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PubMed2002

Leaf stomatal and epidermal cell development: identification of putative quantitative trait loci in relation to elevated carbon dioxide concentration in poplar.

Genetic variation in stomatal initiation and density, and epidermal cell size and number were examined in a hybrid pedigree of Populus trichocarpa T. & G. and P. deltoides Marsh in both ambient ([aCO2]) and elevated ([eCO2]) concentrations of CO2. We aimed to link anatomical traits with the underlying genetic map of F2 Family 331, composed of 350 markers across 19 linkage groups. Leaf stomatal and epidermal cell traits showed pronounced differences between the original parents. We considered the following traits in the F2 population: stomatal density (SD), stomatal index (SI), epidermal cell area (ECA) and the number of epidermal cells per leaf (ECN). In [eCO2], adaxial SD and SI were reduced in the F2 population, whereas ECA increased and ECN remained unchanged. In [aCO2], four putative quantitative trait loci (QTL) with logarithm of the odds ratio (LOD) scores greater than 2.9 were found for stomatal traits on linkage group B: adaxial SI (LOD scores of 5.4 and 5.2); abaxial SI (LOD score of 3.3); and SD (LOD score of 3.2). These results imply that QTL for SI and SD share linkage group B and are under genetic control. More moderate LOD scores (LOD scores >/= 2.5) suggest QTL for SI on linkage groups A and B and for SD on linkage groups B, D and X with a probable co-locating quantitative trait locus for SI and SD on linkage group D (position 46.3 cM). The QTL in both [aCO2] and [eCO2] for adaxial SD were co-located on linkage group X (LOD scores of 3.5 and 2.6, respectively) indicating a similar response across both treatments. Putative QTL were located on linkage group A (position 89.2 cM) for both leaf size and ECN in [aCO2] and for ECA at almost the same position. The data provide preliminary evidence that leaf stomatal and cell traits are amenable to QTL analysis.

Carbon Dioxide↗

PubMed2003

Growth response of cottonwood roots to varied NH4:NO3 ratios in enriched patches.

Maximization of short-rotation forest plantation yield requires frequent applications of nutrients, especially nitrogen (N). Whole-plant growth is known to be sensitive to the proportion of ammonium to nitrate (NH4:NO3). However, the extent to which N form affects root growth, branching and morphology is poorly understood, and these variables may have substantial impacts on plant nutrient and water acquisition. We used rooted cuttings of cottonwood (Populus deltoides Bartr. ex Marsh.) to investigate the effect of various NH4:NO3 ratios on root growth in N-enriched patches. A sand culture study with split-root systems was carried out in which 1-3% of the total root system of each cutting was supplied with 2 mM N at NH4:NO3 ratios of 0:100, 20:80, 40:60, 60:40, 80:20 and 100:0 (molar basis), with the rest of the plant supplied with 0 mM N, resulting in the whole plant becoming N deficient. During the experiment, whole-plant growth was unaffected by the treatments. Of the NH4:NO3 ratios tested, greatest total root length, specific root length, and root N concentration of roots in enriched patches occurred in the 20:80 NH4:NO3 treatment. The largest response of roots in enriched patches was third- and fourth-order root production. We conclude that N form has a profound effect on root development and morphology in enriched patches.

Biomass↗

PubMed2004

Phytohormones and shoot growth in a three-generation hybrid poplar family.

