ITS Trees Research Articles

Morphological and molecular characterization of two species of genus Ageratum.

The species of genus Ageratum (family Asteraceae) are distributed in various parts of the world. Ageratum conyzoides and A. houstonianum are the most commonly occurring species in India. These species are quite similar in their morphology thus creating a challenge in identification during the field survey and taxonomic validation. The accurate identification of the species is highly significant especially when those are of medicinal interest. To overcome the barriers in morphological based identification, DNA barcoding has been employed during the present investigation. Morphological and DNA barcodes matK and ITS genes, were employed to differentiate between Ageratum conyzoides and A. houstonianum. The obtained matK and ITS gene sequences were submitted to GenBank and BOLD system to obtain accession numbers. The DNA sequences were aligned with database sequences using BLAST and phylogenetic trees were constructed through neighbor-joining algorithm in MEGA 11 software. The distinguish features of A. conyzoides include ovate to elliptic-oblong leaves with a cuneate base and inflorescence heads forming domed to flat-topped clusters. However, A. houstonianum has triangular to ovate leaves with a cordate to truncate base, cymose clusters in the inflorescence and stipulate glandular involucre bracts. The matK gene has shown the highest identity percentages (100%) for A. houstonianum and 99.87% for A. conyzoides. The phylogenetic tree analysis has demonstrated a close association of A. conyzoides and A. houstonianum with their respective species, supported by bootstrap values in the matK and ITS trees. This study revealed that morphological and molecular data can be successfully utilized in the identification of A. conyzoides and A. houstonianum. The matK and ITS barcodes provide promising results in the identification of Ageratum species, with their phylogeny supporting classification within the family asteraceae.

Species diversity and taxonomy of Vararia (Russulales, Basidiomycota) with descriptions of six species from Southwestern China.

Vararia is a species-rich genus in the family Peniophoraceae and has been shown to be polyphyletic. In this study, sequences of ITS and LSU rRNA markers of the studied samples were generated and phylogenetic analyses were performed with the maximum likelihood, maximum parsimony, and Bayesian inference methods. Seventeen lineages including six new species from China, i.e., V.fissurata, V.lincangensis, V.punctata, V.isabellina, V.sinensis, and V.yaoshanensis were recognized, in which V.fissurata is characterized by the brittle basidiomata with pruinose and cracking hymenophore having white to olivaceous buff hymenial surface, the clamped generative hyphae, presence of the two types gloeocystidia; V.lincangensis is characterized by the simple-septa generative hyphae, and thick-walled skeletal hyphae, and ellipsoid basidiospores; V.punctata is delimited by its thin to slightly thick-walled generative hyphae, and thick-walled skeletal hyphae, present thick-walled, clavate to cylindrical gloeocystidia; V.isabellina is characterized by having the cream to isabelline to slightly brown hymenial surface, thin to slightly thick-walled generative hyphae, and sub-fusiform to navicular basidiospores; V.sinensis is distinguishable by its white to slightly pink hymenial surface, thick-walled skeletal hyphae, and sub-fusiform to navicular basidiospores; V.yaoshanensis is characterized by cream to pinkish buff to cinnamon-buff hymenial surface, slightly thick-walled generative hyphae, the presence of two types gloeocystidia, and slightly thick-walled, ellipsoid basidiospores. Phylogram based on the ITS+nLSU rDNA gene regions included nine genera within the family Peniophoraceae as Amylostereum, Asterostroma, Baltazaria, Dichostereum, Michenera, Peniophora, Scytinostroma and Vararia, in which the six new wood-inhabiting fungi species were grouped into genus Vararia. The phylogenetic tree inferred from the combined ITS and LSU tree sequences highlighted that V.fissurata was found to be the sister to V.ellipsospora with strong supports. Additionally, V.lincangensis was clustered with V.fragilis. Furthermore, V.punctata was retrieved as a sister to V.ambigua. Moreover, V.sinensis was grouped with five taxa as V.breviphysa, V.pirispora, V.fusispora, V.abortiphysa and V.insolita. The new species V.isabellina formed a monophyletic lineage, in which it was then grouped closely with V.daweishanensis, and V.gracilispora. In addition, V.yaoshanensis was found to be the sister to V.gallica with strong supports. The present results increased the knowledge of Vararia species diversity and taxonomy of corticioid fungi in China. An identification key to 17 species of Vararia in China is provided.

