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PubMed2005

Hybrid poplar clones with Populus maximowiczii parentage demonstrate postoviposition antibiosis to Cryptorhynchus lapathi (Coleoptera: Curculionidae).

Damage caused by the poplar-and-willow borer, Cryptorhynchus lapathi (L.) (Coleoptera: Curculionidae), is reported to vary among hybrid poplar clones. We evaluated oviposition preferences and larval success in four hybrid poplars on potted and field-planted trees. Oviposition occurred somewhat less frequently and abundantly on two clones with Populus maximowiczii Henry parentage in field-planted and potted trees, and significantly fewer larvae survived to adulthood on those clones. No adults emerged from field-planted NM 6 (Populus nigra L. x P. maximowiczii) and four emerged from TM 256-28 (Populus trichocarpa Torrey & Gray x P. maximowiczii) on which male-female pairs of C. lapathi had been caged. In contrast, 50 and 140 adults emerged over the same 2-yr period from two susceptible clones (n = 20), TD 52-226 (P. trichocarpa x Populus deltoides Bartram ex Marshall) and TN 302-9 (P. trichocarpa x P. nigra), respectively. Thus, resistance expressed by clones with P. maximowiczii parentage partially involves decreased levels of oviposition, but more significantly, antibiosis in resistant clones prevents the development of larvae, probably in early spring.

Animals↗

PubMed2000

Leaf age-dependent differences in sulphur assimilation and allocation in poplar (Populus tremula x P. alba) leaves.

35S-sulphate was flap-fed to poplar leaves of different leaf development stages - young developing, expanding, mature, and old mature poplar leaves. (35)S-sulphate was taken up independent of the leaf development stage. Whereas young development leaves did not export the (35)S taken up, export increased with increasing leaf development stage. Expanding leaves allocated the exported (35)S mainly into apical tree parts (73-87%) and only to a minor extent (13-27%) in basipetal direction. Neither lower trunk sections nor the roots were sinks for the exported (35)S. Expanding and developing leaves, but not the shoot apex, were the main sinks for the (35)S allocated in apical direction. In contrast, mature and old mature leaves exported the (35)S taken up mainly in basipetal direction (65-82%) with the roots constituting the main sinks. The (35)S allocated into apical tree parts was found in expanding and developing leaves, but only to a minor extent in the shoot apex. Apical allocated (35)S was identified as sulphate. Apparently the demand of young developing leaves for reduced sulphur was not fulfilled by mature leaves. Therefore, reduced sulphur for growth and development of young developing leaves must be supplied from other sources. In vitro activity of enzymes involved in assimilatory sulphate reduction was measured to investigate whether demand for reduced sulphur by young leaves is met by their own sulphate reduction. ATP sulphurylase and APS reductase activities were not significantly lower in developing than in mature leaves. Sulphite reductase and serine acetyltransferase activities were highest in developing leaves; O:-acetylserine (thiol) lyase activity was similar in all leaf developing stages. Apparently, young developing poplar leaves are able to produce their own reduced sulphur for growth and development. Whether other sources such as storage tissues and/or roots are involved in reduced sulphur supply to developing leaves remains to be elucidated.

Biological Transport↗

PubMed2004

Characterization of full-length enriched expressed sequence tags of stress-treated poplar leaves.

Poplar, whose genome is the first to be sequenced among woody plants, is a favorable model for plant biologists to enable them to understand molecular processes of growth, development and responses to environmental stimuli in trees. The sequence will allow the development of a strategy for improving environmental stress tolerance in forest trees. In this study, we have generated a full-length enriched cDNA library from leaves of axenically grown poplar (Populus nigra var. italica) subjected to environmental stress treatments by dehydration, high salinity, chilling, heat, abscisic acid (ABA) and H2O2. We sequenced >30,000 expressed sequence tags (ESTs) from the cDNA library and consequently collected approximately 4,500 non-redundant clones. We further analyzed cDNAs encoding an ERF/AP2-domain transcription factor which is specific in plants and plays an important role under stress. Thirteen candidates containing the ERF/AP2 domain were found within our EST resource. Some of them showed stress-responsive gene expression. We report here the first collection of full-length enriched stress-related ESTs of poplar and discuss environmental stress responses of forest trees in the light of comparative genomics.

