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Oxidative lime pretreatment of high-lignin biomass: poplar wood and newspaper.

Lime (Ca[OH]2) and oxygen (O2) were used to enhance the enzymatic digestibility of two kinds of high-lignin biomass: poplar wood and newspaper. The recommended pretreatment conditions for poplar wood are 150 degrees C, 6 h, 0.1 g of Ca(OH)2/g of dry biomass, 9 mL of water/g of dry biomass, 14.0 bar absolute oxygen, and a particle size of -10 mesh. Under these conditions, the 3-d reducing sugar yield of poplar wood using a cellulase loading of 5 filter paper units (FPU)/g of raw dry biomass increased from 62 to 565 mg of eq. glucose/g of raw dry biomass, and the 3-d total sugar (glucose + xylose) conversion increased from 6 to 77% of raw total sugars. At high cellulase loadings (e.g., 75 FPU/g of raw dry biomass), the 3-d total sugar conversion reached 97%. In a trial run with newspaper, using conditions of 140 degrees C, 3 h, 0.3 g of Ca(OH)2/g of dry biomass, 16 mL of water/g of dry biomass, and 7.1 bar absolute oxygen, the 3-d reducing sugar yield using a cellulase loading of 5 FPU/g of raw dry biomass increased from 240 to 565 mg of eq. glucose/g of raw dry biomass. A material balance study on poplar wood shows that oxidative lime pretreatment solubilized 38% of total biomass, including 78% of lignin and 49% of xylan; no glucan was removed. Ash increased because calcium was incorporated into biomass during the pretreatment. After oxidative lime pretreatment, about 21% of added lime could be recovered by CO2 carbonation.

Biomass↗

The arbuscular mycorrhizal status of poplar clones selected for phytoremediation of soils contaminated with heavy metals.

The aim of this work was to study the colonization of indigenous arbuscular mycorrhizal fungi (AMF) species in fine-roots of poplar clones. Roots of 7 poplar clones were sampled from a 1-year-old trial established at an industrial site strongly polluted with heavy metals at Balatonfuzfo, Hungary. The poplar clones have shown variable degrees of colonization by AMF, suggesting differential host susceptibility or mycorrhizal dependency. After outplanting the percentage of poplar survival was strongly correlated with the frequency of AMF infection. Two clones that survived at the lowest ratio after outplanting had not been colonized by AMF in contrast to those which survived to a much higher extent.

Biodegradation, Environmental↗

[Nitrogen cycling and its dynamic feature in fine roots of poplar trees and herbs].

The nitrogen cycling and its dynamic feature in fine roots of poplar trees and herbs were studied in Shenyang of Liaoning Province. The results showed that the nitrogen content in the standing biomass of fine roots of poplar trees and herbs was 20.3 kg.hm-2, and its net accumulation was 10.0 kg.hm-2. With regard to the biological cycling of nitrogen, the annual uptake was 14.6 kg.hm-2 of which, the retention and the return was 9.6 and 5.1 kg.hm-2.yr-1 respectively, and the turnover period was 4 year. The relationship between total nitrogen in roots of poplar trees, and herbs and the available nitrogen in soil was also studied, which indicated that there were similar dynamic trends between total nitrogen in herb roots and the NH3-N in soil in growth season. The total nitrogen in fine roots of poplar trees was positively related to the biomass.

Ammonia↗

Differential susceptibility of poplar hybrids to the aphid Chaitophorus leucomelas (Homoptera: Aphididae).

After its recent introduction to Chile, the aphid Chaitophorus leucomelas Koch is becoming a serious pest affecting commercial poplar, Populus spp., plantations. The pattern of natural infestation of C. leucomelas among poplar hybrids with different pedigrees and the aphid intrinsic rate of increase (r(m)), of C. leucomelas were assessed in the field. In most of the hybrids, aphid abundance peaked in March (late summer). Among 12 types of poplar crosses, [(P. trichocarpa Torr. & Gray x P. deltoides Bartram ex Marshall) x (P. trichocarpa x P. deltoides)] and [(P. trichocarpa x P. maximowiczii Henry) x P. maximowiczii] showed the highest and lowest aphid densities, respectively. A trend to find more aphids in branch bases was apparent. The intrinsic rate of C. leucomelas increase was higher in [(P. trichocarpa x P. deltoides) x P. deltoides] hybrids, and lower in [(P. trichocarpa x P. maximowiczii) x P. trichocarpa] hybrids. Aphid density and performance were higher in hybrids with P. deltoides parentage, whereas hybrids with P. maximowiczii parentage showed lower aphid densities and performance. Hybrids with P. nigra L. parentage, namely, [P. trichocarpa x P. nigra], also had high aphid density, but aphid performance was lower compared with hybrids with P. deltoides parentage. These results suggest that among poplar hybrids studied, susceptibility to C. leucomelas is inherited through P. deltoides, whereas resistance seems to be inherited through P. maximowiczii. Thus, P. maximowiczii hybrids are recommended for commercial or ornamental planting programs in zones where there is a high risk of aphid infestation.

