Comparison Of DNA Sequences Research Articles

Oxalate oxidase from Abortiporus biennis – protein localisation and gene sequence analysis

We have described for the first time the localisation of oxalate oxidase (OXO, EC 1.2.3.4) in Abortiporus biennis cells, using histochemical and immunochemical methods coupled with transmission electron microscopy. Rabbit anti-oxalate oxidase immunoglobulins with anti-rabbit secondary antibody conjugated with 10-nm gold particles were used. Moreover, the formation of electron dense precipitation of reaction of diaminobenzidine (DAB) with horseradish peroxidase (HRP) for histochemical localisation of the enzyme was found. OXO was localised close to the membranous structures of the cell membranes, in membranous vesicles located close to the outer cell membrane, and vacuolar membranes surrounding vacuoles. The positive immunoreaction to OXO was also intense in cell wall areas. Moreover, we proved that gene coding for OXO was expressed in the same cultures. Corresponding mRNA was isolated, full length cDNA was synthesized, cloned and sequenced. Two copies of cupin domains were found in the sequence of amino-acids conserved domain coding for the cupin enzyme. Comparison of the genomic DNA and cDNA sequences has revealed the presence of seventeen introns in the gene. The isoelectric point of the protein was estimated at pH 4.5 and several possible N-glycosylation sites were predicted.

Molecular confirmation and morphological reassessment of Udotea geppiorum (Bryopsidales, Chlorophyta) with ecological observations of mesophotic meadows in the Main Hawaiian Islands

ABSTRACTUdotea geppiorum is reported to form meadows in deep-water, soft sediment habitats in the Main Hawaiian Islands. The identification of the species, initially published as U. argentea in the archipelago, is revised and confirmed based on morphological assessment and comparative DNA sequencing with western Pacific specimens. Udotea geppiorum is fully corticated and newly shown to display discrete concentric segments arranged in a ‘tongue and groove’ manner, which are connected by medullary siphons running along the entire segment interface. Udotea geppiorum is phylogenetically nested among a cluster of Udotea spp. recently circumscribed within Udotea sensu stricto. This cluster of species is separate from the remainder of the family Udoteaceae, which includes several genera and Udotea spp. sensu lato that lack or exhibit incomplete cortication. Based on field observations conducted in O‘ahu and Maui, U. geppiorum can be found from 20 m to > 90 m depth with a peak in abundance at mesophotic depths of 60–85 m, where it forms extensive meadows that support associated diversity.

Genome rearrangements induce biofilm formation in Escherichia coli C \u2013 an old model organism with a new application in biofilm research

BackgroundEscherichia coli C forms more robust biofilms than other laboratory strains. Biofilm formation and cell aggregation under a high shear force depend on temperature and salt concentrations. It is the last of five E. coli strains (C, K12, B, W, Crooks) designated as safe for laboratory purposes whose genome has not been sequenced.ResultsHere we present the complete genomic sequence of this strain in which we utilized both long-read PacBio-based sequencing and high resolution optical mapping to confirm a large inversion in comparison to the other laboratory strains. Notably, DNA sequence comparison revealed the absence of several genes thought to be involved in biofilm formation, including antigen 43, waaSBOJYZUL for lipopolysaccharide (LPS) synthesis, and cpsB for curli synthesis. The first main difference we identified that likely affects biofilm formation is the presence of an IS3-like insertion sequence in front of the carbon storage regulator csrA gene. This insertion is located 86 bp upstream of the csrA start codon inside the − 35 region of P4 promoter and blocks the transcription from the sigma32 and sigma70 promoters P1-P3 located further upstream. The second is the presence of an IS5/IS1182 in front of the csgD gene. And finally, E. coli C encodes an additional sigma70 subunit driven by the same IS3-like insertion sequence. Promoter analyses using GFP gene fusions provided insights into understanding this regulatory pathway in E. coli.ConclusionsBiofilms are crucial for bacterial survival, adaptation, and dissemination in natural, industrial, and medical environments. Most laboratory strains of E. coli grown for decades in vitro have evolved and lost their ability to form biofilm, while environmental isolates that can cause infections and diseases are not safe to work with. Here, we show that the historic laboratory strain of E. coli C produces a robust biofilm and can be used as a model organism for multicellular bacterial research. Furthermore, we ascertained the full genomic sequence of this classic strain, which provides for a base level of characterization and makes it useful for many biofilm-based applications.

