Coppicing shifts CO2 stimulation of poplar productivity to above-ground pools: a synthesis of leaf to stand level results from the POP/EUROFACE experiment

A poplar short rotation coppice (SRC) grown for the production of bioenergy can combine carbon (C) storage with fossil fuel substitution. Here, we summarize the responses of a poplar (Populus) plantation to 6 yr of free air CO2 enrichment (POP/EUROFACE consisting of two rotation cycles). We show that a poplar plantation growing in nonlimiting light, nutrient and water conditions will significantly increase its productivity in elevated CO2 concentrations ([CO2]). Increased biomass yield resulted from an early growth enhancement and photosynthesis did not acclimate to elevated [CO2]. Sufficient nutrient availability, increased nitrogen use efficiency (NUE) and the large sink capacity of poplars contributed to the sustained increase in C uptake over 6 yr. Additional C taken up in high [CO2] was mainly invested into woody biomass pools. Coppicing increased yield by 66% and partly shifted the extra C uptake in elevated [CO2] to above-ground pools, as fine root biomass declined and its [CO2] stimulation disappeared. Mineral soil C increased equally in ambient and elevated [CO2] during the 6 yr experiment. However, elevated [CO2] increased the stabilization of C in the mineral soil. Increased productivity of a poplar SRC in elevated [CO2] may allow shorter rotation cycles, enhancing the viability of SRC for biofuel production.

Drivers of increased soil respiration in a poplar coppice exposed to elevated CO2

Background and Aims. The response of soil respiration (SR) to elevated CO2 is driven by a number of processes and feedbacks. This work aims to i) detect the effect of elevated CO2 on soil respiration during the second rotation of a short rotation forest, at two levels of N availability; and ii) identify the main drivers behind any changes in soil respiration. Methods. A poplar plantation (POP-EUROFACE) was grown for two rotations of three years under elevated CO2 maintained by a FACE (Free Air CO2 Enrichment) technique. Root biomass, litter production and soil respiration were followed for two consecutive years after coppice. Results. In the plantation, the stimulation of fine root and litter production under elevated CO2 observed at the beginning of the rotation declined over time. Soil respiration (SR) was continuously stimulated by elevated CO2, with a much larger enhancement during the growing (up to 111 %) than in the dormant season (40 %). The SR increase at first appeared to be due to the increase in fine root biomass, but at the end of the 2nd rotation was supported by litter decomposition and the availability of labile C. Soil respiration increase under elevated CO2 was not affected by N availability. Conclusions. The stimulation of SR by elevated CO2 was sustained by the decomposition of above and belowground litter and by the greater availability of easily decomposable substrates into the soil. C losses through SR were greater in the last year of the plantation due to a lack of effect of elevated CO2 on C allocation to roots, reducing the potential for C accumulation.

The effect of Beauveria bassiana on Brazilian poplar moth Condylorrhiza vestigialis (Lepidoptera: Crambidae)

The Brazilian poplar moth is the most important pest of poplar plantations in Brazil. This research evaluated the effect of Beauveria bassiana Bals. (Vuill.) on the mortality and development of Condylorrhiza vestigialis Guen. (Lepidoptera: Crambidae). The aim was to develop alternative methods for management of this pest. The pathogens were sprayed on poplar leaves and .consequently, the pathogens reached the caterpillars. Bacillus thuringiensis var. kurstaki Berliner was sprayed as the standard treatment. The spray for the control was distilled water. Both pathogens B. bassiana and B. thuringiensis affected insect development with increase mortality at each stage of the insect cycle, reaching a satisfactory control level. Microbial control of Brazilian poplar moth with B. bassiana is promising. Tests with other strains and species of pathogens, mainly under field conditions, were also encouraging. This is the first report about the action of B. bassiana against C. vestigialis.

Report of the Poplar Working Group

Poplars consist of all species of the genus Populus, including cottonwoods, aspens, and the many interspecies hybrids in common use (Dickmann and Stuart 1983). Our working group focused on the fungal pathogens, arthropod herbivores, and weed competitors of Populus in the United States. However, bacterial and viral diseases of Populus are significant in Europe, and genetic engineering approaches toward their control or management are being studied. The key aspects of poplar biology important to understanding the use of pest resistance genes are described below.

Phytoremediation by poplar: polyphenols polyamines and oxidative damage in the response to heavy metals in in vitro and in hydroponic cultures.

