Structural aspects of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] tuberculate ectomycorrhizae

Summary Tubercles of Pseudotsuga menziesii consisted of clusters of ectomycorrhizae surrounded by a peridiumlike rind. Energy dispersive spectroscopy demonstrated that crystals found in the zone of loose hyphae extending from the inner rind to the mantle of each root probably contain calcium oxalate. Inner mantle and Hartig net hyphae showed a labyrinthine branching pattern and stored carbohydrates and protein. The Hartig net formed up to inner cortical cells which had thickened, darkly stained walls. Bacteria were located either along with hyphae within the rind or as colonies on the surface of the tubercle.     

Germination Rate of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] Seeds Affected by Their Orientation

Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] seeds developwith the adaxial surface continuous with the cone scale. Onseparation, the scale side of the seed is lighter coloured andflatter than the side not previously in contact with the scale.Seeds oriented scale-side-up on a moist surface germinate morerapidly than seeds placed scale-side-down. When seeds were germinatedunder laboratory conditions, the response to orientation variedamong seeds of different geographic origin. Most of the geographicvariation in germination rate occurred when seeds were orientedscale-side-up. Pseudotsuga menziesii (Mirb.) Franco, Douglas-fir germination, orientation of seeds     

Transfer conductance in second growth Douglas-fir (Pseudotsuga menziesii (Mirb.)Franco) canopies

The internal conductance from intercellular spaces to the sites of carboxylation (g_i) has only been measured in a few tree species and not in conifers, despite the fact it may impose a large limitation on photosynthesis. The present study provides the first estimates of g_i for a coniferous species, and examines variation in g_i with height and its relationships to anatomical, biochemical and physiological traits. Measurements were made on upper and lower canopy current‐year needles of 50‐year‐old Douglas‐fir (Pseudotsuga menziesii (Mirb.) Franco). Needle thickness and specific leaf area decreased by 30% from the top to bottom of the canopy. These anatomical/morphological changes were accompanied by modest variation in allocation of N to chlorophyll and the chlorophyll a/b ratio. Allocation of N to Rubisco did not vary with height, but the ratio of Rubisco to chlorophyll did owing to the aforementioned changes in allocation to chlorophyll. The value of g_i was estimated in one tree from concurrent measurements of carbon isotope discrimination and net photosynthesis. To examine the variation in g_i among trees a second independent method based on day respiration and the difference between the chloroplastic and intercellular photocompensation points (photocompensation point method) was used. Estimates of g_i obtained by the two methods agreed well with values varying between 0.14 and 0.20 mol m^?2 s^?1. It is estimated that g_i limits photosynthesis by approximately 20% as compared to an approximately 30% stomatal limitation (under well‐watered conditions). The value of g_i scaled approximately with maximum rates of photosynthesis, which were significantly greater in upper canopy needles. Nevertheless, g_i did not vary significantly with canopy height, owing to greater variability in g_i than photosynthesis.     

Mating system and multilocus associations in a natural population of Pseudotsuga menziesii (Mirb.) Franco

Summary Arrays of open-pollinated seeds were assayed for allozyme polymorphisms at ten loci (Aat2, Est1, G6pd, Idh, Mdh2, Mdh3, Pgm, Sod, 6Pgd1, 6Pgd2) to obtain estimates of the outcrossing rate and assess multilocus association in a natural population of coastal Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco. The allele frequencies in the samples of adult trees and pollen-gamete pool were similar. Maximum-likelihood estimators of the outcrossing rate for individual loci and two multilocus models were derived using counting methods. The single-locus maximum likelihood estimates (MLEs) of the outcrossing rate were significantly heterogeneous; they varied over a more than two-fold range from 0.404 to 0.935, with an average MLE of 0.741. Both multilocus MLEs of the outcrossing rate were 0.887. The sample of trees was in random mating equilibrium when assessed on a pairwise-locus basis using Burrows' composite measure of gametic disequilibrium, with one exception (Mdh2 Sod) that was attributable to a rare gametic class. In the sample of pollen gametes, 5 of the 45 pairwise-locus associations were nominally significant at the 0.05 level: Idh Est1, Mdh2 Sod, Aat2 Est1, Aat2 Mdh3, and Est1 Mdh3. These apparent associations were attributable in most cases to the relative excess of uncommon or rare paternal gametes of discernibly outcrossed embryos. An additional two-locus association was identified for Mdh2 Pgm which was marginally significant for the major partition of the contingency table that excluded paternal gametes with the rare allele Mdh2 ² .     

Aphidophagous insect communities in tree crowns of the neophyte Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] and Norway spruce (Picea abies L.)

