Effects of freezing temperatures on early life stages of native trees of different elevational origin: implications for tree recruitment in seasonally dry mountain forests.

In mountain forests, tree regeneration is limited by increasingly frequent frosts with increasing elevation. We investigated the effects of exposure to freezing temperature on early life stages of two native trees of different elevational origin in a seasonally dry mountain forest. We hypothesized that the negative effects of freezing exposure on performance of early life stages increases as freezing temperature decreases, and that frost resistance increases in plants of high elevational origin. We collected seeds of two tree species (Kageneckia lanceolata and Lithraea molleoides) from populations located at different elevations and grew seedlings and saplings in a greenhouse. Dry seeds, imbibed seeds and 1-month-old seedlings were exposed to seven temperature treatments ranging from 4 °C to -20 °C, while 12-month-old saplings were exposed to four temperature treatments from -8 °C to -20 °C. After freezing exposure in a climate chamber, we monitored seed germination and seedling and sapling survival. Germination of K. lanceolata decreased with decreasing temperature only for imbibed seeds from mid- and high elevations, whereas germination of L. molleoides slightly increased with decreasing temperature only for imbibed seeds from high elevations. For both species, seedling survival decreased with decreasing temperature. For K. lanceolata, the negative effects of freezing temperatures were weaker as elevational origin of seeds increased, whereas L. molleoides showed the opposite pattern. For both species, saplings only survived at the mildest applied freezing temperature (-8 °C). We conclude that effects of climatic variation associated with elevation depend on the study species and life stage. The observed patterns could be caused by maternal effects, which are absent at the sapling stage. Moreover, temperatures below -8 °C can limit recruitment since partial mortality of seedlings and saplings occurred at such values.

Sex ratio rather than population size affects genetic diversity in Antennaria dioica.

Habitat fragmentation and small population size can lead to genetic erosion in threatened plant populations. Classical theory implies that dioecy can counteract genetic erosion as it decreases the magnitude of inbreeding and genetic drift due to obligate outcrossing. However, in small populations, sex ratios may be strongly male- or female-biased, leading to substantial reductions in effective population size. This may theoretically result in a unimodal relationship between sex ratios and genetic diversity; yet, empirical studies on this relationship are scarce. Using AFLP markers, we studied genetic diversity, structure and differentiation in 14 highly fragmented Antennaria dioica populations from the Central European lowlands. Our analyses focused on the relationship between sex ratio, population size and genetic diversity. Although most populations were small (mean: 35.5 patches), genetic diversity was moderately high. We found evidence for isolation-by-distance, but overall differentiation of the populations was rather weak. Females dominated 11 populations, which overall resulted in a slightly female-biased sex ratio (61.5%). There was no significant relationship between population size and genetic diversity. The proportion of females was not unimodally but positively linearly related to genetic diversity. The high genetic diversity and low genetic differentiation suggest that A. dioica has been widely distributed in the Central European lowlands in the past, while fragmentation occurred only in the last decades. Sex ratio has more immediate consequences on genetic diversity than population size. An increasing proportion of females can increase genetic diversity in dioecious plants, probably due to a higher amount of sexual reproduction.

Local pre-adaptation to disturbance and inbreeding-environment interactions affect colonisation abilities of diploid and tetraploid Centaurea stoebe.

Primary colonisation in invasive ranges most commonly occurs in disturbed habitats, where anthropogenic disturbance may cause physical damage to plants. The tolerance to such damage may differ between cytotypes and among populations as a result of differing population histories (adaptive differentiation between ruderal verus natural habitats). Moreover, founder populations often experience inbreeding depression, the effects of which may increase through physical damage due to inbreeding-environment interactions. We aimed to understand how such colonisation processes differ between diploid and tetraploid Centaurea stoebe populations, with a view to understanding why only tetraploids are invasive. We conducted a clipping experiment (frequency: zero, once or twice in the growing season) on inbred versus outbred offspring originating from 37 C. stoebe populations of varying cytotype, range and habitat type (natural versus ruderal). Aboveground biomass was harvested at the end of the vegetation period, while re-sprouting success was recorded in the following spring. Clipping reduced re-sprouting success and biomass, which was significantly more pronounced in natural than in ruderal populations. Inbreeding depression was not detected under benign conditions, but became increasingly apparent in biomass when plants were clipped. The effects of clipping and inbreeding did not differ between cytotypes. Adaptive differentiation in disturbance tolerance was higher among populations than between cytotypes, which highlights the potential of pre-adaptation in ruderal populations during early colonisation on anthropogenically disturbed sites. While the consequences of inbreeding increased through clipping-mediated stress, they were comparable between cytotypes, and consequently do not contribute to understanding the cytotype shift in the invasive range.

