Nuclear ITS Region Research Articles

The generic placement of Grimmia teretinervis Limpr.

Kiebacher, T. & Meier, M. 2020. The generic placement of Grimmia teretinervis Limpr. – Herzogia 33: 544 –547.Grimmia teretinervis is a dioicous moss of the Grimmiaceae that has never been found with sporophytes. Consequently, its generic placement is uncertain, and although it is mostly referred to Grimmia, some authors assign it to Schistidium. The phylogenetic analysis of the nuclear ITS region indicates that it is a species of Schistidium and that it is related to the likewise dioicous S. tenerum.

Metabarcoding of Soil Fungal Communities Associated with Alpine Field-Grown Saffron (Crocus sativus L.) Inoculated with AM Fungi.

Soil fungi strongly influence ecosystem structure and functioning, playing a key role in many ecological services as decomposers, plant mutualists and pathogens. Arbuscular mycorrhizal fungi (AMF) establish mutualistic symbiotic associations with plant roots and act as biofertilizers by enhancing plant nutrients and water uptake. Information about the AMF association with Crocus sativus L. (saffron) and their impact on crop performances and spice quality has been increasing in recent years. Instead, there is still little data on the biodiversity of soil microbial communities associated with this crop in the Alpine environments. The aims of this study were to investigate the fungal communities of two Alpine experimental sites cultivated with saffron, and to rank the relative impact of two AMF inocula, applied to soil as single species (R = Rhizophagus intraradices, C. Walker & A. Schüßler) or a mixture of two species (M = R. intraradices and Funneliformis mosseae, C. Walker & A. Schüßler), on the resident fungal communities which might be influenced in their diversity and composition. We used Illumina MiSeq metabarcoding on nuclear ribosomal ITS2 region to characterize the fungal communities associated to Crocus sativus cultivation in two fields, located in the municipalities of Saint Christophe (SC) and Morgex (MG), (Aosta Valley, Italy), treated or not with AMF inocula and sampled for two consecutive years (Y1; Y2). Data analyses consistently indicated that Basidiomycota were particularly abundant in both sites and sampling years (Y1 and Y2). Significant differences in the distribution of fungal taxa assemblages at phylum and class levels between the two sites were also found. The main compositional differences consisted in significant abundance changes of OTUs belonging to Dothideomycetes and Leotiomycetes (Ascomycota), Agaricomycetes and Tremellomycetes (Basidiomycota), Mortierellomycetes and Mucoromycetes. Further differences concerned OTUs, of other classes, significantly represented only in the first or second year of sampling. Concerning Glomeromycota, the most represented genus was Claroideoglomus always detected in both sites and years. Other AMF genera such as Funneliformis, Septoglomus and Microdominikia, were retrieved only in MG site. Results highlighted that neither sites nor inoculation significantly impacted Alpine saffron-field fungal communities; instead, the year of sampling had the most appreciable influence on the resident communities.

Plant DNA Barcoding Principles and Limits: A Case Study in the Genus Vanilla.

Powerful DNA barcodes have been much more difficult to define in plants than in animals. In 2009, the international Consortium for the Barcoding Of Life (CBOL) chose the combination of the chloroplast genes (rbcL + matK) as the proposed official barcode for plants. However, this system has got important limits. First, any barcode system will only be useful if there is a clear barcode gap and if species are monophyletic. Second, chloroplast and mitochondrial (COI gene used for animals) barcodes will not be usable for discriminating hybrid species. Moreover, it was also shown that, using chloroplast regions, maximum species discrimination would be around 70% and very variable among plant groups. This is why many authors have more recently advocated for the addition of the nuclear ITS region to this barcode because it reveals more variations and allows the resolution of hybrid or closely related species. We tested different chloroplast genes (rbcL, matK, psaB, psbC) and the nuclear ITS region in the genus Vanilla, a taxonomically complex group and therefore a good model to test for the efficiency of different barcode systems. We found that the CBOL official barcode system performed relatively poorly in Vanilla (76% species discrimination), and we demonstrate that adding ITS to this barcode system allows to increase resolution (for closely related species and to the subspecies level) and to identify hybrid species. The best species discrimination attained was 96.2% because of one paraphyletic species that could not be resolved.

A phylogeny of species near Agrostis supporting the recognition of two new genera, Agrostula and Alpagrostis (Poaceae, Pooideae, Agrostidinae) from Europe.

