Osservazioni sul comportamento sessuale del Populus tremula L. in Italia

Abstract

OBSERVATIONS ON SEXUAL BEHAVIOUR OF POPULUS TREMULA L. IN ITALY. — A research has been carried on sex distribution of Populus tremula in Italy. On 101 observed populations the sexual ratio resulted of 2:1 in favour of male sex. The analysis of results referred to the two geographycal sectors of the peninsula, North and MiddleSouth, is however showing some discordant data in the sexual composition of several populations. In North-Italy the ratio results of 2,5:1 in favour of male sex, in MiddleSouth 1:1.

These discordant results would show that male sex can be adapted whether in continental or in mountain climate.

The Author deems that the geographycal position of populations and corresponding climatic characteristics can be responsible for the sexual selection, so that the several deviations wich are found in natural populations of P. tremula can get an explanation in an possible correlation between habitat and sex.

The Author dwells particulary upon population discovered at Campiglione, near Pozzuoli (Naples). Stamens, varying in number from 2 to 4, were observed on almost all the pistillate flowers of every single catkin of these individuals and especially from the base to a little over halfway up the catkin.

The high proportion of bisexual catkins found on every individual examined leads to the conclusion that P. tremula is « polygamo-dioecious » in character, that is, it produces bisexual and unisexual character in the case of P. tremuloides, where, however, in the majority of the bisexual individuals, the percentage of hermaphrodite flowers on one catkin was very small (1–2%).

These cases of bisexuality lead one to thing that dioecism is an acquired character in the genus Populus. This condition (wich, at least in the case of P. tremula, is perhaps to be connected with the poor fertility of the species) could in time have lead to the present geographycal distribution in colonies of isolated and sexually anbalanced individuals.

These cold have survived to our times due to their aptitude vegetative propagation by means of suckers, giving rise to scattered clumps of unisexual individuals of the same clone. The finding of these bisexual individuals is of considerable importance for applied genetics, and could initiate new researches in Italy to obtain genotypic and polyploid descendants.     

Genetic linkage mapping in aspen (Populus tremula L. and Populus tremuloides Michx.)

A large number of simple sequence repeat (SSR) marker-containing genetic maps are available for several Populus species. For aspen however, no SSR-containing map has been published so far. In this study, genetic linkage mapping was carried out with an interspecific mapping pedigree of 61 full-sib hybrids of European × quaking aspen (Populus tremula L. × Populus tremuloides Michx.), using the two-way pseudo-testcross strategy. Amplified fragment-length polymorphism (AFLP) and SSR markers were used for mapping, resulting in the first SSR-containing genetic linkage maps for aspen. The maps allow comparisons with a Populus consensus map and other published genetic maps of the genus Populus. The maps showed good collinearity to each other and to the Populus consensus map and provide a direct link to the Populus trichocarpa genomic sequence. Sex as a morphological trait was assessed in the mapping population and mapped on a non-terminal position of linkage group XIX on the male P. tremuloides map.     

Ethylene involvement in in vitro organogenesis and plant growth of Populus tremula L.

The possible involvements of a decrease in medium pH and accumulation of ammonium in ammonium-inhibited growth of rice cells were investigated. Ammonium, applied at concentrations ranging from 20 to 50 mM, markedly inhibited cell growth and decreased medium pH. The accumulation of ammonium in rice cells was observed only when ammonium concentration was 40 mM or higher. Ammonium-inhibited growth was alleviated when medium pH was buffered with MES [2-(N-morpholino)-ethanesulfonic acid]. However, no difference in ammonium level was observed between buffered and unbuffered ammonium-fed rice cells. Succinic acid, -ketoglutaric acid, glutamic acid and glutamine were found to be effective in reversing ammonium-inhibited growth of rice cells and reducing a fall in pH in the ammonium-fed medium. Succinic acid, -ketoglutaric acid and glutamic acid decreased the level of ammonium in ammonium-fed rice cells. However, glutamine was unable to decrease the ammonium level in ammonium-fed rice cells. The current results suggest that a decrease in medium pH is a factor responsible for growth inhibition of ammonium-fed rice cells.     

Chromosome number of Icelandic Populus tremula

Aspen ( Populus tremula ) from two locations in Iceland, representing the north and the east populations, were examined for its chromosome number. The plants all have diploid chromosome number (2n=2x=38), as with most other aspen populations of this species in Europe. No previous record of chromosome count on the Icelandic material is available. The significance of such chromosome study is discussed. — Flora Nordica Notes no. 22.     

Adaptive Introgression Facilitates Adaptation to High Latitudes in European Aspen (Populus tremula L.)

