Is Lotus zhegulensis Klok. (Fabaceae) a separate species endemic of the Middle Volga Region or just a variety of a widespread L. corniculatus L.?

We have studied several samples from Lotus corniculatus s. l. and L. zhegulensis populations with the use of SSR and sequenced nuclear (ITS) and plastid (atpB-rbcL, ndhC-trnV, rpl32-trnL(UAG), trnH-psbA, trnL-trnF) markers. We analysed 9 local populations from the Volga River valley ranging from Ulyanovsk in the north to Volgograd in the south, as well as two local populations from Moscow and Lipetsk Provinces. Analyses of SSR markers using the STRUCRURE program divided the sample into three genetic clusters. Populations from Moscow and Lipetsk Provinces, and three local populations from the ‘locus classicus’ location of L. zhegulensis in Samara Province appeared to be the most genetically differentiated while all the other populations were variously genetically admixed. Low pairwise Fst values indicate low genetic differentiation of Lotus populations and the intraspecific nature of the revealed diversity. The nuclear and plastid DNA sequences analyzed yielded little information. Substitutions and indels revealed were mostly autapomorphies characteristic of separate specimens or small groups of specimens but not of populations. Our study enables to firmly conclude that Lotus populations from the right bank of the Volga River in vicinity of Zhiguli upland described as L. zhegulensis are weakly genetically differentiated from other populations of Lotus corniculatus s. l. and, hence, cannot be regarded as a separate species.

Designation of the neotype of Triatoma dimidiata (Latreille, 1811) (Hemiptera, Reduviidae, Triatominae), with full integrated redescription including mitogenome and nuclear ITS-2 sequences

The taxonomic status of Triatoma dimidiata (Latreille, 1811) is, by far, the most discussed within Triatominae. Molecular studies have recovered at least three independently evolving lineages in T. dimidiata across its range. The original description of T. dimidiata (as Reduvius dimidiatus) included few taxonomic characters, and no types were assigned. To define and describe the cryptic diversity within T. dimidiata sensu lato (s.l.), a neotype must be designated. For this purpose, all 199 specimens identified as T. dimidiata from the collections of the Smithsonian Institution – National Museum of Natural History and the American Museum of Natural History, ranging from Peru to Mexico, were studied. Only one specimen (from Tumbes, Peru) matched the combination of characters as listed in the original description, and it is herein formally designated as the neotype for T. dimidiata. The neotype is morphologically described and DNA sequences of its whole mitochondrial genome and the nuclear second internal transcribed spacer region (ITS2), commonly used in triatomine molecular systematics studies, are presented and compared to other publicly available sequences of T. dimidiata s.l. in GenBank. Our results suggest that T. dimidiata sensu stricto (s.s.) is somewhat rare and, therefore, unlikely to serve as a major vector of Chagas disease.

Complete Mitogenomes of Two Aragoa Species and Phylogeny of Plantagineae (Plantaginaceae, Lamiales) Using Mitochondrial Genes and the Nuclear Ribosomal RNA Repeat

Aragoa, comprising 19 high-altitude North Andean species, is one of three genera in the Plantagineae (Plantaginaceae, Lamiales), along with Littorella and Plantago. Based primarily on plastid data and nuclear ITS, Aragoa is sister to a clade of Littorella + Plantago, but Plantagineae relationships have yet to be assessed using multigene datasets from the nuclear and mitochondrial genomes. Here, complete mitogenomes were assembled for two species of Aragoa (A. abietina and A. cleefii). The mitogenomes of both species have a typical suite of genes for 34 proteins, 17 tRNAs, and three rRNAs. The A. abietina mitogenome assembled into a simple circular map, with no large repeats capable of producing alternative isoforms. The A. cleefii mitogenomic map was more complex, involving two circular maps bridged by a substoichiometric linear fragment. Phylogenetics of three mitochondrial genes or the nuclear rRNA repeat placed Aragoa as sister to Littorella + Plantago, consistent with previous studies. However, P. nubicola, the sole representative of subg. Bougueria, was nested within subg. Psyllium based on the mitochondrial and nuclear data, conflicting with plastid-based analyses. Phylogenetics of the nuclear rRNA repeat provided better resolution overall, whereas relationships from mitochondrial data were hindered by extensive substitution rate variation among lineages.

