Katarzyna Buczkowska

Conocephalum salebrosum (Marchantiopsida, Conocephalaceae) - a new Holarctic liverwort species

Detailed analysis of 270 specimens of Conocephalum conicum (L.) Dumort., mainly from the POZW Herbarium revealed some diagnostic differences between two cryptic species originally detected on the basis of isozyme studies. Several diagnostic characters were found in the structure of the archegoniophore, sporophyte and sterile thallus. The most reliable diagnostic features are size and structure of stomatal apparatus of the archegoniophore ‘heads’, type of junction of the air chamber walls with epidermal cells, and the thallus surface details. On the basis of morphological and anatomical diagnostic characters, two formerly cryptic species are recognized following the rules of formal taxonomy. The lectotype of C. conicum preserved in FI was found to possess characteristics of cryptic species C. conicum- species L. Accordingly, the species previously named as C. conicum–species L must bear the name Conocephalum conicum (L.) Dumort. No published name was related to C. conicum – species S, which is therefore described as a new species: Conocephalum salebrosum Szweykowski, Buczkowska & Odrzykoski. Both species are fully described and their diagnostic characters are illustrated. Distribution maps for both species are given.

Genetic differentiation of Calypogeia fissa Raddi (Hepaticae, Jungermanniales) in Poland

Abstract.The genetic structure of 24 populations of the Calypogeia fissa complex from Poland and Germany has been studied. Fifteen putative loci in eight enzyme systems were resolved. Ten of these loci were polymorphic. The inter-population variation of the examined populations of C. fissa was partitioned into three distinct groups, tentatively called group PS, PB and G. Groups PS and PB occur in Poland, group G in Germany. Fixed alleles were diagnostic of each of the three groups. Obtained values of genetic distances showed a high level of genetic distance (0.664–0.796) between populations from Germany and those of the Polish group PS. A UPGMA dendrogram based on Nei’s genetic distance coefficient was constructed; it showed differentiation of the studied populations in two distinct groups, one consisting of group PS from Poland, the second of plants from Germany and the Polish group PB. Studies of oil bodies in C. fissa showed, as in other Calypogeia species, their high correlation with the isoenzyme phenotypes.

The development of species-specific SCAR markers for delimitation of Calypogeia species

Abstract The genus Calypogeia belongs to one of the most difficult groups of leafy liverworts, with about 90 species. Nine species of the genus are currently known from Europe: C. azurea, C. integristipula, C. neesiana, C. suecica, C. muelleriana, C. sphagnicola, C. fissa, C. arguta, and C. azorica. Recently, another species, morphologically resembling C. muelleriana but genetically distinct from it, was detected using isozyme markers. In the present study, C. muelleriana and C. sp. nov. were further characterized by 13 and 14 specific inter-simple sequence repeat band patterns, respectively. Nei’s genetic distance between the two species was 0·61. Three species-specific bands were converted into sequence characterized amplified region (SCAR) markers. The first SCAR primer pair amplified a product of 360 and 600 bp in C. muelleriana and C. sp. nov., respectively. The two other SCAR markers were diagnostic for other European Calypogeia species.

Allopolyploid speciation of Calypogeia sphagnicola (Jungermanniopsida, Calypogeiaceae) based on isozyme and DNA markers

Calypogeia sphagnicola is one of nine species of the genus Calypogeia known in Europe. Occurrence of the species is closely connected with peat bogs. Nowadays, two forms of this species are distinguished—C. sphagnicola f. sphagnicola and C. sphagnicola f. paludosa. The results of the present study, based on two classes of markers—isozymes and sequences of chloroplast genom (trnH-psbA, rpoC1)—unanimously support the genetic differentiation within the taxon and show that the present-day forms represent genetically distinct species. Phylogenetic analysis resolved two lineages that correspond with the present-day forms with high bootstrap support, which differ in ploidy level: C. sphagnicola f. sphagnicola is haploid, whereas C. sphagnicola f. paludosa is a diploid form. Allopolyploid origin of the diploid form was revealed by the isozyme pattern. Nei’s genetic distance between the two present-day forms of C. sphagnicola was 0.472. The forms in Poland have an allopatric pattern of geographic distribution: C. sphagnicola f. sphagnicola occurs exclusively in the lowlands of the northern part of the country on raised peat bogs, whereas C. sphagnicola f. paludosa is found only in the mountains of southern Poland, mainly in the subalpine zone, where it grows on Sphagnum-Polytrichum hummocks on the upper part of north-facing slopes. Plants regarded in this study as C. sphagnicola f. sphagnicola morphologically correspond to the syntype specimen of C. sphagnicola.

Genetic variation in the liverwort Bazzania trilobata inferred from ISSR markers

Abstract The distribution of Bazzania trilobata in Poland coincides with two parts of the natural distribution range of Norway spruce: the mountains of southern Poland and the northeastern lowlands. The occurrence of this species is connected with primeval forest communities and it was recognized as a relict of them. Genetic variation of 10 populations of B. trilobata from two different regions of Poland (lowlands and mountains) was studied in order to establish the genetic structure of this species and to compare the level of genetic variation within and among populations originating from primeval and managed forests. An analysis based on polymorphic inter-simple sequence repeat (ISSR) loci revealed a high level of total gene diversity in B. trilobata (H T=0.308). A higher amount of polymorphism was found among than within populations. Genetic variation of populations from the primary forest (Tatras and Białowieza National Park) was higher than for those originating from disturbed habitats. The Bayesian method showed consistent grouping of samples corresponding with populations, with the best grouping in 12 different clusters reflecting a geographic pattern. The geographic pattern of genetic differentiation was supported by a neighbour-joining (NJ) dendrogram based on genetic distances. All mountain populations were grouped together in one subcluster, but lowland populations were more differentiated and did not form one clear group.

