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Photoperiod control of poplar bark storage protein accumulation.

Bark storage proteins (BSPs) accumulate in the inner bark parenchyma of many woody plants during autumn and winter. We investigated the effect of a short-day (SD) photoperiod on the accumulation of the 32-kilodalton bark storage protein of poplar (Populus deltoides Bart. ex Marsh.) under controlled environmental and natural growing conditions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein gel blot analysis revealed that 10 days of SD exposure (8 hours of light) resulted in a 20% increase in the relative abundance of the 32-kilodalton bark storage protein of poplar. After 17 days of SD exposure, the 32-kilodalton bark storage protein accounted for nearly one-half of the soluble bark proteins. In natural field conditions, accumulation of the 32-kilodalton bark storage protein was observed to start by August 18 (daylength 14.1 hours). Immunoprecipitation of in vitro translation products with anti-BSP serum revealed that the SD protein accumulation was correlated with changes in the pool of translatable mRNA. A survey of poplar clones from different geographic origins revealed the presence of the 32-kilodalton BSP in the dormant bark of all the clones tested. These results demonstrate that a SD photoperiod induces, whether directly or indirectly, rapid changes in woody plant gene expression, leading to the accumulation of BSP.

Journal Article↗

Exogenous supply of glutamine and active cytokinin to the roots reduces NO3- uptake rates in poplar.

The present study shows for the first time the influence of exogenously applied amino acids and cytokinin on the physiological and molecular aspects of N metabolism in poplar trees. In a short-term feeding experiment, glutamine or trans-zeatin riboside (tZR) was added directly to the nutrient solution. NO3- net uptake declined significantly in response to both treatments. Feeding with glutamine brought about an increase in concentrations of different amino compounds in the roots (glutamine, glutamate, alanine, gamma-amino butyric acid (GABA) and NH4+, which negatively correlated with the net NO3- uptake. The plants showed a reduction of cytosolic glutamine synthetase 1 (GS1) transcript level in the roots. In addition, glutamine feeding changed the root-to-shoot distribution on N assimilation in favour of the leaves and plant internal N cycling. tZR treatment resulted in expansion of zeatin-type (Z-type) cytokinins in the roots and increased nitrate reductase (NR)-mRNA level. The results indicate that both particular amino acids and active cytokinins are involved in the feedback regulation of N uptake and metabolism in poplar. We propose that inhibition of N uptake by cytokinins in poplar is more complex than that mediated by amino compounds, and other effectors are involved in this regulation.

Cytokinins↗

MRI of long-distance water transport: a comparison of the phloem and xylem flow characteristics and dynamics in poplar, castor bean, tomato and tobacco.

We used dedicated magnetic resonance imaging (MRI) equipment and methods to study phloem and xylem transport in large potted plants. Quantitative flow profiles were obtained on a per-pixel basis, giving parameter maps of velocity, flow-conducting area and volume flow (flux). The diurnal xylem and phloem flow dynamics in poplar, castor bean, tomato and tobacco were compared. In poplar, clear diurnal differences in phloem flow profile were found, but phloem flux remained constant. In tomato, only small diurnal differences in flow profile were observed. In castor bean and tobacco, phloem flow remained unchanged. In all plants, xylem flow profiles showed large diurnal variation. Decreases in xylem flux were accompanied by a decrease in velocity and flow-conducting area. The diurnal changes in flow-conducting area of phloem and xylem could not be explained by pressure-dependent elastic changes in conduit diameter. The phloem to xylem flux ratio reflects what fraction of xylem water is used for phloem transport (Münch's counterflow). This ratio was large at night for poplar (0.19), castor bean (0.37) and tobacco (0.55), but low in tomato (0.04). The differences in phloem flow velocity between the four species, as well as within a diurnal cycle, were remarkably small (0.25-0.40 mm s(-1)). We hypothesize that upper and lower bounds for phloem flow velocity may exist: when phloem flow velocity is too high, parietal organelles may be stripped away from sieve tube walls; when sap flow is too slow or is highly variable, phloem-borne signalling could become unpredictable.