Reinstatement of Myelophycus caespitosus (Scytosiphonaceae, Phaeophyceae) from Japan

SUMMARYReinstatement of Myelophycus caespitosus Kjellman (Ectocarpales s.l., Phaeophyceae) described from western Japan is proposed based on the comparisons of DNA sequences of Japanese Myelophycus specimens, the type specimen of Chordaria simplex and lectotypified specimen of Myelophycus caespitosus. In the genetic analyses using mitochondrial cox1 and cox3, chloroplast atpB, psbA and rbcL DNA sequences, the specimens morphologically referable to M. simplex formed two distinct clades (clade‐1 and clade‐2) supported by high statistical values. Clade‐1 was distributed on the western coast of Japan (Honshu, Shikoku and Kyushu) from the Kii Peninsula to Tsushima and the Pacific coast of Aomori, northeastern Honshu, and clade‐2 on the Pacific coast of central Honshu from Sanriku to the Kii Peninsula. Based on the cox3 DNA sequences and the location of the type locality at Shimoda, Izu Peninsula, clade‐2 was concluded to correspond to true M. simplex. There were no recognizable differences in the representative morphological features (height and diameter of sporophytes and gametophytes, number of cells comprising cortical, subcortical and medullary layers, thickness of plurilocular gametangia, or length and diameter of unilocular zoidangia) between the specimens included in the two taxa. Analyses of the specimens of the two taxa, including sympatric populations on the Kii Peninsula, using a nuclear genetic marker ocm3 did not suggest any genetic exchanges between the two taxa. On the other hand, cox3 gene sequence of the voucher specimen of Myelophycus caespitosus collected from Goto, Kyushu, western Japan housed in the UPS herbarium was included in clade‐1. In conclusion, we propose the reinstatement of M. caespitosus and to lectotypify the specimen in UPS. Korean Myelophycus specimens reported from Cheju Island and Wando were considered to belong to M. caespitosus based on the reported rbcL sequences.

DNA BARCODING STUDY OF SHELLED GASTROPODS IN THE INTERTIDAL ROCKY COASTS OF CENTRAL WAKAYAMA PREFECTURE, JAPAN, USING TWO GENE MARKERS

The coasts of Wakayama Prefecture are known to be among the most biologically diverse coastal areas of Japan, and thus have a rich assemblage of shelled gastropods. In this paper, we report the result of our DNA Barcoding of shelled gastropods of the intertidal area of the Nada Coast in central Wakayama, using the mitochondrial genes COI and the nuclear gene Histone H3 as markers. In order to do so, we collected up to five individuals from 12 species of shelled gastropods from the intertidal rocky beach. Collected samples were first identified morphologically and then vouchered at the University Museum of the University of Tokyo. DNA sequence comparisons and phylogenetic analyses indicated that both genes have enough substitutions to differentiate species. We also found that the sequence data for most of our target species are not available on Genbank. Our result presented here indicated that we were not only successful in barcoding/identifying the target gastropod species in the area, but also contributed to the building of a set of reference DNA sequences for future DNA-based environmental and biodiversity monitoring, besides providing sequence data for future systematics studies of this group.

Micropropagation and molecular characterization of Thymus sibthorpii Benth. (Lamiaceae), an aromatic-medicinal thyme with ornamental value and conservation concern

