Gregor Kozlowski

Historical bottlenecks decrease genetic diversity in natural populations of Dryopteris cristata

The reconstruction of recent historical population sizes allowed us to investigate the influence of random evolutionary processes on present-day genetic diversity in populations of Dryopteris cristata. This long-lived, allotetraploid fern is rare and endangered in the study area at the southwestern border of its European distribution. Random amplified polymorphic DNA (RAPD) diversity of 280 individuals from 14 populations of D. cristata was extraordinarily low, suggesting an ancient bottleneck in the species’ history. Analysis of molecular variance (AMOVA) of 25 different RAPD multiband phenotypes revealed significant genetic variation among three geographical regions (15%) and among populations within regions (34%); 51% of total variance was attributed to variation within populations. High population differentiation indicated limited gene flow among populations, and genetic divergence was not correlated with geographical distance. There was no relationship between genetic variation within population, estimated as molecular variance, and present-day population size. Populations with recent historical bottlenecks of fewer than 25 individuals showed a substantial and significant reduction in genetic variation, compared with populations without bottlenecks. Comparatively high levels of genetic variation were still maintained in small remnants (60–110 individuals) of formerly large populations. Average deviations of frequencies of widespread polymorphic markers within populations from their frequencies in the whole dataset were significantly higher in small or recently bottlenecked populations than in constantly large populations, thus providing evidence for random sampling effects during genetic bottlenecks and drift in small populations. The present investigation demonstrates the importance of population history for understanding present-day genetic diversity within natural populations, as well as for conservation biology.

Infection of Norway spruce (Picea abies (L.) Karst.) seedlings with Pythium irregulare Buism. and Pythium ultimum Trow.: histological and biochemical responses

We have studied the reaction of Picea abies seedlings to infection with Pythium. The highly virulent species Pythium ultimum and the less virulent species Pythium irregulare germinated on the root and hypocotyl surface, formed appressoria and penetrated through the stomata as well as through the epidermis. No major differences in the growth of both fungal species were observed during the early events of colonization. The less virulent species formed about 25% more appressoria suggesting that the fungus experienced difficulties with penetration. Differences were observed in the response of the host plant to infection. Autofluorescence, possibly related to deposition of lignin or lignin-like materials increased more in cortical and endodermal tissue colonized with the highly virulent P. ultimum than with the less virulent P. irregulare. Chitinase activity was highest in the tissues most extensively colonized by the fungus. In addition, a systemic increase of chitinase activity was also detected. Interestingly, chitinase activity increased systemically in cotyledons which were never in contact with the pathogen, indicating the translocation of a systemic signal. Salicylic acid was also detected in spruce seedlings; its level increased in roots during infection with the less virulent P. irregulare.

Conservation of threatened relict trees through living ex situ collections: lessons from the global survey of the genus Zelkova (Ulmaceae)

Maintaining living ex situ collections is one of the key conservation methods in botanic gardens worldwide. Despite of the existence of many other conservation approaches used nowadays, it offers for many endangered plants an important insurance policy for the future, especially for rare and threatened relict trees. The aim of this research was to investigate the global extent of living ex situ collections, to assess and discuss their viability and inform the development of conservation approaches that respond to latest global conservation challenges. We used as a model taxon the tree genus Zelkova (Ulmaceae). The genus includes six prominent Tertiary relict trees which survived the last glaciation in disjunct and isolated refugial regions. Our comprehensive worldwide survey shows that the majority of botanic institutions with Zelkova collections are in countries with a strong horticultural tradition and not in locations of their origin. More importantly, the acutely threatened Zelkova species are not the most represented in collections, and thus safeguarded through ex situ conservation. Less than 20% of the ex situ collections surveyed contain plant material of known wild provenance while the majority (90%) of collections are generally very small (1–10 trees). Botanic gardens and arboreta particularly in regions where iconic relict trees naturally occur should play a vital role in the conservation of these species. The coordination of conservation efforts between gardens has to be enhanced to prioritise action for the most threatened relict trees. Large scale genetic studies should be undertaken, ideally at genus level, in order to verify or clarify the provenance of ex situ collections of relict trees in cultivation. For the most threatened relict tree genera, well-coordinated specialist groups should be created.

Is the global conservation status assessment of a threatened taxon a utopia

The signatory countries of the Convention on Biological Diversity agreed to significantly reduce the rate of biodiversity loss by 2010. How will we know, however, if we have achieved this goal? Eight groups of hindrances in evaluating the global conservation status of threatened taxon are identified: (1) the extreme heterogeneity and (2) restricted availability of relevant data; (3) the uncertainty in species number and taxonomic division of the given taxon (Linnean shortfall); (4) the fragmentary distribution knowledge (Wallacean shortfall); (5) the incomplete or incorrect red-listing across the whole distribution area; (6) the lack of homogeneous and exhaustive population trend data; (7) the threat knowledge shortfall; (8) the incomplete general biological knowledge on a given taxon. The Linnean and Wallacean shortfalls lay the foundation of all other hindrances. So long as this dramatic shortfall situation does not change, the adequate assessment of the global status for overwhelming majority of extant taxa will remain a utopia.