Animals↗

Systemically wound-responsive genes in poplar trees encode proteins similar to sweet potato sporamins and legume Kunitz trypsin inhibitors.

When the lower leaves of hybrid poplar trees are mechanically wounded, several novel mRNAs accumulate in the unwounded upper leaves (Parsons TJ, Bradshaw HD, Gordon MP: Systemic accumulation of specific mRNAs in response to wounding in poplar trees, Proc Natl Acad Sci USA, in press). A partial cDNA clone corresponding to a transcript from the wound-responsive gene designated win 3 (wound-inducible) has been cloned by differential hybridization to 32P-labelled cDNA from the leaves of wounded trees. Northern blots show a large accumulation of win 3 transcripts in the unwounded leaves of wounded trees. Southern blot analysis of poplar DNA suggests that win 3 is a member of a multigene family. The nucleotide sequences of several win 3 cDNA clones have been determined, indicating that at least three win 3 gene family members are transcribed. A genomic clone of a win 3 gene family member has been isolated and a 1.5 kb Hind III fragment containing the predicted protein-coding and 5' upstream regions has been sequenced. The putative win 3 gene product is similar to the major soluble proteins of sweet potato tubers, sporamin A and sporamin B. Both Win3 and the sporamins share significant amino acid sequence identity with Kunitz-type trypsin inhibitors from legume seeds. The Kunitz family of proteinase inhibitors thus joints three other proteinase inhibitor families which are systemically responsive to wounding.

Amino Acid Sequence↗

Seasonal changes in the concentration of the major storage protein and its mRNA in xylem ray cells of poplar trees.

In the autumn large amounts of a major storage protein accumulate in the woody stem of poplar trees. This protein is stored in xylem ray cells during the winter season and is degraded in late spring. The accumulation of this protein is preceded by a dramatic but transient appearance of the corresponding mRNA. Thus, the seasonal change in the mRNA content appears to be a crucial event for the storage of nitrogen in the stem. The amino acid sequence of the storage protein has been deduced from the nucleotide sequence of a full-length cDNA. The cDNA shows a nucleotide sequence similarity of approximately 75% with two published cDNA sequences of poplar which represent transcripts that accumulate systemically in leaves of poplar trees in response to wounding.

Amino Acid Sequence↗

Biochemical properties of isoprene synthase in poplar (Populus x canescens).

Isoprene synthase (ISPS) catalyzes the elimination of pyrophosphate from dimethylallyl diphosphate (DMADP) forming isoprene, a volatile hydrocarbon emitted from many plant species to the atmosphere. In the present work, immunological techniques were applied to study and localize ISPS in poplar leaves (Populus x canescens). Immunogold labeling using polyclonal antibodies generated against His-tagged recombinant ISPS protein detected ca. 44% of ISPS in the stroma of the chloroplasts and ca. 56% of gold particles attached to the stromal-facing side of the thylakoid membranes. ISPS isolated from leaves exhibited the same biochemical properties as the recombinant ISPS without the plastid-targeting peptide heterologous expressed in E. coli, whereas an additional C- or N-terminal His-tag changed the biochemical features of the recombinant enzyme with regard to temperature, pH, and substrate dependence. In comparison to the closely related class of monoterpene synthases from angiosperms and ISPS of oaks, the most striking feature of the poplar ISPS is a cooperative substrate dependence which is characteristic to enzymes with positive substrate activation. The detection of four immunoreactive bands in poplar leaf extracts with isoelectric points from 5.0 to 5.5 and a native molecular weight of ca. 51 kDa give reason for future studies on post-translational modifications of ISPS.

Alkyl and Aryl Transferases↗

One-step purification and characterization of a lignin-specific O-methyltransferase from poplar.