A tough nutlet to crack: Resolving the phylogeny of Thesium (Thesiaceae), the largest genus in Santalales

AbstractWith over 350 species, Thesium is the largest genus in Santalales. It is found on all continents except Antarctica; however, its highest diversity is in the Cape Floristic Region of South Africa where approximately half the species occur. Thesium samples of ca. 590 collections from throughout its entire geographic range were obtained and nuclear ribosomal ITS sequenced from 396 accessions representing 196 named taxa and 30 currently unnamed taxa for a total of ca. 230 species. In addition, two chloroplast genome spacers (trnDT, trnLF) were sequenced from 269 and 315 accessions, respectively. Maximum parsimony, maximum likelihood and Bayesian methods were employed to generate gene trees and infer phylogenies. The value of the morphological characters traditionally used in the taxonomy of the genus and previous infrageneric classifications are discussed. Broad scale relationships were generally congruent among the ITS and the chloroplast trees. For example, both the nuclear and chloroplast trees support the presence of Eurasian and African clades. In contrast, major incongruence was detected between nuclear and chloroplast trees for a number of taxa including the recently described T. nautimontanum that is sister to the entire African clade on the ITS tree. Although the causes of this incongruence are currently unknown, a novel form of chloroplast capture is hypothesized. A hypothesis of the biogeographical history of the genus based on our molecular phylogeny is presented.

Dermatophytes and mammalian hair: aspects of the evolution of Arthrodermataceae

AbstractDermatophytes and other members of Onygenales are unique in their ability to degrade keratin, affecting hair and nails, and in the case of human hosts, causing skin infections. Subtillisins are essential proteases in keratin assimilation, and subtilisin-like protease 1 (SUB1) and SUB3–7 are specific for dermatophytes. eIF2α kinases are serine-threonine kinases that perform essential functions in response to infection, proteotoxicity, and nutrient scavenging. The relatively conserved nature of EIF2AK4 among fungi makes them potential evolutionary markers, which may contribute to a deeper understanding of dermatophyte taxonomy and evolution. This study aimed to assess the phylogeny of dermatophytes by examining the EIF2AK4 and SUB1 genes compared to the ITS gene marker. The phylogenetic trees generated from the EIF2AK4 and SUB1 genes exhibited a similar topology, which differed from that observed in the ITS tree. Our preliminary findings with a limited dataset suggest that the EIF2AK4 and SUB1 genes provide a reasonably correct reflection of the evolution of Arthrodermataceae. In addition, the study analyzed in vitro keratinolytic responses of 19 dermatophyte species using hairs of a broad range of mammals, including ancestral as well as derived species, as substrates. Trichophyton mentagrophytes and Nannizzia gypsea were the most active in degrading hair, while Trichophyton verrucosum, Trichophyton tonsurans and Epidermophyton floccosum showed low response. Hairs of Hyracoidea and Rodentia were most affected of all mammal hairs, while in contrast, bat hairs were difficult to degrade by nearly all tested dermatophyte species. Zoophilic species showed more activity than anthropophilic dermatophytes, but hair degradation profiles were not diagnostic for particular dermatophyte species.

