Biometrical study of some individuals chosen from Pinus mugo turra populations in the peat bog "Bór na Czerwonem"

Sixteen individuals were sampled for study of variation in 17 anatomical and morphological characters. Only individuals of low polycormic growth (trait typical for Pinus mugo Turra) connected with incurved one-year-cone stipes (a similar situation exists in Pinus sylvestris L.) were chosen, thus the sample studied cannot be treated as a random one. It has been shown by multivariate statistical analysis that these 16 individuals are quite different from each other, Mahalanobis'generalized distances between them being nearly 50% significantly different from 0. The sample studied in this respect is distinctly different from pure stands of both putative parental species (i.e. Pinus mugo and P. sylvestris). Every plant studied shows a different combination of traits typical (or nearly typical) for both the above-mentioned species and traits that are truly intermediate between them. The results support the frequently expressed opinion that the mountain pine population from the peat bog "Bór na Czerwonem" is, in fact, a hybrid swarm formed by hybridization between Pinus mugo and Pinus sylvestris

Chemical Fingerprinting of Cryptic Species and Genetic Lineages of <i>Aneura pinguis</i> (L.) Dumort. (Marchantiophyta, Metzgeriidae)

Aneura pinguis (L.) Dumort. is a representative of the simple thalloid liverworts, one of the three main types of liverwort gametophytes. According to classical taxonomy, A. pinguis represents one morphologically variable species; however, genetic data reveal that this species is a complex consisting of 10 cryptic species (named by letters from A to J), of which four are further subdivided into two or three evolutionary lineages. The objective of this work was to develop an efficient method for the characterisation of plant material using marker compounds. The volatile chemical constituents of cryptic species within the liverwort A. pinguis were analysed by GC-MS. The compounds were isolated from plant material using the HS-SPME technique. Of the 66 compounds examined, 40 were identified. Of these 40 compounds, nine were selected for use as marker compounds of individual cryptic species of A. pinguis. A guide was then developed that clarified how these markers could be used for the rapid identification of the genetic lineages of A. pinguis. Multivariate statistical analyses (principal component and cluster analysis) revealed that the chemical compounds in A. pinguis made it possible to distinguish individual cryptic species (including genetic lineages), with the exception of cryptic species G and H. The classification of samples based on the volatile compounds by cluster analysis reflected phylogenetic relationships between cryptic species and genetic lineages of A. pinguis revealed based on molecular data

Super-Mitobarcoding in Plant Species Identification? It Can Work! The Case of Leafy Liverworts Belonging to the Genus <i>Calypogeia</i>

Molecular identification of species is especially important where traditional taxonomic methods fail. The genus Calypogeia belongs to one of the tricky taxons. The simple morphology of these species and a tendency towards environmental plasticity make them complicated in identification. The finding of the universal single-locus DNA barcode in plants seems to be ‘the Holy Grail’; therefore, researchers are increasingly looking for multiloci DNA barcodes or super-barcoding. Since the mitochondrial genome has low sequence variation in plants, species delimitation is usually based on the chloroplast genome. Unexpectedly, our research shows that super-mitobarcoding can also work! However, our outcomes showed that a single method of molecular species delimitation should be avoided. Moreover, it is recommended to interpret the results of molecular species delimitation alongside other types of evidence, such as ecology, population genetics or comparative morphology. Here, we also presented genetic data supporting the view that C. suecica is not a homogeneous species

Comparative Analysis of Four Calypogeia Species Revealed Unexpected Change in Evolutionarily-Stable Liverwort Mitogenomes

Liverwort mitogenomes are considered to be evolutionarily stable. A comparative analysis of four Calypogeia species revealed differences compared to previously sequenced liverwort mitogenomes. Such differences involve unexpected structural changes in the two genes, cox1 and atp1, which have lost three and two introns, respectively. The group I introns in the cox1 gene are proposed to have been lost by two-step localized retroprocessing, whereas one-step retroprocessing could be responsible for the disappearance of the group II introns in the atp1 gene. These cases represent the first identified losses of introns in mitogenomes of leafy liverworts (Jungermanniopsida) contrasting the stability of mitochondrial gene order with certain changes in the gene content and intron set in liverworts