Based on a molecular DNA phylogeny of three plastid (rpl32-trnK, rps16 intron, and rps16-trnK) and nuclear ITS regions investigating 32 species of Agrostidinae, we describe two new genera, Agrostulagen. nov. with a single species and Alpagrostisgen. nov. with four species; provide support for five species in a monophyletic Podagrostis; and include a small sample of 12 species of a monophyletic Agrostis s.s. (including the type and most species of Neoschischkinia), that separates into two clades corresponding to A.subg.Agrostis and A.subg.Vilfa. Agrostula differs from Agrostis in having leaf blades with pillars of sclerenchyma which are continuous between the adaxial and abaxial surface of the blades, dorsally rounded glumes with blunt to truncate and erose to denticulate apices, florets ½ the length of the glumes, lemmas equally wide as long, widest at (or near) apex, apices broadly truncate, irregularly 5 to 7 denticulate to erose, awnless, anthers longer than the lemmas, and rugose-papillose caryopses. Alpagrostis differs from Agrostis in having geniculate basally inserted awns and truncate lemma apices with lateral veins prolonged from the apex in (2)4 setae. The following eight new combinations are made: Agrostulatruncatula, Agrostulatruncatulasubsp.durieui, Alpagrostisalpina, Alpagrostisalpinavar.flavescens, Alpagrostisbarceloi, Alpagrostissetacea, Alpagrostissetaceavar.flava, and Alpagrostisschleicheri. In addition, we provide a key separating Agrostula and Alpagrostis from Agrostis s.s. and other genera previously considered as synonyms of Agrostis; lectotypify Agrostisalpina Scop., A.schleicheri Jord. & Verl., A.truncatula Parl., and A.truncatulavar.durieui Henriq.; and neotypify A.setacea Curtis.

Molecular phylogeny and chromosomal evolution of endemic species of Sri Lankan Anacardiaceae

Family Anacardiaceae comprises 70 genera and approximately 985 species distributed worldwide. Sri Lanka harbours 19 species in seven genera, among these 15 are endemics. This study focuses on regionally restricted endemics and native Anacardiaceae species, which have not been investigated before at molecular and cytological level. Nuclear rDNA ITS and plastid matK regions were sequenced for ten species, having nine endemics and one native, and incorporated into the existing sequence data for phylogenetic analyses. The topologies resulting from maximum parsimony, maximum likelihood and Bayesian inference are congruent. Family Anacardiaceae forms a monophyletic group having monophyletic subfamily Anacardioideae and paraphyletic subfamily Spondioideae. Tribes Anacardieae, Semecarpeae and Rhoeae form subclades within the major clade of Anacardioideae. All the endemic species occupy correct position in the molecular phylogeny as per the existing classification except for Campnosperma zeylanica, which shows a close relationship to members of the subfamily Spondioideae. The position of non-native species Trichoscypha acuminata also remains unclear. Chromosome counts and karyograms were constructed for five endemic species. The chromosome numbers incorporated in the tree range from 2n = 28–58. Species of tribe Rhoeae have the lowest chromosome number (2n = 24, 28, 30) while species of tribe Semecarpeae have the higher numbers (2n = 50, 52, 58). Chromosome numbers mapped on the phylogeny shows that dysploidy had played a role in the evolution of the species of the family Anacardiaceae in Sri Lanka.

The macroalgal culture collection in Kobe University (KU-MACC) and a comprehensive molecular phylogeny of macroalgae based on the culture strains