Hybrid vigor for secondary growth in poplar has been linked with increased gibberellin (GA) concentration in cambial tissue, but the relationship between concentrations of GAs and hybrid vigor of primary growth has not been investigated. We explored associations between concentrations of GAs, abscisic acid (ABA) and indoleacetic acid (IAA) and shoot extension in a hybrid family of Populus deltoides Bartr. ex Marsh. (Clone ILL-129) and P. trichocarpa Torr. & Gray (Clone 93-968) and two F1 and 67 F2 genotypes. Rapidly elongating subapical internodes from shoots of 4-year-old clonal saplings were selected for hormone analysis. The F1 hybrids displayed heterosis (hybrid vigor) for primary growth as a result of the complementation of dominance for increased internode length from the P. trichocarpa parent and dominance for increased diameter from the P. deltoides parent. Internodes from the faster-elongating shoots of the P. trichocarpa parent had a fourfold higher concentration of bioactive GA1 and higher concentrations of GA20, GA44, GA29 and GA8 than the P. deltoides parent. However, the two fast-growing F1 hybrids had low concentrations of all five GAs, with concentrations similar to those of the slower-elongating P. deltoides parent. Concentrations of ABA and IAA were correlated with GA concentrations and there was thus no evidence of a consistent promoting or inhibiting effect of ABA or IAA within the F1 family. These results indicate that heterosis for internode growth was not primarily regulated by hormone concentration. The segregating population of F2 hybrids was analyzed to assess the possible utility of hormone screening as a selection tool. The internodes of most of the F2 hybrids were smaller than those of their F1 parents and the larger P. trichocarpa parent. Among the F2 hybrids, mature internode length varied fourfold, and was correlated with lengths of young, subapical internodes from which hormones were analyzed. In these internodes, GA1 concentrations were negatively correlated with length (r = 0.41), diameter (0.33) and mass (0.50). Indoleacetic acid concentration was negatively correlated only with diameter (r = 0.37) and positively correlated with GA1 (r = 0.46), whereas ABA concentration was not correlated with any parameter. Thus, in the F2 population, variation in hormone concentration was not primarily responsible for the variation in shoot growth, indicating that selection for individuals with high GA concentration may not be an effective strategy for identifying vigorous hybrid genotypes.

Abscisic Acid↗

PubMed2004

Component and whole-system respiration fluxes in northern deciduous forests.

We measured component and whole-system respiration fluxes in northern hardwood (Acer saccharum Marsh., Tilia americana L., Fraxinus pennsylvanica Marsh.) and aspen (Populus tremuloides Michx.) forest stands in Price County, northern Wisconsin from 1999 through 2002. Measurements of soil, leaf and stem respiration, stem biomass, leaf area and biomass, and vertical profiles of leaf area were combined with biometric measurements to create site-specific respiration models and to estimate component and whole-system respiration fluxes. Hourly estimates of component respiration were based on site measurements of air, soil and stem temperature, leaf mass, sapwood volume and species composition. We also measured whole-system respiration from an above-canopy eddy flux tower. Measured soil respiration rates varied significantly among sites, but not consistently among dominant species (P < 0.05 and P > 0.1). Annual soil respiration ranged from 8.09 to 11.94 Mg C ha(-1) year(-1). Soil respiration varied linearly with temperature (P < 0.05), but not with soil water content (P > 0.1). Stem respiration rates per unit volume and per unit area differed significantly among species (P < 0.05). Stem respiration per unit volume of sapwood was highest in F. pennsylvanica (up to 300 micro mol m(3) s(-1)) and lowest in T. americana (22 micro mol m(3) s(-1)) when measured at peak summer temperatures (27 to 29 degrees C). In northern hardwood stands, south-side stem temperatures were higher and more variable than north-side temperatures during leaf-off periods, but were not different statistically during leaf-on periods. Cumulative annual stem respiration varied by year and species (P < 0.05) and averaged 1.59 Mg C ha(-1) year(-1). Leaf respiration rates varied significantly among species (P < 0.05). Respiration rates per unit leaf mass measured at 30 degrees C were highest for P. tremuloides (38.8 nmol g(-1) s(-1)), lowest for Ulmus rubra Muhlenb. (13.1 nmol g(-1) s(-1)) and intermediate and similar (30.2 nmol g(-1) s(-1)) for T. americana, F. pennsylvanica and Q. rubra. During the growing season, component respiration estimates were dominated by soil respiration, followed by leaf and then stem respiration. Summed component respiration averaged 11.86 Mg C ha(-1) year(-1). We found strong covariance between whole-ecosystem and summed component respiration measurements, but absolute rates and annual sums differed greatly.

Acer↗

PubMed2004

Physiological responses of wild type and putrescine-overproducing transgenic cells of poplar to variations in the form and concentration of nitrogen in the medium.

