Phylogenetic Re-Evaluation of the Laurencia Complex (Rhodophyta) with a Description of L. Succulenta Sp. Nov. from Korea

Laurencia succulenta sp. nov. (Rhodophyta) is described from Korea. This species exhibits vegetative and reproductive structures typical of the genus, but is distinct from similar species in its epiphytic habitat and the fleshy, robust, thick and subcompressed thalli with basically distichous branching. In addition, it is readily distinguished from the most similar species, such as L. nipponica Yamada and L. okamurae Yamada, by the cystocarps with a somewhat protuberant ostiole. In a phylogenetic analysis of 47 species of the Laurencia Lamouroux complex from various localities around the world based on 49 morphological characters, four major clades (Laurencia, Chondrophycus palisadus (Yamada) Nam group, C. cartilagineus (Yamada) Garbary et Harper group and Osmundea Stackhouse assemblage), each of which forms a monophyletic group, were recognized. Among these, the Laurencia clade is basal to the overall assemblage, and is defined by the vegetative axis with four rather than two pericentral cells. The Osmundea clade is supported by autapomorphic characters for the genus, features associated with spermatangial formation of the filament type and tetrasporangial production from epidermal cells. By contrast, Chondrophycus, a genus characterized by a combination of features (vegetative axis with two pericentral cells, trichoblast-type spermatangial development and tetrasporangial production from pericentral cells), is paraphyletic, and the species were separated into two well-supported clades, the C. palisadus group and C. cartilagineus group. These clades are distinguished from each other by the position of the first pericentral cell relative to the trichoblast, the presence or absence of fertility at the second pericentral cells and number of sterile pericentral cells in the tetrasporangial axis, the pattern of formation of spermatangial branches on trichoblasts, post-fertilization feature associated with the formation time of the auxiliary cell, and, probably, the number of pericentral cells in the procarp-bearing segment. Of these features, the side position of the first pericentral cell in the latter group (a synapomorphy for the C. cartilagineus group plus Osmundea) suggests that the C. cartilagineus group is more closely related to Osmundea than to the C. palisadus group. This cladistic analysis indicates that Chondrophycus is not monophyletic, suggesting that the C. palisadus group should be separated from Chondrophycus at the genus level. Based on this result, Palisada (Yamada) stat. nov. is proposed for the group, together with an emendation of the generic delineation of Chondrophycus, and relevant nomenclatural changes for several Chondrophycus species are also included. In addition, Corynecladia J. Agardh is reinstated for the type species L. clavata Sonder.     

Metabolomics as a Potential Chemotaxonomical Tool: Application in the Genus Vernonia Schreb

The taxonomic classification of the genus Vernonia Schreb is complex and, as yet, unclear. We here report the use of untargeted metabolomics approaches, followed by multivariate analyses methods and a phytochemical characterization of ten Vernonia species. Metabolic fingerprints were obtained by accurate mass measurements and used to determine the phytochemical similarities and differences between species through multivariate analyses approaches. Principal component analysis based on the relative levels of 528 metabolites, indicated that the ten species could be clustered into four groups. Thereby, V. polyanthes was the only species with presence of flavones chrysoeriol-7-O-glycuronyl, acacetin-7-O-glycuronyl and sesquiterpenes lactones piptocarphin A and piptocarphin B, while glaucolide A was detected in both V. brasiliana and V. polyanthes, separating these species from the two other species of the Vernonanthura group. Species from the Lessingianthus group were unique in showing a positive response in the foam test, suggesting the presence of saponins, which could be confirmed by metabolite annotation. V. rufogrisea showed a great variety of sesquiterpene lactones, placing this species into a separate group. Species within the Chrysolaena group were unique in accumulating clovamide. Our results of LC-MS-based profiling combined with multivariate analyses suggest that metabolomics approaches, such as untargeted LC-MS, may be potentially used as a large-scale chemotaxonomical tool, in addition to classical morphological and cytotaxonomical approaches, in order to facilitate taxonomical classifications.     

