Geographical parthenogenesis, genome size variation and pollen production in the arctic-alpine species Hieracium alpinum

Hieracium alpinum L. (Asteraceae) is an arctic-alpine species distributed throughout Europe with both diploid and triploid cytotypes. We determined the ploidy levels of plants from 23 populations from Austria, Bosnia and Herzegovina, Finland, Italy, Norway, Romania, Slovakia, Switzerland and Ukraine. Data showed a non-overlapping pattern of cytotype distribution: sexually reproducing diploids (2n=2x=18) occur solely in the Eastern and Southern Carpathians, while apomictic triploids (2n=3x=27) cover the rest of the range. Such clear-cut allopatry is rather rare in vascular plants with geographical parthenogenesis. Comparison of absolute genome size indicates genome downsizing (by on average 3.7%) of haploid DNA amount in triploids relative to diploids. Genome size further correlated with longitude and latitude in the Alps, with decreasing absolute DNA content from west to east, and from south to north. While previously published data indicated complete male sterility of triploid plants, we found that plants from the Alps and Bosnia and Herzegovina commonly produced some pollen, whereas populations from the Western Carpathians and Scandinavia seemed to be almost completely pollen sterile. Scenarios about the evolution of geographical parthenogenesis in H. alpinum are discusse

Anthropogenic disturbance as a driver of microspatial and microhabitat segregation of cytotypes of Centaurea stoebe and cytotype interactions in secondary contact zones

Background and Aims In a mixed-ploidy population, strong frequency-dependent mating will lead to the elimination of the less common cytotype, unless prezygotic barriers enhance assortative mating. However, such barriers favouring cytotype coexistence have only rarely been explored. Here, an assessment is made of the mechanisms involved in formation of mixed-ploidy populations and coexistence of diploid plants and their closely related allotetraploid derivates from the Centaurea stoebe complex (Asteraceae). Methods An investigation was made of microspatial and microhabitat distribution, life-history and fitness traits, flowering phenology, genetic relatedness of cytotypes and intercytotype gene flow (cpDNA and microsatellites) in six mixed-ploidy populations in Central Europe. Key Results Diploids and tetraploids were genetically differentiated, thus corroborating the secondary origin of contact zones. The cytotypes were spatially segregated at all sites studied, with tetraploids colonizing preferentially drier and open microhabitats created by human-induced disturbances. Conversely, they were rare in more natural microsites and microsites with denser vegetation despite their superior persistence ability (polycarpic life cycle). The seed set of tetraploid plants was strongly influenced by their frequency in mixed-ploidy populations. Triploid hybrids originated from bidirectional hybridizations were extremely rare and almost completely sterile, indicating a strong postzygotic barrier between cytotypes. Conclusions The findings suggest that tetraploids are later immigrants into already established diploid populations and that anthropogenic activities creating open niches favouring propagule introductions were the major factor shaping the non-random distribution and habitat segregation of cytotypes at fine spatial scale. Establishment and spread of tetraploids was further facilitated by their superior persistence through the perennial life cycle. The results highlight the importance of non-adaptive spatio-temporal processes in explaining microhabitat and microspatial segregation of cytotype

Chromosome numbers and mode of reproduction in Picris hieracioides s.l. (Asteraceae), with notes on some other Picris taxa

Chromosome numbers for 88 plants of Picris hieracioides s.l. originating from 32 localities from Austria, Andorra, Croatia, France, Germany, Hungary, Italy, Slovakia and Spain are reported. All analysed plants were diploid, with 2n=2x=10. In addition to P. hieracioides s.l., we also found diploidy (2n=2x=10) in P. nuristanica from Kirgizia and P. japonica from Japan. Our data confirmed previously published results in the literature. We also studied the mode of reproduction in all three taxa. Picris nuristanica and P. japonica s.l. were able to produce some seeds by selfing, and this is the first record of autogamy in this genus. In P. hieracioides s.l. only strict allogamy was found, demonstrating the presence of a self-incompatibility system in this species. Because apomixis, namely apospory, has previously been reported in P. hieracioides, castration experiments were performed in order to confirm or reject this report. However, no evidence of apomictic seed formation was found within this species. In the light of our results, P. hieracioides should be considered a strictly sexual taxon

Biosystematic study of the diploid-polyploid Pilosella alpicola group with variation in breeding system: Patterns and processes

