Genetic differentiation and speciation in prealpine Cochlearia: Allohexaploid Cochlearia bavarica Vogt (Brassicaceae) compared to its diploid ancestor Cochlearia pyrenaica DC. in Germany and Austria

 Significant geographic partitioning of genetic variation within Cochlearia bavarica was found within populations from Allg?u and SE Bavaria (Germany) exhibiting significant genetic differentiation. It has been demonstrated that allohexaploid C. bavarica evolved via hybridization between diploid C. pyrenaica and tetraploid C. officinalis. Presently, only C. pyrenaica is distributed throughout inland Central Europe. It has been concluded that C. bavarica is of inter- or postglacial origin, and its speciation was not influenced by human activities. Isozyme analysis revealed that there is a correlation between interpopulational genetic distances and geographic distances among C. bavarica populations from both regions, and which is not the case for C. pyrenaica in Germany and Austria. Only high alpine C. excelsa is significantly differentiated among the diploid taxa analysed here. Geographically structured distribution of alleles and their frequencies in C. bavarica populations could not be explained with the distribution of these alleles in C. pyrenaica. The presented findings favour disruption of a former wider distribution area rather than migration of C. bavarica or a polytopic origin of this species. Received April 17, 2001 Accepted February 1, 2002     

Cochlearia macrorrhiza (Brassicaceae): A bridging species between Cochlearia taxa from the Eastern Alps and the Carpathians?

Cochlearia macrorrhiza is one of the most highly endangered species in Central Europe and less than five individuals survived at its natural stand in a lowland area between the Eastern Alps and the Carpathians. Amplified fragment length polymorphisms (AFLPs) confirmed the status of C. macrorrhiza as a distinct taxon. Lowland C. macrorrhiza does not bridge the distribution of montainous and alpine Cochlearia species from the Eastern Alps and the Carpathians genetically, and C. macrorrhiza represents a separate lineage which evolved from diploid Cochlearia as C. excelsa in East Austrian high alpine regions did. Another species considered in this study, the Romanian C. borzaeana is more closely related to C. tatrae from the High Tatra mountains than to C. pyrenaica from Slovakia or Austria and the AFLP results suggest a single origin of alpine 2n=42 taxa. Genetic differentiation within and between populations is highly structured geographically, and the AFLP data favour a former widespread distribution of C. pyrenaica in mountainous regions and a parallel evolution of high alpine taxa in the Eastern Alps and the Carpathians, respectively.     

Cytogenetic studies in Cochlearia L. (Cruciferae). The chromosomal constitution of C. danica L.

Genome analysis has been used to investigate the relationship of C. danica L. to the diploid and tetraploid species in the genus. The results of this analysis suggest that C. danica has the genomic constitution A¹ ₇ A¹ ₇ A² ₇ A² ₇ T₇ T₇. Both A₇ genomes in C. danica are segmentally differentiated from the A₇ genome in C. groenlandica L. and from the similar A₆ genomes in C. pyrenaica DC. and C. officinalis L. but, in hybrids, are capable of some pairing with these genomes. A² ₇ is more distantly related to A₇ and A₆ than is A¹ ₇ and in A₇ some of the differentiation has taken place by reciprocal translocations. A¹ ₇ and A² ₇ still retain enough homology for pairing between them. The T₇ genome has no homology with any of the A genomes but may be derived from an ancestral T₆ genome.     

General occurrence of the glucosinolate glucocochlearin within the Cochlearia genus

A natural compound, glucocochlearin, was isolated from the aerial parts of 10 different Cochlearia species. The purification of this compound was achieved through HPLC. The identity of the product was established mainly on the basis of spectroscopic NMR (1H, 13C, COSY, TOCSY, HMQC, HMBC, J-MOD) and high resolution mass spectroscopy data. This compound can be considered as a chemomarker of the genus Cochlearia.     