CURRENT PERSPECTIVES OF DNA BARCODING IN BOSNIA AND HERZEGOVINA THROUGH BOLD DATABASE

UDK: 577.212:574]:007.5(497.6) DNA barcoding is a method designed to provide rapid and precise species identifications by using one or more of short gene sequences called barcodes. In most plant and fungi studies, the standard barcodes of choice are three plastid (rbcL, matK and trnH-psbA) and one nuclear (ITS) gene regions. The relatively high, but comparatively conserved rate of sequence evolution of mtDNA has made COI the marker of choice in animals. BOLD is a freely available cloud-based data storage and analysis platform developed with the aim to advance biodiversity science through DNA barcoding species identification. To date, over 6 million barcodes have been deposited in BOLD with 196,000 animal species, 68,000 plant species and 22,000 species of fungi and other organism entries. In this database, there are currently 447 entries for organisms from Bosnia and Herzegovina, which makes 0.0067% of the total number of BOLD entries. According to BOLD statistics, only 1.11% of all organism entries from B&H were submitted by B&H institutions. Despite the fact that Bosnia and Herzegovina has valuable natural resources with a high percentage of endemic and autochthonous species, BOLD statistics elucidated the lack of coordinated and systematic DNA barcoding research so far. It is necessary to establish continuous progress of molecular-genetic characterization of these resources in the future. It is up to B&H institutions to decide if they want to continue the practice of submitting the data sporadically or if they will animate the research community to actively participate in this global project.

Systematics of Scleranthus (Caryophyllaceae)

<p>Scleranthus is a genus of about 12 species of herbaceous flowering plants or small shrubs with a disjunct Eurasian/Australasian distribution. Monophyly of the genus is supported by the close similarity of gynoecial development of all species and consistent with nuclear ITS DNA sequence analysis. Traditionally the genus had been divided into two sections, section Scleranthus and section Mniarum. Section Mniarum is exclusively Australasian while section Scleranthus has been circumscribed to contain exclusively European species or a combination of European and Australasian species. Pollen and floral characters align the species into Australasian and Eurasian groups also supported by nuclear ITS DNA sequence analysis. Section Scleranthus as more broadly defined (i.e., sensu West and Garnock-Jones, 1986) is therefore at least paraphyletic or at worst polypyhyletic. Phylogenetic reconstructions based on morphological characters differ from those based on ITS sequences in supporting different relationships within the Australasian species of Scleranthus. Hybridisation and introgression within the genus are discussed and suggested as the cause of discordance between morphology and DNA sequence based trees. Low sequence divergence among Scleranthus ITS sequences suggests that the European and Australasian clades within the genus diverged within the last l0 million years. Biogeographic implications of these dating and competing hypotheses explaining the disjunct North-South distribution of the genus are discussed. Nuclear ITS and chloroplast ndhF DNA sequences both suggest that Scleranthus belongs to a clade within the family Caryophyllaceae consisting of members of subfamilies Alsinoideae and Caryophylloideae. Phylogenetic relationships between genera belonging to the three subfamilies of Caryophyllaceae (Alsinoideae, Caryophyloideae, and Paronychioideae) are addressed in this thesis through ndhF sequence analysis, which provides no support for the monophyly of traditionally recognised groups. Morphological character data sets are likely to always encompass multiple incongruent data partitions (sensu Bull et al. 1993). It may therefore be appropriate to combine data from DNA sequence and morphology for parsimony analysis even where the two are significantly incongruent.</p>

Systematics of Scleranthus (Caryophyllaceae)

<p>Scleranthus is a genus of about 12 species of herbaceous flowering plants or small shrubs with a disjunct Eurasian/Australasian distribution. Monophyly of the genus is supported by the close similarity of gynoecial development of all species and consistent with nuclear ITS DNA sequence analysis. Traditionally the genus had been divided into two sections, section Scleranthus and section Mniarum. Section Mniarum is exclusively Australasian while section Scleranthus has been circumscribed to contain exclusively European species or a combination of European and Australasian species. Pollen and floral characters align the species into Australasian and Eurasian groups also supported by nuclear ITS DNA sequence analysis. Section Scleranthus as more broadly defined (i.e., sensu West and Garnock-Jones, 1986) is therefore at least paraphyletic or at worst polypyhyletic. Phylogenetic reconstructions based on morphological characters differ from those based on ITS sequences in supporting different relationships within the Australasian species of Scleranthus. Hybridisation and introgression within the genus are discussed and suggested as the cause of discordance between morphology and DNA sequence based trees. Low sequence divergence among Scleranthus ITS sequences suggests that the European and Australasian clades within the genus diverged within the last l0 million years. Biogeographic implications of these dating and competing hypotheses explaining the disjunct North-South distribution of the genus are discussed. Nuclear ITS and chloroplast ndhF DNA sequences both suggest that Scleranthus belongs to a clade within the family Caryophyllaceae consisting of members of subfamilies Alsinoideae and Caryophylloideae. Phylogenetic relationships between genera belonging to the three subfamilies of Caryophyllaceae (Alsinoideae, Caryophyloideae, and Paronychioideae) are addressed in this thesis through ndhF sequence analysis, which provides no support for the monophyly of traditionally recognised groups. Morphological character data sets are likely to always encompass multiple incongruent data partitions (sensu Bull et al. 1993). It may therefore be appropriate to combine data from DNA sequence and morphology for parsimony analysis even where the two are significantly incongruent.</p>