We determined: (a) the physiological consequences of overproduction of putrescine in transgenic poplar (Populus nigra x maximoviczii) cells expressing an ornithine decarboxylase transgene; and (b) effects of variation in nitrogen (N) concentration of the medium on cellular polyamine concentration in transgenic and non-transgenic cells. Cells grown in the presence of supplemental (to the normal concentrations of N sources in the growth medium) and reduced amounts of NH4NO3 and KNO3 were used to study effects on membrane permeability, mitochondrial respiratory activity, protein accumulation, growth rates and changes in cellular polyamine concentration. The N concentration of the MS medium was not a limiting factor for continued overproduction of putrescine in transgenic cells. However, continued supplies of NH4+ and NO3- were required to maintain homeostatic amounts of putrescine in both cell lines. The presence of high amounts of putrescine in transgenic cells had significant effects on the physiological parameters measured. Compared with non-transgenic cells, transgenic cells had greater plasma membrane permeability, less tolerance to NH4NO3, more tolerance to KNO3, and accumulated higher amounts of soluble protein.

Cells, Cultured↗

PubMed2004

Transgenic poplar characterized by ectopic expression of a pine cytosolic glutamine synthetase gene exhibits enhanced tolerance to water stress.

Physiological responses to water stress in hybrid poplar (INRA 7171-B4, Populus tremula L. x P. alba L.) lines transformed to overexpress a pine cytosolic glutamine synthetase (GS1) gene were compared with those of non-transgenic plants. Before, during and after a drought treatment, net photosynthetic rates (Anet) were higher in transgenic than in non-transgenic plants. Stomatal conductance (gs) was higher in transgenic than in non-transgenic plants before, but not after exposure to drought. Before drought treatment, a sudden reduction in photosynthetic photon flux caused a greater burst of CO2 efflux in transgenic than non-transgenic plants, indicating greater photorespiratory activity. Drought caused greater reductions in photochemical quenching, photosystem II (PSII) antennae transfer efficiency (Fv'/Fm') and light-adapted PSII yield (PhiPSII) in non-transgenic than in transgenic plants, especially at low irradiances. Antennae-based thermal dissipation was higher in transgenic plants than in non-transgenic plants both during the imposition of drought and 1 or 3 days after the relief of drought. Under severe water stress and subsequently, transgenic plants maintained a higher expression of glutamine synthetase, glutamate synthase and Rubisco and higher concentrations of chlorophyll and glycine than non-transgenic plants. These findings indicate that overexpression of pine cytosolic GS1 enhanced sustained photosynthetic electron transport capacity during severe stomatal limitation. The data also suggest that ectopic expression of cytosolic GS increases photorespiratory activity, and that this serves as a protective sink for electrons from photosynthetic reaction centers.

Chlorophyll↗

PubMed2004

Cottonwood growth rate and fine root condensed tannin concentration.

We examined the relationship between trunk diameter and diameter relative growth rate (RGR) and fine root condensed tannin concentration in 12 genotypes of eastern cottonwood (Populus deltoides Bartr. ex Marsh.) planted in three locations across the north central United States. Across genotypes, trunk diameter, diameter RGR and root condensed tannin concentration were negatively correlated at one location (Wisconsin), but showed no significant correlation at the other locations (Iowa and Michigan). The factors responsible for this difference among sites remain unidentified, but may be related to soil fertility.

Nitrogen↗

PubMed2005

Environmental controls on sap flow in a northern hardwood forest.

Our objective was to gain a detailed understanding of how photosynthetically active radiation (PAR), vapor pressure deficit (D) and soil water interact to control transpiration in the dominant canopy species of a mixed hardwood forest in northern Lower Michigan. An improved understanding of how these environmental factors affect whole-tree water use in unmanaged ecosystems is necessary in assessing the consequences of climate change on the terrestrial water cycle. We used continuously heated sap flow sensors to measure transpiration in mature trees of four species during two successive drought events. The measurements were scaled to the stand level for comparison with eddy covariance estimates of ecosystem water flux (Fw). Photosynthetically active radiation and D together explained 82% of the daytime hourly variation in plot-level transpiration, and low soil water content generally resulted in increased stomatal sensitivity to increasing D. There were also species-specific responses to drought. Quercus rubra L. showed low water use during both dry and wet conditions, and during periods of high D. Among the study species, Acer rubrum L. showed the greatest degree of stomatal closure in response to low soil water availability. Moderate increases in stomatal sensitivity to D during dry periods were observed in Populus grandidentata Michx. and Betula papyrifera Marsh. Sap flow scaled to the plot level and Fw demonstrated similar temporal patterns of water loss suggesting that the mechanisms controlling sap flow of an individual tree also control ecosystem evapotranspiration. However, the absolute magnitude of scaled sap flow estimates was consistently lower than Fw. We conclude that species-specific responses to PAR, D and soil water content are key elements to understanding current and future water fluxes in this ecosystem.

