Fitness dynamics within a poplar hybrid zone: II. Impact of exotic sex on native poplars in an urban jungle

Trees bearing novel or exotic gene components are poised to contribute to the bioeconomy for a variety of purposes such as bioenergy production, phytoremediation, and carbon sequestration within the forestry sector, but sustainable release of trees with novel traits in large‐scale plantations requires the quantification of risks posed to native tree populations. Over the last century, exotic hybrid poplars produced through artificial crosses were planted throughout eastern Canada as ornamentals or windbreaks and these exotics provide a proxy by which to examine the fitness of exotic poplar traits within the natural environment to assess risk of exotic gene escape, establishment, and spread into native gene pools. We assessed postzygotic fitness traits of native and exotic poplars within a naturally regenerated stand in eastern Canada (Quebec City, QC). Pure natives (P. balsamifera and P. deltoides spp. deltoides), native hybrids (P. deltoides × P. balsamifera), and exotic hybrids (trees bearing Populus nigra and P. maximowiczii genetic components) were screened for reproductive biomass, yield, seed germination, and fungal disease susceptibility. Exotic hybrids expressed fitness traits intermediate to pure species and were not significantly different from native hybrids. They formed fully viable seed and backcrossed predominantly with P. balsamifera. These data show that exotic hybrids were not unfit and were capable of establishing and competing within the native stand. Future research will seek to examine the impact of exotic gene regions on associated biotic communities to fully quantify the risk exotic poplars pose to native poplar forests.     

Effects of flooding on leaf development,transpiration, and photosynthesis in narrowleaf cottonwood,a willow-like poplar

The narrowleaf cottonwood, Populus angustifolia, occurs in occasionally flooded, low elevation zones along river valleys near the North American Rocky Mountains. This small poplar has narrow leaves and fine branching and thus resembles willows, which are commonly flood-tolerant. We investigated the flood response of narrowleaf cottonwoods and a related native hybrid, jackii cottonwood (P. × jackii = P. balsamifera × P. deltoides), by studying saplings of 24 clones in a greenhouse, with some pots being inundated to provide the flood treatment. Flooding slightly reduced leaf numbers (−10%), and leaf sizes were reduced by about 21% in female P. angustifolia versus a 50% reduction in the female hybrids. Flooding-reduced stomatal conductance and net photosynthetic rate, and reduced transpiration particularly in P. × jackii. The effects on foliar gas exchange declined over a 5-week interval, suggesting compensation. The moderate impact of flooding supports the hypothesis that narrowleaf cottonwoods are flood-tolerant, and we anticipate that these trees could provide traits to increase the flood tolerance of fast-growing hybrid poplars. The results further indicate that female cottonwoods may be more flood-tolerant than males, and females could be more successful in lower, flood-prone sites.     

Allelic ladders and reference genotypes for a rigorous standardization of poplar microsatellite data

A correct identification of members of the poplar hybrid complex Populus × canadensis is essential in breeding programs and studies in introgressive gene flow. Molecular marker protocols have been developed for this purpose. However, due to missing standards, these techniques have so far not been suited to the transfer of results between different laboratories. We present here a powerful system of nuclear microsatellite DNA (nSSR) fingerprints, standardized by allelic ladders and reference genotypes. Seven nSSR loci provided fingerprints of 65 commercial poplar clones. Their alleles were used to construct allelic ladders. Thus, a first standardized register of poplar clones is now available. All procedures were optimized according to simplified DNA extraction protocols, multiplexed PCR and electrophoresis procedures. Corresponding data originating from two different electrophoretic platforms in different laboratories were congruent when the allelic ladder was used. Unambiguous differentiation of the clones was based on a very low probability of identity (PI) of 1.95 × 10⁻⁸. Our results revealed discrepancies between clone denotations and genetic fingerprints. This suggests that, potentially, members of the clone collection could have been mixed up, thus confirming the demand for rigorous standards. The protocol presented can be exploited in a manifold way, e.g. to enlarge the present clonal molecular data base, or to use it for purposes of certification and control. Furthermore, the allelic ladders are recommended for use in poplar population genetic studies across different laboratories. The allelic ladders and single sample reference genotypes can be obtained on demand.     