Patterns of Cryptic Diversity and Phylogeography in Four Freshwater Copepod Crustaceans in European Lakes

Comparative phylogeography has become a powerful approach in exploring hidden or cryptic diversity within widespread species and understanding how historical and biogeographical factors shape the modern patterns of their distribution. Most comparative phylogeographic studies so far focus on terrestrial and vertebrate taxa, while aquatic invertebrates (and especially freshwater invertebrates) remain unstudied. In this article, we explore and compare the patterns of molecular diversity and phylogeographic structure of four widespread freshwater copepod crustaceans in European water bodies: the harpacticoids Attheyella crassa, Canthocamptus staphylinus and Nitokra hibernica, and the cyclopoid Eucyclops serrulatus, using sequence data from mtDNA COI and nuclear ITS/18S rRNA genes. The three taxa A. crassa, C. staphylinus and E. serrulatus each consist of deeply diverged clusters and are deemed to represent complexes of species with largely (but not completely) non-overlapping distributions, while in N. hibernica only little differentiation was found, which may however reflect the geographically more restricted sampling. However, the geographical patterns of subdivision differ. The divisions in A. crassa and E. serrulatus follow an east–west pattern in Northern Europe whereas that in C. staphylinus has more of a north–south pattern, with a distinct Fennoscandian clade. The deep mitochondrial splits among populations of A. crassa, C. staphylinus and E. serrulatus (model-corrected distances 26–36%) suggest that divergence of the lineages predate the Pleistocene glaciations. This study provides an insight into cryptic diversity and biogeographic distribution of freshwater copepods.

Comparative comprehensive analysis on natural infections of Hymenolepis diminuta and Hymenolepis nana in commensal rodents

Summary This first comprehensive report from Punjab province of India relates to patho-physiological alterations alongwith morpho-molecular characterisation and risk assessment of natural infections of Hymenolepis diminuta and Hymenolepis nana in 291commensal rodents including house rat, Rattus rattus (n=201) and lesser bandicoot rat, Bandicota bangalensis (n=90). Small intestine of 53.61 and 64.95 % rats was found infected with H. diminuta and H. nana, respectively with a concurrent infection rate of 50.86 %. There was no association between male and female rats and H. diminuta and H. nana infections (ᵡ2 = 0.016 and 0.08, respectively, d.f.= 1, P>0.05), while the host age had significant effect on prevalence of H. diminuta and H. nana (ᵡ2 = 28.12 and 7.18, respectively, d.f.= 1, P≤0.05) infection. Examination of faecal samples and intestinal contents revealed globular shaped eggs of H. diminuta without polar filaments (76.50 ± 3.01μm x 67.62 ± 2.42 μm), while smaller sized oval eggs of H. nana were with 4 – 8 polar filaments (47.87 ± 1.95 μm x 36.12 ± 3.05 μm). Cestode infection caused enteritis, sloughing of intestinal mucosa, necrosis of villi and inflammatory reaction with infiltration of mononuclear cells in the mucosa and submucosa. Morphometric identification of the adult cestodes recovered from the intestinal lumen was confirmed by molecular characterisation based on nuclear ITS-2 loci which showed a single band of 269 bp and 242 bp for H. diminuta and H. nana, respectively. Pairwise alignment of the ITS-2 regions showed 99.46 % similarity with sequences of H. diminuta from USA and 100 % similarity with sequences of H. nana from Slovakia, Kosice.

Nuclear ITS/ETS sequence data indicate the membership of Senecio racemulifer, but not S. acutipinnus and S. graciliflorus, within the genus Jacobaea (Asteraceae, Senecioneae)

The sectional placement of Senecio racemulifer (Asteraceae, Senecioneae) and the generic affiliation of S. acutipinnus and S. graciliflorus have been in dispute. Senecio racemulifer was tentatively considered a member of S. sect. Doria, and S. acutipinnus and S. graciliflorus were transferred to the genus Jacobaea as J. acutipinna and J. graciliflora, respectively. Our phylogenetic analyses based on nuclear ITS/ETS sequences indicate that S. racemulifer is deeply nested with Jacobaea species in a well-supported clade while S. acutipinnus and S. graciliflorus are nested with other Senecio species in a well-supported clade. We therefore transfer S. racemulifer to Jacobaea as J. racemulifera but retain S. acutipinnus and S. graciliflorus within Senecio. The names Jacobaea acutipinna, J. graciliflora, J. sect. Graciliflorae and S. ser. Racemuliferae are all synonymized. Karyological characters are reported for the three species. Jacobaea racemulifera, earlier known from the Western Tian-Shan (Kazakhstan, Kyrgyzstan, Uzbekistan), is reported here as new to China.