Italian neo-endemism in a widespread group of leafhoppers insects: A revision of the Eupteryx aurata group (Auchenorrhyncha: Cicadellidae: Typhlocybinae) using morphology,ecology and genetics

The Eupteryx aurata group is characterised by several features, primarily by colouration and aedeagus morphology. It is distributed mainly in Europe, with some species also in Asia and one species perhaps in North Africa. At least one species is introduced in North America. Until now 12 taxa were known. Two new species, Eupteryx divulsa from central Italy and Eupteryx oscorum from central and southern Italy, are described in this paper. Eupteryx petasitidis Ferrari, 1882, described as Eupteryx carpini petasitidis and later synonymised with Eupteryx atropunctata (Goeze, 1778), is reestablished on species level and redescribed including genital morphology. Ten taxa of the species group are studied in relation to their morphological and molecular characters. The colouration, aedeagus, and pygofer appendage are figured for all of these taxa. Molecular study showed that the E. aurata group is monophyletic. The little or moderate distances in sequences (below 6.0% for mtDNA and up to 2.5% for nuclear ITS2) between the different taxa of the E. aurata group and incongruence of genetic signals from mtDNA and nuclear DNA could be the result of incomplete lineage sorting in young species and/or hybridisation and introgression of DNA among taxa. Of particular importance in this study is material from Italy, a peripheral region in relation to the distribution of the species group and due to its geomorphological features favouring isolation and speciation. A distribution map for the E. aurata species group in Italy and an identification key for all 15 taxa of the aurata group are given. Phylogenetic relationships within the aurata group are discussed in relation to morphological, molecular, ecological, and zoogeographic aspects.     

Complete Denitration of Nitroglycerin by Bacteria Isolated from a Washwater Soakaway

Four axenic bacterial species capable of biodegrading nitroglycerin (glycerol trinitrate [GTN]) were isolated from soil samples taken from a washwater soakaway at a disused GTN manufacturing plant. The isolates were identified by 16S rRNA gene sequence homology as Pseudomonas putida, an Arthrobacter species, a Klebsiella species, and a Rhodococcus species. Each of the isolates utilized GTN as its sole nitrogen source and removed nitro groups sequentially from GTN to produce glycerol dinitrates and mononitrates (GMN), with the exception of the Arthrobacter strain, which achieved removal of only the first nitro group within the time course of the experiment. The Klebsiella strain exhibited a distinct preference for removal of the central nitro group from GTN, while the other five strains exhibited no such regioselectivity. All strains which removed a second nitro group from glycerol 1,2-dinitrate showed regiospecific removal of the end nitro group, thereby producing glycerol 2-mononitrate. Most significant was the finding that the Rhodococcus species was capable of removing the final nitro group from GMN and thus achieved complete biodegradation of GTN. Such complete denitration of GTN has previously been shown only in mixed bacterial populations and in cultures of Penicillium corylophilum Dierckx supplemented with an additional carbon and nitrogen source. Hence, to the best of our knowledge, this is the first report of a microorganism that can achieve complete denitration of GTN.     

A new genus and species from Borneo and a new species from Java and Sumatra of Cicadini (Hemiptera: Cicadidae)

A new cicada genus and species, Minilomia triquetra, gen. and sp. nov., is described from Borneo. The new genus Minilomia is closely allied to Platylomia Stål and is placed in the subtribe Dundubiina of the tribe Cicadini. Another new cicada species, Pomponia brevicaudata, sp. nov. (subtribe Psithyristriina), is described from Java and Sumatra. The new species belongs to the Pomponia linearis species group and the P. linearis species complex but is distinguished from its congeners by the unusually short male abdomen.     

In the shadow of a megalopolis,a new Flexamia from a threatened grass species in the New Jersey Pine Barrens (Hemiptera,Cicadellidae, Deltocephalinae,Paralimnini)

A previously unknown species of the North American leafhopper genus Flexamia, Flexamia whitcombi sp. n., is described from pinebarren smokegrass (Muhlenbergia torreyana (Schult.) Hitchc.), a state-listed threatened grass in the New Jersey Pine Barrens. The serrata species group, to which it belongs, is redefined and a key to the species of the group is provided. This is the first reported insect association with Muhlenbergia torreyana.     