Abscisic Acid↗

PubMed1994

Differentiation of poplar and willow clones using RAPD fingerprints.

The technique of random amplified polymorphic DNA (RAPD) fingerprinting was used to differentiate species and identify clones of 15 poplar and 15 willow clones (ramets of three poplar clones were also included to verify the stability of the results). Four random DNA primers (Deca-11, Chl-1, Chl-4 and Chl-10) and an M13 universal primer were used to determine DNA polymorphism among the clones. Based on the DNA banding pattern obtained with the four DNA primers by polymerase chain reaction (PCR) amplification we conclude that RAPD fingerprinting with properly selected primers can be used for clonal and species differentiation of poplar and willow. The technique is simple, accurate and inexpensive.

Journal Article↗

PubMed1994

ELISA Quantitation and GC-MS identification of abscisic acid in stigma, ovary and pedicel of pollinated poplar flowers (Populus nigra L.).

A method of fractionation, quantitation and identification of abscisic acid (ABA) in poplar flowers is presented in the context of intra- and interspecific pollination studies in Populus nigra x P. nigra and P. nigra x P. deltoides, respectively. Poplar flowers were dissected into stigma, ovary and pedicel. Abscisic acid concentrations were measured at receptivity and 5 days after pollination in each part of the flower. Abscisic acid was fractionated by reverse-phase HPLC followed by an enzyme-linked immunosorbent assay (ELISA) with anti-ABA polyclonal antibody and GC-MS identification of the immunoreactive ABA fractions. Samples were fractionated and ABA was identified by the presence of a cross-reactive metabolite of ABA in the extracts. The interspecific cross (P. nigra x P. deltoides) had a high concentration of ABA in the pedicel, where abscission occurred, whereas in the intraspecific cross, ABA concentration in the pedicel was low. The results are discussed with reference to the control ABA exerts on the abscission of poplar flowers after interspecific incompatible pollinations.

Journal Article↗

PubMed1997

Simulating the response of mature yellow poplar and loblolly pine trees to shifts in peak ozone periods during the growing season using the TREGRO model.

Multiple TREGRO simulations were conducted with meteorological data files containing different growing season peak ozone (O(3)) episodes at O(3) exposures of 1.0 and 2.0 x ambient O(3) to assess the relationship between O(3) response and the phenology of mature yellow poplar (Liriodendron tulipifera L.) and loblolly pine (Pinus taeda L.) trees. Regardless of O(3) exposure and peak O(3) episode occurrence, a peak O(3) episode in August caused the greatest reduction in carbon (C) gain in yellow poplar, whereas a peak O(3) episode in July caused the greatest reduction in C gain of loblolly pine. In both species, timing of the greatest simulated O(3) effect corresponded with the completion of the annual foliage production phenophase. Simulated C gain of yellow poplar (total tree, coarse root, and total nonstructural carbohydrate) was reduced by O(3) to a greater extent than the corresponding compartments in loblolly pine, but the opposite was true for fine roots. This differential sensitivity to O(3) reflects the fact that both C assimilation and the O(3) response of the species were parameterized according to observed field measurements of each species. The differential sensitivity to O(3) of these species may have long-term implications for species composition in southeastern USA forests.

Journal Article↗

PubMed2004

Nitrogen mobilization, nitrogen uptake and growth of cuttings obtained from poplar stock plants grown in different N regimes and sprayed with urea in autumn.

