Jaromír Kučera

Intraspecific Variation in Viola suavis in Europe: Parallel Evolution of White-flowered Morphotypes

BACKGROUND AND AIMS Viola species are commonly grown for their ornamental flowers, but their evolutionary history and taxonomy are often complicated and have been poorly explored so far. This is a study of the polymorphic, typically blue-flowered species Viola suavis, concentrating on the white-flowered populations of uncertain taxonomic assignment that occur in Spain and central and south-eastern Europe. The aim was to resolve their origin and taxonomic status and to study the intraspecific structure and (post)glacial history of this species. METHODS Viola suavis and five close relatives were sampled from multiple locations and subjected to molecular (AFLP, sequencing of nrDNA ITS) and morphometric analyses. Data on ploidy level and pollen fertility were also obtained, to address an assumed hybrid origin of the white-flowered populations. KEY RESULTS In V. suavis a strong intraspecific genetic split into two groups was observed, indicating that there has been a long-term isolation and survival in distinct glacial refugia. The white-flowered populations could be placed within the variation range of this species, and it is clear that they evolved independently in two distant areas. Their parallel evolution is supported by both morphological and genetic differentiation. The strongly reduced genetic variation and absence of unique AFLP fragments suggest their derived status and origin from the typical, blue-flowered populations. CONCLUSIONS These results suggest that intraspecific variation in V. suavis has been largely shaped by population isolations during the last glaciation and subsequent recolonizations, although cultivation and vegetative spread by humans have affected the present picture as well.

Molecular study of the Cardamine maritima group (Brassicaceae) from the Balkan and Apennine Peninsulas based on amplified fragment length polymorphism

The amphi-Adriatic region, and especially the Western Balkan Peninsula, belongs to the most important biodiversity hotspots in the temperate region. Nevertheless, detailed phylogeographic and molecular systematic studies in the Western Balkan are rare due to sporadic sampling in regions, where access has been, until recently, restricted by war. The Cardamine maritima group, which is the focus of this study, comprises not only the currently recognised species C. maritima and C. monteluccii, but also other taxa, which have been rendered to synonymy by most of the national floras and checklists. Molecular data acquired by the amplified fragment length polymorphism method showed a clear pattern within the group. Italian populations of C. monteluccii are well separated from Balkan taxa. In a step forward from previous taxonomic confusion surrounding Balkan populations, the present study confirms that five allopatric units—each with a clearly delimited and a rather restricted distribution range—can be easily recognised here. They correspond to C. fialae, C. serbica, C. rupestris, and two genetically distant and allopatric units within C. maritima. While individual taxa gained high bootstrap support in the neighbour-joining tree, there is low support for the internal nodes and it is hard to infer any relationships among taxa based on this information. The majority of Balkan populations of the C. maritima group exhibit features of genetic variability that enable us to hypothesise that these populations are relic ones.

The Morphological and Genetic Variation in the Polymorphic Species Picris hieracioides (Compositae, Lactuceae) in Europe Strongly Contrasts with Traditional Taxonomical Concepts

Abstract The present paper provides a large-scale taxonomic revision of Picris hieracioides, a highly polymorphic and taxonomically controversial species in Europe. Altogether, 104 populations were sampled and examined using multivariate morphometrics and genetic amplified fragment length polymorphism (AFLP) data. Two morphotypes are delimited, the ‘higher altitude’ and the ‘lower altitude’ morphotypes, which are congruent with the two main genetic groupings revealed by AFLP markers. It is concluded that two subspecies should be recognized within this species, P. hieracioides subsp. hieracioides, comprising annual to biennial plants occupying dry, sunny, often man-made habitats at low altitudes, and P. hieracioides subsp. umbellata, being mostly a short-lived perennial that grows in mesic, semi-natural or natural habitats at higher altitudes. This infraspecific treatment strongly contrasts with the traditional taxonomic concepts, which recognize up to 10 subspecies of P. hieracioides in Europe. An identification key and a nomenclatural account are presented, including the designation of lectotypes. Picris hieracioides harbors large genetic variation, and two lineages can be recognized within each subspecies, most likely reflecting their glacial survival and postglacial colonization routes. Long-distance dispersals, anthropogenic introductions and recent spread are suggested to have shaped their genetic structure as well. A Balkan endemic P. hispidissima, although morphologically and ecologically well defined, appears to be genetically close to P. hieracioides.