Allometric Biomass,Biomass Expansion Factor and Wood Density Models for the OP42 Hybrid Poplar in Southern Scandinavia

Biomass and biomass expansion factor functions are important in wood resource assessment, especially with regards to bioenergy feedstocks and carbon pools. We sampled 48 poplar trees in seven stands with the purpose of estimating allometric models for predicting biomass of individual tree components, stem-to-aboveground biomass expansion factors (BEF) and stem basic densities of the OP42 hybrid poplar clone in southern Scandinavia. Stand age ranged from 3 to 31 years, individual tree diameter at breast height (dbh) from 1.2 to 41 cm and aboveground tree biomass from 0.39 to 670 kg. Models for predicting total aboveground leafless, stem and branch biomass included dbh and tree height as predictor variables and explained more than 97 % of the total variation. The BEF was approaching 2.0 for the smallest trees but declined with increasing tree size and stabilized around 1.2 for trees with dbh >10 cm. Average stem basic density was more than 400 kg m^?3 for the smallest trees but declined with increasing tree height and stabilized around 355 kg m^?3, at a tree height of about 20 m. Existing biomass functions from the literature all underestimated the measured sample tree biomass. Possible explanations include not only differences in competition among trees in the examined stands and site conditions but also differences in sampling procedures. We observed that basic density increased with height above the ground. This trend may have led to the observed underestimation by existing biomass functions including only few samples from the lower end of the stems.     

Above- and below-ground characteristics associated with wind toppling in a young Populus plantation

 Damage from a dormant-season windstorm in a 3-year-old Populus research trial differed among four clones and three spacings and between monoclonal and polyclonal plots. Clonal differences in susceptibility to toppling (or leaning) were associated with both above- and below-ground characteristics. Susceptible clones had less taper in the lower stem and more weight in branches on the upper stem. The most susceptible clone also had the most above-ground biomass per unit of cross-sectional root area. The other susceptible clone had the least root system development in the windward quadrants. Wind toppling was least at the closest spacing. Apparently, mutual support was more important than individual tree characteristics from which the most damage would be expected at the closest spacing. Differences between paired trees of the same clone and spacing which did or did not topple were primarily associated with distribution of root systems by compass quadrant or depth. At the closest spacing where crown sway would have been minimized, trees which did not topple had greater cross-sectional root area in the windward direction than trees which did topple. At the widest spacing where crown sway would have been greatest, windfirm trees had greater cross-sectional root area than non-windfirm trees in both the windward and leeward directions. Toppling was reduced in polyclonal plots; this reduction may have been the result of more rapid stand differentiation in the polyclonal plots or reduction in the “domino effect” by inclusion of more windfirm clones in the mixture. Received: 23 October 1995 / Accepted: 22 February 1996     

Variation of heartwood and sapwood in 18-year-old Eucalyptus globulus trees grown with different spacings

Heartwood and sapwood development was studied in 18-year-old Eucalyptus globulus trees from pulpwood plantations with different spacings (3 × 2, 3 × 3, 4 × 3, 4 × 4 and 4 × 5 m), on cross-sectional discs taken at breast height. The trees possessed a large proportion of heartwood, on average 60% of the wood cross-sectional surface. Spacing was a statistically significant source of variation of heartwood area, which ranged between 99 and 206 cm² for the closer (3 × 2) and wider (4 × 5) spacings, respectively. There was a positive and high statistical significant correlation between heartwood diameter and tree diameter (heartwood diameter = −0.272 + 0.616 dbh; r ² = 0.77; P < 0.001), and larger trees contained more heartwood regardless of spacing. Heartwood proportion in cross-section remained practically constant between spacings but increased with tree diameter class: 55.1, 62.2, 65.0 and 69.5% for diameter at breast height classes <15, 15–20, 20–25 and >25 cm, respectively. The sapwood width did not depend on tree diameter growth and remained practically constant at an average of 18 mm (range 15–21 mm), but sapwood area showed a good linear regression with tree diameter. Therefore, tree growth enhancement factors, such as wide spacings, will induce formation of larger heartwoods that can negatively impact raw-material quality for pulping. The increase in heartwood in relation with tree dimensions should therefore be taken into account when designing forest management guidelines.     