Biological Transport↗

Molecular analysis of poplar defense against herbivory: comparison of wound- and insect elicitor-induced gene expression.

In order to characterize defense responses of hybrid poplar (Populus trichocarpax P. deltoides), we profiled leaf transcript patterns elicited by wounding and by regurgitant from forest tent caterpillar (FTC; Malacosoma disstria), a Lepidopteran defoliator of poplars. Macroarrays were used to compare transcript profiles. Both FTC-regurgitant (FTC-R) and mechanical wounding with pliers elicited expression of a variety of genes, and for these genes our analysis indicated that these treatments induced qualitatively similar responses. Similarly, a comparison of responses of directly treated and systemically induced leaves indicated extensive overlap in the sets of induced genes. FTC-R was found to contain the insect-derived elicitor volicitin. The simulated herbivory treatments resulted in the induction of genes involved in poplar defense and secondary metabolism. We also identified wound-responsive genes with roles in primary metabolism, including a putative invertase, lipase, and acyl-activating enzyme; some of these genes may have roles in defense signaling. In addition, we found three unknown genes containing a ZIM motif which may represent novel transcription factors.

Animals↗

Assessing the use of poplar tree systems as a landfill evapotranspiration barrier with the SHAW model.

The use of poplar tree systems (PTS) as evapotranspiration barriers on decommissioned landfills is gaining attention as an option for leachate management. This study involved field-testing the Simultaneous Heat and Water (SHAW) model for its ability to reliably estimate poplar transpiration, volumetric soil water content, and soil temperature at a landfill located in southern Ontario, Canada. The model was then used to estimate deep drainage and to ascertain the influence of a young PTS on the soil water balance of the landfill cover. The SHAW model tended to underestimate poplar transpiration [mean difference (MD) ranged from 0.33 to 3.55 mm on a daily total basis] and overestimate volumetric soil water content by up to 0.10 m3 m(-3). The model estimated soil temperature very well, particularly in the upper 1 m of the landfill cover (MD ranged from -0.1 to 1.6 x degrees C in this layer). The SHAW model simulations showed that deep drainage decreased appreciably with the presence of a young PTS largely through increased interception of rainfall, and that PTS have a good potential to act as effective evapotranspiration barriers in northern temperate climate zones.

Biodegradation, Environmental↗

Hydrothermal pretreatment conditions to enhance ethanol production from poplar biomass.

Pretreatment has been recognized as a key step in enzyme-based conversion processes of lignocellulose biomass to ethanol. The aim of this study is to evaluate two hydrothermal pretreatments (steam explosion and liquid hot water) to enhance ethanol production from poplar (Populus nigra) biomass by a simultaneous saccharification and fermentation (SSF) process. The composition of liquid and solid fractions obtained after pretreatment, enzymatic digestibility, and ethanol production of poplar biomass pretreated at different experimental conditions was analyzed. The best results were obtained in steam explosion pretreatment at 210 C and 4 min, taking into account cellulose recovery above 95%, enzymatic hydrolysis yield of about 60%, SSF yield of 60% of theoretical, and 41% xylose recovery in the liquid fraction. Large particles can be used for poplar biomass in both pretreatments, since no significant effect of particle size on enzymatic hydrolysis and SSF was obtained.

Biomass↗

[Insect-resistant transgenic poplar expressing AaIT gene].

Insect-specific scorpion neurotoxin AaIT gene inserted into a binary vector was transferred into a hybrid poplar clone N-106(P. deltoides x P. simonii) growing in the Southern of China. We obtained sixty-two regenerated plants by Agrobacterium tumefaciens transferring system. PCR and PCR-Southern analysis showed that AaIT gene was incorporated into the genome of some recovered poplar plants. One of the transformed plants named A5 was significantly resistant to feeding by first instar larvae of Lymantria dispar, compared with the untransformed control plant. It caused a decrease in leaf consumption by larvae, a lower larval weight gain and a higher larval motality rate of Lymantria dispar. ELISA analysis proved that AaIT gene was expressed in this transfomed poplar plant.

Animals↗

[A review on the eco-physiological study of poplars in oasis and its prospect].