Abstract:  Number of specimens and species as well as structure of aphidophagous communities in tree crowns of Douglas-fir ( Pseudotsuga menziesii ) and Norway spruce ( Picea abies ) were studied in different forest stand types (Douglas-fir-, spruce- and beech-dominated) in southern Germany in the year 2000, using flight-interception traps. Total as well as average numbers of specimens and species of aphid predators were higher on Douglas-fir in all stand types. This was particularly evident in Douglas-fir-dominated stand type. The aphidophagous community revealed a low β -diversity on spruce but a high β -diversity on Douglas-fir between stand types. Both species, with a broad and narrow ecological niche, respectively, were responsible for a community on Douglas-fir conspicuous different from that on spruce.     

Hydraulic control of stomatal conductance in Douglas fir [Pseudotsuga menziesii (Mirb.) Franco] and alder [Alnus rubra (Bong)] seedlings

Experiments were conducted on 1-year-old Douglas fir [Pseudotsuga menziesii (Mirb.) Franco] and 2- to 3-month-old alder [Alnus rubra (Bong)] seedlings growing in drying soils to determine the relative influence of root and leaf water status on stomatal conductance (g_c). The water status of shoots was manipulated independently of that of the roots using a pressure chamber that enclosed the root system. Pressurizing the chamber increases the turgor of cells in the shoot but not in the roots. Seedling shoots were enclosed in a whole-plant cuvette and transpiration and net photosynthesis rates measured continuously. In both species, stomatal closure in response to soil drying was progressively reversed with increasing pressurization. Responses occurred within minutes of pressurization and measurements almost immediately returned to pre-pressurization levels when the pressure was released. Even in wet soils there was a significant increase in g_c with pressurization. In Douglas fir, the stomatal response to pressurization was the same for seedlings grown in dry soils for up to 120 d as for those subjected to drought stress over 40 to 60 d. The stomatal conductance of both Douglas fir and alder seedlings was less sensitive to root chamber pressure at higher vapour pressure deficits (D), and stomatal closure in response to increasing D from 1.04 to 2.06 kPa was only partially reversed by pressurization. Our results are in contrast to those of other studies on herbaceous species, even though we followed the same experimental approach. They suggest that it is not always appropriate to invoke a ‘feedforward’ model of short-term stomatal response to soil drying, whereby chemical messengers from the roots bring about stomatal closure.     

Impact of common European tree species and Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) on the physicochemical properties of the rhizosphere

Trees play a crucial role in nutrient cycling and ecosystem fertility, notably through rhizosphere processes. The aim of this study was to compare soil physicochemical properties between bulk soil and rhizosphere of several tree species, and to compare rhizosphere properties between fertilized and non-fertilized conditions. The soil sampling was performed in Breuil-Chenue forest (North-East of France) in seven stands: native forest (old beech (Fagus sylvatica L.) and oak (Quercus sessiliflora Smith) coppice with standards; CwS), beech, oak (Quercus petraea [Matt.] Liebl.), Douglas-fir and fertilised Douglas-fir, Norway spruce (Picea abies Karst.) and fertilised Norway spruce. Systematic soil sampling was performed at 0–3, 3–10, and 10–23 cm in 20 calibrated pits. The rhizosphere of the different species was generally enriched in C, N, Ca, Mg, and K. Interestingly, the same positive effect was observed in the fertilised plots. The rhizosphere effect varied between tree species for C, “base” cations, pH_water and cation exchange capacity. This study reveals that interactions between roots, microorganisms and soil can enrich the pool of nutrients in the rhizosphere compared to bulk soil whatever the soil fertility conditions, and that the magnitude of the rhizosphere effect depends on tree species.     

Nuclear bodies in Douglas fir (Pseudotsuga menziesii Mirb.) microspores

The identification of nucleolar proteins and immunocytochemical localization of small nuclear ribonucleoprotein (snRNP) elements revealed the presence of three types of nuclear bodies in Douglas fir microspore nuclei. One type consists of structures resembling Cajal bodies (CBs) and contains nucleolar proteins as well as snRNPs and U2 snRNA. The second type is bizonal bodies, which are nuclear bodies also linked with the splicing system. The bizonal body comprises two parts: the first contains Sm proteins and stains strongly with silver stain, and the second resembles CBs in terms of the degree of silver staining and molecular composition. Douglas fir is the second species after larch where the presence of bizonal bodies has been demonstrated. Pseudotsuga menziesii Mirb and Larix decidua Mill are species with one of the longest microsporogenesis processes known in plants. The presence of bizonal bodies in both species may be linked to the intensification of the splicing processes in microspores with an exceptionally long cell cycle. The third type of structure is dense bodies, whose morphology and degree of silver staining strongly indicate their functional and spatial relationship to the dense part of bizonal bodies.     

Wood density proxies of adaptive traits linked with resistance to drought in Douglas fir (Pseudotsuga menziesii (Mirb.) Franco)

Key message

Proxies of adaptive traits for resistance to drought were discovered among original annual ring density variables in Douglas fir.