Four new species of Russula from the Xizang Autonomous Region and other provinces of China

ABSTRACT Russula is the largest genus in the Russulales and is widespread throughout the world. Almost all Russula species are known to be ectomycorrhizal with high ecological and edible values, and some are lethal poisonous. In this study, four new species belonging to the subgenus Russula crown clade are identified based on morphological and phylogenetic evidence from the Xizang Autonomous Region and other provinces of China. Morphologically, Russula paragraveolens (sect. Polychromae, subsect. Xerampelinae) is mainly characterised by a cherry red to blood red pileus centre, a reddish orange pileus margin; R. pseudograveolens (sect. Polychromae, subsect. Xerampelinae) is characterised by a violet brown to brownish red pileus centre, a pale red to pastel red pileus margin and short basidia; R. shigatseensis (sect. Flavisiccantes, subsect. Lepidinae) is characterised by a brownish orange to madder red pileus centre, pinkish red pileus margin, and having lateral branches or branches of hyphal terminations in pileipellis; R. yadongensis (sect. Tenellae, subsect. Laricinae) is characterised by a dark purplish red pileus centre with brownish purple tints and having isolated to clustered spines of spore ornamentations. Their distinct taxonomic status is confirmed by the positions of the four new species in both the ITS and 4-locus (nucLSU, mtSSU, rpb2, tef1) phylogenetic trees.

Phylogenetic Relationships among some Species of the Genus Hypericum (Hypericaceae) in Kurdistan-Rgeion Iraq

Phylogenetic hypotheses for the large cosmopolitan genus Hypericum (St. John’s wort) have previously been based on morphology, and molecular studies have thus far included only a few species. In this study, we used Eight taxa representing 2 sections of the genus Hypericum were analyzed along with one representatives of Thornea calcicola using nuclear internal transcribed spacer ITS and 28SrRNA sequences in order to assess the scope of phylogenetic relationships in this species-rich genus. The resulting parsimony analysis and Bayesian Inference corroborates the actual classification and interspecific affinities. The molecular data provide support for the monophyletic assemblage of the sections in both ITS and 28SrRNA regions trees shows four different clades with minimum changes between species location in both tree. We used Maximum Parsimony and Bayesian approach to reconstruct ancestral states of selected characters, which resulted in recognition of characters that support major clades within the genus Hypericum. The obtained results encourage the use of ITS and 28SrRNA sequence analysis in resolving the phylogenetic relationships within the member rich genus Hypericum. Our results differ somewhat from those of recent morphological and cytological studies. The phylogenetic relationships among Hypericum species have now been mostly resolved via 28SrRNA phylogeny.

Molecular Phylogeny of Holarctic Aeshnidae with a Focus on the West Palaearctic and Some Remarks on Its Genera Worldwide (Aeshnidae, Odonata)

Aeshnidae Rambur, 1842 are impressive large insects distributed worldwide. Currently, over 500 species are recognized. Nevertheless, the phylogeny of this family is not completely understood. We applied molecular phylogenetic analysis using two popular phylogenetic markers, the mitochondrial COI gene fragment (barcoding sequence) and the nucleic ITS region, containing the ITS1, 5.8S rRNA, and ITS2 sequences. We used available and credible published sequences and 96 newly sequenced specimens. Our analysis involved all West Palaearctic species, all but one genera of the Holarctic Aeshnidae, and most genera worldwide, and is by far the largest molecular study of this family. The topology of all trees created with different algorithms and genes is in favour of the current taxonomic concept, with some remarkable outcomes. Aeshna Fabricius, 1775, was found to be diverged into several branches, especially with respect to the COI gene. Although it appeared not monophyletic in phylogenetic reconstructions based on the ITS region, the analysis of COI and joint analysis suggest its monophyly in the current taxonomical sense, with one notable exception. Aeshna isoceles (Müller, 1767) has fallen out of Aeshna in all analyses, so a new monophyletic genus, Isoaeschna gen. nov. is introduced for it. The genus Brachytron Evans, 1845 tightly clustered with Aeschnophlebia Selys, 1883, Epiaeschna Hagen in Selys, 1883, and Nasiaeschna Selys in Förster, 1900. Thus, we suggest subsuming these four genera under the priority name Brachytron. Tetracanthagyna Selys, 1883 clusters as expected with Brachytron in the ITS tree, but is an independent ancient clade of its own in all COI trees. The genus Polycanthagyna Fraser, 1933 syn. nov. is synonymised to Indaeschna Fraser, 1926. On the species level, we suggest that the American Aeshna septentrionalis Burmeister, 1839 be treated as a subspecies of A. caerulea (Ström, 1783), Aeshna caerulea septentrionalis. We synonymize Gynacantha hyalina Selys, 1882 with Gynacantha subinterrupta Rambur, 1842. Our analysis provides new insights on the tight relationships of the circumboreal species Aeshna juncea and A. subarctica and the intraspecies phylogeny of Aeshna juncea.