Efficiency of inventory plot patterns for the estimation of woody vegetation recruit density in a tropical dense forest in Bénin

This study assessed the effectiveness of plot patterns for estimating recruit density of woody species in the dense forest of Lama Reserve (Bénin). The experimental design consisted of thirty 0.04 ha plots randomly settled in the forest and each subdivided into four hundred 1‐m² quadrats. Within each quadrat, recruits (dbh ≤10 cm) were counted and saplings (h ≥ 2 m and 2 cm ≤ dbh < 7 cm) and young trees (h ≥ 2 m and 7 cm ≤ dbh < 10 cm) were measured in dbh. In each 0.04 ha plot, seven different plot shapes and sizes were considered by grouping adjacent 1‐m² quadrats. Relationship between mean square error of the estimation of the density of recruitments and the plot sizes was modelled using the Smith law. Results obtained showed an average value of density of recruitments of 10.7 plants/m² with Green index value of 0.01. Shape and size of plots highly influenced the estimation of the density of recruitments. Rectangular plots of length/width = 2 and size of 72 m² (12 m × 6 m) were most efficient for the estimation of the density of recruitments in tropical dense forest with standard error of 0.79 plants/m².     

Comparison of growth, biomass and nutrient distribution in five promising clones of Populus deltoides under an agrisilviculture system

Variations in growth, above- and below-ground biomass and nutrient distribution were examined in five clones (G3, G48, 65/27, D121 and S7C1) of Populus deltoides grown under agrisilviculture system in sub-humid tropics of Central India. The monoclonal blocks were planted at 4x5 m in a randomized block design with three replications. Diameter at breast height (dbh) and tree height were consistently higher in clone 65/27 and lowest in clone S7C1. Mean annual increments (MAI) in dbh and height were 1.6 and 1.3 times higher in clone 65/27 compared to clone S7C1. Total biomass varied from 48.5 to 62.2 Mg ha(-1) in six-year-old clones. In rank order, the total biomass of clones was: 65/27>D121>G48>G3>S7C1. Stem wood accounted 60.4-68.9% to total biomass followed by coarse roots (12.2-18.9%), branches (12.3-15%), leaves (3.02-6.9%) and fine roots (1.5-2.7%). Root-shoot ratio ranged from 0.2 to 0.35. It was highest in clone G48 and lowest in clone S7C1. In six-year-old clones, total N ranged from 184.3 to 266.3 kg ha(-1), P from 16.8 to 31.1 kg ha(-1) and K from 81.9 to 128.7 kg ha(-1). Total N and P were highest in clone 65/27, while K in clone G48. Nutrients were lowest in clone S7C1. In general, maximum nutrients (N, P and K) were allocated to above-ground components (leaves>stem>branches) than below-ground components. Available N, P and K in the soil improved significantly after six years of planting. It was higher in 0-20 cm and decreased with soil depth. At 0-20 cm soil depth, N increased from 14.9% to 24.1%, P from 17.2% to 23.3% and K from 3.1% to 5.1% under different clones. The yield of both soybean and wheat reduced under poplar clones. Yield losses in soybean ranged from 10.1% to 33% and wheat from 15% to 30.3% under different clones. The management strategies for reducing tree-crop competition and nutrient export from the site under P. deltoides based agrisilviculture system for achieving sustainable production are discussed.     