Description of a new species of the genus Anochetus Mayr (Hymenoptera: Formicidae) from Orchid Island,Taiwan

A trap-jaw ant Anochetus lanyuensis sp. nov., belonging to the graeffei species group Brown, 1978, is described on the basis of worker and gyne castes from Orchid Island (Lanyu), Taiwan. A. lanyuensis sp. nov. is easily distinguishable from the other species in the graeffei species group by the large size of its eye, longitudinal striae on the pronotal disc, and concave dorsal margin of the petiolar node in the frontal view.     

A new species of Bembidion ( Ecuadion) from Ecuador (Coleoptera,Carabidae, Bembidiini), with a key to members of the georgeballi species group

A new species of ground beetle, Bembidion ricei, is described from the Andes mountains of Ecuador east of Quito. It belongs to the georgeballi species group of subgenus Ecuadion, and is most similar to Bembidion georgeballi. A key to the species of the group is provided.     

Identification of a Bacillus anthracis specific indel in the yeaC gene and development of a rapid pyrosequencing assay for distinguishing B. anthracis from the B. cereus group

Bacillus anthracis, the causative agent of anthrax, is a potential source of bioterrorism. The existing assays for its identification lack specificity due to the close genetic relationship it exhibits to other members of the B. cereus group. Our comparative analyses of protein sequences from Bacillus species have identified a 24 amino acid deletion in a conserved region of the YeaC protein that is uniquely present in B. anthracis. PCR primers based on conserved regions flanking this indel in the Bacillus cereus group of species (viz. Bacillus cereus, B. anthracis, B. thuringiensis, B. mycoides, B. weihenstephnensis and B. pseudomycoides) specifically amplified a 282 bp fragment from all six reference B. anthracis strains, whereas a 354 bp fragment was amplified from 15 other B. cereus group of species/strains. These fragments, due to large size difference, are readily distinguished by means of agarose gel electrophoresis. In contrast to the B. cereus group, no PCR amplification was observed with any of the non-B. cereus group of species/strains. This indel was also used for developing a rapid pyrosequencing assay for the identification of B. anthracis. Its performance was evaluated by examining the presence or absence of this indel in a panel of 81 B. cereus-like isolates from various sources that included 39 B. anthracis strains. Based upon the sequence data from the pyrograms, the yeaC indel was found to be a distinctive characteristic of various B. anthracis strains tested and not found in any other species/strains from these samples. Therefore, this B. anthracis specific indel provides a robust and highly-specific chromosomal marker for the identification of this high-risk pathogen from other members of the B. cereus group independent of a strain's virulence. The pyrosequencing platform also allows for the rapid and simultaneous screening of multiple samples for the presence of this B. anthracis-specific marker.     

Description of new taxa of Halenia Borkh. (Gentianaceae) from Colombia and Venezuela with significance for testing a key innovation hypothesis

As a result of an ongoing complete taxonomic revision of Halenia (Gentianaceae), the new taxa Halenia perijana K.B. Hagen and H. major subsp. meridensis K.B. Hagen are described from Colombia and Venezuela, respectively. Flower morphology suggests that both taxa belong to the Halenia viridis group. H. perijana is vegetatively well separated from its probable closest relatives, and the only member of Halenia from a small and rather isolated mountain range. The new subspecies meridensis has small remnants of nectary spurs, and probably links the unspurred viridis group to its prominently spurred Central American sister group. Previous molecular clock approaches showed that the viridis group arrived in South America approximately at the same time as the weddelliana group, the distantly related second major South American group, which contains species with prominent spurs. Based on the new extensive knowledge of all species, the distribution patterns of both groups in Colombia and Venezuela are compared. The existence of marked differences—weddelliana group species grow in sympatry more often than species of the viridis group—is consistent with a key innovation effect of the presence/absence of nectary spurs, i.e., the reduction of pollinator overlap mediated by the presence of nectary spurs may allow sympatric speciation or remigration after a shorter phase of allopatric differentiation. However, this new evidence is relatively weak due to lack of statistical support and several other unresolved problems.     