ABSTRACT The Kobe University Macroalgal Culture Collection (KU-MACC), as a section of the Kobe University Research Center for Inland Seas, distributes cultures of macroalgae to the public under the umbrella of the National BioResource Project of Ministry of Education, Culture, Sports, Science and Technology and Japan Agency for Medical Research and Development, in collaboration with the Microbial Culture Collection of the National Institute of Environmental Studies. KU-MACC principally holds macroalgae (Phaeophyceae, Rhodophyta and Ulvophyceae), but also holds some important related taxa of Schizocladiophyceae and Charophyceae. They are unialgal strains maintained as vegetative thalli and by cryo-preservation. Stock cultures are grown in culture chambers under various temperature conditions in transparent polypropylene vials containing Provasoli’s Enriched Seawater (PES) or PESI medium. The collection includes the “genome strains” of Ectocarpus siliculosus, Chondrus crispus, Chara braunii and their related stains. Depending on the phylogenetic group, DNA sequences of the nuclear rDNA ITS region, chloroplast rbcL, and mitochondrial cox3 genes are determined and used for taxonomy and as genetic tags. Based on the genetic data, molecular phylogenies of Ulvophyceae, Phaeophyceae and Rhodophyta are presented. In the rbcL molecular phylogeny of Ulvophyceae, due to rather limited coverage of culture strains, only members of Ulothricales, Ulvales and Bryopsidales are represented by KU-MACC strains, whereas 10 orders are recognized in the class. The molecular phylogeny of Phaeophyceae based on the cox3 gene showed a tree topology similar to the published multigene molecular phylogeny except for the position of Ishige okamurae, although the bootstrap supports of most nodes were low: Ishigeales branched near the base of Phaeophyceae next to Discosporangiales in the multigene molecular phylogeny, but in the cox3 tree, it showed a long branch and was sister to Fucales. In the molecular phylogeny of Rhodophyta, 14 orders are represented by KU-MACC strains covering about half of currently recognized Bangiophycean and Florideophycean orders.

Distribution and genetic diversity of the toxic benthic dinoflagellate genus Ostreopsis in Korea

Species belonging to the toxic dinoflagellate genus Ostreopsis are widespread, occurring from tropical to temperate waters. As mainly benthic/epiphytic species, they would be expected to show distinct geographical patterns. In this study, ribosomal DNA (rDNA) sequences from partial nuclear LSU D8-D10, 5.8S, and ITS regions were determined for 169 isolates of Ostreopsis species collected from three coastal sites (i.e., Jeju Island, Chuja Island, and Pohang) within Korea. The phylogenetic tree inferred from the LSU rDNA D8-D10 sequences showed that Korean Ostreopsis species corresponded to either Ostreopsis sp. 1 or sp. 6, with Ostreopsis sp. 1 being relatively predominant regarding their distribution. While Ostreopsis sp. 1 occurred throughout all the three sampling sites within Korea, Ostreopsis sp. 6 was confined to the northern part of Jeju Island. When further investigated, the genetic diversity of Ostreopsis sp. 1 in Korea based on ITS sequences showed a total of 21 haplotypes. The presumed ancestral haplotype H3, was also present in the Japanese and Russian populations of Ostreopsis sp. 1. Although the overall demographic history of all the Korean populations of Ostreopsis sp. 1 could not be clearly identified, probably due to a mixture of different regional demographic patterns within Korea, each Ostreopsis sp. 1 population showed a characteristic demographic pattern at a regional scale. While the Jeju Island Ostreopsis sp. 1 population showed a signal in agreement with population equilibrium, the Chuja Island and Pohang Ostreopsis sp. 1 populations showed distribution patterns that are expected in a sudden population expansion model. The results from this study provide a basis for a better understanding of the distribution and genetic structure of the Asian Ostreopsis sp. 1 populations.

Evolutionary history of the Eurasian steppe plant Schivereckia podolica (Brassicaceae) and its close relatives

The genus Schivereckia (Brassicaceae) – presently included in the genus Draba – comprises two species, S. podolica and S. doerfleri. Schivereckia podolica is an eastern European species with disjunct distribution in Russia, Ukraine, and Podolian-Galician outposts. Schivereckia doerfleri is reported from alpine fields of the Balkan Peninsula and from mountain chains of northern Anatolia, Turkey. Schivereckia podolica shares its ecological and biogeographic characteristics with a number of other steppe plants. The ‘relic hypothesis’ interprets the present locations of Schivereckia and its associates as relics of a formerly continuous distribution belt of a cold periglacial steppe vegetation.We employed nuclear ITS and chloroplast trnL-trnF region, and DNA fingerprinting (RAPD) to elucidate the evolutionary history and historical biogeography of Schivereckia. RAPD data prove isolation by distance and mirror the present disjunct geographical distribution pattern of Schivereckia podolica. Phylogenetic analyses revealed that Schivereckia might be polyphyletic and nested within a highly supported clade of central Asian, Caucasian and North American Draba species. Time divergence estimation and ancestral area reconstruction support the interpretation of species of the Schivereckia subclade as periglacial relic. Our results argue for a continuous persistence of certain steppe species since the Pleistocene, and support the view that the modern steppes of Europe have derived partly from Pleistocene glacial steppes.