Thymus sibthorpii Benth. (Lamiaceae), with accession number 01,1796-22, is a biotype of native Greek thyme with ascending stems and potential use as a new medicinal-aromatic crop and ornamental plant. An efficient and reliable protocol for in vitro clonal propagation of T. sibthorpii from nodes and meristem tip explants was developed. Shoot proliferation succeeded on a new basal medium (BB) without plant growth regulators, as prior experiments with 6-benzyladenine generated hyperhydricity. Eight different basal media were compared; on two formulations using the new BB 5.9 and 5.6 shoots per explant were produced. Regenerated single shoots were rooted in the BB medium, supplemented with 5 μM of indole-3-butyric acid, and produced 3.1 roots along with 2.5 adventitious shoots. Three types of acclimatization were assessed: in vitro, using two different systems (no significant differences); ex vitro, using eight soil substrates under greenhouse and outdoor nursery conditions (in two of them, 100% of plantlets survived); and in field cultivations, established at eight geographically distant areas of Greece (100% survival rate at all locations). Molecular characterization of T. sibthorpii was evaluated with one nuclear ribosomal DNA and seven chloroplast DNA markers, followed by DNA sequence comparisons with a total of 30 different Thymus species, subspecies, and varieties. The trnH/psbA, trnL/trnF, and matK genes were the most efficient markers for molecular characterization of T. sibthorpii. The molecular markers rpoC1 and petB/petD did not match to any Thymus species and therefore, these DNA sequences provide new sequence information for entire Thymus taxa.

Metagenomic Classification using Centrifuge

To assess the quality of food is a major challenge the food industry faces today. It is of utmost importance to test it for contaminates and non-edible material that may be present. To overcome these challenges metagenomic classification is majorly useful. Several researches involve various classification techniques and their studies. Difficulties in metagenomic classification include increasing number of genomes thereby requirement of computational methods to have high speed as well as high accuracy so as to compare DNA sequences to genomes. Centrifuge is a classification tool for quantification of species present in a sample so as to monitor the quality of the same. Given a food sample Centrifuge effectively classifies the species present in it enabling a timely and accurate analysis.

A comparison of complete mitochondrial DNA sequences of Mnais costalis Selys, 1869 (Odonata: Calopterygidae) from three different populations (one allopatric and two sympatric)

In Japan, two closely-related damselflies, Mnais costalis Selys, 1869 (Odonata: Calopterygidae) and M. pruinosa Selys-Longchamps (Odonata: Calopterygidae), 1853, coexist, and they exhibit geographic variations in wing color, body size, and habitat preference. In this study, we analyzed the complete mitochondrial genome of M. costalis from Saga Prefecture, Japan (sympatric populations that exhibit wing color polymorphism), and compared the genome with M. costalis that exhibit monomorphic orange wing color. The mitochondrial genome of M. costalis from Saga Prefecture was identified as a circular molecule of 15,488 bp, similar to that found in other M. costalis populations. It was predicted to contain 13 protein-coding (PCG), 22 tRNA, and two rRNA genes, along with one A + T-rich control region. Among the PCGs, ATP8 and ATP6, ATP6 and COIII, ND4 and ND4L, and ND6 and Cytb shared seven, one, seven, and one nucleotides, respectively. The initiation codon ATG was found in eight genes, ATC in four, and ATT in one, while the termination codons TAA, TAG, TA, and T were observed in seven, one, two, and three genes, respectively. All the tRNA genes possessed a cloverleaf secondary structure, except for tRNA-His that lacks the TΨC loop. The average AT content of mitochondrial genome was 66.06%. From a phylogenetic analysis, the loss of wing color polymorphism in monomorphic sympatric populations is likely to occur with the coexistence of two Mnais species.

Ophiostomatales associated with wounds on hardwood trees in Poland

Wounds on trees provide entrance for many members of the Ophiostomatales (Ascomycota), including economically damaging tree pathogens. Because very little is known regarding wound‐associated fungi on hardwoods in Europe or their potential nitidulid beetle vectors (Coleoptera), the aim of this survey was to explore the diversity of these fungi in Poland. Wound samples associated with 26 tree species were collected from 18 stands in Poland. Fungi belonging to the Ophiostomatales were isolated and identified based on morphology and DNA sequence comparisons of five gene regions, combined with phylogenetic analyses. In total, 32 taxa were identified, including 20 known and 12 currently undescribed species. In addition, Grosmannia cainii, Ophiostoma cf. ponderosae, O. sparsiannulatum and Sporothrix aurorae were reported for the first time outside of the USA. The wound‐associated members of the Ophiostomatales recovered from hardwoods showed high diversity and specificity. Among the species recovered, 16 belonged to Sporothrix, 11 represented Ophiostoma s. l., five could be assigned to Leptographium s. l., and one Graphilbum species was detected. Among the studied tree species, wounds on Quercus robur appeared to be the most vulnerable to infection by members of the Ophiostomatales. All the nitidulid beetle species were found in association with species assignable to Ophiostoma and Sporothrix. This study detected a significant association between the occurrence of species belonging to the Ophiostomatales and the health status of the wounded trees. Some of the isolated species were host specific although Ophiostoma quercus, the most frequently encountered species, had a broad host range.