O-Methyltransferases (OMT; EC 2.1.1.6) play an important role in the synthesis of lignin precursors by catalyzing the O-methylation of o-diphenolic substrates such as caffeic acid (CA) and 5-hydroxyferulic acid (5OH). Here, we report on the purification of a lignin-specific OMT (38 kDa) from poplar (Populus trichocarpa x P. deltoides). The OMT was purified from xylem by a single affinity chromatography step on adenosine agarose. The enzyme uses both CA and 5OH as substrates. We previously have reported the cloning of a corresponding OMT cDNA [Dumas et al., Plant Physiol. 98 (1992) 796-797]. Expression of this OMT cDNA in Escherichia coli further confirmed the identity of the clone. Genomic hybridization demonstrates the presence of one or two OMT genes per haploid poplar genome. RNA gel blot hybridization shows high levels of steady-state OMT mRNA in the xylem of young poplar trees, as compared to the levels in leaves.

Blotting, Northern↗

The transient complex of poplar plastocyanin with cytochrome f: effects of ionic strength and pH.

The orientation of poplar plastocyanin in the complex with turnip cytochrome f has been determined by rigid-body calculations using restraints from paramagnetic NMR measurements. The results show that poplar plastocyanin interacts with cytochrome f with the hydrophobic patch of plastocyanin close to the heme region on cytochrome f and via electrostatic interactions between the charged patches on both proteins. Plastocyanin is tilted relative to the orientation reported for spinach plastocyanin, resulting in a longer distance between iron and copper (13.9 A). With increasing ionic strength, from 0.01 to 0.11 M, all observed chemical-shift changes decrease uniformly, supporting the idea that electrostatic forces contribute to complex formation. There is no indication for a rearrangement of the transient complex in this ionic strength range, contrary to what had been proposed earlier on the basis of kinetic data. By decreasing the pH from pH 7.7 to pH 5.5, the complex is destabilized. This may be attributed to the protonation of the conserved acidic patches or the copper ligand His87 in poplar plastocyanin, which are shown to have similar pK(a) values. The results are interpreted in a two-step model for complex formation.

Brassica napus↗

Transport and fate of dieldrin in poplar and willow trees analyzed by SPME.

Dieldrin is a hydrophobic organochlorine insecticide that is persistent in the environment. The fate and transport of dieldrin in trees is important both in the context of potential remediation, as well as food chain impacts through dieldrin transport to shoots and leaves. Experiments were conducted to measure the degree of dieldrin partitioning to plant tissue and the potential for biodegradation of dieldrin in the microbe rich tree rhizosphere. Dieldrin was analyzed in water and plant tissue using headspace solid-phase microextraction (SPME) coupled with gas chromatography. Poplar and willow saplings planted in soil and watered with 10 microgl(-1) dieldrin for up to 9 months showed no adverse effects due to dieldrin exposure and no dieldrin was observed in plant shoots with a method detection limit (MDL) of 7 ngg(-1). One-week hydroponic tests of poplar saplings exposed to aqueous dieldrin also showed no detection of dieldrin in shoots, with an average of 66% of the dieldrin partitioned to the plant roots and an overall mass balance recovery of 76% in the plant-water system. The root concentration factor (RCF) was found to be 30+/-3 ml water g(-1) root. Biodegradation of dieldrin was not observed in an aqueous batch bioreactor containing 8 microgl(-1) dieldrin, nutrients and bacteria from the root zone of a poplar sapling that had been exposed to dieldrin for 9 months. These results show that planting trees is likely to be safe and potentially useful at sites containing low-levels of dieldrin in groundwater.

Biodegradation, Environmental↗

White poplar (Populus alba) as a biomonitor of trace elements in contaminated riparian forests.

Trees can be used to monitor the level of pollution of trace elements in the soil and atmosphere. In this paper, we surveyed the content of eight trace elements (As, Cd, Cu, Fe, Mn, Ni, Pb and Zn) in leaves and stems of white poplar (Populus alba) trees. We selected 25 trees in the riparian forest of the Guadiamar River (S. Spain), one year after this area was contaminated by a mine spill, and 10 trees in non-affected sites. The spill-affected soils had significantly higher levels of available cadmium (mean of 1.25 mg kg(-1)), zinc (117 mg kg(-1)), lead (63.3 mg kg(-1)), copper (58.0 mg kg(-1)) and arsenic (1.70 mg kg(-1)), than non-affected sites. The concentration of trace element in poplar leaves was positively and significantly correlated with the soil availability for cadmium and zinc, and to a lesser extent for arsenic (log-log relationship). Thus, poplar leaves could be used as biomonitors for soil pollution of Cd and Zn, and moderately for As.

Arsenic↗

Characterization of a poplar NIMA-related kinase PNek1 and its potential role in meristematic activity.