Members of the Pilosella alpicola species group (Asteraceae) are distributed throughout the alpine region of the European mountains (Alps, Carpathians, Balkan mountains). Like other Pilosella species groups (Hieracium subg. Pilosella ), the taxonomy and species' relationships are poorly understood mostly due to widespread facultative apomixis, frequent hybridization and polyploidization—the most important phenomena substantially involved in the evolutionary history of the genus. We assessed morphology, ploidy level, variation in breeding system and molecular variation within the P. alpicola group to provide a new taxonomic concept and to clarify evolutionary relationships among species and origin of polyploids. Multivariate morphometric analyses (UPGMA, CDA, PCA) applied on 324 plants originated from 21 populations revealed existence of four well-separated clusters corresponding to four allopatric taxa: P. alpicola s. str. (Alps), P. rhodopea (Balkan Peninsula, Southern Carpathians), P. serbica (Serbia and Montenegro) and P. ullepitschii (Carpathians). In total, four ploidy levels (2 x, 3 x, 4 x, 5 x ) were detected among 557 plants analysed from 19 populations by classical chromosome counting and flow cytometric analysis. The tetra- and pentaploids of P. alpicola s. str. have an allopatric distribution (Wallis Alps vs. Dolomites, respectively). Four ploidy levels with complex cytogeographic pattern and high frequency of mixed ploidy populations (75%) indicating a primary contact zone were recorded inP. rhodopea. Pilosella ullepitschii andP. serbica are exclusively diploid and are both sexually reproducing. In spite of a clear morphological separation, a molecular analysis (ITS and cpDNA sequences) suggests close relationships and rather recent origin of all studied taxa, except P. alpicola s. str. The latter taxon is an agamospermic allopolyploid that likely originated polytopi cally from a hybridization between P. rhodopea from the Balkans and P. glacialis from the Alps. In contrast to P. alpicola s. str., our data strongly support an autopolyploid origin of P. rhodopea polyploids which reproduced strictly sexually. Pilosella petraea , sometimes treated as a member of the P. alpicola group, differs from the remaining taxa by its conspicuous morphology, ecology and ITS polymorphism and should be removed from the group. Range shifts and extinctions were likely involved in shaping the evolutionary and modern distributional pattern of the group. Our combined methodological approach enabled us to propose a new taxonomic circumscription for the P. alpicola group and revealed auto- and allopolyploidization events

Allopatric distribution, ecology and conservation status of the Pilosella alpicola group (Asteraceae)

The Pilosella alpicola group comprises four morphologically distinct and geographically vicariant alpine taxa. We performed a thorough herbarium revision and literature survey to infer their distributional pattern(s). Pilosella alpicola s.s. occurs in the Alps in two disjunct areas: the Swiss Valais Alps and the Italian Dolomites. Historical records come also from the Austrian Alps (Gurktaler Alps and Hohe Tauern) and from one site from the Alpes Maritimes (Col de Larche), but the localities have not been recently confirmed. Pilosella rhodopea, a Balkan subendemic taxon, is quite widespread in Bulgaria (Stara planina Mts, Rila Mts and Pirin Mts), but is more rare in Albania, Greece and Macedonia. Interestingly, this species has also been recorded at two isolated sites in the Romanian southern Carpathians (the Căpăţînii and Cozia Mts). This occurrence underlines the floristic affinities of this part of the Carpathians to the Balkan flora. Only two localities of P. serbica, based on voucher specimens, have been recorded so far; Kopaonik Mts in Serbia and the Prokletije Mts in Montenegro. The records from other ranges are related to P. rhodopea. Pilosella ullepitschii, the detailed distribution of which has already been published, is a Carpathian endemic with its core area of distribution in the western Carpathians (Slovakia and Poland). Three isolated localities are also known in the eastern Carpathians (Nemira Mts) and one locality in the southern Carpathians (Bucegi Mts). The possible causes of disjunctions between and within species ranges are briefly discussed. Based on the distributional data, population sizes and ecology, we evaluate the conservation status of the P. alpicola taxa and propose their inclusion in national Red Lists

A taxonomic revision of the Pilosella alpicola group in the Carpathians

A taxonomic study of the Pilosella alpicola group growing in the Carpathians revealed the presence of two morphologically distinguishable taxa: P. ullepitschii (Błocki) Szeląg and P. rhodopea (Griseb.) Szeląg. While P. ullepitschii is endemic to the Carpathians, P. rhodopea is a Balkan subendemic with two isolated localities in the Southern Carpathians (Mt Cozia and Mt Zmeuretu). The core area of distribution of P. ullepitchii is the natural subalpine and alpine meadows of the Western Carpathians (the Vysoké and Západné Tatry Mts in Slovakia and Poland). In addition, only three isolated localities are known from the Nemira Mts (Romanian Eastern Carpathians) and one from the Bucegi Mts (Romanian Southern Carpathians). Interestingly, the Romanian populations occur in man-made habitats (secondary pastures). Karyological and flow cytometric analyses of 305 plants from 13 populations of P. ullepitschii revealed only diploid plants (2n = 2x = 18). One Carpathian population of P. rhodopea from Mt Cozia is also diploid. This is the first report of diploidy in this species. However, the populations from the main part of the distribution of this taxon in the Balkan mountains include other cytotypes. Detailed morphological descriptions and distributions for both taxa are given