Abstract

A comparison of dead and surviving Douglas fir trees following the 2003 drought was made to define proxies of adaptive traits for resistance to drought. Increment cores were sampled from trees from three French regions: Centre, Midi-Pyrénées and Burgundy. Original tree-ring variables were calculated, based on a sliding density criterion dividing the microdensity profile into high- and low-density segments. Tree rings were analysed at each site in a number of consecutive annual rings before the 2003 drought event. Comparison between pairs of surviving and dead trees and between pairs of randomly selected trees (whether dead or alive) supports the evidence of systematic dissimilarities between surviving and dead trees in a number of original density variables. Correlation analysis between original and conventional ring density variables indicates a weak association. We found that the surviving trees were denser than the dead trees in all three sites, but that the denser part of the ring varied from region to region. We identified several original density variables intended to be used as proxies of adaptive traits in future studies of genetic determinism of Douglas fir resistance to drought.     

Xeromorphy increases in shoots of Pseudotsuga menziesii (Mirb.) Franco seedlings with exposure to elevated temperature but not elevated CO2

Seedling structure influences tree structure and function, ultimately determining the potential productivity of trees and their competitiveness for resources. We investigated changes in shoot structure for seedlings of Pseudotsuga menziesii (Douglas-fir) grown under climate change scenarios of ambient or elevated CO₂ (+180 mol mol^–1) plus ambient or elevated temperature (+3.5°C), for 4 years in outdoor, sunlit chambers. Mass allocation and allometry were measured for buds, leaves, branches, and stems, and anatomy was evaluated for leaves and stems. Seedlings became more xeromorphic with elevated temperature: allocation of total mass to branches over stems and leaves increased, sapwood area to height ratio increased, number of growing points relative to seedling size increased, and stem and branch length and mass decreased for sections initiated during the three full CO₂ and temperature seasons. Neither stem nor leaf anatomy was affected by elevated temperature. Elevated CO₂ increased specific mass of leaves, but had few other effects on mass allocation, allometry, or anatomy for any shoot organ. There were no CO₂ × temperature interactions for any important parameter. Thus, under realistic simulated field environmental conditions representative for in at least some P. menziesii forests (i.e., OR, USA, forests with limited soil nitrogen and summer soil moisture), elevated temperature, but not elevated CO₂, may affect seedling shoot structure and, hence, function.     

Morphotype community structure of ectomycorrhizas on Douglas fir (Pseudotsuga menziesii Mirb. Franco) seedlings grown under elevated atmospheric CO2 and temperature

Mycorrhizas alter the acquisition of carbon and nutrients, thereby affecting numerous plant and ecosystem processes. It is important, therefore, to determine how mycorrhizal populations will change under possible future climate conditions. Individual and interactive effects of elevated atmospheric CO₂ concentration and atmospheric temperature were assessed in a 2×2 factorial design [ambient and elevated (200 ppm above ambient) CO₂ concentrations, and ambient and elevated (4°C above ambient) temperatures]. In June 1993, 2-year-old Douglas fir (Pseudotsuga menziesii Mirb. Franco) seedlings were planted in 12 environment-tracking chambers (n=3) containing reconstructed, low-nitrogen, native forest soil. Climate treatments were imposed shortly thereafter, and the seedlings grew until June 1997. Soil cores were taken twice per year during the exposure period. We present findings on changes in the community structure of ectomycorrhizal (ECM) root tips, categorized into morphotypes using gross morphological traits. A diverse and stable community of morphotypes (a total of 40) was encountered; no more than 30 of which were seen at any sampling time. In the first sample, there were only 15 morphotypes found in the 12 chambers. Morphotype numbers increased during the first half of the experiment, remaining fairly constant thereafter. Near the end of the exposure, elevated-temperature treatments maintained more morphotypes than ambient treatments. However, overall, absolute measures (number of ECM tips) were affected primarily by CO₂ treatment, whereas proportional measures (e.g., Simpson’s index) were affected primarily by temperature. While some morphotypes were negatively affected seasonally by higher temperatures (putative Rhizopogon group), others (Cenococcum) seemed to thrive. Underlying the dominant patterns of change in diversity, driven by the Rhizopogon group, subdominant populations responded slightly differently. Community diversity through time tended to increase at a greater rate for all subdominant populations compared with the rate when dominant populations were included. Received: 16 August 1999 / Accepted: 2 March 2000     

Ethanol synthesis, nitrogen, carbohydrates, and growth in tissues from nitrogen fertilized Pseudotsuga menziesii (Mirb.) Franco and Pinus ponderosa Dougl. ex Laws. seedlings