Morphological and molecular study on Yininemertespratensis (Nemertea, Pilidiophora, Heteronemertea) from the Han River Estuary, South Korea, and its phylogenetic position within the family Lineidae.

Outbreaks of ribbon worms observed in 2013, 2015, and 2017–2019 in the Han River Estuary, South Korea, have caused damage to local glass-eel fisheries. The Han River ribbon worms have been identified as Yininemertespratensis (Sun & Lu, 1998) based on not only morphological characteristics compared with the holotype and paratype specimens, but also DNA sequence comparison with topotypes freshly collected near the Yangtze River mouth, China. Using sequences of six gene markers (18S rRNA, 28S rRNA, histone H3, histone H4, 16S rRNA, and COI), the phylogenetic position of Y.pratensis was inferred among other heteronemerteans based on their sequences obtained from public databases. This analysis firmly placed Y.pratensis as a close relative to Apatronemertesalbimaculosa Wilfert & Gibson, 1974, which has been reported from aquarium tanks containing tropical freshwater plants in various parts of the world as well as a wild environment in Panama.

Cutaneotrichosporon suis sp. nov., a lipolytic yeast species from food and food-related environment.

Two conspecific yeast strains, which based on DNA sequence comparisons represented an undescribed species in the order Trichosporonales were isolated during two independent studies in Hungary and France. One of them (NCAIM Y.02224) was recovered from minced pork in Hungary while the other one (UBOCC-A-218003) was isolated from the air of a dairy plant in France. The two strains shared identical nucleotide sequences in the D1/D2 domain of the nuclear large subunit (LSU) rRNA gene and in the internal transcribed spacer (ITS) region. Analysis of the concatenated DNA sequences for the ITS region and D1/D2 domain of the LSU rRNA gene indicated that the novel species belongs to the recently erected genus Cutaneotrichosporon. According to sequence comparisons and phylogenetic analysis, the novel species is most closely related to Cutaneotrichosporon curvatum (formerly Cryptococcus curvatus), which is often associated with humans and warm-blooded animals. The physiological characteristics of this novel species are also very similar to that of Cutaneotrichosporon curvatum. The only clear-cut difference is that, unlike C. curvatum, the novel species does not utilize imidazole as a nitrogen-source. The species name Cutaneotrichosporon suis sp. nov. is proposed to accommodate the above-noted two strains.

Four new Ophiostoma species associated with conifer- and hardwood-infesting bark and ambrosia beetles from the Czech Republic and Poland

Fungi under the order Ophiostomatales (Ascomycota) are known to associate with various species of bark beetles (Coleoptera: Curculionidae: Scolytinae). In addition this group of fungi contains many taxa that can impart blue-stain on sapwood and some are important tree pathogens. A recent survey that focussed on the diversity of the Ophiostomatales in the forest ecosystems of the Czech Republic and Poland uncovered four putative new species. Phylogenetic analyses of four gene regions (ITS1-5.8S-ITS2 region, ß-tubulin, calmodulin, and translation elongation factor 1-α) indicated that these four species are members of the genus Ophiostoma. All four newly described species can be distinguished from each other and from closely related species based on DNA sequence comparisons, morphological characters, growth rates, and their insect associations. Based on this study four new taxa can be circumscribed and the following names are provided: Ophiostoma pityokteinis sp. nov., Ophiostoma rufum sp. nov., Ophiostoma solheimii sp. nov., and Ophiostoma taphrorychi sp. nov. O. rufum sp. nov. is a member of the Ophiostoma piceae species complex, while O. pityokteinis sp. nov. resides in a discrete lineage within Ophiostoma s. stricto. O. taphrorychi sp. nov. together with O. distortum formed a well-supported clade in Ophiostoma s. stricto close to O. pityokteinis sp. nov. O. solheimii sp. nov. groups within a currently undefined lineage A, which also includes Ophiostoma grandicarpum and Ophiostoma microsporum. This study highlights the need for more intensive surveys that should include additional countries of Central Europe, insect vectors and host tree species in order to elucidate Ophiostoma species diversity in this region.