Genetic and morphological variation in Viola suavis s.l. (Violaceae) in the western Balkan Peninsula: two endemic subspecies revealed

The Balkan Peninsula, with many endemic species, is known as one of the most important speciation and diversification centres in Europe. Here, we present a study of the western Balkan populations of the polymorphic European species, V. suavis s.l., which have been reported under the name V. adriatica, but their taxonomic status and position within the genus have remained uncertain. Viola suavis s.l. and nine close relatives sampled across Europe were subjected to molecular (sequencing of nuclear ribosomal internal transcribed spacers and amplified fragment length polymorphism), karyological and morphometric analyses. Our results revealed the presence of four allopatric, genetically and morphologically differentiated lineages within V. suavis s.l. in Europe, which are suggested here to be recognized at the subspecific rank. Populations from the western Balkans were segregated into two distinct entities: (1) those from north-western Croatia correspond to the previously recognized taxon, V. suavis subsp. adriatica and (2) those from southern Dalmatia (southern Croatia, southern Bosnia and Herzegovina, and south-western Montenegro) are described here as V. suavis subsp. austrodalmatica subsp. nov. The other two lineages of V. suavis s.l., which both harbour blue- and white-flowered morphotypes, occur in central and eastern Europe (V. suavis subsp. suavis) and in north-eastern Spain (plants provisionally treated as V. suavis ‘Spain’). The AFLP and morphological data indicate gene flow between the nominate subspecies and V. suavis subsp. adriatica in a few localities. The distribution of the two western Balkan subspecies is discussed and an identification key to the V. suavis subspecies in Europe is presented.

Dealing with discordant genetic signal caused by hybridisation, incomplete lineage sorting and paucity of primary nucleotide homologies: a case study of closely related members of the genus Picris subsection Hieracioides (Compositae).

We investigated genetic variation and evolutionary history of closely related taxa of Picris subsect. Hieracioides with major focus on the widely distributed P. hieracioides and its closely related congeners, P. hispidissima, P. japonica, P. olympica, and P. nuristanica. Accessions from 140 sample sites of the investigated Picris taxa were analyzed on the infra- and the inter-specific level using nuclear (ITS1-5.8S-ITS2 region) and chloroplast (rpl32-trnL (UAG) region) DNA sequences. Genetic patterns of P. hieracioides, P. hispidissima, and P. olympica were shown to be incongruent and, in several cases, both plastid and nuclear alleles transcended borders of the taxa and genetic lineages. The widespread P. hieracioides was genetically highly variable and non-monophyletic across both markers, with allele groups having particular geographic distributions. Generally, all gene trees and networks displayed only a limited and statistically rather unsupported resolution among ingroup taxa causing their phylogenetic relationships to remain rather unresolved. More light on these intricate evolutionary relationships was cast by the Bayesian coalescent-based analysis, although some relationships were still left unresolved. A combination of suite of phylogenetic analyses revealed the ingroup taxa to represent a complex of genetically closely related and morphologically similar entities that have undergone a highly dynamic and recent evolution. This has been especially affected by the extensive and recurrent gene flow among and within the studied taxa and/or by the maintenance of ancestral variation. Paucity of phylogenetically informative signal further hampers the reconstruction of relationships on the infra- as well as on the inter-specific level. In the present study, we have demonstrated that a combination of various phylogenetic analyses of datasets with extremely complex and incongruent phylogenetic signal may shed more light on the interrelationships and evolutionary history of analysed species groups.

Cardamine pancicii (Brassicaceae), a neglected species of the Balkan Peninsula

Abstract Lakušić, D., Novčić, R., Kučera, J. & Marhold, K.: Cardamine pancicii (Brassicaceae), a neglected species of the Balkan Peninsula. — Willdenowia 36 (Special Issue): 177–191. — ISSN 0511-9618;

Re-evaluation of the morphological variability of Microglossum viride and M. griseoviride sp. nov.

Studies in Microglossum viride (Pers.) Gillet revealed that the name was used incorrectly for two similar but different taxa. Analyses of morphological, ecological and molecular (sequences of ITS and LSU region of rRNA gene) characters of type and voucher specimens of M. viride and related taxa resulted in delimitation and description of a new species, Microglossum griseoviride V. Kučera, Lizoň & M. Tomšovský. Lectotypes of Geoglossum viride Pers., and epitype of Geoglossum viride are designated. Species Microglossum minus Velen. and Microglossum lutescens Boud. are confirmed to be conspecific to M. viride.