Nitrogen mobilization, nitrogen uptake and growth of cuttings obtained from poplar stock plants fertigated with different nitrogen (N) treatments and sprayed with urea in autumn were studied. Stock plants propagated from poplar cuttings were trained to a single shoot and fertigated with 0, 5, 10, 15 or 20 mmol l(-1) N during the first growing season. In October, a subset of stock plants from each N fertigation treatment was sprayed twice with either 3% urea or water, and overwintered outside. In March, total tree biomass and total N concentration and content of stems were estimated for stock plants in each treatment, and cuttings were taken from the middle of each stock plant and stored in plastic bags at 2 degrees C. In mid-April, cuttings were planted in 7.5-l pots containing N-free medium and grown outdoors with a weekly fertigation with nutrient solution containing 0 or 10 mmol l(-1) 15NH4 15NO3. In mid-July, cuttings were harvested, and new shoot (new stems and leaves), shank (old cutting stem) and roots were analyzed for new biomass growth and total N and 15N content. Growth of stock plants was positively related to N supply in the previous growing season. Foliar urea application in autumn had no effect on subsequent stock plant growth even though urea sprays increased both N concentration and content in stem tissues. Biomass growth of cuttings obtained from stock plants was closely related to their N content when the cuttings were grown in an N-free medium regardless of previous treatments applied to the stock plants. When N was supplied in the growth medium, the strength of the relationship between regrowth and N content of cuttings was significantly reduced. Cuttings from stock plants treated with foliar urea and grown in a N-free medium remobilized between 75 and 82% of their total N for new growth, whereas cuttings from plants receiving no urea spray remobilized only between 60 and 69% of their total N for new growth. Current N fertilization of the cuttings reduced the percentage of N remobilized. We conclude that new growth of poplar cuttings in spring was more dependent on currently applied N than on reserve N, and urea N applied as a spray in autumn was more easily remobilized than N taken up by roots during the previous season.

Nitrogen↗

PubMed2005

Differential expression of a poplar copper chaperone gene in response to various abiotic stresses.

Copper chaperone (CCH) is upregulated during Arabidopsis (Arabidopsis thaliana L. Columbia) leaf senescence, suggesting that it mobilizes certain metal ions in leaves and transports them to other growing parts of the plants. The CCHs are also involved in defense mechanisms against oxidative stress in Arabidopsis and tomato (Lycopersicon esculentum Mill. cv. 'Ailsa Craig'). To elucidate the functions of CCH in poplar, we cloned a CCH gene (PoCCH) from Populus (Populus alba x P. tremula var. glandulosa) suspension cells and tested its expression in response to various treatments including heavy metals, plant growth regulators and abiotic stresses. The PoCCH cDNA is 540 bp in length, including a 55-bp 5' noncoding domain, a 258-bp open reading frame (ORF), and 227-bp 3' termination region. The coding region of PoCCH represents a putative 85-amino-acid protein with a molecular weight of 8.9 kDa. The deduced amino acid sequence of the PoCCH gene product is 87 and 78% identical to those of tomato and Arabidopsis, respectively, with a high degree of conservation in both the metal-binding and lysine-rich regions. However, the PoCCH gene product lacks the C-terminal extension identified in Arabidopsis CCH. Southern blot analysis suggested that the PoCCH gene is present in low copy numbers in poplar. The expression of PoCCH increased under copper deprivation conditions. The expression of PoCCH was down-regulated by high concentrations of copper, whereas some metals, such as aluminum and zinc, markedly induced PoCCH, and others including cadmium, cobalt and lead had little effect on PoCCH expression. The plant growth regulator jasmonic acid caused an increase in PoCCH transcript whereas abscisic acid, salicylic acid and gibberellic acid did not. The gene was highly induced when cells were exposed to physical stress by high-speed agitation on a gyratory shaker. Other effective inducers of PoCCH expression in suspension culture were methyl viologen and NaCl. Thus, PoCCH does not respond to all stresses, but responds specifically to certain metals and abiotic stresses that induce oxidative damage. Our results suggest that JA is involved in regulating PoCCH expression in poplar cells.

Adaptation, Physiological↗

PubMed1991

A device for studying the influence of declining water table on poplar growth and survival.