Understanding local adaptation has become a key research area given the ongoing climate challenge and the concomitant requirement to conserve genetic resources. Perennial plants, such as forest trees, are good models to study local adaptation given their wide geographic distribution, largely outcrossing mating systems, and demographic histories. We evaluated signatures of local adaptation in European aspen (Populus tremula) across Europe by means of whole-genome resequencing of a collection of 411 individual trees. We dissected admixture patterns between aspen lineages and observed a strong genomic mosaicism in Scandinavian trees, evidencing different colonization trajectories into the peninsula from Russia, Central and Western Europe. As a consequence of the secondary contacts between populations after the last glacial maximum, we detected an adaptive introgression event in a genome region of ∼500 kb in chromosome 10, harboring a large-effect locus that has previously been shown to contribute to adaptation to the short growing seasons characteristic of Northern Scandinavia. Demographic simulations and ancestry inference suggest an Eastern origin—probably Russian—of the adaptive Nordic allele which nowadays is present in a homozygous state at the north of Scandinavia. The strength of introgression and positive selection signatures in this region is a unique feature in the genome. Furthermore, we detected signals of balancing selection, shared across regional populations, that highlight the importance of standing variation as a primary source of alleles that facilitate local adaptation. Our results, therefore, emphasize the importance of migration–selection balance underlying the genetic architecture of key adaptive quantitative traits.     

Adverse Effects of Epiphytic Crustose Lichens upon Stem Photosynthesis and Chlorophyll of Populus tremula L.

Dry cork layer (phellem) in stems of Populus tremula transmitted 35–55 percent of incident irradiation, depending upon moisture content. A cover of crustose Lecanora lichens reduced transmission through phellem to 10 percent or less of incident irradiation. The bark contains photosynthetically active cells. Apparent quantum yield for photosynthetic O₂-evolution was 0.017 in bark covered with dry Lecanora compared with 0.070 in naked bark. The capacity for gross photosynthesis in high light (1090 μmol photons m^?2 s^?1) was reduced by 50 percent in Lecanora-covered bark. Lecanora did not reduce the ratio between variable and maximal chlorophyll a fluorescence (F_v/F_m). Chlorophyll content per unit area was similar in leaves and naked bark of Populus tremula. The chlorophyll content in the bark decreased with increasing chlorophyll content in Lecanora. Chlorophyll a/b ratio was 2.5 in the bark compared with 4.0 in leaves and in Lecanora, and the ratio decreased down the stems. The a/b ratio was 2.3 in Lecanora covered bark compared with 2.6 in naked bark. The changes in bark photosynthesis below a Lecanora crust were probably due to acclimation of bark photosynthesis to shade, since the lichen acids in the measured lichens neither suppressed photosynthetic O₂-evolution nor changed the F_v/F_m in bark disks.     

Media formulation influences in vitro ectomycorrhizal synthesis on the European aspen Populus tremula L.

The effect of various media formulations on in vitro ectomycorrhizal synthesis of identified fungal strains with European aspen (Populus tremula L.) was tested in Petri dishes. Pre-grown seedlings were transferred to various nutrient media and inoculated with Paxillus involutus isolates using modified sandwich techniques. Mycorrhiza formation was evaluated macroscopically and further confirmed by microscopic examination of semi-thin sections for anatomical features of the mantle and the Hartig net. Standard media formulations did not support successful ectomycorrhiza formation because of either very poor plant survival (below 20%) or impaired fungal growth. The inclusion of micronutrients and vitamins in a Melin Norkrans (MMN)-based medium increased plant survival rate to above 60% and supported successful mycorrhizal synthesis. P. involutus isolates formed mycorrhizas with a characteristic Hartig net restricted to the epidermis. Mantle density and thickness varied depending on the isolate. In a follow-up experiment, the adapted medium supported successful ectomycorrhiza formation by various Laccaria and Hebeloma isolates. Our results show that an exogenous supply of vitamins and micronutrients in the medium was a prerequisite for successful mycorrhization of P. tremula in vitro in Petri dishes.     