Allozyme variation in diploid, polyploid and mixed-ploidy populations of the Pilosella alpicola group (Asteraceae): relation to morphology, origin of polyploids and breeding system

The Pilosella alpicola group includes four species (P. alpicola s.str., P. ullepitschii, P. rhodopea and P. serbica) with allopatric distributions (Alps, Balkans, Carpathians) and contrasting cytotype patterns (diploid, diploid-polyploid and polyploid species). Whereas diploid taxa (P. ullepitschii and P. serbica) reproduce sexually, the mode of reproduction of polyploid cytotypes reflects their origin: autopolyploids of P. rhodopea reproduce sexually, while allopolyploid cytotypes of P. alpicola s.str. apomictically. We used allozymes to elucidate overall genetic variation within the group and to test their utility for taxon discrimination, assessment of polyploid origin and possible correlations with breeding systems. Variation of five allozyme systems encoded by eight polymorphic loci and 29 alleles was studied in 20 populations and 298 plants representing all taxa. Allozymes were proved to be only of limited usefulness for the taxonomic classification within the P. alpicola group. The Western Carpathian populations of P. ullepitschii formed the only genetically well-differentiated group. The same allele suite shared by all cytotypes of P. rhodopea and presence of both balanced and unbalanced heterozygotes in tetraploids was consistent with autopolyploid origins of polyploids and provided further evidence for a primary contact zone. An isolated relic population of P. rhodopea from the Southern Carpathians exhibited lowered values of genetic diversity when compared to the core area. Pronounced fixed heterozygosity was found in P. alpicola s.str., supporting its allopolyploid origin. In accordance with assumptions, genotypic variability was significantly higher in sexually reproducing diploid and diploid-polyploid taxa than in apomictic P. alpicola s.str.</i

Geographical parthenogenesis, genome size variation and pollen production in the arctic-alpine species Hieracium alpinum

Hieracium alpinum L. (Asteraceae) is an arctic-alpine species distributed throughout Europe with both diploid and triploid cytotypes. We determined the ploidy levels of plants from 23 populations from Austria, Bosnia and Herzegovina, Finland, Italy, Norway, Romania, Slovakia, Switzerland and Ukraine. Data showed a non- overlapping pattern of cytotype distribution: sexually reproducing diploids (2n = 2x = 18) occur solely in the Eastern and Southern Carpathians, while apomictic triploids (2n = 3x = 27) cover the rest of the range. Such clear-cut allopatry is rather rare in vascular plants with geographical parthenogenesis. Comparison of absolute genome size indicates genome downsizing (by on average 3.7%) of haploid DNA amount in triploids relative to diploids. Genome size further correlated with longitude and latitude in the Alps, with decreasing absolute DNA content from west to east, and from south to north. While previously published data indicated complete male sterility of triploid plants, we found that plants from the Alps and Bosnia and Herzegovina commonly produced some pollen, whereas populations from the Western Carpathians and Scandinavia seemed to be almost completely pollen sterile. Scenarios about the evolution of geographical parthenogenesis in H. alpinum are discussed

Loss of genetic diversity in isolated populations of an alpine endemic Pilosella alpicola subsp. ullepitschii : effect of long-term vicariance or long-distance

Pilosella alpicola subsp. ullepitschii (Asteraceae) is a strictly allogamous, diploid Carpathian endemic. Its distribution range comprises two areas separated by about 600 km. While in the Western Carpathians (Slovakia and Poland) the taxon occurs in numerous sites, only four localities of man-made origin are known from the Eastern and Southern Carpathians (Romania). We used allozyme markers to test two likely possible scenarios for the origin of this disjunction: long distance dispersal and vicariance. Our data indicate a significant loss of genetic diversity in the isolated Eastern and Southern Carpathian populations in following genetic parameters (averaged per region): percentage of polymorphic loci (38.9% found in the Eastern and Southern Carpathians versus 58.3% in the Western Carpathians), allelic richness (1.4 vs. 1.6), expected heterozygosity (0.134 vs. 0.235), mean number of distinguishable multilocus genotypes (4.3 vs. 10.6) and proportion of distinguishable multilocus genotypes (0.34 vs. 0.68). Higher proportion of homozygous loci found in the Eastern and Southern Carpathian populations might indicate a higher rate of inbreeding due to non-random mating. We assume that these genetically depauperate populations have experienced a very strong genetic bottleneck, probably due to a founder effect. Although our data suggest that the long-distance dispersal model is most likely, more discriminate genetic markers should be used to test this further