<Emphasis Type="Italic">Dasytricha</Emphasis> Dominance in Surti Buffalo Rumen Revealed by 18S rRNA Sequences and Real-Time PCR Assay

The genetic diversity of protozoa in Surti buffalo rumen was studied by amplified ribosomal DNA restriction analysis, 18S rDNA sequence homology and phylogenetic and Real-time PCR analysis methods. Three animals were fed diet comprised green fodder Napier bajra 21 (Pennisetum purpureum), mature pasture grass (Dicanthium annulatum) and concentrate mixture (20% crude protein, 65% total digestible nutrients). A protozoa-specific primer (P-SSU-342f) and a eukarya-specific primer (Medlin B) were used to amplify a 1,360 bp fragment of DNA encoding protozoal small subunit (SSU) ribosomal RNA from rumen fluid. A total of 91 clones were examined and identified 14 different 18S RNA sequences based on PCR–RFLP pattern. These 14 phylotypes were distributed into four genera-based 18S rDNA database sequences and identified as Dasytricha (57 clones), Isotricha (14 clones), Ostracodinium (11 clones) and Polyplastron (9 clones). Phylogenetic analyses were also used to infer the makeup of protozoa communities in the rumen of Surti buffalo. Out of 14 sequences, 8 sequences (69 clones) clustered with the Dasytricha ruminantium-like clone and 4 sequences (13 clones) were also phylogenetically placed with the Isotricha prostoma-like clone. Moreover, 2 phylotypes (9 clones) were related to Polyplastron multivesiculatum-like clone. In addition, the number of 18S rDNA gene copies of Dasytricha ruminantium (0.05% to ciliate protozoa) was higher than Entodinium sp. (2.0 × 10⁵ vs. 1.3 × 10⁴) in per ml ruminal fluid.     

A fast and simple genetic survey reveals the spread of poplar hybrids at a natural Elbe river site

It is known that various poplar species and cultivated poplar hybrids have the potential to interbreed and produce fertile offspring. Conservation strategies for the genetic resources of the endangered Eurasian black poplar (Populus nigra L.) thus rely on a monitoring which enables the identification and verification of the pure species status. At the same time, the risk of hybrid dispersal and introgressive gene flow has to be estimated. In the present study a combination of two molecular markers, one from chloroplast DNA and the other from nuclear DNA, was applied to evaluate a large P. nigra population on the Elbe River. Hybrid clones of P. × canadensis are scattered within this population and also occur as plantations in the surrounding landscape. By means of the DNA markers the taxonomic status of 208 adult trees in the population and 140 young poplars along the riverbank was monitored. From the analysed young poplars, almost 20 percent were found to exhibit at least one of the two P. deltoides or P. × canadensis diagnostic alleles or genotypes, respectively. Possible vegetative spreads of F1 hybrids and precedent mating scenarios are discussed. Most interestingly we found clear evidence for a small number of backcross hybrids where P.  × canadensis acted as pollen donor. This case had long been debated and thought to be less probable, so far.     

Clonal identification by microsatellite loci in sporadic flowering of a dwarf bamboo species, <Emphasis Type="Italic">Sasa cernua</Emphasis>

Dwarf bamboo species are monocarpic. They flower simultaneously and die after several decades. The type of flowering in the genus Sasa ranges from sporadic to gregarious. In order to determine whether or not the sporadic flowering of dwarf bamboo is fixed genetically, we investigated the distribution of clones using eight microsatellite (SSR) loci in a sporadic flowering patch of Sasa cernua Makino, a major dwarf bamboo species found in central Hokkaido. In May 2006, flowering occurred on 60.5% of living culms in a 1600 m² patch. We established a 50 × 10 m study plot in this patch and noted 1267 clumps consisting of 2529 living culms. We investigated all 1267 clumps and identified six multilocus genotypes as clones using five variable SSR loci. All flowering clumps belonged to the same clone. On the other hand, non-flowering vegetative clumps were also discovered to be of the same clone. These data suggest that all flowering culms originated from a single clone of a sporadic flowering patch of S. cernua. Clonal analysis for investigation of sporadic flowering of S. cernua revealed that only a portion of a clone flowers and dies instead of the whole clone.     