Molecular Detection and Phylogenetic Analysis of Endosymbiont Wolbachia pipientis (Rickettsiales: Anaplasmataceae) Isolated from Dirofilaria immitis in Northwest of Iran

The purpose of this study was molecular detection and phylogenetic analysis of Wolbachia species of Dirofilaria immitis. Adult filarial nematodes were collected from the cardiovascular and pulmonary arterial systems of naturally infected dogs, which caught in different geographical areas of Meshkin Shahr in Ardabil Province, Iran, during 2017. Dirofilaria immitis genomic DNA were extracted. Phylogenetic analysis for proofing of D. immitis was carried out using cytochrome oxidase I (COI) gene. Afterward, the purified DNA was used to determine the molecular pattern of the Wolbachia surface protein (WSP) gene sequence by PCR. Phylogeny and homology studies showed high consistency of the COI gene with the previously-registered sequences for D. immitis. Comparison of DNA sequences revealed no nucleotide variation between them. PCR showed that all of the collected parasites were infected with W. pipientis. The sequence of the WSP gene in Wolbachia species from D. immitis was significantly different from other species of Dirofilaria as well as other filarial species. The maximum homology was observed with the Wolbachia isolated from D. immitis. The greatest distance between WSP nucleotides of Wolbachia species found between D. immitis and those isolated from Onchocerca lupi. PCR could be a simple but suitable method for detection of Wolbachia species. There is a pattern of host specificity between Wolbachia and Dirofilaria that can be related to ancestral evolutions. The results of this phylogenetic analysis and molecular characterization may help us for better identification of Wolbachia species and understanding of their coevolution.

Identification and pathogenicity of Cryphonectriaceae species associated with chestnut canker in China

Cryphonectriaceae species cause serious canker diseases on chestnut, oak and eucalypt trees. Recently, canker symptoms with typical orange fruiting bodies were observed on Chinese chestnut and oak trees in Hebei, Hubei, Shaanxi and Shandong Provinces in China. In the present study, isolates of these fungi were identified based on phylogenetic and morphological evidence, and their pathogenicities were tested on detached chestnut (Castanea mollissima) branches. DNA sequence comparisons of the internal transcribed spacer (ITS) regions and two regions of β‐tubulin (TUB1/TUB2) indicate that these isolates represent five species in the Cryphonectriaceae, viz. Cryphonectria japonica, Cryphonectria parasitica, Aurantiosacculus castaneae sp. nov., Cryphonectria neoparasitica sp. nov. and Endothia chinensis sp. nov. The sexual morph of Aurantiosacculus is discovered for the first time and can be distinguished from the other genera in Cryphonectriaceae by dark brown ascospores and tubiform appendages at both ends. Cryphonectria neoparasitica sp. nov. is different from the other Cryphonectria species by its aseptate ascospores. Endothia chinensis sp. nov. is the sole species in Endothia infecting the host genus Castanea. Additionally, it is much smaller than E. gyrosa and narrower than E. singularis in ascospores. The inoculation results showed that these five Cryphonectriaceae fungi isolated from chestnut or oak are all pathogenic to tested chestnut branches. Cryphonectria parasitica appears to be the most aggressive fungus, followed by C. neoparasitica sp. nov., C. japonica, E. chinensis sp. nov. and A. castaneae sp. nov.

Cloning, characterization and tissue-specific expression of the antimicrobial peptide hepcidin from caspian trout (Salmo caspius) and the antibacterial activity of the synthetic peptide