Genetic and Ecological Consequences of Transgene Flow to the Wild Flora

Gene flow from crops to wild relatives by sexual reproduction is one of the major issues in risk assessment for the cultivation of genetically engineered (GE) plants. The main factors which influence hybridization and introgression, the two processes of gene flow, as well as the accompanying containment measures of the transgene, are reviewed. The comparison of risks between Switzerland and Europe highlights the importance of regional studies. Differences were assessed for barley, beet and wheat. Moreover, transgene flow through several wild species acting as bridge (bridge species) has been up to now poorly investigated. Indeed, transgene flow may go beyond the closest wild relative, as in nature several wild species complexes hybridize. Its importance is assessed by several examples in Poaceae. Finally, the transgene itself has genetic and ecological consequences that are reviewed. Transgenic hybrids between crops and wild relatives may have lower fitness than the wild relatives, but in several cases, no cost was detected. On the other hand, the transgene provides advantages to the hybrids, in the case of selective value as a Bt transgene in the presence of herbivores. Genetic and ecological consequences of a transgene in a wild species are complex and depend on the type of transgene, its insertion site, the density of plants and ecological factors. More studies are needed for understanding the short and long term consequences of escape of a transgene in the wild.

Phylogenetics and phylogeography of the monocot genus Baldellia (Alismataceae): Mediterranean refugia, suture zones and implications for conservation

Aquatic plants, and especially the emblematic genus Baldellia (Alismataceae), are among the most threatened organisms, due to unprecedented human-driven habitat destructions. Therefore protection plans are crucially needed and call for thoroughly documenting the genetic diversity and clarifying the taxonomy of this endangered genus. Our sampling included 282 individuals from 42 natural populations and covered the whole geographical range of the genus, across Europe and the Mediterranean. We combined sequencing of nuclear internal transcribed spacer (ITS) and chloroplastic trnL-ndhF regions with amplified fragment length polymorphism (AFLP) genotyping to investigate the Alismataceae phylogeny, and produce a phylogeography of Baldellia. Our phylogeny strongly supported the monophyly of Baldellia and placed it as the sister clade to Luronium and Alisma, therefore excluding, as previously supposed, a close genetic relatedness to the predominantly neotropical genus Echinodorus. The phylogeography of Baldellia outlined patterns consistent with a hypothesis considering glacial refugia located in the Iberian Peninsula and the Italy/Balkan region from which two distinct genetic lineages re-colonized Europe. These two lineages corresponded respectively to Baldellia ranunculoides (Italy/Balkan derived populations) and Baldellia repens (populations recovered from the Iberian Peninsula refuge), therefore supporting differences outlined between the two taxa in previous ecological and morphological studies. These results allowed clarifying taxonomic uncertainties by confirming the genetic distinctness of B. repens according to B. ranunculoides. A third lineage, Baldellia alpestris, originated and remained endemic to the mountainous regions of the Iberian Peninsula. Unexpectedly, B. repens populations collected in northern Africa, appeared to be genetically distinct from their European counterparts, this calls for further investigation to fully address their genetic and conservation status. Finally, we detected a large hybridization zone in northwestern Europe between B. repens and B. ranunculoides. These results were discussed in light of conservation approaches for Baldellia populations.

The Tertiary relict tree Zelkova abelicea (Ulmaceae): distribution, population structure and conservation status on Crete

Relict species provide a unique opportunity to understand past and recent biogeographical and evolutionary processes. Zelkova abelicea (Ulmaceae), which is endemic to the island of Crete (Greece), is one of the most prominent Tertiary relict trees of the Mediterranean region. We collected distribution, threat and population structure data by reviewing literature and herbaria and through field surveys at 14 study plots throughout the range of the species. The present distribution of Z. abelicea is extremely fragmented. Although the total estimated number of individuals is relatively high, the populations are dominated by dwarf, severely browsed, non-flowering individuals. The population structure is asymmetric. At most, 5% of a plots trees are large and fruit-bearing. The asymmetric structure is particularly pronounced in isolated and small populations. Based on its limited geographical range, the fragmented spatial pattern, and data on distribution and population structure, our study confirms that Z. abelicea is a threatened species (IUCN category Endangered). Our research aim is to promote the development of new approaches for the improvement of conservation strategies for Tertiary relict trees characterized by major local disjunctions.