Morphological and molecular characterisation of a new cryptic species of Criconemoides informis group, C. neoinformis n. sp., and C. persicus n. sp., with notes on C. avicenniae

Summary Criconemoides neoinformis n. sp., a cryptic species of C. informis group, and C. persicus n. sp. were recovered from the rhizosphere of pear and grass, in Zanjan and West Azerbaijan provinces, northwestern Iran, respectively. Criconemoides neoinformis n. sp. is characterised by 522-585 μm long females, having four separated pseudolips, anteriorly flattened lip region comprised of two annuli, 57-63 retrorse body annuli with smooth to slightly crenate margins, without anastomoses, 86.9-90.4 μm long stylet, having 6-10 annuli from vulva to posterior end and tail conical rounded with two, three or multilobed terminus and presence of males. Criconemoides persicus n. sp. could be characterised by flatly rounded lip region, having fused rectangular pseudolips, 515-615 μm body length, stylet 60-65 μm, body annuli 120-132 with smooth to slightly crenate margins, without anastomoses, 9-12 annuli from vulva to posterior end and tail conical rounded with two trilobed terminus. In inferred phylogenies, these two new species clustered with known Criconemoides species in 18S, 28S, ITS and COI trees. These species are clearly separated molecularly by all the studied markers. The use of molecular markers, mainly D2-D3 expansion segments of 28S rRNA, ITS region and COI, are very promising in the case of cryptic species found in this group of nematodes. We provided molecular markers (two sequences per marker) for these new species with morphological/morphometrical data using an integrative taxonomy approach for the unequivocal identification of these species.

Crataegus bretschneideri (Rosaceae), a separate species endemic to northeast China

Crataegus bretschneideri has been considered a variety of Crataegus pinnatifida, but has also been proposed as a separate species. We used ITS and three cpDNA sequence fragments (psbA‐trnH, trnG‐trnS, trnH‐trnK) to perform maximum parsimony, maximum likelihood, Bayesian inference and neighbor‐joining molecular phylogenetic analysis on 108 individuals of 19 hawthorn (Crataegus) species and 4 outgroups (Malus). The ITS, cpDNA and cpDNA‐ITS molecular phylogenetic trees constructed using 4 methods had similar consistency.In the cpDNA‐ITS tree, C. bretschneideri was resolved as an independent clade separated from Crataegus maximowiczii and C. pinnatifida. In the ITS (biparental inheritance) tree, C. bretschneideri and C. pinnatifida were resolved as closely related and only distantly related to C. maximowiczii. In contrast, the cpDNA (maternal inheritance) tree indicated that C. bretschneideri is closely related to C. maximowiczii but distantly related to C. pinnatifida. In total, 15 chloroplast haplotypes were revealed in the 108 Crataegus individuals, as determined by DNASP analysis. Of these, C. bretschneideri and C. maximowiczii share the same chloroplast haplotype, H1, while C. pinnatifida contains three haplotypes, H13–H15. Taken together with previous results, we confirm that C. bretschneideri is not a variety of C. pinnatifida, but a separate species that gradually developed from natural hybrid offspring in mixed forest areas with C. maximowiczii as the female parent and C. pinnatifida as the male parent.

The Amazonian luminescent Mycena cristinae sp. nov. from Brazil.