Acer↗

PubMed2005

Mechanical perturbation affects conductivity, mechanical properties and aboveground biomass of hybrid poplars.

Xylem development in trees is affected by dynamic mechanical stresses imposed on stems by wind. To assess clonal differences in response to mechanical perturbation (MP), we subjected seven greenhouse-grown F1 hybrids of Populus trichocarpa Torr. and A. Gray. x P. deltoides Bartr. ex Marsh. to a standard MP treatment consisting of 20 manually imposed stem flexures per day for 70-90 days. Effects of MP on aboveground biomass, hydraulic conductivity (k(h)), specific conductivity (k(s)), flexural stiffness (EI), modulus of elasticity (MOE) and modulus of rupture (MOR) were determined. Treatment increased stem radial growth and decreased height growth, leaf area and total aboveground biomass. It also significantly decreased k(s), MOE and MOR, but significantly increased EI and wood specific gravity in most clones. Mechanical perturbation caused greater stem rigidity, without having a significant effect on whole-stem k(h) or percent loss of conductivity due to embolism. Maximum k(h) was positively correlated with EI in both control (r(2) = 0.54, P < 0.0001) and MP-treated (r(2) = 0.61, P < 0.0001) plants, and k(s) and MOE were positively correlated with percent vessel lumen area (r(2) = 0.45, P < 0.0001 and r(2) = 0.28, P = 0.002, respectively). Thus, contrary to our expectation of a trade-off between conductivity and wood strength, there may be an opportunity to select clones for woody biomass production that are superior in both mechanical strength and hydraulic conductivity, as is the triploid Clone 19-61.

Biomass↗

PubMed2005

Net carbon storage in a poplar plantation (POPFACE) after three years of free-air CO2 enrichment.

A high-density plantation of three genotypes of Populus was exposed to an elevated concentration of carbon dioxide ([CO(2)]; 550 micromol mol(-1)) from planting through canopy closure using a free-air CO(2) enrichment (FACE) technique. The FACE treatment stimulated gross primary productivity by 22 and 11% in the second and third years, respectively. Partitioning of extra carbon (C) among C pools of different turnover rates is of critical interest; thus, we calculated net ecosystem productivity (NEP) to determine whether elevated atmospheric [CO(2)] will enhance net plantation C storage capacity. Free-air CO(2) enrichment increased net primary productivity (NPP) of all genotypes by 21% in the second year and by 26% in the third year, mainly because of an increase in the size of C pools with relatively slow turnover rates (i.e., wood). In all genotypes in the FACE treatment, more new soil C was added to the total soil C pool compared with the control treatment. However, more old soil C loss was observed in the FACE treatment compared with the control treatment, possibly due to a priming effect from newly incorporated root litter. FACE did not significantly increase NEP, probably as a result of this priming effect.

Biomass↗

PubMed2005

Functional genomics analysis of foliar condensed tannin and phenolic glycoside regulation in natural cottonwood hybrids.