Identification of a Bacillus anthracis specific indel in the yeaC gene and development of a rapid pyrosequencing assay for distinguishing B. anthracis from the B. cereus group

Bacillus anthracis, the causative agent of anthrax, is a potential source of bioterrorism. The existing assays for its identification lack specificity due to the close genetic relationship it exhibits to other members of the B. cereus group. Our comparative analyses of protein sequences from Bacillus species have identified a 24 amino acid deletion in a conserved region of the YeaC protein that is uniquely present in B. anthracis. PCR primers based on conserved regions flanking this indel in the Bacillus cereus group of species (viz. Bacillus cereus, B. anthracis, B. thuringiensis, B. mycoides, B. weihenstephnensis and B. pseudomycoides) specifically amplified a 282 bp fragment from all six reference B. anthracis strains, whereas a 354 bp fragment was amplified from 15 other B. cereus group of species/strains. These fragments, due to large size difference, are readily distinguished by means of agarose gel electrophoresis. In contrast to the B. cereus group, no PCR amplification was observed with any of the non-B. cereus group of species/strains. This indel was also used for developing a rapid pyrosequencing assay for the identification of B. anthracis. Its performance was evaluated by examining the presence or absence of this indel in a panel of 81 B. cereus-like isolates from various sources that included 39 B. anthracis strains. Based upon the sequence data from the pyrograms, the yeaC indel was found to be a distinctive characteristic of various B. anthracis strains tested and not found in any other species/strains from these samples. Therefore, this B. anthracis specific indel provides a robust and highly-specific chromosomal marker for the identification of this high-risk pathogen from other members of the B. cereus group independent of a strain's virulence. The pyrosequencing platform also allows for the rapid and simultaneous screening of multiple samples for the presence of this B. anthracis-specific marker.     

Milnesium berladnicorum sp. n. (Eutardigrada,Apochela, Milnesiidae), a new species of water bear from Romania

In a lichen sample collected from a tree in Bârlad town (Vaslui County, Romania), a new tardigrade species belonging to the genus Milnesium (granulatum group) was found. Milnesium berladnicorum sp. n. is most similar (in the type of dorsal sculpture) to Milnesium beasleyi Kaczmarek et al., 2012 but differs from it mainly by having a different claw configuration and some morphometric characters. Additionally, the new species differs from other congeners of the granulatum group by the different type of dorsal sculpture, claw configuration and some morphometric characters.     

Inonotus tenuicontextus sp. nov. (Hymenochaetaceae) from Guizhou, southwest China with a preliminary discussion on the phylogeny of its kin

Inonotus tenuicontextus collected from Guizhou, southwest China was described and illustrated as a new species based on a combination of phylogenetic and morphological evidence. It is characterized by perennial and effused-reflexed to pileate basidiocarps; duplex and very thin context; a monomitic hyphal system in context; a dimitic hyphal system in trama; and broadly ellipsoid, hyaline and thick-walled basidiospores. Phylogenetically I. tenuicontextus clustered within Inonotus s. s. clade; moreover, it formed a well supported monophyletic subclade with Inonotus baumii, I. linteus, I. lonicericola, I. vaninii and I. weirianus. In morphology I. tenuicontextus distinguishes from I. linteus, also a species with duplex context, by its smaller basidiospores, while its duplex context makes it different from the other four species with homogeneous context. We proposed this subclade as a medicinal group for most of its members with medicinal functions. The phylogeny of the six species in this medicinal group was briefly discussed based on our results. An identification key to them is also provided.     

Identification of Lactic Acid Bacteria from Chili Bo, a Malaysian Food Ingredient