DNA barcoding, ecology and geography of the cryptic species of <i>Aneura pinguis</i> and their relationships with <i>Aneura maxima</i> and <i>Aneura mirabilis</i> (Metzgeriales, Marchantiophyta)

<div><p><i>Aneura pinguis</i> is a thalloid liverwort species with broad geographical distribution. It is composed of cryptic species, however, the number of cryptic species within <i>A</i>. <i>pinguis</i> is not known. Five cpDNA regions (<i>matK</i>, <i>rbcL</i>, <i>rpoC1</i>, <i>trnH-psbA</i> and <i>trnL-trnF</i>) and the entire nuclear ITS region were studied in 130 samples of <i>A</i>. <i>pinguis</i> from different geographical regions. The relationships between the cryptic species of <i>A</i>. <i>pinguis</i>, <i>A</i>. <i>maxima</i> and <i>A</i>. <i>mirabilis</i> were analyzed. All of the examined samples were clustered into 10 clades corresponding to 10 cryptic species of <i>A</i>. <i>pinguis</i> (marked A to J). <i>Aneura mirabilis</i> and <i>A</i>. <i>maxima</i> were nested among different cryptic species of <i>A</i>. <i>pinguis</i>, which indicates that <i>A</i>. <i>pinguis</i> is a paraphyletic taxon. Subgroups were found in cryptic species A, B, C and E. As single barcodes, all tested DNA regions had 100% discriminant power and fulfilled DNA barcode criteria for species identification; however, the only combination detected in all subgroups was <i>trnL</i>-<i>trnF</i> with <i>trnH</i>-<i>psbA</i> or ITS2. The distances between cryptic species were 11- to 35-fold higher than intraspecific distances. In all analyzed DNA regions, the distances between most pairs of cryptic <i>A</i>. <i>pinguis</i> species were higher than between <i>A</i>. <i>maxima</i> and <i>A</i>. <i>mirabilis</i>. All cryptic species of <i>A</i>. <i>pinguis</i> clearly differed in their habitat preferences, which suggests that habitat adaptation could be the main driving force behind cryptic speciation within this taxon.</p></div

Does Calypogeia azurea (Calypogeiaceae, Marchantiophyta) occur outside Europe? Molecular and morphological evidence.

Oil bodies are the unique feature of most liverworts. Their shape, color and distribution pattern in leaf and underleaf cells are important taxonomic features of the genus Calypogeia. Most species of the genus Calypogeia have pellucid and colorless oil bodies, whereas colored, including gray to pale brown, purple-brown or blue oil bodies, are rare. To date, C. azurea was the only species with blue oil bodies to have been considered as a species of the Holarctic range. This species has been noted in various parts of the northern hemisphere-from North America, through Europe to the Far East. The aim of this study was to determine the genetic diversity of C. azurea from different parts of its distribution range and to ascertain whether blue oil bodies appeared once or several times in the evolution of the genus Calypogeia. The phylogenetic analyses based on four plastid regions (rbcL, trnG, trnL, trnH-psbA) and one nuclear region (ITS2) revealed that C. azurea is presently a paraphyletic taxon, with other Calypogeia species nested among C. azurea accessions that were clustered into four different clades. Based on the level of genetic divergence (1.03-2.17%) and the observed morphological, ecological and geographical differences, the evaluated clades could be regarded as previously unrecognized species. Four species were identified: C. azurea Stotler & Crotz (a European species corresponding to the holotype), two new species from Pacific Asia-C. orientalis Buczkowska & Bakalin and C. sinensis Bakalin & Buczkowska, and a North American species which, due to the lack of identifiable morphological features, must be regarded as the cryptic species of C. azurea with a provisional name of C. azurea species NA

K2P (%) genetic variation in the DNA sequences of studied groups of <i>Aneura pinguis</i>.

<p>K2P (%) genetic variation in the DNA sequences of studied groups of <i>Aneura pinguis</i>.</p