Regulation of leaf condensed tannins (CT) and salicylate-derived phenolic glycosides (PG) in fast- and slow-growing cottonwood backcrosses was analyzed by metabolic profiling and cDNA microarray hybridization. Seven hybrid lines of Populus fremontii L. and P. angustifolia James exhibiting growth/CT-PG phenotypes ranging from fast/low (Lines 18 and 1979) to slow/high (Lines 1012 and RL2) and intermediate (Lines NUL, 3200 and RM5) were investigated. Methanol-extractable leaf metabolites were analyzed by gas chromatography-mass spectrometry, and the results evaluated by principal component analysis. The hybrid lines formed separate clusters based on their primary metabolite profiles, with cluster arrangement also reflecting differences in CT-PG phenotype. Nitrogen (N) supply was manipulated to alter CT-PG partitioning and to obtain molecular insights into how primary metabolism interfaces with CT-PG accumulation. Three backcross lines (RM5, 1012, 18) exhibiting differential CT-PG responses to a 10-day hydroponic N-deprivation treatment were chosen for metabolite and gene expression analyses. The fast- growing Line 18 showed a minimal CT-PG response to N deprivation, and a reduction in photosynthetic gene expression. Line 1012 exhibited a strong phenylpropanoid response to N deprivation, including a doubling in phenylalanine ammonia-lyase (PAL) gene expression, and a shift from CT accumulation in the absence of stress toward PG accumulation under N-deprivation conditions. Amino acid concentrations were depressed in Lines 18 and 1012, as was expression of nitrate-sensitive genes coding for transketolase (TK), and malate dehydrogenase (MDH). Genes associated with protein synthesis and fate were down-regulated in Line 1012 but not in Line 18. Line RM5 exhibited a comparatively large increase in CT in response to N deprivation, but did not sustain decreases in amino acid concentrations, or changes in PAL, TK or MDH gene expression. Molecular characterization of the variable CT-PG responses shows promise for the identification and future testing of candidate genes for CT-PG trait selection or manipulation.

Gas Chromatography-Mass Spectrometry↗

PubMed2005

Production physiology of three fast-growing hardwood species along a soil resource gradient.

We determined how specific leaf area (SLA), specific leaf nitrogen (SLN), leaf area index (LAI), light-saturated photosynthesis (Amax) and aboveground net primary productivity (ANPP) of three commercially important hardwood species, eastern cottonwood (Populus deltoides Bartr.), American sycamore (Platanus occidentalis L.) and cherrybark oak (Quercus falcata var.pagodafolia Ell.), vary across a soil resource gradient. Five treatments were applied in a randomized block design (control, irrigation only (IRR), and irrigation plus fertilization with 56, 112 or 224 kg N ha-1 year-1 (N56, N112 and N224)) with four replications per species. When trees were 6 years old, Amax, SLA, SLN, LAI and ANPP were quantified during peak leaf production within a single growing season. In all species, Amax for sun leaves was significantly higher than for shade leaves (34, 32 and 29 micromol m2 s-1 versus 27, 23 and 23 micromol m2 s-1 for cottonwood, cherrybark oak and sycamore sun and shade leaves, respectively) and tended to plateau in the N112 treatment. The SLA was significantly lower in sun than in shade leaves and reached a plateau in IRR-treated cottonwood and sycamore, and in N56-treated oak. Values of SLN peaked in the N122 treatment for cottonwood sun leaves (1.73 g N m2) and in the N56 treatment for sycamore and oak (1.54 and 1.90 g N m2, respectively). In sun and shade leaves of all species, Amax increased with increasing SLN. Cherrybark oak LAI reached a plateau across the resource gradient in the N56 treatment, whereas cottonwood and sycamore LAI reached a plateau in the IRR treatment. All species exhibited significant curvilinear relationships between canopy Amax and ANPP. These findings indicate that nutrients and water regulate leaf-level traits such as SLA and SLN, which in turn influence LAI and canopy photosynthesis, thereby affecting ANPP at the tree and stand levels.

Biomass↗

PubMed2005

Effects of elevated concentrations of atmospheric CO2 and tropospheric O3 on decomposition of fine roots.