Isozymes,Speciation and Evolution in the Polyploid Complex Cochlearia L. (Brassicaceae)

Within the genus Cochlearia L. sect. Cochlearia three isozyme systems (aspartate aminotransferase, leucine aminopeptidase and phosphoglucomutase) have been analysed at the populational level for eleven species with different ploidy levels. Nine loci complexes were resolved with a total of 33 alleles from 89 populations. In general, polyploids displayed higher numbers of alleles per species and per population. A notable exception is diploid C. pyrenaica with as many alleles as some polyploids. Data support: (1) the origin of tetraploid C. officinalis from C. pyrenaica s.l., particularly from C. aestuaria; (2) the allopolyploid origin of octoploid C. anglica from C. officinalis; (3) the allopolyploid origin of hexaploid inland taxa C. polonica, C. tatrae and C. bavarica via hybridization between C. pyrenaica and C. officinalis; (4) a putative allopolyploid origin of hexaploid coastal species C. danica by hybridization between C. pyrenaica s.l. and C. officinalis accompanied by a shift in chromosome complement. A scenario summarizing our present understanding of the evolution within Cochlearia sect. Cochlearia is presented.     

Cytogenetic studies in Cochlearia L. (Cruciferae). The origins of C. officinalis L. and C. micacea Marshall

Genome analysis has been used to investigate the evolutionary relationships of the tetraploid species in the genus Cochlearia. The results indicate that both C. officinalis L. (2n=24) and C. micacea Marshal (2n=26) are essentially autotetraploid in origin and that C. scotica Druce is simply a morphological variant of C. officinalis. The chromosomal relationships of the tetraploids to each other and to the diploids in the genus are discussed and the possible routes for the formation of all the species from a single, 2n=12, basic taxon are given. Evidence for the existence of a genic mechanism causing C. officinalis to form only bivalents is given and the mode of evolution of such a mechanism discussed.     

Ecotypic differentiation in relation to soil nitrogen in northern Scandinavian Cochlearia officinalis

Ecotypic differentiation of Cochlearia officinalis L. was studied in four populations, roughly described as "beach", "bird cliff", "spring" and "estuary", which occupy different habitats in the northern part of Norway. The plants were cultivated under controlled conditions on soil to which nitrogen was applied at various amounts (0, 0.2 and 1.0 g N pot⁻¹). The above ground part of the "bird cliff" population showed the highest dry weight and nitrogen uptake at the highest nitrogen concentration. No effect of nitrogen treatment and the lowest dry weight were found in the "estuary" population. The differences in population response to added nitrogen were correlated to the nitrogen content of the soil in their natural habitat. Morphological features of the populations were also significantly different. The "estuary", "beach" and "bird cliff" populations showed differences in all characteristics measured – ratio of length/width of the leaves, shape of the leaf base, and length of the petiole. "Beach" and "spring" differed significantly in petiole length, while "bird cliff" and "spring" differed significantly in both leaf ratio and shape of the leaf base. These morphological features were not significantly altered by the application of nitrogen to the soil. This study shows that the four populations of Cochlearia officinalis are discrete biotypes characteristic of distinct habitats, and therefore, should be considered ecotypes.     

Morphology and taxonomy of the genus Cochlearia (Brassicaceae) in Northern Scandinavia

The morphological variation between populations of Cochlearia in Northern Scandinavia has been analysed. The variation is correlated to ecology, and four ecotypes are recognized. Based on the morphological differentiation of the ecotypes and the results of crossing experiments, the populations are referred to the following three taxa: C. officinalis ssp. officinalis which includes the common beach ecotype and the bird cliff ecotype; C. officinalis ssp. integrifolia stat. nov. which includes the spring ecotype growing in inland localities up to about 600 m a.s.l. in cold more or less eutrophic springs, and C. officinalis ssp. norvegica ssp. nov. which includes the estuary ecotype, growing in poor, often inundated habitats near river outlets at the fjord ends. All the differentiation has taken place at the tetraploid level (2n = 24). The subspecies are kept more or less distinct due to ecological and seasonal isolating mechanisms rather than internal, reproductive barriers. A key to the subspecies and distribution maps for ssp. integrifolia and ssp. norvegica are presented.     

Chloroplast DNA restriction site variation and RAPD-analyses in Cochlearia (Brassicaceae): Biosystematics and speciation