A new luminescent lignicolous fungal species, Mycena cristinae sp. nov., is proposed from the Central Amazon forest. This is unique and supported by morphological evaluation along with LSU- and ITS-based phylogenetic analyses. The basidiomata have mostly fuscous olivaceous brown pileus, adnate to subdecurrent and distant lamellae, and stipe with slightly bulbous base (basal mycelium absent). It also has inamyloid and/or weakly amyloid basidiospores, ramose cheilocystidia and a pileipellis composed of an aerated tangle of slender, diverticulate hyphae forming a coralloid pellicle overlaying the hypodermium. The luminescence is evident in the basidiomata (especially the stipe) and in the mycelium on the substrate. The LSU phylogenetic trees reveal that M. cristinae is sister to M. coralliformis within the Mycenaceae clade. In the ITS trees, it forms a unique lineage grouping with undetermined Mycena taxa. Morphological data support M. cristinae as a different species compared to previously described taxa.

Cymbidium motuoense (Orchidaceae; Epidendroideae), a new species from China: evidence from morphological and molecular data

This study describes a new orchid species, Cymbidium motuoense, from Xizang, China based on morphological and molecular analyses. Several unique morphological features distinguish this new orchid from all other species of Cymbidium. Based on its morphology, this orchid is similar to C. tracyanum but differs in its yellow-green flower with dark purple-red spots on the veins, oblong-elliptic sepals, side lobes of the lip with reddish brown hairs along the veins and disc with three purple-red lamellae. Phylogenetic analyses based on plastid DNA (matK and rbcL) supported C. motuoense as a new species. However, the low support in nuclear ribosomal ITS tree provide the possibility that the new orchid may be a natural hybrid of C. tracyanum and C. iridioides or another closely related species.

Biogeographical patterns and speciation of the genus Pinguicula (Lentibulariaceae) inferred by phylogenetic analyses.

Earlier phylogenetic studies in the genus Pinguicua (Lentibulariaceae) suggested that the species within a geographical region was rather monophyletic, although the sampling was limited or was restricted to specific regions. Those results conflicted with the floral morphology-based classification, which has been widely accepted to date. In the current study, one nuclear ribosomal DNA (internal transcribed spacer; ITS) and two regions of chloroplast DNA (matK and rpl32-trnL), from up to ca. 80% of the taxa in the genus Pinguicula, covering all three subgenera, were sequenced to demonstrate the inconsistency and explore a possible evolutionary history of the genus. Some incongruence was observed between nuclear and chloroplast topologies and the results from each of the three DNA analyses conflicted with the morphology-based subgeneric divisions. Both the ITS tree and network, however, corresponded with the biogeographical patterns of the genus supported by life-forms (winter rosette or hibernaculum formation) and basic chromosome numbers (haploidy). The dormant strategy evolved in a specific geographical region is a phylogenetic constraint and a synapomorphic characteristic within a lineage. Therefore, the results denied the idea that the Mexican group, morphologically divided into the three subgenera, independently acquired winter rosette formations. Topological incongruence among the trees or reticulations, indicated by parallel edges in phylogenetic networks, implied that some taxa originated by introgressive hybridisation. Although there are exceptions, species within the same geographical region arose from a common ancestor. Therefore, the classification by the floral characteristics is rather unreliable. The results obtained from this study suggest that evolution within the genus Pinguicula has involved; 1) ancient expansions to geographical regions with gene flow and subsequent vicariance with genetic drift, 2) acquirement of a common dormant strategy within a specific lineage to adapt a local climate (i.e., synapomorphic characteristic), 3) recent speciation in a short time span linked to introgressive hybridisation or multiplying the ploidy level (i.e., divergence), and 4) parallel evolution in floral traits among lineages found in different geographical regions (i.e., convergence). As such, the floral morphology masks and obscures the phylogenetic relationships among species in the genus.

Clarifying Recent Adaptive Diversification of the Chrysanthemum-Group on the Basis of an Updated Multilocus Phylogeny of Subtribe Artemisiinae (Asteraceae: Anthemideae).