Influence of altitude on local adaptation in upland tree species from central Argentina.

Steep climatic gradients boost morphological and physiological adjustments in plants, with consequences on performance. The three principal woody species of the Sierras Grandes Mountains of central Argentina have marked differences in sapling performance along their altitudinal distribution. We hypothesize that the steep gradient of climatic conditions across the species' altitudinal distribution promotes trait differences between populations of different altitudes that are inherited by the following generation. Seeds from different altitudes were exposed to three temperature regimes to assess differential germination responses. Saplings were then transplanted to a greenhouse to assess possible variations in attributes and performance after 18 months. The three species showed differences in germination responses to temperature among altitudes and/or in sapling attributes and performance. In Maytenus boaria and Escallonia cordobensis, germination success was higher under high temperatures for the highest-altitude, whereas lower temperatures boosted germination of the lowest altitudes. Polylepis australis showed no differences in germination among temperature treatments. In the greenhouse, saplings of the three species from intermediate altitudes showed high performance, whereas the upper and lower populations seemed to be adjusted to tolerating more stressful conditions (i.e., lower temperatures at the upper end and water stress at the lower end), showing lower performance toward both altitudinal limits. These patterns agree with those described for saplings growing under field conditions, suggesting adjustments in response to environmental changes undergone by populations along the altitudinal range. The marked adjustments of populations to the local environment suggest a potentially high impact of climatic change on species distribution.

Environmental gradients shape the genetic structure of two medicinal Salvia species in Jordan.

Environmental gradients, and particularly climatic variables, exert a strong influence on plant distribution and, potentially, population genetic diversity and differentiation. Differences in water availability can cause among-population variation in ecological processes and can thus interrupt populations' connectivity and isolate them environmentally. The present study examines the effect of environmental heterogeneity on plant populations due to environmental isolation unrelated to geographic distance. Using AFLP markers, we analyzed genetic diversity and differentiation among 12 Salvia spinosa populations and 13 Salvia syriaca populations from three phytogeographical regions (Mediterranean, Irano-Turanian and Saharo-Arabian) representing the extent of the species' geographic range in Jordan. Differences in geographic location and climate were considered in the analyses. For both species, flowering phenology varied among populations and regions. Irano-Turanian and Saharo-Arabian populations had higher genetic diversity than Mediterranean populations, and genetic diversity increased significantly with increasing temperature. Genetic diversity in Salvia syriaca was affected by population size, while genetic diversity responded to drought in S. spinosa. For both species, high levels of genetic differentiation were found as well as two well-supported phytogeographical groups of populations, with Mediterranean populations clustering in one group and the Irano-Turanian and Saharo-Arabian populations in another. Genetic distance was significantly correlated to environmental distance, but not to geographic distance. Our data indicate that populations from moist vs. arid environments are environmentally isolated, where environmental gradients affect their flowering phenology, limit gene flow and shape their genetic structure. We conclude that environmental heterogeneity may act as driver for the observed variation in genetic diversity.

High genetic diversity declines towards the geographic range periphery of Adonis vernalis, a Eurasian dry grassland plant.

Genetic diversity is important for species' fitness and evolutionary processes but our knowledge on how it varies across a species' distribution range is limited. The abundant centre hypothesis (ACH) predicts that populations become smaller and more isolated towards the geographic range periphery - a pattern that in turn should be associated with decreasing genetic diversity and increasing genetic differentiation. We tested this hypothesis in Adonis vernalis, a dry grassland plant with an extensive Eurasian distribution. Its life-history traits and distribution characteristics suggest a low genetic diversity that decreases and a high genetic differentiation that increases towards the range edge. We analysed AFLP fingerprints in 28 populations along a 4698-km transect from the geographic range core in Russia to the western range periphery in Central and Western Europe. Contrary to our expectation, our analysis revealed high genetic diversity (range of proportion of polymorphic bands = 56-81%, He = 0.168-0.238) and low genetic differentiation across populations (Φ(ST) = 0.18). However, in congruence with the genetic predictions of the ACH, genetic diversity decreased and genetic differentiation increased towards the range periphery. Spanish populations were genetically distinct, suggesting a divergent post-glacial history in this region. The high genetic diversity and low genetic differentiation in the remaining A. vernalis populations is surprising given the species' life-history traits and points to the possibility that the species has been widely distributed in the studied region or that it has migrated from a diverse source in an East-West direction, in the past.

Performance and responses to competition in two congeneric annual species: does seed heteromorphism matter?