Riparian poplar forests are declining downstream from many dams in the western prairie regions of North America. Drought-induced seedling and sapling mortality caused by abrupt drops in water table level following river flow regulation may be contributing to this forest decline. Here we describe a device, the rhizopod, for studying the influence of a changing water table on poplar seedling survival and growth. Seeds of a natural poplar hybrid (Populus deltoides x P. balsamifera) were planted in five rhizopods and treated to water table declines of 0, 1, 2, 4, or 8 cm day(-1). Each rhizopod consisted of 15 growth tubes connected to a central water reservoir through which the water level was regulated. Seedling survival was over 90% in rhizopods in which water table declined by 0, 1 or 2 cm day(-1), but was reduced to about 40% and less than 25% in rhizopods in which the water table level declined by 4 and 8 cm day(-1), respectively. Maximum shoot height, leaf number, and leaf area were observed in the rhizopod with a constant water table and decreased progressively with increasing rates of water table decline. Maximum root mass and length were observed in plants subjected to declines in water table level of 0 and 1 cm day(-1), respectively.

Journal Article↗

PubMed2026

A Leucine-Rich Repeat Receptor-Like Protein Associated with a QTL for Septoria Stem Canker in Populus trichocarpa × Populus deltoides Hybrid Poplar.

The fungal plant pathogen Sphaerulina musiva (Ascomycota) causes Septoria stem canker, the most economically damaging disease of Populus plantations in North America, yet the genetic determinants of host resistance remain uncharacterized in hybrid poplar. Using an inoculation experiment with the 52-124 pseudo-backcross family of Populus trichocarpa × Populus deltoides (TD × D) hybrid poplar, a single significant QTL was identified on Chromosome 16 (LOD = 4.93) associated with both stem canker count and disease severity score. Transcriptomic analysis of two resistant and two susceptible genotypes across a 72-hour infection time course identified a single differentially expressed gene within the QTL candidate gene window: Podel.16G125900, a putative leucine-rich repeat receptor-like protein (LRR-RLP) with homology to receptor-like protein 33 in Arabidopsis thaliana. Podel.16G125900 is located 3001 bp (0.019 cM) upstream of the QTL peak and showed a strong infection-induced upregulation in susceptible genotype 852 (log2 fold-change = 20.47) and higher baseline expression in resistant genotypes relative to susceptible genotypes across all infection time points, consistent with a resistance mechanism in which expression level contributes to the degree of resistance conferred. Two P. trichocarpa homologs were not differentially expressed and differ substantially in sequence content, suggesting the resistance function is specific to the resistant P. deltoides lineage. These findings identify Podel.16G125900 as a strong candidate gene underlying quantitative resistance mechanisms modulating Septoria stem canker resistance in the 52-124 family of TD × D hybrid poplar and provide a target for future functional validation and marker-assisted resistance breeding.

Disease Resistance↗

PubMed1993

Modification of Rubisco and Altered Proteolytic Activity in O3-Stressed Hybrid Poplar (Populus maximowizii x trichocarpa).

Exposing hybrid poplar (Populus maximowizii x trichocarpa) plants to ozone (O3) resulted in an acceleration of the visual symptoms of senescence and a decrease in the activity and quantity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Whole plants, crude leaf extracts, and isolated intact chloroplasts of hybrid poplar clone 245 were used to test the hypothesis that O3-induced structural modifications of Rubisco affect the activity of this key photosynthetic enzyme. Proteolytic activity, per se, could not account for losses in Rubisco; acidic and alkaline protease activities declined or were unaffected in foliage of O3-treated poplar saplings. In vitro treatment of leaf extracts with O3 decreased total Rubisco activity and binding of the enzyme's transition-state analog, 2-carboxyarabinitol bisphosphate. Additionally, O3 increased the loss of Rubisco large subunit (LSU) when extracts were incubated at 37[deg]C. Treatment of isolated intact chloroplasts with O3 accelerated both the loss of the 55-kD Rubisco LSU and the accumulation of Rubisco LSU aggregates, as visualized by immunoblotting. The time-dependent modification in Rubisco structure was the primary response of the isolated organelles to O3 treatment, with little proteolytic degradation of the LSU detected.

Journal Article↗

PubMed1993

Structure, inheritance, and expression of hybrid poplar (Populus trichocarpa x Populus deltoides) phenylalanine ammonia-lyase genes.