Rising atmospheric carbon dioxide (CO2) concentration ([CO2]) could alter terrestrial carbon (C) cycling by affecting plant growth, litter chemistry and decomposition. How the concurrent increase in tropospheric ozone (O3) concentration ([O3]) will interact with rising atmospheric [CO2] to affect C cycling is unknown. A major component of carbon cycling in forests is fine root production, mortality and decomposition. To better understand the effects of elevated [CO2] and [O3] on the dynamics of fine root C, we conducted a combined field and laboratory incubation experiment to monitor decomposition dynamics and changes in fine root litter chemistry. Free-air CO2 enrichment (FACE) technology at the FACTS-II Aspen FACE project in Rhinelander, Wisconsin, elevated [CO2] (535 microl 1-1) and [O3] (53 nl 1-1) in intact stands of pure trembling aspen (Populus tremuloides Michx.) and in mixed stands of trembling aspen plus paper birch (Betula papyrifera Marsh.) and trembling aspen plus sugar maple (Acer saccharum Marsh.). We hypothesized that the trees would react to increased C availability (elevated [CO2]) by increasing allocation to C-based secondary compounds (CBSCs), thereby decreasing rates of decomposition. Because of its lower growth potential, we reasoned this effect would be greatest in the aspen-maple community relative to the aspen and aspen-birch communities. As a result of decreased C availability, we expected elevated [O3] to counteract shifts in C allocation induced by elevated [CO2]. Concentrations of CBSCs were rarely significantly affected by the CO2 and O3 treatments in decomposing fine roots. Rates of microbial respiration and mass loss from fine roots were unaffected by the treatments, although the production of dissolved organic C differed among communities. We conclude that elevated [CO2] and [O3] induce only small changes in fine root chemistry that are insufficient to significantly influence fine root decomposition. If changes in soil C cycling occur in the future, they will most likely be brought about by changes in litter production.

Acer↗

PubMed2005

Effects of elevated concentrations of atmospheric CO2 and tropospheric O3 on leaf litter production and chemistry in trembling aspen and paper birch communities.

Human activities are increasing the concentrations of atmospheric carbon dioxide ([CO2]) and tropospheric ozone ([O3]), potentially leading to changes in the quantity and chemical quality of leaf litter inputs to forest soils. Because the quality and quantity of labile and recalcitrant carbon (C) compounds influence forest productivity through changes in soil organic matter content, characterizing changes in leaf litter in response to environmental change is critical to understanding the effects of global change on forests. We assessed the independent and combined effects of elevated [CO2] and elevated [O3] on foliar litter production and chemistry in aspen (Populus tremuloides Michx.) and birch-(Betula papyrifera Marsh.) aspen communities at the Aspen free-air CO2 enrichment (FACE) experiment in Rhinelander, WI. Litter was analyzed for concentrations of C, nitrogen (N), soluble sugars, lipids, lignin, cellulose, hemicellulose and C-based defensive compounds (soluble phenolics and condensed tannins). Concentrations of these chemical compounds in naturally senesced litter were similar in aspen and birch-aspen communities among treatments, except for N, the C:N ratio and lipids. Elevated [CO2] significantly increased C:N (+8.7%), lowered mean litter N concentration (-10.7%) but had no effect on the concentrations of soluble sugars, soluble phenolics and condensed tannins. Elevated [CO2] significantly increased litter biomass production (+33.3%), resulting in significant increases in fluxes of N, soluble sugars, soluble phenolics and condensed tannins to the soil. Elevated [O3] significantly increased litter concentrations of soluble sugars (+78.1%), soluble phenolics (+53.1%) and condensed tannins (+77.2%). There were no significant effects of elevated [CO2] or elevated [O3] on the concentrations of individual C structural carbohydrates (cellulose, hemicellulose and lignin). Elevated [CO2] significantly increased cellulose (+37.4%) input to soil, whereas elevated [O3] significantly reduced hemicellulose and lignin inputs to soil (-22.3 and -31.5%, respectively). The small changes in litter chemistry in response to elevated [CO2] and tropospheric [O3] that we observed, combined with changes in litter biomass production, could significantly alter the inputs of N, soluble sugars, condensed tannins, soluble phenolics, cellulose and lignin to forest soils in the future.

Betula↗

PubMed2005

Elevated CO2 concentration, fertilization and their interaction: growth stimulation in a short-rotation poplar coppice (EUROFACE).

We investigated the individual and combined effects of elevated CO2 concentration and fertilization on aboveground growth of three poplar species (Populus alba L. Clone 2AS-11, P. nigra L. Clone Jean Pourtet and P. x euramericana Clone I-214) growing in a short-rotation coppice culture for two growing seasons after coppicing. Free-air carbon dioxide enrichment (FACE) stimulated the number of shoots per stool, leaf area index measured with a fish-eye-type plant canopy analyzer (LAIoptical), and annual leaf production, but did not affect dominant shoot height or canopy productivity index. Comparison of LAIoptical with LAI estimates from litter collections and from allometric relationships showed considerable differences. The increase in biomass in response to FACE was caused by an initial stimulation of absolute and relative growth rates, which disappeared after the first growing season following coppicing. An ontogenetic decline in growth in the FACE treatment, together with strong competition inside the dense plantation, may have caused this decrease. Fertilization did not influence aboveground growth, although some FACE responses were more pronounced in fertilized trees. A species effect was observed for most parameters.