Development of high-resolution chloroplast markers for intraspecific phylogeographic studies of Phaeocystis globosa

Phaeocystis globosa is an important harmful algal bloom causative species distributing widely in temperate and tropical coastal waters in the world. The morphological, physiological, and biochemical characteristics are different among geographic strains, which can not be distinguished with nuclear ribosomal DNA markers at present. Therefore, the genetic distance and phylogeographic relationships of nuclear 28S rDNA D1-D2 and ITS regions, and three chloroplast intergenic spacers (petN-trnS1, trnM1-psbA, and rbcS-rpl27) were analyzed and compared among 13 strains of P. globosa isolated from the Pacific Ocean and Atlantic Ocean in this study. In addition, the nucleotide polymorphisms of 28S rDNA D1-D2, ITS, and rbcS-rpl27 regions were evaluated in two P. globosa strains. The various levels of nucleotide polymorphism were in the nuclear 28S rDNA D1-D2 region and ITS region, but no polymorphism was in the chloroplast rbcS-rpl27 intergenic spacer. A reasonable intraspecific phylogeographic relationship was presented by rbcS-rpl27 intergenic spacer, which had the strongest distinction to geographic strains compared to those of 28S rDNA D1-D2 and ITS regions. In the phylogenetic tree of rbcS-rpl27 intergenic spacer, the two strains from the North Sea of the Atlantic Ocean were divided firstly from the species of P. globosa, and then formed an independent clade, while the other Atlantic strains and all of Pacific strains joined up to build the other clade. It was implied that at least two genetically distant populations of P. globosa existed in the Atlantic coastal regions. This study provided a high-resolution chloroplast marker to analyze intraspecific phylogeographic populations of P. globosa, and preliminarily clarified the genetic relationships of the Pacific and Atlantic strains of P. globosa.

New insight into the molecular phylogeny of the genus Liparis s.l. (Orchidaceae: Malaxideae) with a new generic segregate: Blepharoglossum

Molecular studies have shown that Liparis s.l. is polyphyletic, with the epiphytic Liparis alliance well separated from the clade representing Liparis in its restricted sense and further forming a phylogenetically heterogeneous assemblage. The generic status of Cestichis, as the key member of the epiphytic Liparis alliance, remains contentious, and the relationships between Cestichis, Stichorkis and Platystyliparis are still unclear and in need of further studies. We herein employ DNA sequence data from nuclear ITS and plastid matK regions to clarify the generic boundaries of Cestichis and its relationship to the allied genera. Results from both Bayesian inference and maximum likelihood analysis congruently indicate that Cestichis as currently circumscribed is not monophyletic. The Blepharoglossum clade, corresponding to commonly recognized sect. Blepharoglossum, forms a monophyletic lineage closely related to the genus Oberonia instead of the core Cestichis clade. A re-evaluation of the diagnostic morphological characters reveals that this lineage represents a morphologically distinct group. Based on these results, Blepharoglossum is removed from Cestichis and elevated to generic status. Our study also sheds light on the homoplastic nature of some diagnostic morphological characters used traditionally in the taxonomy of the Malaxidinae.

Tracking the evolutionary history of the Allium ampeloprasum L. complex (section Allium) provides evidence of the contribution of North African diploids to the formation of allopolyploid horticultural groups

The economically important Allium ampeloprasum L. represents a polyploid complex, comprising hexa- and octoploid Great Headed Garlic horticultural cultivars (6x–8x GHG) and several traditional varieties of the tetraploid (4x) leeks (Leek, Bulbous leek, Kurrat and Pearl onion). Its wild representatives were indicated as rare in the Mediterranean region. This study aims to explore the diversity and origin of polyploidy in this complex, including its wild relatives A. baeticum Bossier and A. guttatum Steven with particular focus on the poorly investigated North-African region. Natural populations were sampled in Algeria in various bioclimatic conditions, then subjected to karyological and molecular phylogenetic analyses based on nuclear rDNA ITS region and chloroplast trnL–trnF and trnD–trnT intergenic spacers. Comparative analyses included available Genbank accession sequences representing old-world relatives. Chromosome count surveys revealed an unexpected higher occurrence of diploid (2n = 16) than tetraploid (2n = 32) cytotypes. The phylogenetic analyses first allowed positioning the Algerian material within the A. ampeloprasum complex. Interestingly, all the Algerian diploid and tetraploid populations from A. ampeloprasum and A. baeticum form a distinct monophyletic group. The results provide novel and robust evidence demonstrating that the North African diploid A. ampeloprasum genetic pool widely contributed as a source of progenitors not only for the A. ampeloprasum and A. baeticum Algerian tetraploids, but also in the formation of the GHG and Leek cultivated allopolyploids. Therefore, the North African populations emerge as an important reservoir of new wild genetic resources of great interest for tracing the origin of crop domestication and for breeding programs of cultivated varieties.