Variations in seed characteristics observed in heteromorphic species may affect various stages of their life cycles, e.g. seed dormancy, germination characteristics or even adult plant performance. Highly specialised seed morphs - described as colonisers and maintainers - exhibit a trade-off between colonisation capacity and competitive traits. The performance of distinct seed morph progeny under competitive conditions, and especially in multi-species arrangements, had previously not been given much attention. In this study, we compared performance and response to competition among distinct seed morph progenies in two congeneric, co-occurring species: the invasive Bidens frondosa and the non-invasive Bidens tripartita. We hypothesised that maintainer seed morphs of both species would perform better under increased plant densities and within inter-morphic mixtures, while coloniser morphs would show stronger responses to increased densities and perform relatively poorly in inter-morphic mixtures. We conducted a growth trial and a greenhouse experiment, which revealed that seed morph progeny differed significantly in plant height when grown without competition, while under competitive conditions such differences became less apparent. The observed pattern was more strongly pronounced in B. frondosa, which showed a general predominance in stature and biomass over its non-invasive congener. Although seed morphs performed equally well under competitive conditions, increased plant height and more rapid germination can favour the maintainer seed morph on sites where vegetation is already present.

Bioclimatic regions influence genetic structure of four Jordanian Stipa species.

Strong environmental gradients can affect the genetic structure of plant populations, but little is known as to whether closely related species respond similarly or idiosyncratically to ecogeographic variation. We analysed the extent to which gradients in temperature and rainfall shape the genetic structure of four Stipa species in four bioclimatic regions in Jordan. Genetic diversity, differentiation and structure of Stipa species were investigated using amplified fragment length polymorphism (AFLP) molecular markers. For each of the four study species, we sampled 120 individuals from ten populations situated in distinct bioclimatic regions and assessed the degree of genetic diversity and genetic differentiation within and among populations. The widespread ruderals Stipa capensis and S. parviflora had higher genetic diversity than the geographically restricted semi-desert species S. arabica and S. lagascae. In three of the four species, genetic diversity strongly decreased with precipitation, while genetic diversity increased with temperature in S. capensis. Most genetic diversity resided among populations in the semi-desert species (Φ(ST) = 0.572/0.595 in S. arabica/lagascae) but within populations in the ruderal species (Φ(ST) = 0.355/0.387 S. capensis/parviflora). Principal coordinate analysis (PCoA) and STRUCTURE analysis showed that Stipa populations of all species clustered ecogeographically. A genome scan revealed that divergent selection at particular AFLP loci contributed to genetic differentiation. Irrespective of their different life histories, Stipa species responded similarly to the bioclimatic gradient in Jordan. We conclude that, in addition to predominant random processes, steep climatic gradients might shape the genetic structure of plant populations.

Ex situ cultivation affects genetic structure and diversity in arable plants.

Worldwide, botanical gardens cultivate around 80,000 taxa, corresponding to approximately one-quarter of all vascular plants. Most cultivated taxa are, however, held in a small number of collections, and mostly only in small populations. Lack of genetic exchange and stochastic processes in small populations make them susceptible to detrimental genetic effects, which should be most severe in annual species, as sowing cycles are often short. In order to assess whether ex situ cultivation affects genetic diversity of annuals, five annual arable species with similar breeding systems were assessed with 42 in situ populations being compared to 20 ex situ populations using a random amplified polymorphic DNA (RAPD) analysis approach. Population sizes tended to be lower under ex situ cultivation and levels of genetic diversity also tended to be lower in four of the five species, with differences being significant in only two. Ex situ populations showed incomplete representation of alleles found in the wild. The duration of cultivation did not indicate any effect on genetic diversity. This implies that cultivation strategies resulted in different genetic structures in the garden populations. Although not unequivocally pronounced, differences nonetheless imply that conservation strategies in the involved gardens may need improvement. One option is cold storage of seeds, a practice that is not currently followed in the studied ex situ collections. This may reflect that the respective gardens focus on displaying living plant populations.

Low genetic variability and strong differentiation among isolated populations of the rare steppe grass Stipa capillata L. in Central Europe.

Stipa capillata L. (Poaceae) is a rare grassland species in Central Europe that is thought to have once been widespread in post-glacial times. Such relict species are expected to show low genetic diversity within populations and high genetic differentiation between populations due to bottlenecks, long-term isolation and ongoing habitat fragmentation. These patterns should be particularly pronounced in selfing species. We analysed patterns of random amplified polymorphic DNA (RAPD) variation in the facultatively cleistogamous S. capillata to examine whether genetic diversity is associated with population size, and to draw initial conclusions on the migration history of this species in Central Europe. We analysed 31 S. capillata populations distributed in northeastern, central and western Germany, Switzerland and Slovakia. Estimates of genetic diversity at the population level were low and not related to population size. Among all populations, extraordinarily high levels of genetic differentiation (amova: phi(ST) = 0.86; Bayesian analysis: theta(B) = 0.758) and isolation-by-distance were detected. Hierarchical amova indicated that most of the variability was partitioned among geographic regions (59%), or among populations between regions when the genetically distinct Slovakian populations were excluded. These findings are supported by results of a multivariate ordination analysis. We also found two different groups in an UPGMA cluster analysis: one that contained the populations from Slovakia, and the other that combined the populations from Germany and Switzerland. Our findings imply that S. capillata is indeed a relict species that experienced strong bottlenecks in Central Europe, enhanced by isolation and selfing. Most likely, populations in Slovakia were not the main genetic source for the post-glacial colonization of Central Europe.