Antimicrobial peptides have a wide range of antimicrobial activity and widely occur in different organisms including mollusks, crustaceans and vertebrates. Hepcidins are a group of cysteine-rich antimicrobial peptides that are active against a variety of pathogens including gram-positive and gram-negative bacteria, as well as viruses. In this study, the hepcidin gene of Caspian trout (CtHep) was identified and characterized. Our results showed that CtHep cDNA has a 267-bp Open Reading Frame (ORF), which is translated to 88 amino acids. The CtHep was classified in the HAMP1 class of hepcidins. Comparison of DNA and cDNA sequences showed that CtHep has 3 exons and 2 introns. The signal, prodomain and mature part of CtHep have 24, 39 and 25 amino acids, respectively. The mature peptide has a molecular weight of 2881.43 Da and a theoretical isoelectric point of 8.53. The expression of CtHep mRNA was detected in different tissues of healthy and infected fish. CtHep expression in the liver, head kidney, spleen and skin was significantly enhanced after bacterial challenge. Expression of CtHep in different embryonic development stages was also substantial. Antibacterial activity of synthetic CtHep peptides was investigated against a number of Gram-positive and Gram-negative bacteria. CtHep inhibited some pathogenic bacteria such as Streptococcus iniae and Aeromonas hydrophila. In the in vivo experiment, CtHep upregulated the cytokines IL-6 and TNF-α in both kidney and spleen tissues after 24 h of the peptide injection. In conclusion, our study showed that CtHep plays an important role in the immune system of Caspian trout and also in the embryonic stages. Moreover, CtHep peptide has a potential to be used as an antimicrobial therapeutic agent as well as an immunostimulant in aquaculture.

Genome-wide effects on Escherichia coli transcription from ppGpp binding to its two sites on RNA polymerase

The second messenger nucleotide ppGpp dramatically alters gene expression in bacteria to adjust cellular metabolism to nutrient availability. ppGpp binds to two sites on RNA polymerase (RNAP) in Escherichia coli, but it has also been reported to bind to many other proteins. To determine the role of the RNAP binding sites in the genome-wide effects of ppGpp on transcription, we used RNA-seq to analyze transcripts produced in response to elevated ppGpp levels in strains with/without the ppGpp binding sites on RNAP. We examined RNAs rapidly after ppGpp production without an accompanying nutrient starvation. This procedure enriched for direct effects of ppGpp on RNAP rather than for indirect effects on transcription resulting from starvation-induced changes in metabolism or on secondary events from the initial effects on RNAP. The transcriptional responses of all 757 genes identified after 5 minutes of ppGpp induction depended on ppGpp binding to RNAP. Most (>75%) were not reported in earlier studies. The regulated transcripts encode products involved not only in translation but also in many other cellular processes. In vitro transcription analysis of more than 100 promoters from the in vivo dataset identified a large collection of directly regulated promoters, unambiguously demonstrated that most effects of ppGpp on transcription in vivo were direct, and allowed comparison of DNA sequences from inhibited, activated, and unaffected promoter classes. Our analysis greatly expands our understanding of the breadth of the stringent response and suggests promoter sequence features that contribute to the specific effects of ppGpp.

Differential patterns of ophiostomatoid fungal communities associated with three sympatric Tomicus species infesting pines in south-western China, with a description of four new species.

Bark beetles and their associated fungi, which cause forest decline and sometimes high mortality in large areas around the world, are of increasing concern in terms of forest health. Three Tomicus spp. (T.brevipilosus, T.minor and T.yunnanensis) infect branches and trunks of Pinusyunnanensis and P.kesiya in Yunnan Province, in south-western China. Tomicus spp. are well known as vectors of ophiostomatoid fungi and their co-occurrence could result in serious ecological and economic impact on local forest ecosystems. Nonetheless, knowledge about their diversity, ecology, including pathogenicity and potential economic importance is still quite rudimentary. Therefore, an extensive survey of ophiostomatoid fungi associated with these Tomicus species infesting P.yunnanensis and P.kesiya was carried out in Yunnan. Seven hundred and seventy-two strains of ophiostomatoid fungi were isolated from the adult beetles and their galleries. The strains were identified based on comparisons of multiple DNA sequences, including the nuclear ribosomal large subunit (LSU) region, the internal transcribed spacer regions 1 and 2, together with the intervening 5.8S gene (ITS) and the partial genes of β-tubulin (TUB2), elongation factor 1α (TEF1-α) and calmodulin (CAL). Phylogenetic analyses were performed using maximum parsimony (MP) as well as maximum likelihood (ML). Combinations of culture features, morphological characters and temperature-dependent growth rates were also employed for species identification. Eleven species belonging to five genera were identified. These included six known species, Esteyavermicola, Leptographiumyunnanense, Ophiostomabrevipilosi, O.canum, O.minus and O.tingens and four novel taxa, described as Graphilbumanningense, O.aggregatum, Sporothrixpseudoabietina and S.macroconidia. A residual strain was left unidentified as Ophiostoma sp. 1. The overall ophiostomatoid community was by far dominated by three species, representing 87.3% of the total isolates; in decreasing order, these were O.canum, O.brevipilosi and O.minus. Furthermore, the ophiostomatoid community of each beetle, although harbouring a diversity of ophiostomatoid species, was differentially dominated by a single fungal species; Ophiostomacanum was preferentially associated with and dominated the ophiostomatoid community of T.minor, whereas O.brevipilosi and O.minus were exclusively associated with and dominated the ophiostomatoid communities of T.brevipilosus and T.yunnanensis, respectively. Eight additional species, representing the remaining 12.7% of the total isolates, were marginal or sporadic. These results suggested that sympatric Tomicus populations are dominated by distinct species showing some level of specificity or even exclusivity.