Poplar is considered a good candidate for phytoremediation, but its tolerance to heavy metals has not been fully investigated yet. In the present work, two different culture systems (in vitro and aeroponic/hydroponic) and two different stress tolerant clones of Populus alba (AL22 and Villafranca) were investigated for their total polyphenol and flavonoid content, individual phenolic compounds, polyamine, lipid peroxidation and hydrogen peroxide levels in response to Cu. In AL22 poplar plants cultured in vitro in the presence or absence of 50 μM Cu, total leaves polyphenol and flavonoid content was higher in treated samples than in controls but unaltered in the roots. Equally the same clone, grown under aeroponic conditions and hydroponically treated for 72 h with 100 μM Cu, displayed increased amount of polyphenols and flavonoids in the leaves, in particular chlorogenic acid and quercetin, and no differences in the roots. In exudates from treated roots total polyphenols and flavonoids, in particular catechin and epicatechin, were more abundant than in controls. Polyamine levels show an increase in conjugated putrescine (Put) and spermidine (Spd) was found. In the Villafranca clone, treated with 100 μM Cu for 6, 24 and 72 h, the pattern of polyphenol and flavonoid accumulation was the same as in AL22; in Cu-treated roots these compounds decreased compared with controls while they increased in root exudates. Free polyamine levels rose at 24 and 72 h while only conjugated Put increased at 24 h. Cu-treated Villafranca plants exhibited a higher malondialdehyde production than controls indicative of membrane lipid peroxidation and, therefore, oxidative stress. An in vitro experiment was carried to investigate the antioxidant effect of the polyamine spermidine (Spd). Exogenous Spd, supplied together with 100 μM Cu, reduced the accumulation of polyphenols and flavonoids, MDA and hydrogen peroxide induced by Cu.

Converting Land from Poplar Tree Bioenergy Planting to Cropland: An Example

Many landowners ask about the process and costs for returning land to crop production after trees are cut for biomass. A field on the Squaw Creek bottom, Story County, Iowa was planted to hybrid poplar trees in spring 2000. The trees were planted in rows with a 10-ft spacing. The trees were cut in spring 2010. The resulting field was four acres, and this is the account of the first corn crop in 2011 on the area.

Increased glutamine in leaves of poplar transgenic with pine GS1a caused greater anthranilate synthetase a-subunit (ASA1) transcript and protein abundances: an auxin-related mechanism for enhanced growth in GS transgenics?

The initial reaction in the pathway leading to the production of indole-3-acetic acid (IAA) in plants is the reaction between chorismate and glutamine to produce anthranilate, catalysed by the enzyme anthranilate synthase (ASA; EC 4.1.3.27). Compared with non-transgenic controls, leaves of transgenic poplar with ectopic expression of the pine cytosolic glutamine synthetase (GS1a; EC 6.3.1.2) produced significantly greater glutamine and significantly enhanced ASA a-subunit (ASA1) transcript and protein (approximately 130% and 120% higher than in the untransformed controls, respectively). Similarly, tobacco leaves fed with 30 mM glutamine and 2 mM chorismate showed enhanced ASA1 transcript and protein (175% and 90% higher than controls, respectively). Furthermore, free IAA was significantly elevated both in leaves of GS1a transgenic poplar and in tobacco leaves fed with 30 mM glutamine and 2 mM chorismate. These results indicated that enhanced cellular glutamine may account for the enhanced growth in GS transgenic poplars through the regulation of auxin biosynthesis

Uncovering cold disruption of the circadian clock in poplar

Dormancy is an adaptive mechanism that allows woody plants to survive at low temperatures during the winter. Disruption of circadian clock genes in winter or under low temperatures, both in long days as in short days, were described in our group few years ago (Ramos et al., 2005). Basic mechanisms of the circadian clock function are similar in herbaceous as well as in woody plants although there are differences in their response to low temperatures (Bieniawska et al., 2008). Woody plants growing in daylight conditions should have a specific transcriptional control above the circadian clock genes, which is responsible of their constitutive transcriptional activation observed under low temperatures conditions. In order to understand this regulatory process, we are analyzing the behavior of a circadian clock gene in poplar. To this aim, we have isolated its promoter region and fused to the luciferase reporter gene. This construct has been transformed into Populus tremula x P. alba 717-1B4 INRA clone. Here we present the characterization of these transgenic lines under different conditions of light and temperature.

Gene discovery in the wood-forming tissues of poplar: Analysis of 5,692 expressed sequence tags

A rapidly growing area of genome research is the generation of expressed sequence tags (ESTs) in which large numbers of randomly selected cDNA clones are partially sequenced. The collection of ESTs reflects the level and complexity of gene expression in the sampled tissue. To date, the majority of plant ESTs are from nonwoody plants such as Arabidopsis, Brassica, maize, and rice. Here, we present a large-scale production of ESTs from the wood-forming tissues of two poplars, Populus tremula L. × tremuloides Michx. and Populus trichocarpa ‘Trichobel.’ The 5,692 ESTs analyzed represented a total of 3,719 unique transcripts for the two cDNA libraries. Putative functions could be assigned to 2,245 of these transcripts that corresponded to 820 protein functions. Of specific interest to forest biotechnology are the 4% of ESTs involved in various processes of cell wall formation, such as lignin and cellulose synthesis, 5% similar to developmental regulators and members of known signal transduction pathways, and 2% involved in hormone biosynthesis. An additional 12% of the ESTs showed no significant similarity to any other DNA or protein sequences in existing databases. The absence of these sequences from public databases may indicate a specific role for these proteins in wood formation. The cDNA libraries and the accompanying database are valuable resources for forest research directed toward understanding the genetic control of wood formation and future endeavors to modify wood and fiber properties for industrial use.