Chloroplast DNA (cpDNA) restriction site variation (RFLP), has been analysed in the genus Cochlearia L. sections Cochlearia O.E. Schulz and Glaucocochlearia O.E. Schulz. Members of section Glaucocochlearia are clearly separated from those of section Cochlearia by 28 site mutations, thus supporting taxonomic arrangements into the two sections mentioned. RAPD studies provide further strong evidence for closer relationships among tax a within each section than between sections. CpDNA sequence divergence values in section Cochlearia are extremely low. Although 91 accessions representing 11 species have been analysed by 25 restriction enzymes, only four restriction site mutations were detected characterising six different cp genome types. Low levels of chloroplast DNA divergence would suggest that taxa of section Cochlearia are closely related and most likely have diversified recently. Polyploid Cochlearia offcinalis (2n=24), C. anglica (2n=48) and C. danica (2n=42) displayed infraspecific variation for cp genomes, which could be partly explained by multiple origin and by reticulate evolution. RAPD analyses (22 primers, 140 informative characters, ten taxa from 17 accessions) do not only strongly support these arguments but also helped to clarify speciation processes and biogeographic aspects in more detail. It is argued that South West Europe was the primary center of origin of the polyploids, second centers may be the British Isles, Iceland and locally restricted parts of Middle Europe.     

Variability of polar scurvygrass Cochlearia groenlandica individual traits along a seabird influenced gradient across Spitsbergen tundra

In the resource-limited Arctic environment, vegetation developing near seabird colonies is exceptionally luxuriant. Nevertheless, there are very few detailed quantitative studies of any specific plant species responses to ornithogenic manuring. Therefore, we studied variability of polar scurvygrass Cochlearia groe nlandica individual biomass and leaf width along a seabird influenced gradient determining environmental conditions for vegetation in south-west Spitsbergen. We found seabird colony effect being a paramount factor responsible for augmented growth of C. groenlandica. The species predominated close to the colony and reached the highest mean values of individual biomass (1.4 g) and leaf width (26.6 mm) 10 m below the colony. Its abundance and size declined towards the coast. Both C. groenlandica individual traits significantly decreased with distance from the colony, soil water and organic matter content and increased with guano deposition, soil δ ¹⁵N, conductivity, acidity and nitrate, phosphate and potassium ion content. Our study supports the hypothesis that seabirds have fundamental importance for vegetation growth in poor Arctic environment. Highly plastic species such as C. groenlandica may be a useful instrument in detecting habitat condition changes, for instance resulting from climate change.     

Reintroduction of the endangered and endemic plant species Cochlearia bavarica—Implications from conservation genetics

Population reintroduction is a common practice in conservation, but often fails, also due to the effects of inbreeding or outbreeding depression. Cochlearia bavarica is a strongly endangered plant species endemic to Bavaria in Germany, constantly declining since the late 1980s. Therefore, population reintroduction is intended. In this study, we analyzed genetic diversity within and genetic differentiation between all 32 remnant populations of the species in Swabia and Upper Bavaria using amplified fragment length polymorphisms. Our aim was to increase reintroduction success by providing data to avoid negative effects of inbreeding and outbreeding and to preserve the natural genetic pattern of the species. Genetic diversity within populations was low but similar to other rare and endemic species and varied strongly between populations but did not depend on population size. Our analysis revealed a strong geographic pattern of genetic variation. Genetic differentiation was strongest between Swabia and Upper Bavaria and at the population level, whereas differentiation between subpopulations was comparatively low. Isolation by distance and genetic differentiation was stronger among populations from Upper Bavaria than from Swabia. From the results of our study, we derived recommendations for a successful reintroduction of the species. We suggest using rather genetically variable than large populations as reintroduction sources. Moreover, the exchange of plant material between Swabia and Upper Bavaria should be completely avoided. Within these regions, plant material from genetically similar populations should preferably be used for reintroduction, whereas the exchange among subpopulations seems to be possible without a negative impact on genetic variation due to natural gene flow.     

Suppression of multivalent formation by B chromosomes in natural and artificial autopolyploids of scurvy-grass (Cochlearia L.)

Summary In diploid Cochlearia pyrenaica, its established natural autotetraploid C. officinalis, and their newly induced autotetraploid and auto-octoploid derivatives, B chromosomes change the normal pattern of chromosome association by suppressing homologous pairing. Frequency of bivalents increases at the expense of multivalents from lower to higher numbers of B chromosomes. The reduction of multivalents due to the direct influence of the B chromosomes, independent of pollen mother cell chiasma frequency, is suggested as being related to the mechanism that prevents A/B chromosome pairing.     