A Novel Approach to Clustering Genome Sequences Using Inter-nucleotide Covariance.

Classification of DNA sequences is an important issue in the bioinformatics study, yet most existing methods for phylogenetic analysis including Multiple Sequence Alignment (MSA) are time-consuming and computationally expensive. The alignment-free methods are popular nowadays, whereas the manual intervention in those methods usually decreases the accuracy. Also, the interactions among nucleotides are neglected in most methods. Here we propose a new Accumulated Natural Vector (ANV) method which represents each DNA sequence by a point in ℝ18. By calculating the Accumulated Indicator Functions of nucleotides, we can further find an Accumulated Natural Vector for each sequence. This new Accumulated Natural Vector not only can capture the distribution of each nucleotide, but also provide the covariance among nucleotides. Thus global comparison of DNA sequences or genomes can be done easily in ℝ18. The tests of ANV of datasets of different sizes and types have proved the accuracy and time-efficiency of the new proposed ANV method.

A redescription of Antispastis clarkei Pastrana (Lepidoptera, Glyphipterigidae) immature stages, with notes on the life history and phylogenetic placement of the genus

Antispastis Meyrick, 1926 is a poorly known genus of leaf-mining micromoths endemic to the Neotropics, with still uncertain taxonomic position within the Yponomeutoidea. In the present study, the egg, larva and pupa of A. clarkei Pastrana, previously known only from Argentina, are described and illustrated with the aid of optical and scanning electron microscopy. Data on life history, including histology of the mine, are also provided. Family placement of the genus is reassessed based on comparison of morphology and DNA sequences with closely related lineages. The larvae form blotch mines on the upper surface of Solanum L. (Solanaceae) leaves, feeding on palisade parenchyma in all instars. Pupation occurs outside the mine, in an inverted basket-like, large-meshed cocoon constructed on the plant surface. DNA analysis of Cytochrome oxidase I gene of A. clarkei revealed interspecific differences averaging 10% with A. xylophragma, which provided species separation matching morphological differences. Antispastis was closely related phylogenetically to Digitivalva, clustering in the Acrolepiinae together with the genera Acrolepia and Acrolepiopsis, and ultimately placed within Glyphipterigidae. The geographical distribution of A. clarkei is expanded to the Southern Atlantic forest (Rio Grande do Sul and Paraná states, Brazil).

A cryptic new species of bulbul from Borneo

Cream-vented Bulbul Pycnonotus simplex of Borneo was previously considered to be polymorphic in iris colour, having either red or white (creamy-yellow) irides. Mitochondrial DNA sequence comparisons, however, indicate that white- and red-eyed Bornean individuals are not closely related to one another. Instead, white-eyed birds are sister to Ashy-fronted Bulbul P. cinereifrons of Palawan Island, in the south-west Philippines, and red-eyed birds are sister to white-eyed P. simplex of the Thai-Malay Peninsula. Consequently, we elect to treat the white-eyed Bornean population as a distinct, previously overlooked species. In respect to plumage, white- and red-eyed individuals are almost identical, varying only slightly in the amount of yellow coloration in their feathers. The two taxa are sympatric at some localities, but white-eyed individuals are rarer and more consistently associated with mature forest than red-eyed birds.