Ozone Sensitivity in Hybrid Poplar Is Correlated with a Lack of Defense-Gene Activation1

Ozone is a major gaseous pollutant thought to contribute to forest decline. Although the physiological and morphological responses of forest trees to ozone have been well characterized, little is known about the molecular basis for these responses. Our studies compared the response to ozone of ozone-sensitive and ozone-tolerant clones of hybrid poplar (Populus maximowizii × Populus trichocarpa) at the physiological and molecular levels. Gas-exchange analyses demonstrated clear differences between the ozone-sensitive clone 388 and the ozone-tolerant clone 245. Although ozone induced a decrease in photosynthetic rate and stomatal conductance in both clones, the magnitude of the decrease in stomatal conductance was significantly greater in the ozone-tolerant clone. RNA-blot analysis established that ozone-induced mRNA levels for phenylalanine ammonia-lyase, O-methyltransferase, a pathogenesis-related protein, and a wound-inducible gene were significantly higher in the ozone-tolerant than in the ozone-sensitive plants. Wound- and pathogen-induced levels of these mRNAs were also higher in the ozone-tolerant compared with the ozone-sensitive plants. The different physiological and molecular responses to ozone exposure exhibited by clones 245 and 388 suggest that ozone tolerance involves the activation of salicylic-acid- and jasmonic-acid-mediated signaling pathways, which may be important in triggering defense responses against oxidative stress.

Purification and Characterization of Peroxidases Correlated with Lignification in Poplar Xylem1

Lignin is an integral cell wall component of all vascular plants. Peroxidases are widely believed to catalyze the last enzymatic step in the biosynthesis of lignin, the dehydrogenation of the p-coumaryl alcohols. As the first stage in identifying lignin-specific peroxidase isoenzymes, the classical anionic peroxidases found in the xylem of poplar (Populus trichocarpa Trichobel) were purified and characterized. Five different poplar xylem peroxidases (PXP 1, PXP 2, PXP 3–4, PXP 5, and PXP 6) were isolated. All five peroxidases were strongly glycosylated (3.6% to 4.9% N-glucosamine), with apparent molecular masses between 46 and 54 kD and pI values between pH 3.1 and 3.8. Two of the five isolated peroxidases (PXP 3–4 and PXP 5) could oxidize the lignin monomer analog syringaldazine, an activity previously correlated with lignification in poplar. Because these isoenzymes were specifically or preferentially expressed in xylem, PXP 3–4 and PXP 5 are suggested to be involved in lignin polymerization.

Overexpression of Iron Superoxide Dismutase in Transformed Poplar Modifies the Regulation of Photosynthesis at Low CO2 Partial Pressures or Following Exposure to the Prooxidant Herbicide Methyl Viologen1

Chloroplast-targeted overexpression of an Fe superoxide dismutase (SOD) from Arabidopsis thaliana resulted in substantially increased foliar SOD activities. Ascorbate peroxidase, glutathione reductase, and monodehydroascorbate reductase activities were similar in the leaves from all of the lines, but dehydroascorbate reductase activity was increased in the leaves of the FeSOD transformants relative to untransformed controls. Foliar H2O2, ascorbate, and glutathione contents were comparable in all lines of plants. Irradiance-dependent changes in net CO2 assimilation and chlorophyll a fluorescence quenching parameters were similar in all lines both in air (21% O2) and at low (1%) O2. CO2-response curves for photosynthesis showed similar net CO2-exchange characteristics in all lines. In contrast, values of photochemical quenching declined in leaves from untransformed controls at intercellular CO2 (Ci) values below 200 μL L−1 but remained constant with decreasing Ci in leaves of FeSOD transformants. When the O2 concentration was decreased from 21 to 1%, the effect of FeSOD overexpression on photochemical quenching at limiting Ci was abolished. At high light (1000 μmol m−2 s−1) a progressive decrease in the ratio of variable (Fv) to maximal (Fm) fluorescence was observed with decreasing temperature. At 6oC the high-light-induced decrease in the Fv/Fm ratio was partially prevented by low O2 but values were comparable in all lines. Methyl viologen caused decreased Fv/Fm ratios, but this was less marked in the FeSOD transformants than in the untransformed controls. These observations suggest that the rate of superoxide dismutation limits flux through the Mehler-peroxidase cycle in certain conditions.

Silvical characteristics of balsam poplar (Populus balsamifera).

"This is the thirteenth of the silvical reports being prepared by the Lake States Forest Experiment Station."