Poplar is the main component species in shelter forest subsystem, which forms a special kind of ecosystem in oasis with particular structure and function. Based on the studies on the ecophysiological characteristics of poplars in the Cele oasis, a preliminary comment was given on the experimental methods used at home and abroad, aim ed to facilitate the further studies on the characteristics of ecophysiology of poplars in the oasis. In order to sustain the oasis ecosystem management, the theoretical bases were also discussed on the optimization of shelter-forest structure and the selection of proper tree species.

China↗

[Effect of flooding stress on transpiration of poplar I-69/55].

The rates of transpiration(Tr) and photosynthesis(Pn) of poplar I-69/55 trees were measured under flooding stress. Flooding stress caused Pn and Tr decreasing, while in the period of measurement, Pn almost didn't change. The longer the period of flooding stress was, the more the Tr and stomatal conductance (Gs) decreased. Undre no stress, Pn and Tr of test trees displayed daily double-peak curves and seasonal variations, with the maximum in summer. Water use efficiency also displayed seasonal variation, with the maximum in autumn. Under no stress, Tr obviously changed with tree ages, with the order of 1 year-old > 3 year-old > 7 year-old. The rate of Tr/Pn in spring, summer and autumn was higher than that of other deciduous broad-leaved trees in the same region, especially in summer. The high Tr/Pn ratio means a high water consumption, which is beneficial to water loss of poplar I-69/55. In shoal sites, the temperature of leaf surface in summer and autumn is the most important factor that affects the transpiration rate of poplar I-69/55, suggesting that decreasing the temperature of leaf suface is beneficial to increase the transpiration rate. During the two days after flooding, the temperature of leaf surface is still the important factor that affects transpiration rate.

Disasters↗

[Influence of temperature on insect-resistance of transgenic hybrid poplar 741].

The study showed that the insect-resistance of transgenic hybrid poplar 741 was closely related with temperature. The total mortality and accumulative mortality of Gypsy moth (Lmantriadispar L.) larvae increased with increasing temperature, and the sensitivity of larvae to the clones of transgenic hybrid poplar 741 and temperature decreased gradually with the increasing instars. Temperature had a promotion to the development of larvae fed on CK. When the temperature was high, the development of larvae quickened, the development period shortened, and the weight increasing rate and the frass increasing quantity quickened, but the development rate of larvae fed on transgenic hybrid poplar 741 was not obvious with varied temperature. The regressive equation of temperature and Clostera anachoreta (Fabricius) mortality during 1998-2000 year indicated that high resistant and medium resistant clones' temperature-insect mortality curve presented an obvious regressive relation.

Animals↗

[Effects of exogenous nitric oxide on photosynthetic characteristics of poplar leaves under water stress].

Nitric oxide (NO) is an active molecule involved in many biological pathways, but its effects on photosynthesis of tree leaves have not been established yet. This paper studied the effects of exogenous NO, sodium nitroprusside (SNP), on the water status, photosynthesis and scavenging enzyme activities in poplar leaves. Different levels of SNP treatments had remarkable effects on the water content of leaves, which increased with increasing SNP levels. When the SNP level exceeded 500 micromol x L(-1), differences in leaf water content were no longer significant between different SNP treatments. Exogenous NO increased the photosynthesis rate, photochemical efficiency of PSII Fv/Fm, and Fm/Fo and Fv/Fo ratios, and the effects decreased with increasing duration of water stress. The SOD and POD activities in poplar leaves were higher in 1 hour water stress treatment than in 3 h treatment. Treating with SNP could markedly increase POD activity, but SOD activity did not change much. POD and SOD activities increased initially, and then decreased with increasing SNP levels. The results indicated that exogenous NO delayed the accumulation of active oxygen by increasing POD and SOD activities, and thereby, alleviated the effects of water stress on photosynthetic organization of poplar leaves.

Nitric Oxide↗

[Effects of exogenous nitric oxide on drought-resistance of poplar].