Biomass↗

PubMed2005

Visualizing water-conduction pathways of living trees: selection of dyes and tissue preparation methods.

To visualize water-conduction pathways in living trees, we introduced aqueous solutions of safranin and acid fuchsin into stems of Populus sieboldii Miquel. To examine the spread of each dye in the trees, we compared several techniques for preparing tissue for light microscopy. Acid fuchsin was distributed more rapidly and more widely than safranin, reflecting differences between the dye molecules in state of ionization. We prepared some sections without allowing the dye to redissolve after it had been stabilized by freeze-drying. In these sections, the dye was observed in vessels and in some of the adjacent ray parenchyma cells. Other sections were prepared without stabilizing the dye. In these sections, acid fuchsin in the sap stream left cell walls unstained, whereas safranin stained wood fibers in the vicinity of vessels, as well as the vessels themselves, provided that the sections were mounted in glycerin, which dissolves safranin. Although stained with safranin, the wood fibers contained no water. The results indicate that stabilization of the introduced dye and subsequent preparation of tissues under conditions that avoid dye resolublilization allow accurate visualization of water-conduction pathways at the cellular level.

Phenazines↗

PubMed2006

Molecular cloning of a peroxidase gene from poplar and its expression in response to stress.

To elucidate the precise functions of peroxidase in poplar (Populus alba x P. tremula var. glandulosa), we cloned a peroxidase gene (PoPOD1) from poplar suspension culture cells and examined its expression pattern in response to various stresses. PoPOD1 showed the highest homology with a bacterial-induced peroxidase gene from cotton (Gossypium hirsutum L.). The PoPOD1 gene encodes a putative 316 amino acid protein with an N-terminal signal peptide of 23 residues. The DNA blot analysis indicated that PoPOD1 is a single copy gene in the poplar genome. The RNA blot analyses indicated that PoPOD1 shows cell-culture-specific expression. Expression of PoPOD1 is down-regulated by various treatments including treatment with some metals, NaCl, methyl viologen and polyethylene glycol, and by the plant growth regulators, jasmonic acid (JA) and gibberellic acid (GA(3)). The gene is significantly up-regulated by the bacterial-elicitor laminarin and by wounding. Thus, PoPOD1 gene expression is sensitively and specifically regulated at the transcription level. Because both JA and GA3 appear to be involved in the regulation of PoPOD1 expression in poplar cells, we postulate that the peroxidase encoded by PoPOD1 plays a pivotal role in defense against pathogen invasion, possibly through the formation of a cell wall barrier over the wound.

Amino Acid Sequence↗

PubMed2006

Responses of condensed tannins in poplar roots to fertilization and gypsy moth defoliation.

We examined the effects of fertilization and gypsy moth defoliation on condensed tannin concentration (%CT) of hybrid poplar (Populus x canadensis cv 'Eugeneii') fine roots in the summers of 1997 and 1998. This factorial experiment included two defoliation treatments (defoliated and a foliated control) and fertilization treatments (100 kg nitrogen (N) ha(-1) and an unfertilized control). Gypsy moth (Lymantria dispar L.) populations were experimentally increased to obtain defoliation in the summers of 1996, 1997 and 1998; fertilization subplots were supplemented with NH4NO3 (100 kg N ha(-1)) in the spring of each year. Despite the severity of defoliation, the effects were small, and significant on only two sampling dates: in May 1997, when fine root %CT was 23% lower in the defoliated trees, and in November 1997, when trees in the defoliated unfertilized plots had 35% higher root %CT than trees in all other plots. Defoliation effects on root %CT did not follow the same seasonal pattern as defoliation effects on root starch content, N uptake capacity or leaf %CT. Regulation of root condensed tannin concentration appeared to be partially uncoupled from these traits. The small transient effects on root defense reflect the resilience of this early successional tree to severe early season defoliation.

Animals↗