Effect of climate change on bud phenology of young aspen plants (Populus tremula. L)

Boreal tree species are excellent tools for studying tolerance to climate change. Bud phenology is a trait, which is highly sensitive to environmental fluctuations and thus useful for climate change investigations. However, experimental studies of bud phenology under simulated climate change outdoors are deficient. We conducted a multifactorial field experiment with single (T, UVA, UVB) and combined treatments (UVA+T, UVB+T) of elevated temperature (T, +2°C) and ultraviolet‐B radiation (+30% UVB) in order to examine their impact on both male and female genotypes of aspen (Populus tremula L.). This study focuses on the effect of the treatments in years 2 and 3 after planting (2013, 2014) and follows how bud phenology is adapting in year 4 (2015), when the treatments were discontinued. Moreover, the effect of bud removal was recorded. We found that elevated temperature played a key role in delaying bud set and forcing bud break in intact individuals, as well as slightly delaying bud break in bud‐removed individuals. UVB delayed the bud break in bud‐removed males. In addition, both UVA and UVB interacted with temperature in year 3 and even in year 4, when the treatments were off, but only in male individuals. Axillary bud removal forced both bud break and bud set under combined treatments (UVA+T, UVB+T) and delayed both under individual treatments (T, UVB). In conclusion, male aspens were more responsive to the treatments than females and that effect of elevated temperature and UV radiation on bud set and bud break of aspen is not disappearing over 4‐year study period.     

Responses of Populus tremula to Picloram and Other Translocated Herbicides

Whole aspen plants and isolated aspen root segments were three to ten times more sensitive to the growth-inhibitory and toxic effects of picloram than to those of 2,4-D, 2,4,5-T and dicamba. The activity of picloram in the inhibition of root growth was about ten times higher than that of 2,4-D and dicamba when tbe substances were added to the nutrient solution. Epinastic responses indicated a very rapid translocation of picioram from the roots to the growing shoot parts. When the herbicides were applied to the mature leaves dicamba rapidly caused a complete inhibition of root growth indicating a rapid translocation of this compound from the leaves to the root tips. Leaf-applied picloram, 2,4-D and 2,4,5-T affected root growth considerably more slowly. Dicamba, 2,4-D and 2,4,5-T rapidly killed the directly treated leaf tissue due to high acute toxicity while picloram did not show this type of toxicity. It is concluded that the higher activity of picloram in killing the plant and in inhibition of root and shoot growth can only partly be explained as a result of greater uptake and translocation of this compound.     

Response of Populus tremula to heterogeneous B distributions in soil

Background

Poplars accumulate inordinate amounts of B in their leaves and are candidate plants for the remediation of B contaminated soil. We aimed to determine the effect of heterogeneous B distribution in soil by comparing the growth and B accumulation of young Populus tremula trees growing in soil with heterogeneous and homogeneous B distributions.

Methods

The first of two experiments focused on the tolerance and B accumulation of P. tremula under heterogeneous soil B distributions, while the second was designed to study fine root growth under such conditions in detail.

Results

Growth and B accumulation of P. tremula were unaffected by the spatial distribution of B. Root and shoot growth were both reduced simultaneously when leaf B concentrations increased above 800 mg kg^?1. In the heterogeneous soil B treatments, root growth was more reduced in spiked soil portions with B concentrations >20 mg kg^?1. Fine root length growth was stronger inhibited by B stress than secondary growth.

Conclusions

The root growth responses of P. tremula to B are primarily a systemic effect induced by shoot B toxicity and local toxicity effects on roots become dominant only at rather high soil B concentrations. Local heterogeneity in soil B should have little influence on the phytoremediation of contaminated sites.     

Adaptive evolution of the Populus tremula photoperiod pathway

Perennial plants monitor seasonal changes through changes in environmental conditions such as the quantity and quality of light and genes in the photoperiodic pathway are known to be involved in controlling these processes. Here, we examine 25 of genes from the photoperiod pathway in Populus tremula (Salicaceae) for signatures of adaptive evolution. Overall, levels of synonymous polymorphism in the 25 genes are lower than at control loci selected randomly from the genome. This appears primarily to be caused by lower levels of synonymous polymorphism in genes associated with the circadian clock. Natural selection appears to play an important role in shaping protein evolution at several of the genes in the photoperiod pathways, which is highlighted by the fact that approximately 40% of the genes from the photoperiod pathway have estimates of selection on nonsynonymous polymorphisms that are significantly different from zero. A surprising observation we make is that circadian clock-associated genes appear to be over-represented among the genes showing elevated rates of protein evolution; seven genes are evolving under positive selection and all but one of these genes are involved in the circadian clock of Populus.     