<p><strong>Notes on <em>Crocus</em> L. Series <em>Flavi</em> Mathew (Iridaceae) and a new species with unique corm structure</strong></p>

Crocus asymmetricus (Iridaceae) is described as a new species endemic to the southern part of the Anatolian diagonal in Turkey. It is phylogenetically related to C. vitellinus and morphologically to C. antalyensis, but differs from these species in showing an asymmetric corm and a single point of root emergence. Both C. antalyensis and C. asymmetricus are illustrated and compared in this paper. A Bayesian phylogenetic tree of the nuclear rDNA ITS region confirms the affiliation of C. asymmetricus to C. ser. Flavi, and its close relationship to C. vitellinus. A new identification key to the species of C. ser. Flavi occurring in Turkey is also presented.

Description, molecular characteristics and Wolbachia endosymbionts of Onchocerca borneensis Uni, Mat Udin & Takaoka n. sp. (Nematoda: Filarioidea) from the Bornean bearded pig Sus barbatus M\xfcller (Cetartiodactyla: Suidae) of Sarawak, Malaysia

BackgroundThe genus Onchocerca Diesing, 1841 includes species of medical importance, such as O. volvulus (Leuckart, 1893), which causes river blindness in the tropics. Recently, zoonotic onchocercosis has been reported in humans worldwide. In Japan, O. dewittei japonica Uni, Bain & Takaoka, 2001 from wild boars is a causative agent for this zoonosis. Many filarioid nematodes are infected with Wolbachia endosymbionts which exhibit various evolutionary relationships with their hosts. While investigating the filarial fauna of Borneo, we discovered an undescribed Onchocerca species in the bearded pig Sus barbatus Müller (Cetartiodactyla: Suidae).MethodsWe isolated Onchocerca specimens from bearded pigs and examined their morphology. For comparative material, we collected fresh specimens of O. d. dewittei Bain, Ramachandran, Petter & Mak, 1977 from banded pigs (S. scrofa vittatus Boie) in Peninsular Malaysia. Partial sequences of three different genes (two mitochondrial genes, cox1 and 12S rRNA, and one nuclear ITS region) of these filarioids were analysed. By multi-locus sequence analyses based on six genes (16S rDNA, ftsZ, dnaA, coxA, fbpA and gatB) of Wolbachia, we determined the supergroups in the specimens from bearded pigs and those of O. d. dewittei.ResultsOnchocerca borneensis Uni, Mat Udin & Takaoka n. sp. is described on the basis of morphological characteristics and its genetic divergence from congeners. Molecular characteristics of the new species revealed its close evolutionary relationship with O. d. dewittei. Calculated p-distance for the cox1 gene sequences between O. borneensis n. sp. and O. d. dewittei was 5.9%, while that between O. d. dewittei and O. d. japonica was 7.6%. No intraspecific genetic variation was found for the new species. Wolbachia strains identified in the new species and O. d. dewittei belonged to supergroup C and are closely related.ConclusionsOur molecular analyses of filarioids from Asian suids indicate that the new species is sister to O. d. dewittei. On the basis of its morphological and molecular characteristics, we propose to elevate O. d. japonica to species level as O. japonica Uni, Bain & Takaoka, 2001. Coevolutionary relationships exist between the Wolbachia strains and their filarial hosts in Borneo and Peninsular Malaysia.