Meristems are sites of undifferentiated cell division, which carry on developing into functional organs. Using the two-hybrid system with a poplar 14-3-3, we uncovered poplar NIMA-related kinase 1 (PNek1) as an interacting protein. PNek1 shows high homology to the mammalian NIMA-related kinases, which are thought to be involved in cell cycle progression. Using a synchronized poplar cell suspension, we observed an accumulation of PNek1 mRNA at the G1/S transition and throughout the G2-to-M progression. Moreover, PNek1-GFP fusion protein localized in the cytoplasm and in both the nuclear and nucleolar regions. Overexpression of PNek1-GFP in Arabidopsis caused morphological abnormalities in flower and siliques. Overall, these results suggest that PNek1 is involved in plant development.

14-3-3 Proteins↗

Impact of flooding on soil bacterial communities associated with poplar (Populus sp.) trees.

Soil bacterial communities were analyzed in different habitats (bulk soil, rhizosphere, rhizoplane) of poplar tree microcosms (Populus tremulaxP. alba) using cultivation-independent methods. The roots of poplar trees regularly experience flooded and anoxic conditions. Therefore, we also determined the effect of flooding on microbial communities in microcosm experiments. Total community DNA was extracted and bacterial 16S rRNA genes were amplified by PCR and analyzed by terminal restriction fragment length polymorphism (T-RFLP) analysis, cloning and sequencing. Clone libraries were created from all three habitats under both unflooded and flooded conditions resulting in a total of 281 sequences. Numbers of different sequences (<97% similarity) in the different habitats represented 16-55% of total bacterial species richness determined from the nonparametric richness estimator Chao1. According to the number of different terminal restriction fragments (T-RFs), all of the different habitats contained approximately 20 different operational taxonomic units (OTUs), except the flooded rhizoplane habitat whose community contained less OTUs. Results of cloning and T-RFLP analysis generally supported each other. Correspondence analysis of T-RFLP patterns showed that the bacterial communities were different in bulk soil, rhizosphere and rhizoplane and changed upon flooding. For example OTUs representing Bacillus sp. were highest in the unflooded bulk soil and rhizosphere. Sequences related to Aquaspirillum, in contrast, were predominant on the poplar roots and in the rhizosphere of flooded microcosms but were rarely found in the other habitats.

Bacteria↗

Biomass production and carbon sequestration potential in poplar plantations with different management patterns.

Biomass production and carbon storage in short-rotation poplar plantations over 10 years were evaluated at the Hanyuan Forestry Farm, Baoying County, China. Experimental treatments applied in a split-plot design included four planting densities (1111, 833, 625 and 500 stems ha(-1)) and three poplar clones (NL-80351, I-69 and I-72). Based on the model of total biomass production developed, total plantation biomass production was significantly different in the plantations. The ranking of the plantation biomass production by planting density was 1111>833 more more than 625>500 stems ha(-1), and by components was stem>root>or=branch>leaf for all plantations. At 10 years, the highest total biomass in the plantation of 1111 stems ha(-1) reached about 146 t ha(-1), which was 5.3%, 11.6% and 24.2% higher than the plantations of 833, 625 and 500 stems ha(-1), respectively. The annual increment of biomass production over 10 years differed significantly among initial planting densities and stand ages (p<0.01), but no significant difference was observed from age 7 to 10. Mean carbon concentration among all biomass components ranged from 42-50%, with the highest carbon concentrations in stems and the lowest in leaves. Over the study period, the dynamic pattern of total plantation carbon storage by planting density was similar to that of total biomass production. At age 10, the highest total plantation carbon storage in the plantation of 1111 stems ha(-1) reached about 72.0 t ha(-1), which was 5.4%, 11.9% and 24.8% higher than in the plantations of 833, 625 and 500 stems ha(-1), respectively. The annual carbon storage increment over 10 years differed significantly among initial planting densities and stand ages (p<0.01), and it showed a pattern similar to the annual biomass production increment of the plantations. The results suggest that biomass production and carbon storage potential were highest for planting densities of 1111 and 833 stems ha(-1) grown over 5- and 6-year cutting cycles, respectively. If 3- or 4-year cutting cycles are used, the planting density should be higher than 1111 stems ha(-1) (e.g., 1667 or 2500 stems ha(-1)). Based on the mean annual carbon storage for the plantation of 625 stems ha(-1), as an estimation, the mean carbon storage in the biomass of poplar plantations (excluding leaves) amounts to 3.75x10(7) t ha(-1)yr(-1) in China.

Biomass↗

Testing applicability of black poplar (Populus nigra L.) bark to heavy metal air pollution monitoring in urban and industrial regions.