Enhanced bud regeneration in aspen (Populus tremula L.) roots cultured in liquid media

 The regeneration potential of excised aspen (Populus tremula L.) roots cultivated in liquid medium, as affected by plant growth regulators and by the position of the isolated root explant on the main root, was investigated. The effect of various levels of benzyladenine (BA) and thidiazuron (TDZ) on bud regeneration in root explants was studied. TDZ in the medium had a marked effect on bud development as compared with BA, inducing a tenfold increase in the number of buds regenerated from various root explants. TDZ enhanced both root and root-borne shoot biomass production but reduced further shoot development and elongation. The position of the isolated root sections on the main root affected regeneration, the proximal sections further away from the root tip producing the highest number of buds per explant in both BA and TDZ treatments. Buds regenerated in close proximity to the site of lateral roots in BA-treated roots, while in TDZ-treated root sections, the buds formed all over the root regardless of the presence of lateral roots. The buds developed from inner cortical and sub-epidermal cell layers, disrupting the epidermis and the inner layers. Root biomass production and growth was greatly enhanced in well-aerated bioreactor culture in the presence of 4.5×10^–2 μM TDZ. A high number of the root-borne shoots could be rooted and converted to plantlets. However, while shoots regenerated in a medium with BA rooted well in a growth regulator-free medium, shoots formed in a medium with TDZ required auxin for rooting. Roots cultured in the presence of ancymidol, a gibberellin biosynthesis inhibitor, regenerated non-hyperhydric bud clusters and hyperhydric shoots. These were separated mechanically, subcultured to growth and rooting medium and transplanted ex vitro resulting in phenotypically true-to-type plantlets. The potential of liquid cultures for aspen shoot biomass production from roots is discussed. Received: 24 January 2000 / Revision received: 6 March 2000 / Accepted: 7 March 2000     

Spatial distribution of epiphytes on Populus tremula in relation to dispersal mode

Abstract. We examined if the spatial distribution of epiphytes in a forest stand differs between two sexually and three asexually dispersed species. As study species we have chosen two sexually dispersed species, the lichen Collema curtisporum and the bryophyte Orthotrichum speciosum, and three asexually dispersed species, the lichens C. furfuraceum and Leptogium saturninum and the bryophyte O. obtusifolium. All of these species grow on Populus tremula. In P. tremula stands, individual trees may be regarded as distinct patches for the studied species, while the stand represents a “landscape” of discrete patches. The study was performed in two relatively homogeneous forest stands. The study revealed that the asexually dispersed species were more common than the sexually dispersed. It was also shown that the sexually dispersed species were more aggregated than P. tremula in both stands. In contrast, the distribution of the asexually dispersed species mirrors, more or less, the distribution of their host, i.e. the spatial pattern shown by P. tremula. It is unlikely that the measured environmental variables strongly influenced the observed spatial distribution. Thus, we suggest that the different dispersal and establishment strategies were important in shaping the local distribution of the species.     

Dependence of Root Growth on Photosynthesis in Populus tremula

Hoot elongation of rooted cuttings of aspen, Populus tremula L., was followed on moist filter paper. The time course of root growth during treatments which decrease or interrupt the supply of carbohydrates to the roots was studied. At light intensities below 4000 lux the rate of root elongation was found to berelated to the light intensity over the leaves. In the dark, root growth decreased within 24 hours and the roots stopped growing completely during the second or third day. Excision of the leaves or steam-killing of the tissue in a stem section below the leaves was followed by root growth stoppage within 24 hours. These results imply that the rate of root growth under certain conditions is determined by the supply of carbohydrates from the leaves.     

Phylogeography of Populus alba (L.) and Populus tremula (L.) in Central Europe: secondary contact and hybridisation during recolonisation from disconnected refugia

The central aim of this paper is to clarify the picture of postglacial recolonisation and the reconstruction of refugia of Populus alba (L.) and Populus tremula (L.) in the light of hybridisation of the two species. We focussed our study on Central and Southeastern Europe including reference samples from Spain, Sweden and Northern Africa.We investigated 414 individuals of 26 populations using restriction fragment length polymorphisms (PCR-RFLPs) in six maternally inherited chloroplast markers. Altogether, 57 haplotypes were analysed of which four indicated hybridisation events in the past. Phylogeographic structure was found for P. alba with low diversity in Eastern Europe versus high diversity in Italy and Central Europe. A lack of phylogeographic structure was assessed for P. tremula as expected for a boreal forest tree, and diversity was evenly distributed in the studied populations. Two main refugia were identified for P. alba in Italy and Romania. A previously described hybrid zone between species in Central Europe turned out also to be a zone of contact between southern and eastern chloroplast lineages in P. alba. In contrast, P. tremula recolonised its present habitats in Central Europe from several refugia near the former ice cap. We assume separate disconnected refugia for P. alba and P. tremula and suggest an immigration scenario involving the mixing of colonisation routes and interspecific introgression to be responsible for the observed patterns.     