 Seedlings of Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, and ponderosa pine, Pinus ponderosa Dougl. ex Laws., were grown in a controlled environment and fertilized with nutrient solutions containing 150 ppm (+N), or 0 ppm nitrogen (−N). These treatments greatly altered seedling growth, and the concentrations of N and carbohydrates in their tissues. Metabolically active tissues, such as roots, incubated with a limited supply of O₂ became hypoxic faster and synthesized more ethanol than less active tissues, such as needles. All tissues that were incubated for 4 h in N₂ synthesized ethanol. Needles incubated in N₂ and light had much lower quantities of ethanol than needles in N₂ and dark, suggesting that O₂ from photosynthetsis limited internal anoxia. Most tissues from +N seedlings synthesized greater quantities of ethanol in N₂ anoxia than tissues from −N seedlings, probably because they were able to produce more enzymes with a greater availability of N. However, this increase in ethanol synthesis between N treatments was most pronounced in the phloem. Ethanol and soluble sugar concentrations were negatively related in needles and positively related in roots of N+ seedlings, but not −N seedlings. Starch concentrations had no effect on the amount of ethanol produced by any tissue. Regardless of N treatments, all tissues from ponderosa pine produced more N₂-induced ethanol than Douglas-fir, in part because its tissues contained different concentrations of soluble sugars and N as a consequence of phenological differences between the species. However, ponderosa pine tissues may also maintain greater quantities of anaerobic enzymes, or their isozymes than Douglas-fir. Received: 22 February 1998 / Accepted 23 June 1998     

Chloroplast DNA restriction fragment length polymorphism in Sequoia sempervirens D. Don Endl., Pseudotsuga menziesii (Mirb.) Franco,Calocedrus decurrens (Torr.), and Pinus taeda L.

Summary The extent and type of chloroplast DNA restriction fragment length polymorphism was determined among individual tree samples of coast redwood, Douglas fir, incense-cedar, and loblolly pine. A total of 107 trees was surveyed for three restriction enzymes (BamHI, EcoRI, HindIII) and six chloroplast DNA probes from petunia (P3, P4, P6, P8, P10, S8). The probes comprise 64% of the petunia chloroplast genome. Polymorphisms were detected in all species but loblolly pine. Coast redwood and incense-cedar had a small number of rare variants, whereas Douglas fir had one highly polymorphic region of insertions/deletions in sequences revealed by the P6 probe from petunia. The mutation hotspot is currently being studied by DNA sequence analysis.     

The effect of stratification on the endogenous levels of gibberellins and cytokinins in seeds of douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and sugar pine (Pinus lambertiana Dougl.)

Abstract Seeds of Douglas-fir and sugar pine which had been exposed to various periods of moist chilling (stratification) were extracted and bioassayed for gibberellin and cytokinin activity. In Douglas-fir a peak of gibberellin activity increased progressively during stratification and was seven times higher after 7 weeks, but declined subsequently. No change in the level of cytokinin activity was observed during stratification. However, a slight increase (1.7-fold) occurred in stratified seeds placed at 22°C for 3 days. In sugar pine a high level of gibberellin activity, approximately 20-fold higher than in unstratified controls, was present in extracts of seeds which had been stratified for 8 weeks. After 10 weeks of stratification cytokinin levels had increased approximately 200-fold. Four cytokinin-like substances were detected in seeds. Three of the substances are similar in their chromatographic properties to zeatin glucoside, zeatin riboside and zeatin, respectively. Both gibberellin and cytokinin activity declined rapidly after reaching maximum levels. In seeds with the coats removed, germination was rapid and complete in the absence of stratification. Prior to radicle emergence in coatless seeds cytokinin activity increased to a maximum (approximately a 1-7-fold increase) within 24 h. Significant levels of gibberellin-like substances were present after 8 h. Gibberellin activity could not be detected in coatless seeds which were imbibed in AMO-1618, and germination was inhibited significantly.     

Assessing population genetic structure of sorghum landraces from North-western Morocco using allozyme and microsatellite markers

 The level of genetic diversity and the population genetic structure of sorghum landraces from North-western Morocco have been investigated based on direct field-sampling using both allozyme and microsatellite markers. As expected, microsatellite markers showed a much higher degree of polymorphism than allozymes, but relative measures of genetic structure such as Wright’s inbreeding coefficient F _IS and Nei’s coefficient of genetic differentiation G _ST were similar for the two sets of markers. Substantial inbreeding was found to occur within fields, which confirms that sorghum is predominantly selfing under cultivation. Most of the genetic diversity in Moroccan landraces occurs within fields (more than 85%), as opposed to among fields or among regions, a result which contrasts to those of studies based on accessions from germplasm collections. It is suggested that individual fields of sorghum constitute valuable units of conservation in the context of in situ conservation practices. Received: 8 December 1998 / Accepted: 28 December 1998