This paper studied the effects of NO donor sodium nitroprusside (SNP) on the photosynthesis and active oxygen-scavenging enzyme activities of poplar leaves under drought stress. The results showed that SNP had a remarkable positive effect on leaf water content. When the SNP level was > 500 micromol.L(-1), no significant difference in leaf water content was found between SNP treatments. SNP could increase photosynthesis rate, photochemical efficiency of PSII (F(v)/F(m)), and ratios of F(m)/F(o) and F(v)/F(o), but the effects decreased with the prolongation of drought stress. The SOD and POD activities of poplar leaves were higher in short time (1 h) drought stress than in longer time (3 h) one. The activity of POD was increased by SNP, while that of SOD was changed little. With increasing SNP level, the POD and SOD activities first increased and then decreased. The NO donor could retard the accumulation of active oxygen through inducing POD and SOD activities, alleviate the effects of drought stress on photosynthesis, and increase the drought-resistance of poplar.

Disasters↗

Isolation of a poplar and an Arabidopsis thaliana dihydrodipicolinate synthase cDNA clone.

A poplar DHDPS cDNA clone has been isolated by functional rescue of the dapA-deficient AT997 mutant of Escherichia coli. By sequence comparison between the poplar and maize DHDPS cDNAs, two oligonucleotides were designed to perform polymerase chain reaction (PCR) on Arabidopsis thaliana genomic DNA. The PCR fragment was subsequently used to isolate an Arabidopsis DHDPS genomic and cDNA clone.

Amino Acid Sequence↗

A poplar tree proteinase inhibitor-like gene promoter is responsive to wounding in transgenic tobacco.

Wounding of poplar trees leads to the accumulation of several mRNA species that encode proteins with putative defensive function. One class of wound-induced poplar RNA (win3) has amino acid sequence similarity to Kunitz-type trypsin inhibitors. Northern blots and cDNA sequencing show that several win3 mRNAs accumulate in the uninjured leaves of wounded trees. We report further characterization of the win3 family including sequence comparisons, gene family organization, and the identification of one win3 member that is transcriptionally activated in response to mechanical wounding. We also show that 1.5 kb of 5'-flanking sequence of one win3 member (win3.12) is sufficient to confer wound-regulated expression of a beta-D-glucuronidase (GUS) reporter gene in transgenic tobacco. Annual herbaceous plants such as tobacco can thus be used to study the expression of genes from a perennial woody angiosperm.

Amino Acid Sequence↗

Polar-localised poplar K+ channel capable of controlling electrical properties of wood-forming cells.

In previous studies, we have shown that annual expression profiles of cambial and wood tissue with respect to the Shaker K+ channel PTORK correlate with cambial activity. To follow PTORK-gene activity on the cellular level, we isolated the respective promoter regions and generated transgenic Arabidopsis plants expressing the GUS gene under the control of the PTORK promoter. Cross-sections of petioles showed PTORK-driven signals predominantly in the xylem parenchyma surrounding the vessels and in the phloem. Antibodies raised against a unique N-terminal region of PTORK in histo-immunochemical analyses recognised this K+-release channel in growth-active poplar plants only. PTORK labelling was found in differentiating xylem cells (young fibres) and mature xylem (vessel-associated cells of the ray parenchyma). Patch-clamp measurements on fibre cell protoplasts, derived from young poplar twigs, identified outward-rectifying K+ channels as the major K+ conductance of this cell type, which resembled the biophysical properties of PTORK when expressed in Xenopus oocytes.

Amino Acid Sequence↗

Aquaporins in poplar: what a difference a symbiont makes!

The formation of ectomycorrhizas, a tight association between fine roots of trees and certain soil fungi, improves plant nutrition in a nutrient-limited environment and may increase plant survival under water stress conditions. To investigate the impact of mycorrhiza formation on plant water uptake, seven genes coding for putative water channel proteins (aquaporins) were isolated from a poplar ectomycorrhizal cDNA library. Four out of the seven genes were preferentially expressed in roots. Mycorrhiza formation resulted in an increased transcript level for three of these genes, two of which are the most prominently expressed aquaporins in roots. When expressed in Xenopus laevis oocytes, the corresponding proteins of both genes were able to transport water. Together, these data indicate, that the water transport capacity of the plasma membrane of root cells is strongly increased in mycorrhized plants. Measurements of the hydraulic conductance of intact root systems revealed an increased water transport capacity of mycorrhized poplar roots. These data, however, also indicate that changes in the properties of the plasma membrane as well as those of the apoplast are responsible for the increased root hydraulic conductance in ectomycorrhizal symbiosis.