A heterologous probe encoding phenylalanine ammonia-lyase (PAL) was used to identify PAL clones in cDNA libraries made with RNA from young leaf tissue of two Populus deltoides x P. trichocarpa F1 hybrid clones. Sequence analysis of a 2.4-kb cDNA confirmed its identity as a full-length PAl clone. The predicted amino acid sequence is conserved in comparison with that of PAL genes from several other plants. Southern blot analysis of popular genomic DNA from parental and hybrid individuals, restriction site polymorphism in PAL cDNA clones, and sequence heterogeneity in the 3' ends of several cDNA clones suggested that PAL is encoded by at least two genes that can be distinguished by HindIII restriction site polymorphisms. Clones containing each type of PAL gene were isolated from a poplar genomic library. Analysis of the segregation of PAL-specific HindIII restriction fragment-length polymorphisms demonstrated the existence of two independently segregating PAL loci, one of which was mapped to a linkage group of the poplar genetic map. Developmentally regulated PAL expression in poplar was analyzed using RNA blots. Highest expression was observed in young stems, apical buds, and young leaves. Expression was lower in older stems and undetectable in mature leaves. Cellular localization of PAL expression by in situ hybridization showed very high levels of expression in subepidermal cells of leaves early during leaf development. In stems and petioles, expression was associated with subepidermal cells and vascular tissues.

Amino Acid Sequence↗

PubMed1996

Red Xylem and Higher Lignin Extractability by Down-Regulating a Cinnamyl Alcohol Dehydrogenase in Poplar.

Cinnamyl alcohol dehydrogenase (CAD) catalyzes the last step in the biosynthesis of the lignin precursors, the monolignols. We have down-regulated CAD in transgenic poplar (Populus tremula X Populus alba) by both antisense and co-suppression strategies. Several antisense and sense CAD transgenic poplars had an approximately 70% reduced CAD activity that was associated with a red coloration of the xylem tissue. Neither the lignin amount nor the lignin monomeric composition (syringyl/guaiacyl) were significantly modified. However, phloroglucinol-HCl staining was different in the down-regulated CAD plants, suggesting changes in the number of aldehyde units in the lignin. Furthermore, the reactivity of the cell wall toward alkali treatment was altered: a lower amount of lignin was found in the insoluble, saponified residue and more lignin could be precipitated from the soluble alkali fraction. Moreover, large amounts of phenolic compounds, vanillin and especially syringaldehyde, were detected in the soluble alkali fraction of the CAD down-regulated poplars. Alkaline pulping experiments on 3-month-old trees showed a reduction of the kappa number without affecting the degree of cellulose degradation. These results indicate that reducing the CAD activity in trees might be a valuable strategy to optimize certain processes of the wood industry, especially those of the pulp and paper industry.

Journal Article↗

PubMed1998

4-Coumarate:coenzyme A ligase in hybrid poplar. Properties of native enzymes, cDNA cloning, and analysis of recombinant enzymes.

The enzyme 4-coumarate:coenzyme A ligase (4CL) is important in providing activated thioester substrates for phenylpropanoid natural product biosynthesis. We tested different hybrid poplar (Populus trichocarpa x Populus deltoides) tissues for the presence of 4CL isoforms by fast-protein liquid chromatography and detected a minimum of three 4CL isoforms. These isoforms shared similar hydroxycinnamic acid substrate-utilization profiles and were all inactive against sinapic acid, but instability of the native forms precluded extensive further analysis. 4CL cDNA clones were isolated and grouped into two major classes, the predicted amino acid sequences of which were 86% identical. Genomic Southern blots showed that the cDNA classes represent two poplar 4CL genes, and northern blots provided evidence for their differential expression. Recombinant enzymes corresponding to the two genes were expressed using a baculovirus system. The two recombinant proteins had substrate utilization profiles similar to each other and to the native poplar 4CL isoforms (4-coumaric acid > ferulic acid > caffeic acid; there was no conversion of sinapic acid), except that both had relatively high activity toward cinnamic acid. These results are discussed with respect to the role of 4CL in the partitioning of carbon in phenylpropanoid metabolism.