Phylogenetic placement of Ivodea and biogeographic affinities of Malagasy Rutaceae

The genus Ivodea is endemic to Madagascar and the Comoros and consists of 30 species. This study is the first to include the genus in a molecular phylogenetic analysis. We sequenced the plastid trnL–trnF and the nuclear ITS regions for three Ivodea species and revealed that the genus is monophyletic and most closely related to the African and Malagasy Vepris, refuting earlier suggestions of a close relationship between Ivodea and the Asian, Malesian, Australasian and Pacific genera Euodia and Melicope. Ivodea and Vepris provide another example of closely related pairs of Rutaceous groups that have drupaceous and capsular/follicular fruits, respectively, thus further confirming that fruit types are not suited to delimit subfamilies in Rutaceae, as has often been done in the past. Ivodea was the last of the seven Malagasy genera to be included in the Rutaceae phylogeny, making it possible to conduct an assessment of biogeographic affinities of the genera that occur on the island. Our assessments based on sister-group relationships suggest that the eight lineages (representing seven genera) of Malagasy Rutaceae either have African or have Asian affinities. Two lineages have an African origin, and one lineage has an Asian origin. Taxon sampling is insufficient to interpret the directionality of dispersal events in the remaining lineages.

Relationships of Conoclinium, a Recently Diverged Genus (Asteraceae, Eupatorieae) and Description of a New Species from Western Mexico.

Molecular phylogenetic studies were conducted to clarify the phylogenetic placement of Conoclinium within Eupatorieae and to analyze interspecific relationships in the genus. Analysis of a six gene data set placed Conoclinium sister to Ageratum and closely related to Fleischmannia and Paneroa, a result that is consistent with the overall appearance and the phytochemistry of these genera but not with their current subtribal placement. Results of an analysis of noncoding DNA, including the chloroplast trnH-psbA spacer and the nuclear ITS and ETS regions, was consistent with the close relationship of Conoclinium to Ageratum, Fleischmannia, and Paneroa, and showed Conoclinium to be monophyletic. Within Conoclinium, the results of the phylogenetic analyses were slightly at variance with current classification, and it is suggested that C. dichotomum (populations from peninsular Florida previously included within C. coelestinum) and C. oligolepis (formerly C. betonicifolium var. integrifolium) be recognized as distinct species. The low overall sequence variability suggests that Conoclinium is a relatively recent arrival in eastern North America. Conoclinium mayfieldii is not monophyletic and the Sierra Madre Occidental populations of the species are herein recognized as a new species, C. gonzaleziae. Neotypes for C. dichotomum and C. oligolepis are designated.

Silene sunhangii (Caryophyllaceae), a new species from China.

Silene sunhangii, a new species of Caryophyllaceae known from only three populations in Hubei and Hunan provinces of central China, is described. Both morphological and molecular data were used to assess the taxonomic status and relationships of this species. Morphologically, S. sunhangii is most similar to S. platyphylla Franch. from which it differs most readily in having 3-veined elliptical leaves without pubescence, tasseled catacorolla, pale purple to red petals without a linear lobe or narrow tooth and lanceolate, bifid to one third. A phylogenetic analysis based on nuclear ITS region identified the new species as a well-supported, independent lineage. Our new species is nested within a grade that encompasses species representing a polyphyletic Silene sect. Physolychnis (Benth.) Bocquet. Both the genetic and morphological data support the recognition of Silene sunhangii as a distinct species, although there is inconsistency between these two datasets as to the relationships of the new species.

The Anatolian Diagonal and Paleoclimatic Changes Shaped the Phylogeography of Cynips quercus (Hymenoptera, Cynipidae)

Cynips quercus is a with widespread oak-gall wasp species distributed across the Western Palearctic from Europe, the northern Caucasus and North Africa to Iran. In this study, we used 175 asexual-generation individuals of C. quercus collected from 30 localities in Turkey to investigate the effects of topography and climatic factors on the phylogeographic structure of the species. Sequence analysis of the mitochondrial cytochrome-b gene (MT-CYB) and the nuclear ITS2 region supported a genetic break between the eastern and the western sides of the Anatolian Diagonal. The ITS2 results indicated that the populations underwent expansion, while in contrast the MT-CYB data implied a stable population structure for C. quercus. Molecular clock applied to the MT-CYB data suggested that the climatic and environmental changes during the Pliocene and Pleistocene had a profound effect on the phylogeography of the species.