Understanding the roles played by geography and ecology in driving species diversification and in the maintenance of species cohesion is the central objective of evolutionary and ecological studies. The multi-phased orogenesis of Qinghai-Tibetan Plateau (QTP) and global climate changes over late-Miocene has profoundly influenced the environments and evolution of organisms in this region and the vast areas of Asia. In this study, we investigate the lineage diversification of Chrysanthemum-group in subtribe Artemisiinae (tribe Anthemideae, Asteraceae) likely under the effects of climate changes during this period. Using DNA sequences of seven low-copy nuclear loci and nrITS and the coalescent analytical methods, a time-calibrated phylogeny of subtribe Artemisiinae was reconstructed with emphasis on Chrysanthemum-group. The monophyletic Chrysanthemum-group was well resolved into two major clades corresponding to Chrysanthemum and Ajania, two genera which can be well identified by capitulum morphology but have been intermingled in previous plastid and ITS trees. Within Chrysanthemum, a later divergence between Ch. indicum-complex and Ch. zawadskii-complex can be recognized. The time frames of these sequential divergences coincide with the late Cenozoic uplift of the Northern QTP and the concomitant climatic heterogeneity between eastern and inland Asia. Reconstruction of historical biogeography suggested the origin of Chrysanthemum-group in Central Asia, followed by eastward migration of Chrysanthemum and in situ diversification of Ajania. Within Chrysanthemum, Ch. indicum-complex and Ch. zawadskii-complex exhibited contemporary distributional division, the former in more southern and the latter in more northern regions. The geographic structure of the three lineages in Chrysanthemum-group have been associated with the niche differentiation, and environmental heterogenization in Asia interior.

<p><strong>Two new species of <em>Hydnophlebia </em>(Meruliaceae, Polyporales) from China based on morphological and molecular evidence</strong></p>

Two new corticioid species, Hydnophlebia sinensis and H. subchrysorhiza, are described and illustrated from China. Hydnophlebia sinensis was collected from northern China (Beijing, Jilin and Shaanxi Provinces), while H. subchrysorhiza was collected from southern China (Hainan, Hunan and Yunnan Provinces). The two new species belong to the H. chrysorhiza species complex and are similar to each other with few morphological differences. However, they are phylogenetically independent in both the ITS-LSU tree of Meruliaceae and the ITS tree of Hydnophlebia. In the trees, H. sinensis was sister to H. sp., whilst H. subchrysorhiza was closely related to H. chrysorhiza. Our analyses show that we do not have H. chrysorhiza in China but possess two undescribed sibling species

Hidden in plain sight: Morphological and phylogenetic evidence for Bouteloua arizonica, a species distinct from Bouteloua aristidoides (Poaceae, Chloridoideae)

Two varieties of Bouteloua aristidoides have been recognized, the widespread var. aristidoides and the more narrowly distributed var. arizonica. The two varieties differ in inflorescence form even more than that seen between many other closely related species of Bouteloua. We therefore asked whether these taxa might be better regarded as distinct species. A total of 93 vouchers were studied by using morphometry (principal components analysis and statistical tests), leaf micromorphology, ancestral state reconstruction, and/or molecular (ITS, trnC-rpoB and trnT-L-F) phylogenetic analyses. Except from the ITS tree, all results supported elevation of B. aristidoides var. arizonica to the rank of species, thus the new combination Bouteloua arizonica (M.E. Jones) L.F. Cuellar & Columbus comb. nov. et stat. nov., is proposed. Chloroplast and combined chloroplast-nuclear molecular trees depicted var. arizonica as monophyletic (even in sympatric populations with var. aristidoides) and reveals phylogenetic structure within var. aristidoides for which the presence of new undescribed varieties of B. aristidoides (different from B. arizonica) is addressed. B. arizonica differs from B. aristidoides in having fewer branches per inflorescence, a bigger branch with more spikelets, and a shorter branch extension. Scanning electron microscopy revealed the presence of papillae on leaves of B. arizonica as a clear synapomorphy. Growing mature plants of B. arizonica from seeds in a greenhouse revealed a strong cleistogamous nature for this species for which gene flow in sympatric populations with B. aristidoides seems unlikely. A taxonomic treatment and distribution map for identification of B. arizonica is provided.