Amino Acid Sequence↗

PubMed1998

Purification and characterization of peroxidases correlated with lignification in poplar xylem.

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.

Amino Acid Sequence↗

PubMed2000

Polyphenol oxidase from hybrid poplar. Cloning and expression in response to wounding and herbivory.

The inducible expression of polyphenol oxidase (PPO), a presumed antiherbivore enzyme, was examined in hybrid poplar (Populus trichocarpa x Populus deltoides). Following mechanical wounding simulating insect damage, PPO activity increased dramatically in wounded and unwounded leaves on wounded plants beginning at 24 and 48 h, respectively. A hybrid poplar PPO cDNA was isolated and its nucleotide sequence determined. On northern blots, PPO transcripts were detected within 8 h of wounding, and reached peak levels at 16 and 24 h in wounded and unwounded leaves, respectively. Methyl jasmonate spray and feeding by forest tent caterpillar also induced PPO expression. The induction of PPO was strongest in the youngest four leaves, which were generally avoided by caterpillars in free feeding experiments. This wound- and herbivore-induced expression of PPO in hybrid poplar supports the defensive role of this protein against insect pests.

Acetates↗

PubMed2000

Essential role of caffeoyl coenzyme A O-methyltransferase in lignin biosynthesis in woody poplar plants.

Caffeoyl coenzyme A O-methyltransferase (CCoAOMT) has recently been shown to participate in lignin biosynthesis in herbacious tobacco plants. Here, we demonstrate that CCoAOMT is essential in lignin biosynthesis in woody poplar (Populus tremula x Populus alba) plants. In poplar stems, CCoAOMT was found to be expressed in all lignifying cells including vessel elements and fibers as well as in xylem ray parenchyma cells. Repression of CCoAOMT expression by the antisense approach in transgenic poplar plants caused a significant decrease in total lignin content as detected by both Klason lignin assay and Fourier-transform infrared spectroscopy. The reduction in lignin content was the result of a decrease in both guaiacyl and syringyl lignins as determined by in-source pyrolysis mass spectrometry. Fourier-transform infrared spectroscopy indicated that the reduction in lignin content resulted in a less condensed and less cross-linked lignin structure in wood. Repression of CCoAOMT expression also led to coloration of wood and an elevation of wall-bound p-hydroxybenzoic acid. Taken together, these results indicate that CCoAOMT plays a dominant role in the methylation of the 3-hydroxyl group of caffeoyl CoA, and the CCoAOMT-mediated methylation reaction is essential to channel substrates for 5-methoxylation of hydroxycinnamates. They also suggest that antisense repression of CCoAOMT is an efficient means for genetic engineering of trees with low lignin content.

Base Sequence↗

PubMed1990

Factors Influencing the Tissue Culture and the Agrobacterium tumefaciens-Mediated Transformation of Hybrid Aspen and Poplar Clones.

Tissue culture conditions and transformation have been established for both aspen and poplar. The use of previously described culture conditions resulted in shoot tip necrosis in the shoot cultures and necrosis of stem and leaf explants. Shoot tip necrosis could be overcome by buffering the medium with 2-(N-morpholino)ethanesulfonic acid and Ca-gluconate and by growing the shoots below 25 degrees C. Necrosis of the explants was probably due to an accumulation of ammonium in the explants and could be overcome by adapting the NO(3) (-)/NH(4) (+) ratio of the media. Stem explants of established shoot cultures of the aspen hybrid Populus alba x P. tremula and of the poplar hybrid Populus trichocarpa x P. deltoides were cocultivated with Agrobacterium strains having chimeric bar and neo genes on their disarmed tDNAs. Transformed aspen shoots were obtained from 30 to 40% of the explants, while transformed poplar shoots were obtained from 10% of the explants. Extracts from the transformed trees contained high phosphinotricin acetyltransferase and neomycin phosphotransferase activities, and the trees contained one to three copies of the chimeric genes. The transformed trees were completely resistant to the commercial preparations of the herbicide phosphinotricin (glufosinate), while control trees were not.

Journal Article↗