Genetic structure in peripheral Western European populations of the endangered species Cochlearia pyrenaica (Brassicaceae)

Cochlearia pyrenaica is one of the most endangered plant species in Europe, listed in many European and regional conservation policy documents (e.g. Spain, France, Belgium, Switzerland). To study its genetic structure, define its conservation units and propose a management strategy for this species, amplified fragment length polymorphism markers were used to analyse the genetic diversity within and between five representative populations of the species distribution in Western Europe (Cantabrian Range, North of Spain; Pyrenees, France; Wallonia, Belgium). Low levels of genetic diversity were revealed by the population percentage of polymorphic bands (PPB?=?36.56%), average within-population diversity (H _S?=?0.0990) and genetic diversity within populations (H _pop?=?0.1541), although high levels were reported at species level (PPB?=?81.16%; total genetic diversity for the species, H _T?=?0.0990; and genetic diversity within whole species, H _sp?=?0.2515). The coefficient of genetic differentiation among populations (G _ST) was 0.3869. The analysis of Shannon diversity index in population and for the total data set partitioned (38.72%) and AMOVA (53%) detected a high level of interpopulation diversity, in broad agreement with the result of genetic differentiation analysis. NeighborNet network and principal coordinate analyses clustered the populations in three major groups congruent with geographical regions. Bayesian clustering also confirmed these three distinct genetic clusters. The level of gene flow (Nm) was estimated as 0.3961 individuals per generation among populations, with the genetic identity (I) and genetic distance (D) among populations ranging from 0.8679 to 0.9651 and from 0.0355 to 0.1417, respectively. Therefore, the low levels of genetic variation and high divergence of regional gene pools indicate that there is a need to protect each disjunct region of Western Europe.     

Evolution de I'amidon et des sucres solubles chez des feuilles isoléaes d' Aster tripolium et de Cochlearia tniglica. Action du NaCI Par

Starch and soluble sugars in leaves of Aster tripolium L. and Cochlearia angliea L. Effect of NaCI. - The limbs or foliar disks of Aster tripolium L. and Cochlearia anglica L. synthelize starch when exposed to light at 25°C, and in the presence of glucose or saceharose together with 0.25 to 0.61 % of NaCI.     

Relationship between population size, allozyme variation, and plant performance in the narrow endemic Cochlearia bavarica

We studied population viability in relation topopulation size and allelic variation in thenarrowly-endemic, monocarpic perennial plantCochlearia bavarica in Bavaria. In 1996,we analysed allelic variation by allozymeelectrophoresis in 24 populations ranging from8–2000 flowering individuals. Fitness-relatedcharacters were investigated in 22 of the 24populations in the field in 1996 (reproductiveand vegetative traits) and 1998 (reproductivetraits only). Differences in allozyme patternwere large between a south-eastern and awestern population group. Genetic diversity,assessed by the Shannon-Wiener diversity index,was low within but high among populations.Small populations had fewer alleles per locus,fewer polymorphic loci, lower observedheterozygosity, and lower genetic diversitythan large populations. Environmentalvariables were not significantly correlatedwith population size or fitness with theexception of light availability, indicatingthat habitat quality was similar for large andsmall populations. Population size showedpositive correlations with number of flowers,fruit set per plant, number of seeds per fruit,and total seed output per plant. Fruit set andnumber of seeds per fruit were positivelycorrelated with the observed heterozygosity andthe proportion of polymorphic loci. We usedpath analyses to study the possible causalrelationships among population size, allelicvariation, and reproductive characters. Thesemodels showed that allelic variation had nodirect influence on reproductive characters,whereas population size did. We conclude thatat present population size reduces viabilityand also reduces allelic variation; but thereduced allelic variation may in the longerterm have negative feed-backs on bothpopulation size and viability.     

Genetic structure of the critically endangered endemic Cochlearia polonica (Brassicaceae): efficiency of the last-chance transplantation

Cochlearia polonica , a narrow endemic of southern Poland, is one of the rarest and most endangered species of the European flora. All natural populations are extinct and the species has survived in only one transplanted population derived from 14 original individuals. Using AFLPs, the genetic variation and spatial structure of this population were analysed approximately 30 years after transplantation. The incidence of polymorphic AFLP bands (30.46%) is low compared with data from a natural population of another Cochlearia species, C. tatrae . Principal co-ordinates and spatial autocorrelation analyses demonstrated the presence of significant genetic structure. It is recommended that conservation efforts on C. polonica should preserve the complete population area, because local extinctions may lead to a loss of genetic information. The presence of genetic structure should also be taken into account during the sampling of material (plants or seeds) for ex situ conservation measures.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 527–532.