Amino Acid Sequence↗

Wood formation in poplar: identification, characterization, and seasonal variation of xylem proteins.

Proteins that are preferentially produced in developing xylem may play a substantial role in xylogenesis. To reveal the identity of these proteins, comparative two-dimensional polyacrylamide gel electrophoresis was performed on young differentiating xylem, mature xylem, and bark of poplar (Populus trichocarpa Hook. cv. 'Trichobel') harvested at different times of the year. The most-abundant xylem proteins were identified by microsequence analysis. For 17 of these proteins a putative function could be assigned based on similarity with previously characterized proteins, and for 15 out of these corresponding expressed sequence tags (ESTs) were found in the poplar EST database. The identified xylem-preferential proteins, defined by comparing the protein patterns from xylem and bark, were all involved in the phenylpropanoid pathway: two caffeoyl-coenzyme A O-methyltransferases (CCoAOMT), one phenylcoumaran benzylic ether reductase (PCBER), one bispecific caffeic acid/5-hydroxyferulic acid O-methyltransferase (COMT), five S-adenosyl-L-methionine synthetases, and one homologue of glycine hydroxymethyltransferase (GHMT). Remarkably, the biological function of the two most-abundant xylem-preferential proteins (PCBER and a GHMT homologue) remains unclear. In addition, several housekeeping enzymes were identified: two enolases, two glutamine synthetases, one 70-kDa heat-shock cognate, one calreticulin, and one alpha-tubulin. In comparison to the xylem-preferential proteins, the housekeeping proteins were expressed at significant levels in the bark as well. Also, several additional protein spots were detected for CCoAOMT, PCBER, and COMT by immunoblot. Our data show that for the study of xylogenesis, two-dimensional protein gel comparisons combined with systematic protein sequencing may yield information complementary to that from EST sequencing strategies.

Amino Acid Sequence↗

A vegetative storage protein homolog is expressed in the growing shoot apex of hybrid poplar.

The ability of poplars (Populus deltoides Bartr. ex Marsh., and Populus trichocarpa Torr. and Gray) to sequester nitrogen in stems in preparation for winter has been associated with the massive accumulation of protein bodies in the bark and xylem ray parenchyma. These protein bodies contain a bark storage protein (BSP) that can account for up to 30% of the total soluble bark protein during the winter months. Perhaps the plant's ability to efficiently cycle nitrogen through BSP is an important aspect of its growth potential. Sequence analysis of BSP led to the identification of a leaf-associated homolog, win4, which was initially isolated because its transcript increased in abundance upon mechanical wounding. The goal of this work was to characterize this putative leaf-associated vegetative storage protein, and determine whether it might perform a storage role in vivo. Antibodies, produced against protein synthesized upon over-expression of the win4 coding region in Escherichia coli, were used to examine the relative abundance of WIN4 protein in response to supplemental nitrogen, and during development. The transcript and protein were most abundant in the youngest leaves and also increased with nitrogen fertilization. Immunolocalization of the protein was performed and showed that WIN4 was associated with cells surrounding the vasculature, and cells of the lower epidermis and stipules of immature leaves. Under moderate nitrogen fertilization regimes, WIN4 accounted for only about 2% of total soluble leaf protein; however, given the cellular specificity and enhancement with nitrogen, the protein is regulated in a manner similar to other vegetative storage proteins. Since poplar is amenable to DNA transformation and regeneration, it is now possible to ask direct questions about the role these proteins play in nitrogen storage in rapidly expanding or in dormant tissue. This type of analysis could determine whether these proteins mainly ameliorate the toxic effects of excess nitrogen, if they are instrumental in controlling nitrogen allocation or if they simply represent an efficient method for sequestering this valuable nutrient.

Cloning, Molecular↗