Volatile compounds in cryptic species of the Aneura pinguis complex and Aneura maxima (Marchantiophyta, Metzgeriidae).

Aneura pinguis is one of the liverwort species complexes that consist of several cryptic species. Ten samples collected from different regions in Poland are in the focus of our research. Eight of the A. pinguis complex belonging to four cryptic species (A, B, C, E) and two samples of closely related species Aneura maxima were tested for the composition of volatile compounds. The HS-SPME technique coupled to GC/FID and GC/MS analysis has been applied. The fiber coated with DVB/CAR/PDMS has been used. The results of the present study, revealed the qualitative and quantitative differences in the composition of the volatile compounds between the studied species. Mainly they are from the group of sesquiterpenoids, oxygenated sesquiterpenoids and aliphatic hydrocarbons. The statistical methods (CA and PCA) showed that detected volatile compounds allow to distinguish cryptic species of A. pinguis. All examined cryptic species of the A. pinguis complex differ from A. maxima. Species A and E of A. pinguis, in CA and PCA, form separate clusters remote from two remaining cryptic species of A. pinguis (B and C) and A. maxima. Relationship between the cryptic species appeared from the chemical studies are in accordance with that revealed on the basis of DNA sequences.

Morphological differentiation of Calypogeia muelleriana (Jungermanniales, Hepaticae) in Poland

Morphological differentiation of Calypogeia muelleriana (Jungermanniales, Hepaticae) in Poland Morphological and anatomical characters and oil bodies were studied in 52 samples from Poland: 31 samples of typical Calypogeia muelleriana and 21 of a newly detected taxon with the use of isozyme markers. Plants of the new taxon morphologically resemble C. muelleriana, but differ from it significantly in 16 morphological traits as well as oil body characters. The greatest morphometric differences were found in the features connected with underleaves. The Mahalanobis distance based on the 47 quantitative traits, between the newly detected taxon and C. muelleriana was larger than that between C. muelleriana and C. azurea - a generally accepted species. The forward stepwise method of discriminant analysis showed that the set of diagnostic characters could be limited to five. A dendrogram constructed on the basis of the Euclidean distance, using the set of diagnostic characters, divided the examined samples into two groups that correlated with groups detected by genetic markers. Results of a multivariate analysis showed that five morphological traits are sufficient for a proper classification of plants to these two taxa.

The extraordinary variation of the organellar genomes of the Aneura pinguis revealed advanced cryptic speciation of the early land plants

Aneura pinguis is known as a species complex with several morphologically indiscernible species, which are often reproductively isolated from each other and show distinguishable genetic differences. Genetic dissimilarity of cryptic species may be detected by genomes comparison. This study presents the first complete sequences of chloroplast and mitochondrial genomes of six cryptic species of A. pinguis complex: A. pinguis A, B, C, E, F, J. These genomes have been compared to each other in order to reconstruct phylogenetic relationships and to gain better understanding of the evolutionary process of cryptic speciation in this complex. The chloroplast genome with the nucleotide diversity 0.05111 and 1537 indels is by far more variable than mitogenome with π value 0.00233 and number of indels 1526. Tests of selection evidenced that on about 36% of chloroplast genes and on 10% of mitochondrial genes of A. pinguis acts positive selection. It suggests an advanced speciation of species. The phylogenetic analyses based on genomes show that A. pinguis is differentiated and forms three distinct clades. Moreover, on the cpDNA trees, Aneura mirabilis is nested among the cryptic species of A. pinguis. This indicates that the A. pinguis cryptic species do not derive directly from one common ancestor.

Differentiation and Genetic Variability of Three Cryptic Species within the Aneura pinguis Complex (Jungermanniidae, Marchantiophyta)

ABSTRACT 1652 individuals of Aneura pinguis from Poland were surveyed for variation in 12 putative gene loci. Based on isozyme data, we distinguished three cryptic species. No evidence for gene flow between these species was found. To date, no qualitative morphological characters are available, which would allow delimitation of the cryptic species of A. pinguis. Hence, these species are not formally described, but assigned as cryptic species A, B, and C. The mean genetic distance (D) between them is 1.3393. The highest genetic variation within populations (Hs ) was found in species A, and the lowest in species B. Individual species of A. pinguis differ in their habitat preferences. Species A is the most common, it occurs mostly in the Western Carpathians, grows mainly on calcareous rocks and humus. Species B is the most frequent in the Eastern Carpathians on clay soil. Species C is the rarest, it can be found both in lowlands and mountains, but mainly in lowlands and on various substrata. All studied cryptic species occur partly sympatrically.

Two ploidy levels of genetically delimited groups of the Calypogeia fissa complex (Jungermanniopsida, Calypogeiaceae)

Abstract Calypogeia fissa is a suboceanic-mediterrean and amphiatlantic species, which comprises two subspecies: C. fissa subsp. fissa occurring in Europe and C. fissa subsp. neogea Schust. known from North America. Recently, within the European part of distribution, three groups (PS, PB and G) were distinguished with the aid of genetic and molecular markers. The flow cytometry results revealed that two of the detected groups of the European C. fissa, which are frequent in Poland (PS and PB), differ in ploidy level: the PS group is haploid, whereas the PB group is diploid. Isozyme pattern at two loci may suggest an allopolyploid origin of the diploid PB group.