A comparative study of the capabilities of black poplar-tree (Populus nigra L.) bark as a biomonitor of atmospheric heavy-metal pollution is reported. Performance indicators (concentrations and enrichment factors) of heavy metal bioaccumulation of bark were compared to the corresponding indicators of epiphytic lichens Xanthoria parietina (L.) Th. Fr. and Physcia adscendens (Fr.) H. Oliver, collected simultaneously with bark samples within the Kiev urban-industrial conurbation. The concentrations of 40 minor and trace elements in the samples were measured by a combination of epithermal and instrumental neutron activation analysis (NAA) using a 10 MW nuclear research reactor WWR-M as the neutron source. Statistical analysis of the data was carried out using non-parametric tests. It was shown that for the majority of the elements determined a good correlation exists between their concentrations in bark and in the lichen species. The accumulation capability of the bark was found to be as effective as, and in some cases better, for both types of lichens. Based on the background levels and variations of the elemental concentration in black poplar-tree bark, threshold values for the enrichment factors were established. For a number of elements (As, Au, Ce, Co, Cr, Cu, La, Mn, Mo, Ni, Sb, Sm, Ti, Th, U, V, W) an interspecies calibration was performed. An optimized pre-irradiation treatment of the bark sample was employed which efficiently separated the most informative external layer from the deeper layers of the bark and thus minimized variations of the element concentrations. Results of this study support black poplar-tree bark as an alternative to epiphytic lichens for heavy metal air pollution monitoring in urban and industrial regions, where severe environmental conditions may result in scarcity or even lack of the indicator species.

Air Pollutants↗

Endophytic bacterial diversity in poplar trees growing on a BTEX-contaminated site: the characterisation of isolates with potential to enhance phytoremediation.

The diversity of endophytic bacteria found in association with poplar was investigated as part of a larger study to assess the possibility and practicality of using endophytic bacteria to enhance in situ phytoremediation. Endophytic bacteria were isolated from the root, stem and leaf of two cultivars of poplar tree growing on a site contaminated with BTEX compounds. They were further characterised genotypically by comparative sequence analysis of partial 16S rRNA genes and BOX-PCR genomic DNA fingerprinting, and phenotypically by their tolerance to a range of target pollutants, heavy metals and antibiotics. One hundred and 21 stable, morphologically distinct isolates were obtained, belonging to 21 genera, although six isolates could not be identified with confidence to a genus. The endophytic bacteria exhibited marked spatial compartmentalisation within the plant, suggesting there are likely to be species-specific and non-specific associations between bacteria and plants. A number of isolates demonstrated the ability to degrade BTEX compounds or to grow in the presence of TCE. This study demonstrates that within the diverse bacterial communities found in poplar several endophytic strains are present that have the potential to enhance phytoremediation strategies.

Biodegradation, Environmental↗

Poplar genome sequence: functional genomics in an ecologically dominant plant species.

In addition to their value for wood products, members of the genus Populus (poplars) provide a range of ecological services, including carbon sequestration, bioremediation, nutrient cycling, biofiltration and diverse habitats. They are also widely used model organisms for tree molecular biology and biotechnology. The sequencing of the poplar genome to an approximately 6x depth adds to a long list of important attributes for research. These include facile transformation, vegetative propagation, rapid growth, modest genome size and extensive expressed sequence tags. Here, we discuss how the genome sequence and transformability of poplar, together with its high levels of genetic and ecological diversity, are enabling new insights into the genetic programs controlling ontogeny, ecological adaptation and environmental physiology of trees.

Ecosystem↗

Biochemical characterization of two differentially expressed polyphenol oxidases from hybrid poplar.

Two polyphenol oxidase isoforms with distinct expression patterns were identified in hybrid poplar (Populus trichocarpaxP. deltoides). PPO-1, corresponding to the previously cloned PtdPPO (Constabel et al., Plant Physiol. 124: 285-295) was primarily leaf tissue-specific and detected only after wounding. PPO-2 was expressed constitutively in all tissue types tested except mature leaves, with highest expression in very young leaves and conducting tissues such as roots, stems and petioles. These two PPO isoforms were partially purified from hybrid poplar by ammonium sulfate fractionation followed by hydrophobic interaction chromatography. They were found to differ in stability, pH optimum, and activation by SDS. Tests with common phenolic substrates showed that PPO-1 had a broader substrate specificity than PPO-2. The distinct enzymatic properties and expression patterns of these two PPO isoforms suggest that they may have different physiological functions in hybrid poplar.

Catechol Oxidase↗