The transferability of microsatellite loci from a homoploid to a polyploid hybrid complex: an example from fine-leaved Festuca species (Poaceae).

BackgroundMicrosatellite loci, or single sequence repeats (SSR), are widely used as powerful markers in population genetics. They represent an attractive tool for studying plants such as grasses, whose evolution is driven by hybridisation and polyploidization. However, the development of microsatellite markers has been challenging and time-consuming, especially for non-model organisms lacking available genome-wide sequence data. One straightforward and low-cost approach is to transfer the SSR loci developed for one species, or complex, to another closely-related one. This work evaluates the transferability of microsatellite loci from homoploid to allopolyploid complexes of fine-leaved Festuca species and to assess their use in two new species. The studied complex (F. amethystina—F. tatrae) is a useful model for research on the local adaptability of grasses with different ploidy levels. Since both species can be considered as rare or threatened (F. tatrae—as a mountain and narrow endemic species and F. amethystina—a mountain species with relict lowland populations), any tool enabling studies on genetic diversity and population genetics, such as SSR markers, could also be very useful in a conservation context.MethodsThe ploidy level within populations was estimated using flow cytometry. One diploid and one tetraploid population of F. amethystina and a diploid population of F. tatrae were chosen to test the transferability of SSR loci. Because our work describes the transfer of SSR nuclear markers designed originally for F. gautieri, a phylogenetic tree was prepared based on the ITS marker to assess the genetic distance between the studied complexes. The PCR products were separated on a high-resolution agarose gel, intended for SSR marker analysis. Appropriate solutions for the allotetraploid population and whole mixed-ploidy complex were implemented.ResultsFlow cytometry confirmed earlier data regarding DNA content in the investigated species and cytotypes. The phylogenetic ITS tree indicated a small genetic distance between F. gautieri complexes and the studied species. Ten microsatellite markers were successfully transferred. All markers were polymorphic. In total, 163 different alleles were scored from the 10 SSR loci. PCoA of accessions revealed well-separated groups corresponding to studied populations. Over 60% of the total variance is explained by differentiation within populations and one third among them.ConclusionsThe transferred markers are valid tools for the study of population genetics and inheritance relationships within cytotypes and species and between them. The presented markers can be used to study inbreeding depression in the Festuca species, and variations in the degrees of genetic diversity between different cytotypes in mountain and lowland areas. Our findings can also be applied to study conservation strategies for ensuring biodiversity at the genetic level in polyploid complexes.

Biogeographic history and diversification patterns in the Irano-Turanian genus Acanthophyllum s.l. (Caryophyllaceae)

The Irano-Turanian (IT) floristic region in Asia contains three biodiversity hotspots and has been a source of xerophytic taxa for neighboring regions. Despite its species richness and large territory, the evolutionary history and biogeography of this floristic region is poorly understood. Acanthophyllum, encompassing ca. 70 subshrubby mostly thorny-cushion species, is one of the important components of the steppe and mountain vegetation of this region. In this study, we investigate the tempo and mode of diversification as well as the biogeographic patterns of this genus. The ancestral area analysis suggests that Acanthophyllum originated east of the Zagros Mountains in the Miocene, from where it expanded westwards. A shift in the speciation rate in the late Pliocene (ca. 3.24 Ma) was detected, based on a nrDNA ITS tree, affecting the lineage of the largest section in the genus, Acanthophyllym sect. Oligosperma. Parallelisms with another IT species-rich genera in aspects such as largely coincident ranges and habitats, presence of a thorny-cushion life-form, similar areas reconstructed for their MRCA, and the inference of a shift in speciation in the region suggest common underlying abiotic and biotic factors or even drivers for diversification and speciation in the high elevation dry continental landscape of IT region.