Scale and direction of adaptive introgression between black cottonwood (Populus trichocarpa) and balsam poplar (P. balsamifera)

Introgression can introduce novel genetic variation at a faster rate than mutation alone and result in adaptive introgression when adaptive alleles are maintained in the recipient genome over time by natural selection. A previous study from our group demonstrated adaptive introgression from Populus balsamifera into P. trichocarpa in a target genomic region. Here we expand our local ancestry analysis to the whole genome of both parents to provide a comprehensive view of introgression patterns and to identify additional candidate regions for adaptive introgression genomewide. Populus trichocarpa is a large, fast‐growing tree of mild coastal regions of the Pacific Northwest, whereas P. balsamifera is a smaller stature tree of continental and boreal regions with intense winter cold. The species hybridize where they are parapatric. We detected asymmetric patterns of introgression across the whole genome of these two poplar species adapted to contrasting environments, with stronger introgression from P. balsamifera to P. trichocarpa than vice versa. Admixed P. trichocarpa individuals contained more genomic regions with unusually high levels of introgression (19 regions) and also the largest introgressed genome fragment (1.02 Mb) compared with admixed P. balsamifera (nine regions). Our analysis also revealed numerous candidate regions for adaptive introgression with strong signals of selection, notably related to disease resistance, and enriched for genes that may play crucial roles in survival and adaptation. Furthermore, we detected a potential overrepresentation of subtelomeric regions in P. balsamifera introgressed into P. trichocarpa and possible protection of sex‐determining regions from interspecific gene flow.     

NH4 : NO3 nutrition influence on biomass productivity and root respiration of poplar and willow clones

Nitrogen fertilization often improves the yield of intensively managed, short‐rotation coppices. However, information of N nutrition form on the growth of common species and clones used for biomass production is limited. Thus, this study aims at evaluating N form effects on the growth of two Salicaceae clones. Cuttings of the poplar clone Max 4 (Populus maximovizcii × P. nigra) and the willow clone Inger (Salix triandra × S. viminialis) were fertilized in a pot experiment with four ratios of nitrate (NO₃^?) to ammonium (50%, 62.5%, 75% and 87.5% NO₃^? balanced with ammonium (NH₄⁺) to constant total N) for one growing season and under stable soil pH. Plants were harvested for analysis of biomass and morphology of leaves, stem and roots. Respiration of fine and coarse roots (RR) was determined and related to biomass growth. Salix cv. Inger accumulated more total dry matter than Populus cv. Max 4. In both Salicaceae clones, the total biomass was significantly influenced by the nitrate ratio and greatest in plants fertilized with 50% NO₃^? of the total N supply. Both clones possess a different leaf and root morphology, but no significant influence of the NO₃^? ratio on the morphology was found. Fine RR rates differed significantly between clones, with significantly greater fine RR in Max 4; 87.5% NO₃^? fertilization increased the fine RR. Fine RR and total accumulated plant biomass were closely related. Our study is the first to show the tremendous influence of fine root respiration, especially including the carbon‐intensive reduction of NO₃^? to NH₄⁺, on the aboveground growth of Salicaceae clones. Ways to improve yield in SRC are thus to lower the assimilate consumption by fine roots and to match fertilization regimes to the used clones or vice versa.     

Relationship between genotype and soil environment during colonization of poplar roots by mycorrhizal and endophytic fungi

Poplars are among the few tree genera that can develop both ectomycorrhizal (ECM) and arbuscular (AM) associations; however, variable ratios of ECM/AM in dual mycorrhizal colonizations were observed in the roots of a variety of poplar species and hybrids. The objective of our study was to analyze the effect of internal and external factors on growth and dual AM and ECM colonization of poplar roots in three 12–15-year-old common gardens in Poland. We also analyzed the abundance of nonmycorrhizal fungal endophytes in the poplar roots. The Populus clones comprised black poplars (Populus deltoides and P. deltoides × Populus nigra), balsam poplars (Populus maximowiczii × Populus trichocarpa), and a hybrid of black and balsam poplars (P. deltoides × P. trichocarpa). Of the three sites that we studied, one was located in the vicinity of a copper smelter, where soil was contaminated with copper and lead. Poplar root tip abundance, mycorrhizal colonization, and soil fungi biomass were lower at this heavily polluted site. The total mycorrhizal colonization and the ratio of ECM and AM colonization differed among the study sites and according to soil depth. The influence of Populus genotype was significantly pronounced only within the individual study sites. The contribution of nonmycorrhizal fungal endophytes differed among the poplar clones and was higher at the polluted site than at the sites free of pollution. Our results indicate that poplar fine root abundance and AM and ECM symbiosis are influenced by environmental conditions. Further studies of different site conditions are required to characterize the utility of poplars for purposes such as the phytoremediation of polluted sites.     