Ninety-two strains of lactic acid bacteria (LAB) were isolated from a Malaysian food ingredient, chili bo, stored for up to 25 days at 28°C with no benzoic acid (product A) or with 7,000 mg of benzoic acid kg⁻¹ (product B). The strains were divided into eight groups by traditional phenotypic tests. A total of 43 strains were selected for comparison of their sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) whole-cell protein patterns with a SDS-PAGE database of LAB. Isolates from product A were identified as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus farciminis, Pediococcus acidilactici, Enterococcus faecalis, and Weissella confusa. Five strains belonging to clusters which could not be allocated to existing species by SDS-PAGE were further identified by 16S rRNA sequence comparison. One strain was distantly related to the Lactobacillus casei/Pediococcus group. Two strains were related to Weissella at the genus or species level. Two other strains did not belong to any previously described 16S rRNA group of LAB and occupied an intermediate position between the L. casei/Pediococcus group and the Weissella group and species of Carnobacterium. The latter two strains belong to the cluster of LAB that predominated in product B. The incidence of new species and subspecies of LAB in chili bo indicate the high probability of isolation of new LAB from certain Southeast Asian foods. None of the isolates exhibited bacteriocin activity against L. plantarum ATCC 14917 and LMG 17682.     

Dorsipes caspius n. sp. (Acari: Podapolipidae), a subelytral parasite of Pterostichus caspius (Menetries) (Coleoptera: Carabidae) with notes on host range of the genus and the distribution of the platysmae group

The ectoparasitic mite Dorsipes caspius n. sp. (Heterostigmata: Podapolipidae) belonging to the platysmae species group collected from the beneath elytra of Pterostichus (Lyrothorax) caspius (Menetries) (Coleoptera: Carabidae) in northern Iran, is described. This is the first record of species of the platysmae group from the Middle East. Keys to adult and larval female stages of world species of the platysmae group are provided. The host range of all species of the genus and the distribution of all representatives of the group are discussed.     

The endemic Genista versicolor from Sierra Nevada National Park in Spain is nodulated by putative new Bradyrhizobium species and a novel symbiovar (sierranevadense)

Genista versicolor is an endemic legume from Sierra Nevada National Park which constitutes one of the UNESCO-recognized Biosphere Reserves. In the present study, a collection of strains nodulating this legume was analysed in characteristic soils of this ecosystem. Most strains nodulating G. versicolor belonged to rrs group I within the genus Bradyrhizobium and only one strain, named GV137, belonged to rrs group II from which only a single species, B. retamae, has been described in Europe to date. Strain GV137, and some strains from rrs group I, belonged to putative new species of Bradyrhizobium, although most strains from group I belonged to B. canariense, according to the ITS fragment and atpD gene analysis. This result contrasted with those obtained in Genista tinctoria in Northeast Europe whose endosymbionts were identified as B. japonicum. The analysis of the symbiotic nodC and nifH genes carried by G. versicolor-nodulating strains showed that most of them belonged to symbiovar genistearum, as did those isolated from G. tinctoria. Nevertheless, strain GV137, belonging to rrs group II, formed a divergent lineage that constituted a novel symbiovar within the genus Bradyrhizobium for which the name sierranevadense is proposed. This finding showed that the Genisteae are not restrictive legumes only nodulated by symbiovar genistearum, since Genista is a promiscuous legume nodulated by at least two symbiovars of Bradyrhizobium, as occurs in Retama species.     

Vigna unguiculata is nodulated in Spain by endosymbionts of Genisteae legumes and by a new symbiovar (vignae) of the genus Bradyrhizobium

Vigna unguiculata was introduced into Europe from its distribution centre in Africa, and it is currently being cultivated in Mediterranean regions with adequate edapho-climatic conditions where the slow growing rhizobia nodulating this legume have not yet been studied. Previous studies based on rrs gene and ITS region analyses have shown that Bradyrhizobium yuanmingense and B. elkanii nodulated V. unguiculata in Africa, but these two species were not found in this study. Using the same phylogenetic markers it was shown that V. unguiculata, a legume from the tribe Phaseolae, was nodulated in Spain by two species of group I, B. cytisi and B. canariense, which are common endosymbionts of Genisteae in both Europe and Africa. These species have not been found to date in V. unguiculata nodules in its African distribution centres. All strains from Bradyrhizobium group I isolated in Spain belonged to the symbiovar genistearum, which is found at present only in Genisteae legumes in both Africa and Europe. V. unguiculata was also nodulated in Spain by a strain from Bradyrhizobium group II that belonged to a novel symbiovar (vignae). Some African V. unguiculata-nodulating strains also belonged to this proposed new symbiovar.