Do living ex situ collections capture the genetic variation of wild populations? A molecular analysis of two relict tree species, Zelkova abelica and Zelkova carpinifolia

Botanic gardens and arboreta, particularly in regions where iconic relict trees naturally occur, play a vital role in the conservation of these species. Maintaining well-managed living ex situ collections of rare and threatened relict tree species provides an immediate insurance policy for the future species conservation. The aim of this research was to investigate the origin, representativeness and genetic diversity of relict trees kept in botanic gardens and arboreta. We used as a model two ecologically and biogeographically distinct members of the prominent relict genus Zelkova (Ulmaceae), which survived the last glaciation in disjunct and isolated refugial regions: Z. carpinifolia in Transcaucasia and Z. abelicea endemic to Crete (Greece) in the Mediterranean. Our study revealed substantial differences in the genetic diversity and the origin of living ex situ collections of the two investigated taxa. The living ex situ collections of Z. carpinifolia have relatively high representativeness compared with the global genetic variability of natural stands identified in a previous study. In contrast, Z. abelicea, which possesses an extraordinarily high genetic variability in natural populations, is clearly underrepresented in botanic garden collections. Moreover, all Z. abelicea trees investigated in this study most probably originated from a single region, the Levka Ori in western Crete. Thus, the ex situ conservation of Z. abelicea requires major planning and coordination efforts, including the establishment of well-documented collections in botanic gardens in Greece and especially on Crete. New living ex situ collections should be created using plant material collected from all of the mountain regions where Z. abelicea still occurs. Our study highlights the need for re-evaluating the existing living ex situ collections of trees and the development of new strategies for future conservation efforts in botanic gardens and arboreta. The coordination of conservation efforts between gardens must be enhanced to prioritize actions for the most threatened relict tree species.

Global conservation status assessment of the threatened aquatic plant genus Baldellia (Alismataceae): challenges and limitations

In this study, the aquatic monocot Baldellia (Alismataceae) is used as a model for evaluating the general hindrances and shortfalls in the global conservation status assessment of a threatened taxon. Our study clearly shows that Linnean shortfalls (uncertainty in the number of species and taxonomy) and the Wallacean shortfall (fragmentary knowledge regarding distribution) form the basis for all other hindrances. We demonstrate that even in Europe, which has traditionally been very well investigated, between 60 and 75% of regions or countries possess no detailed distribution maps and/or data banks for Baldellia spp. Furthermore, between 50 and 60% of regions do not have any published red list category. Thus, only general conclusions concerning the global conservation status of the three Baldellia taxa are possible—a global assessment of conservation status for B. ranunculoides subsp. repens is nearly impossible. Baldellia ranunculoides s.str. shows a strong decline in practically all regions of its natural range, and thus it is probably the most threatened species in the genus. Baldellia alpestris is the least threatened species in the genus, even though it is a narrow endemic. Our case study clearly shows the need for reinforced coordination of research and conservation activities as well as an urgent need for data accessibility regarding taxonomic, chorological and conservation studies of endangered species.

Analysis of nuclear microsatellites reveals limited differentiation between Colchic and Hyrcanian populations of the wind-pollinated relict tree Zelkova carpinifolia (Ulmaceae)

UNLABELLED • PREMISE OF THE STUDY The Caucasus represents one of the worlds biodiversity hotspots and includes the climatic refugia Hyrcan on the southern coast of the Caspian Sea and Colchis on the eastern coast of the Black Sea, where different species survived during the Quaternary climatic oscillations. We evaluated the genetic diversity of the relict tree Zelkova carpinifolia shared between the two refugia and distributed throughout the Caucasus and adjacent areas.• METHODS Specimens were collected from 30 geographical sites in Azerbaijan, Georgia, Iran, and Turkey and screened for variability at eight nuclear microsatellite loci. The genetic diversity among and within populations was assessed using a set of statistical measures.• KEY RESULTS We detected 379 different genotypes from a total of 495 individuals with varying degrees of clonal reproduction at the different sites. Low to intermediate levels of genetic diversity were observed at all sites, and strong differentiation between sampling sites was absent. In addition, we observed no clear genetic differentiation between the Colchis and Hyrcan. Bayesian clustering of the genotypes revealed three populations with high levels of admixture between the sampling sites.• CONCLUSIONS The lack of strong genetic structure of studied populations of Z. carpinifolia contrasts with a previous study based on chloroplast markers and suggests that long-distance pollen dispersal is an important factor of gene flow among populations of Z. carpinifolia. The present study does not reveal any particular site with particularly isolated genotypes that would deserve more attention for conservation purposes than others, although some sites should be considered for further investigation.