Phylogenetic relationships, biogeography and taxonomic delimitation of Astragalus sect. Acanthophace (Fabaceae) using cpDNA and nrDNA ITS sequences analyses

Phylogenetic relationships within Astragalus sect. Acanthophace and several spiny Astragalus species were studied using nuclear ribosomal DNA internal transcribed spacer (nrDNA ITS) and two plastid intergenic spacers (rpl32-trnL(UAG) and trnS(GCU)–trnG(UCC)). The strongly congruent expression using maximum likelihood and Bayesian inference methods was the non-monophyly of sect. Acanthophace at the current status suggesting the exclusion of A. ovigerus and A. lamprocarpus as a single clade. This study also shed light on taxonomic controversies on the synonymy of A. lamprocarpus under A. ovigerus and indicated the non-monophyly of A. lycioides complex, since one accession of A. lycioides (sect. Acanthophace) united with members of A. sect. Aegacantha (resurrected as A. pseudoangustifolius). Another interesting finding was the unexpected placement of A. montis-parrowii (sect. Aegacantha) within the bulk of spiny Astragalus (Astracantha clade). The reconstructed plastid and nrDNA ITS trees showed considerable conflict in the position of several species. Biogeographical analyses indicated that sect. Acanthophace was most probably originated in western Iran.

Species diversity of ring nematodes of the genus Criconemoides (Nematoda: Criconematidae) based on three new species from China, using integrative taxonomy

Ring nematodes of the genus Criconemoides are a group of polyphagous, migratory root-ectoparasites of plants. In a nematological survey of Zhejiang Province, three new Criconemoides species were isolated from the rhizosphere of Citrus sp., Euonymus alatus and Pterocarya stenoptera trees. In the present study, the new species characterized using an integrative taxonomic approach and described as Criconemoides parainformis n. sp., C. geraerti n. sp. and C. rotundicaudatus n. sp. Three of the species shares truncated anteriorly flattened lip region comprised of two annuli, body annuli margin smooth with few anastomoses, (except C. rotundicaudatus n. sp. it has crenated annuli margins and numerous anastomoses), excretory pore 1–2 annuli posterior to pharyngeal bulb base, (except C. geraerti n. sp. it has excretory pore at the same level, or 1–2 annuli anterior to pharyngeal bulb base), vulva closed, anus hardly visible 1–3 annuli posterior to vulva. The new species can be distinguished based on their en face view pattern under scanning electron microscope (SEM) and the tail shapes. i.e. C. parainformis n. sp. has an ellipsoid oral disc, slightly fused pseudolips and tail ending in a bilobed terminus. C. geraerti n. sp. has a hexagonal oral disc, separated pseudolips, and a bluntly conoid tail. C. rotundicaudatus n. sp. has a rounded oral disc, well developed pseudolips, and a short broadly rounded tail. Besides detailed morphological studies using SEM, genomic and mitochondrial molecular data were obtained for the three described new species. In inferred phylogenies, these three species clustered with known Criconemoides species in 18S, 28S, ITS and coxI trees. Currently, only four known Criconemoides species occur in China and the discovery of the three new Criconemoides species in present study has increased our knowledge on the diversity of Criconemoides in China.

Laserocarpum, a new genus of Apiaceae endemic to Greece

<em>Laserpitium pseudomeum</em> is an endemic umbellifer of Greece occurring in the mountains of Sterea Ellas and northern Peloponnese. Molecular data indicate that it is not related to its putative congeners, but instead constitutes an isolated lineage. The nuclear ribosomal DNA ITS tree places it as a weakly supported sister group to <em>Portenschlagiella ramosissima</em>, the sole species in its genus, whereas the tree inferred from three noncoding cpDNA loci does not confirm this relationship. <em>Portenschlagiella ramosissima</em> is sometimes placed in <em>Athamanta</em>, an affinity supported neither by molecular data nor by fruit morphology. <em>Laserpitium pseudomeum</em> notably differs from <em>P. ramosissima</em> in vegetative, flower, and fruit characteristics, and no obvious morphological synapomorphies indicate their close relationship. Therefore, we place <em>L. pseudomeum</em> in the newly described genus <em>Laserocarpum</em> and lectotypify the species with the specimen <em>Orphanides 2019</em> (G00766460) at G-Boiss.