Diversification dynamics and transoceanic Eurasian-Australian disjunction in the genus Picris (Compositae) induced by the interplay of shifts in intrinsic/extrinsic traits and paleoclimatic oscillations

Understanding transcontinental biogeographic patterns has been one of the main foci of the field of biogeography. While multiple explanations for transcontinental disjunctions have been proposed, little is still known about the relative importance of intrinsic and extrinsic traits for the diversification dynamics of disjunct taxa. Here, we study the evolutionary history of the genus Picris L. (Compositae), a great model for investigating the diversification dynamics of transoceanic bipolar disjunct organisms. Ancestral state reconstructions indicate that the most recent common ancestor (MRCA) of Picris was a semelparous and heterocarpic herb that lived in unpredictable environments of North Africa and West Asia. Diversification analyses suggest a significant shift in speciation ca. 1 million years ago, likely associated with the onset of the mid-Pleistocene revolution. Longevity characters are correlated with the evolution of particular fruit types and with environmental conditions. Heterocarpic species are mostly semelparous herbs strongly linked with unpredictable habitats, while homocarpic taxa are mostly iteroparous plants occurring in predictable environments. Binary-state speciation and extinction analyses suggest that homocarpy, iteroparity, and habitats predictability accelerate diversification. Although the combination of homocarpy and iteroparity evolved in several lineages, only members of the P. hieracioides group were able to colonise Eurasia and expand to Australia by transoceanic dispersal. Those findings indicate that large-scale colonisation events depend on a complex interplay of intrinsic and extrinsic factors.

Karyological patterns in the European endemic genus Soldanella L.: Absolute genome size variation uncorrelated with cytotype chromosome numbers

PREMISE OF THE STUDY Detailed knowledge about the karyological diversity of organisms undoubtedly represents one of the crucial steps toward a better understanding of their evolutionary trends and history. We investigated the cytotype and absolute genome size (AGS) patterns in the European mountain-dwelling genus Soldanella (Primulaceae) in light of its geographic distribution and ecological diversification. METHODS Our chromosome number survey was based on 34 newly determined and 125 previously published chromosome counts. AGS was estimated on the basis of propidium iodide (PI) flow cytometry (299 individuals, 110 populations). KEY RESULTS We confirmed the existence of two cytotypes with the same ploidy level, i.e., euploid 2n = 40 and dysploid 2n = 38. The overall infrageneric AGS variation ranged between 2.97 and 3.99 pg (25.6% variation). The 2n = 40 cytotype harbors a modest amount of continuous AGS variation. With regard to its distribution area and ecology, the cytotype is ubiquitous. By contrast, the 2n = 38 cytotype was detected only in six forest-dwelling taxa with AGS variation segregated into three discrete, geographically separated groups. The AGS variation of the 2n = 38 cytotype was strongly correlated with elevation and longitude. CONCLUSIONS Despite the apparent morphological and ecological variation, members of the genus Soldanella have not undergone any pronounced cytotype and AGS diversification during their evolutionary history. The lack of correlation between chromosome numbers and AGS indicates that the evolutionary mechanism behind the origin of the dysploid cytotype 2n = 38 was a chromosomal fusion.

Disentangling identity of species of the genus Taphrina parasitizing herbaceous Rosaceae, with proposal of Taphrina gei-montani sp. nov.

Five strains (CCY 058-007-001T, CCY 058-007-002, CCY 058-007-003, CCY 058-007-004 and CCY 058-007-005) of a novel parasitic yeast belonging to the genus Taphrina were isolated from leaf tissues of Geum montanum L. (Rosaceae), collected from the Vysoké Tatry Mountains, Slovakia. Genetic analyses revealed that these isolates differ by 15 unique substitutions in the ITS region and by six substitutions in the rns gene from all other species of the genus Taphrina analysed hitherto. The novel strains are also distinguished from all other species of the genus Taphrina by their morphology, biochemical properties and ecology. These strains represent a novel species, for which the name Taphrina gei-montani sp. nov. is proposed. The type strain is CCY 058-007-001T (=CBS 14159=BU001). The MycoBank number is MB815677. The present study also demonstrates that two distinct species of the genus Taphrina parasitize the herbaceous Rosaceae: Taphrina gei-montani sp. nov. on Geum montanum and Taphrina tormentillae on Potentilla species.