pine, Pinus contorta

In sterile-cultured explants of stems of the pine Pinus contorta Dougl., fusiform cambial cells differentiated entirely into axial parenchyma cells when exogenous indol-3yl-acetic acid (IAA) was omitted. The normal appearance of the cambial zone was maintained when IAA was included in the medium. The IAA-maintained stability of cambial structure suggests physiological rather than epigenetic control over vascular cambium structure. IAA was essential for the occurrence of callus growth in stem explants. Callus growth was similar in appearance and extent in winter- and summer-explanted material. Tracheids differentiated in explants only when actively differentiating tracheids were already present at the moment of explanting, suggesting the absence of factors necessary for tracheid differentiation in over-wintering tissues. Sclereid differentiation, which normally does not occur in phloem or xylem development in P. contorta, occurred in callus derived from active cambial explants. The sclereids were identical to sclereids which differentiated in pith of intact stems. The possibility that sclereid and tracheid differentiation may be fundamentally similar types of gene expression is discussed. Growth of P. contorta trees in continuous darkness resulted in extensive compression-wood tracheid differentiation in the upright main stem. Normal-wood tracheids differentiated in similar trees grown in light. More tracheids differentiated in light than in darkness. This apparently is the first report of induction of compression-wood tracheid differentiation in the absence of hormone treatment or tilting of trees. Different types and numbers of tracheids differentiated at different position in two-year-old disbudded defoliated stem cuttings of P. contorta in response to apically supplied IAA. No evidence for new tracheid differentiation was seen in control cuttings; however, the results suggest that neither cambial cell division nor tracheid differentiation were actually initiated by IAA. Directed transport of additional regulatory factors toward areas of high IAA concentration is formulated as a hypothesis to explain these observations. Gibberellic acid, (S)-abscisic acid and IAA inhibited tracheid differentiation when individually supplied to basal ends of P. contorta cuttings predisposed to differentiate new tracheids. Experiments with single intact needles on Pinus cembroides var. monophylla cuttings confirmed a previous interpretation that the mature pine needle, rather than the short-shoot apical meristem at its base, promotes tracheid differentiation in the stem.