Growth-phase-dependent gene expression profiling of poplar (Populus alba x Populus tremula var. glandulosa) suspension cells

Complex sequences of morphological and biochemical changes occur during the developmental course of a batch plant cell culture. However, little information is available about the changes in gene expression that could explain these changes, because of the difficulties involved in isolating specific cellular events or developmental phases in the overlapping phases of cell growth. In an attempt to obtain such information we have examined the global growth phase-dependent gene expression of poplar cells in suspension cultures by cDNA microarray analysis. Our results reveal that significant changes occur in the expression of genes with functions related to protein synthesis, cell cycling, hormonal responses and cell wall biosynthesis, as cultures progress from initiation to senescence, that are highly correlated with observed developmental and physiological changes in the cells. Genes encoding protein kinases, calmodulin and proteins involved in both ascorbate metabolism and water-limited stress responses also showed strong stage-specific expression patterns. Our report provides fundamental information on molecular mechanisms that control cellular changes throughout the developmental course of poplar cell cultures.     

Fading of wound-induced volatile release during Populus tremula leaf expansion

Journal of Plant Research - The release of stress-driven volatiles throughout leaf development has been little studied. Therefore, we subjected poplar leaves during their developmental stage (from...     

Growth Regulators in Populus tremula I. Distribution of Auxin and Growth Inhibitors

The content of acid ether-soluble auxins and inhibitors in vegetative tissues of aspen (Populus tremula L.) was determined with the Avena coleoptile straightgrowth assay. A growth promotor, tentativety identified as indol-3-yl-acetic acid, occurred in highest concentration in the growing stem parts. It was also detected in various parts of the roots. Large amounts of inhibitors (inhihitor β) were found in growing stem tissue and in upper mature stem parts. Considerably lower content of inhibitors was found in leaves, in the basal stem part and in the roots. Nonwoody growing roots and the wood of older root segments, especially, contained very smalt quantities of inhibitors. The elution volume of a part of the inhibitory activity on a Sephadex column was similar to that of abscisic acid.     

Genetic variation in functional traits influences arthropod community composition in aspen (Populus tremula L.)

We conducted a study of natural variation in functional leaf traits and herbivory in 116 clones of European aspen, Populus tremula L., the Swedish Aspen (SwAsp) collection, originating from ten degrees of latitude across Sweden and grown in a common garden. In surveys of phytophagous arthropods over two years, we found the aspen canopy supports nearly 100 morphospecies. We identified significant broad-sense heritability of plant functional traits, basic plant defence chemistry, and arthropod community traits. The majority of arthropods were specialists, those coevolved with P. tremula to tolerate and even utilize leaf defence compounds. Arthropod abundance and richness were more closely related to plant growth rates than general chemical defences and relationships were identified between the arthropod community and stem growth, leaf and petiole morphology, anthocyanins, and condensed tannins. Heritable genetic variation in plant traits in young aspen was found to structure arthropod community; however no single trait drives the preferences of arthropod folivores among young aspen genotypes. The influence of natural variation in plant traits on the arthropod community indicates the importance of maintaining genetic variation in wild trees as keystone species for biodiversity. It further suggests that aspen can be a resource for the study of mechanisms of natural resistance to herbivores.     

Pathway position constrains the evolution of an ecologically important pathway in aspens (Populus tremula L.)

Many ecological interactions of aspens and their relatives (Populus spp.) are affected by products of the phenylpropanoid pathway synthesizing condensed tannins (CTs), whose production involves trade‐offs with other ecologically important compounds and with growth. Genes of this pathway are candidates for investigating the role of selection on ecologically important, polygenic traits. We analysed sequences from 25 genes representing 10 steps of the CT synthesis pathway, which produces CTs used in defence and lignins used for growth, in 12 individuals of European aspen (Populus tremula). We compared these to homologs from P. trichocarpa, to a control set of 77 P. tremula genes, to genome‐wide resequencing data and to RNA‐seq expression levels, in order to identify signatures of selection distinct from those of demography. In Populus, pathway position exerts a strong influence on the evolution of these genes. Nonsynonymous diversity, divergence and allele frequency shifts (Tajima's D) were much lower than for synonymous measures. Expression levels were higher, and the direction of selection more negative, for upstream genes than for those downstream. Selective constraints act with increasing intensity on upstream genes, despite the presence of multiple paralogs in most gene families. Pleiotropy, expression level, flux control and codon bias appear to interact in determining levels and patterns of variation in genes of this pathway, whose products mediate a wide array of ecological interactions for this widely distributed species.