The effects of agroforestry practices on fiber attributes in <Emphasis Type="Italic">Populus nigra</Emphasis> var. <Emphasis Type="Italic">betulifolia</Emphasis>

The effects of intercropping with alfalfa and initial tree spacing on fiber properties in Populus nigra var. betulifolia are reported on the basis of a study of 15 trees. The trees were harvested from an agroforestry trial near Karaj city, Iran. Treatments included four initial spacings of 3 m × 4 m, 3 m × 6.7 m, 3 m × 8 m, and 3 m × 10 m intercropped with alfalfa as nitrogen-fixing plant, as well as one 3 m × 4 m treatment without alfalfa. Results showed that initial spacing and intercropping with alfalfa increased the growth rate. Also, alfalfa-intercropping had increasing effects on all fiber dimensions, especially on diameter and wall thickness. The effect of initial spacing on fiber attributes based on ring numbers was not statistically significant; however, whole-disk fiber length, weighted by ring area, was increased with stem diameter. The reason for this increase may be traced to the fact that much of the basal area of fast-growing trees is concentrated in farther rings from the pith that have longer fibers. Clear correlation was not found between fiber length and growth rate and therefore, no firm conclusion can be made in this regard.     

Stoichiometric differences in food quality: impacts on genetic diversity and the coexistence of aquatic herbivores in a <Emphasis Type="Italic">Daphnia</Emphasis> hybrid complex

The maintenance of genetic and species diversity in an assemblage of genotypes (clones) in the Daphnia pulex species complex (Cladocera: Anomopoda) in response to variation in the carbon:phosphorus ratio (quantity and quality) of the green alga, Scenedesmus acutus, was examined in a 90-day microcosm competition experiment. Results indicated that mixed assemblages of seven distinct genotypes (representing clonal lineages of D. pulex, D. pulicaria and interspecific hybrids) showed rapid loss of genetic diversity in all treatments (2 × 2 factorial design, high vs. low quantity, and high vs. low quality). However, the erosion of diversity (measured as the effective number of clones) was slowest under the poorest food conditions (i.e., low quantity, low quality) and by the conclusion of the experiment (90 days) had resulted in the (low, low) treatment having significantly greater genetic diversity than the other three treatments. In addition, significant genotype (clone) × (food) environment interactions were observed, with a different predominant species/clone found under low food quality versus high food quality (no significant differences were detected for the two food quantities). A clone of D. pulex displaced the other clones under low food quality conditions, while a clone of D. pulicaria displaced the other clones in the high food quality treatments. Subsequent life-history experiments were not sufficient to predict the outcome of competitive interactions among members of this clonal assemblage. Our results suggest that genetic diversity among herbivore species such as Daphnia may be impacted not only by differences in food quantity but also by those in food quality and could be important in the overall maintenance of genetic diversity in natural populations. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Successful hybridisation of normally incompatible hybrid aspen (Populus tremula?��?P. tremuloides) and eastern cottonwood (P. deltoides)

Hybrid aspen (Populus tremula × P. tremuloides) belong to the section Populus. Eastern cottonwood (P. deltoides) is a member of the section Aigeiros within the genus Populus. These poplar sections are generally considered to be incompatible. Here, we describe successful hybridisation between these parents, producing an offspring family with 27 individuals. The hybrid character of individuals was proven by genotypes at 16 nuclear microsatellite loci. One individual was suspected to have more than the diploid chromosome number of 2n = 38 due to the observation of more than two alleles at several loci. This individual is a triploid, ascertained by flow cytometry. Two distinct growth classes of tall and dwarf plants were observed in the progeny, reflecting different degrees of postzygotic incompatibility. Two loci linked to the tested microsatellites have an effect on height growth. Some fast-growing individuals were micropropagated to test them for biomass performance together with other clones in field trials.