Genetic structure of coastal and inland populations of the annual halophyte Suaeda maritima (L.) dumort. in Central Europe, inferred from amplified fragment length polymorphism markers.

Naturally occurring inland salt habitats are highly threatened due to increasing fragmentation and area reduction, while the surroundings of former potash mining dumps have experienced a massive invasion by halophytes over the last 20 years. We reconstructed colonisation patterns of these purely anthropogenic inland salt sites using molecular markers in the obligate halophyte Suaeda maritima (L.) dumort. (Chenopodiaceae), a typical plant in such areas. In the present study, 120 individual plants from 40 coastal and inland populations in Central Europe were subjected to AFLP analysis with nine primer combinations. A total of 243 AFLP band positions were scored as presence/absence characters. Genetic diversity values were not significantly different in populations from natural and anthropogenic inland salt sites as compared to coastal habitats. Results from principal coordinate analysis, neighbour-joining analysis and analysis of molecular variance (amova) all indicated that most of the genetic variation is preserved within populations, while genetic differentiation among populations is comparatively low. We conclude that S. maritima has repeatedly and independently colonised the surroundings of former potash mining dumps in Central Germany. However, the absence of founder effects and the lack of phylogeographic structure prevented us from identifying putative donor populations.

Extensive clonality of the endemic Calamagrostis pseudopurpurea Gerstl. ex O.R. Heine in central Germany revealed by RAPD markers.

Calamagrostis pseudopurpurea is one of only a few endemic species in Germany and is confined to the catchment area of the River Mulde in the states of Saxony and Saxony-Anhalt. We studied the genetic structure and seed viability across its entire distribution area. Patterns of random amplified polymorphic DNA (RAPD) variation were analysed using 183 individuals from 43 stands in order to assess the overall genetic structure and the extent of clonality. In addition, four related Calamagrostis species (C. canescens, C. epigejos, C. phragmitoides and C. villosa) were included in our study to consider the probable phylogenetic origin of C. pseudopurpurea. We detected two clearly different RAPD phenotypes of C. pseudopurpurea, each distributed along the river banks of two spatially isolated stream courses. Both phenotypes are present downstream of the confluence. Our results indicate that C. pseudopurpurea originates from two distinct periods of hybridisation between the same parental taxa, and that clonal propagation is most likely the main reproduction method. In line with its hybrid origin, embryos of sampled C. pseudopurpurea caryopses were found to be mostly degraded or unviable over several years. Calamagrostis pseudopurpurea is genetically closer to C. canescens and C. phragmitoides than it is to other studied species, but C. canescens and C. phragmitoides have not been proven to be direct parental taxa of C. pseudopurpurea. Calamagrostis pseudopurpurea should therefore still be treated as a separate species that needs special attention from a conservation point of view.

Genetic structure of Galitzkya macrocarpa and G. potaninii, two closely related endemics of central Asian mountain ranges.

UNLABELLED: BACKGROUND AND AIMS Habitats in mountains are often isolated. Plants growing in these sites face severe dispersal limitations, but also difficulties for recruitment. The focus was laid on the magnitude of genetic differences among populations but also on the size of potentially occurring clones. METHODS: RAPD fingerprints were obtained from 23 populations in southern Mongolia. Sampling covered the entire distribution range of Galitzkya macrocarpa; samples of G. potaninii represented only the Mongolian part of its mainly northern Chinese range. KEY RESULTS: The Mongolian endemic G. macrocarpa showed moderately strong population differentiation (Phi ST = 0.251), and limited evidence for isolation by distance. Local genetic diversity was not positively correlated to habitat size, and not reduced in peripheral populations. Clonal growth is possible, but most plants originate from sexual reproduction. In contrast, populations of G. potaninii were highly differentiated (Phi ST = 0.550); and the most remote outposts had reduced genetic diversity. In these areas, isolation is expected to date back to glacial times. CONCLUSIONS: Effects of natural fragmentation differ among species. Both are rare, but G. macrocarpa appears to be able to maintain genetic diversity over its range. Clonal growth is an option in its mixed reproduction strategy and allows survival under harsh conditions. In contrast, genetic structure in G. potaninii gives reason for concern, and further studies on population dynamics are needed.