Taxonomy of the Verrucaria kalenskyi – V. xyloxena species complex in Finland

The taxonomy of species related to Verrucaria kalenskyi and V. xyloxena in Finland was studied using morphological characters and analyses of the ITS region. Nine species are accepted as part of the V. kalenskyi – V. xyloxena -complex: V. danica, V. inverecundula sp. nov., V. juankoskiensis sp. nov., V. kalenskyi, V. kiskoensis sp. nov., V. raesaenenii sp. nov., V. tallbackaensis sp. nov., V. aff. trabicola and V. xyloxena. The species group is characterised by relatively small perithecia and spores, a predominantly brown non-areolate thallus and their similarity in the nuclear ITS region. Only V. inverecundula differs from all other species in this group by a pale endolithic thallus and shorter involucrellum. All species are restricted to calcareous rocks and/or pebbles except V. xyloxena, which occurs mainly on calcareous soils, and V. aff. trabicola, which is epiphytic. Although morphologically similar V. modica sp. nov. is not part of the V. kalenskyi – V. xyloxena complex according to our ITS-genetree. In the ITS all other taxa of the complex form a strongly supported monophyletic clade. Verrucaria kalenskyi and V. xyloxena share almost identical ITS sequences, but are morphologically distinct. Most species treated in this study resemble V. kalenskyi but differ from that species in the following features: Verrucaria danica has a fimbriate prothallus, smaller perithecia and thinner involucrellum, V. juankoskiensis has broader spores, a poorly developed thallus and more sparsely occurring perithecia, V. kiskoensis has a thicker thallus and narrower spores, V. modica has a thicker involucrellum, V. raesaenenii has a fimbriate prothallus and perithecia often leaving pits in the substratum, and V. tallbackaensis has thalline covered, more sparsely occurring and slightly larger perithecia. Occurrences of V. xyloxena on calcareous pebbles are confirmed. Most studied species are very rare in Finland except for V. kalenskyi and V. xyloxena. Occurrences of V. danica, V. juankoskiensis, V. kalenskyi and V. raesaenenii are also confirmed from Norway. Verrucaria amylacea and V. anceps are excluded from the Finnish lichen biota. A lectotype is selected for V. danica.

<p><strong>A new autumn flowering <em>Crocus</em> (Iridaceae) from SW Turkey: <em>C. terzioghluii</em></strong></p>

Crocus terzioghluii (Iridaceae) is described as a new species endemic to southwestern Anatolia, Turkey. It is related to Crocus babadagensis and differs from this species in having corm tunics with well-developed rings, and prominent stripes on outer segments, as well as in leaf number and flowering time. The corms and tunics of both species are illustrated. A phylogenetic tree obtained by Bayesian phylogenetic inference of the combined sequences of the nuclear rDNA ITS and ETS regions confirms the affiliation of C. terzioghluii to C. ser. Biflori and its close relationship to C. babadagensis. A microphotograph of the metaphase chromosomes (2n = 2x = 8) along with the idiogram and a leaf cross section of the new species are given.

Phylogeny of Tacca (Taccaceae) and traits in reproductive structures, with description of a new Bornean species

Abstract. Wong SY, Chua KS. 2019. Phylogeny of Tacca (Taccaceae) and traits in reproductive structures, with description of a new Bornean species. Biodiversitas 20: 3096-3118. A phylogenetic study of 16 of the c. 20 Tacca species is presented using nuclear ITS and plastid matK gene regions. Tacca leontopetaloides and T. maculata are a sister clade to the rest of the Tacca species. Tacca palmata and T. palmatifida are strongly supported together in a clade with this clade sister to T. bibracteata and T. plantaginea. All endemic Bornean taxa except T. bibracteata formed a derived clade together with non-Bornean T. cristata and T. sumatrana. The Bornean taxa are separated into three clades with T. borneensis sister to the other two clades. Twelve morphological characteristics are coded and mapped onto the Maximum Parsimony tree to elucidate potential evolutionary patterns. Results indicated shifts from a geophytic (irregularly-) seasonally dormant to an evergreen mesophytic habit, a decompound to a simple leaf blade, a long to a short peduncle, green to dark-colored inflorescences, numerous to few inner bracts, bracteoles, and flowers, and a low to an increased number of ovules. Within the Bornean taxa, there is a reversal from showy bracts and elaborate inflorescences to less showy bracts and simplified inflorescences. Tacca cristata Jack is resurrected from within T. integrifolia Ker Gawl. sensu Drenth. A new species, Tacca havilandii S.Y.Wong & K.S.Chua from Borneo is described.