Isolation and partial characterization of antimicrobial compounds from a new strain <Emphasis Type="Italic">Nonomuraea</Emphasis> sp. NM94

An actinomycete strain NM94 was isolated from a Saharan soil sample by a dilution agar plating method using chitin-vitamins B medium supplemented with penicillin. The strain presented the morphological and chemical characteristics of the genus Nonomuraea. On the basis of 16S rDNA analysis and physiological tests, this isolate was found to be quite different from the known species of Nonomuraea and might be new. The strain NM94 secreted several antibiotics on yeast extract malt extract glucose medium that were active against some Gram-positive bacteria, yeast, and fungi. The antibiotics were extracted with dichloromethane and detected by bioautography on silica gel plates using Mucor ramannianus and Bacillus subtilis as the test organisms. Among these antibiotics, a complex called 94A showed interesting antifungal activity. It was selected and purified by reverse-phase HPLC. This complex was composed of five compounds. Spectroscopic studies by infrared, mass, and ¹H NMR of the compounds were carried out. Initial results showed that these molecules differed from the known antibiotics produced by other Nonomuraea species.     

Methods for the genetic manipulation of <Emphasis Type="Italic">Nonomuraea</Emphasis> sp. ATCC 39727

Nonomuraea sp. ATCC 39727 belongs to the Streptosporangiaceae family of filamentous actinomycetes. This microorganism produces the teicoplanin-like glycopeptide A40926, which is the starting material for the synthesis of the second-generation glycopeptide dalbavancin. Notwithstanding the strain’s pharmaceutical relevance, the lack or poor efficiency of genetic tools to manipulate Nonomuraea sp. ATCC 39727 has hampered strain and product improvement. Here we report the development of gene transfer systems based on protoplast transformation and intergeneric conjugation from Escherichia coli. Efficiency of transformation and conjugation, followed by site specific or homologous recombination with the Nonomuraea sp. genome, were determined using the integrative plasmid pSET152 (5.7 kb), and the Supercos1 derivative cosmid A40ΔY (30 kb). To our knowledge, this is the first report of the transformation of protoplasts of Nonomuraea sp. ATCC 39727, even though the improved procedure for intergeneric conjugation makes it the method of choice for introducing large segments of DNA into Nonomuraea sp. ATCC 39727.     

The taxonomic relationships within the genus Excoecaria L. (Euphorbiaceae) based on leaf morphology and rDNA sequence data

The tropical Indo-Pacific genus Excoecaria L. (Euphorbiaceae) has several closely related species in Australia whose taxonomic relationships are unclear. The most widely reported species in Australia is the mangrove species Excoecaria agallocha L. (type species), whose taxonomic and geographic limits are difficult to define from its closely related species or sub-species. Two additional taxa have also been described but not clearly differentiated from the type species: Excoecaria dallachyana Baillon and Excoecaria ovalis Endl. This project aimed to determine the taxonomic relationships of the Australian Excoecaria species using both leaf morphological data and DNA sequence data from the internal transcribed spacer (ITS) region of ribosomal genes. The nucleotide differences in the examined ITS1 region show that E. agallocha from eastern Australia and E. ovalis from Western Australia respectively, are genetically uniform within species but differ from each other consistently, thus supporting species status. The leaf morphological data also support this view: single factor analysis of variance consistently separated E. ovalis from E. agallocha on the basis of leaf width, leaf length and length of petiole. In contrast, E. ovalis from the Gulf of Carpentaria differs only slightly from E. ovalis in Western Australia, but no evidence was found to suggest any leaf morphological differentiation within this species. The analysis also suggests that E. dallachyana is not closely related to either mangrove species E. agallocha or E. ovalis, despite superficial morphological similarities.     

Numerical and chemical classification ofStreptosporangium and some related actinomycetes

One hundred and seventeen streptosporangia from soil were compared with marker strains of the familyStreptosporangiaceae for many phenotypic properties. The data were examined using the Jaccard, pattern and simple matching coefficients with clustering achieved using average, complete and single linkage algorithms. Particular confidence was placed in the product of the pattern, average linkage analysis given the sharp definition of aggregate groups and clusters and a combination of low test error and high cophenetic correlation values. The test strains were assigned to five aggregate groups that were equated with the generaStreptosporangium (group A),Microbispora (group B),Planobispora andPlanomonospora (Group C),Kutzneria (neéStreptosporangium viridogriseum (group D), andMicrotetraspora (group E). The streptosporangia, both isolates and marker strains, were assigned to 5 major, 7 minor and 18 single membered clusters. Representative streptosporangia examined for chemical markers were characterised by the presence ofmeso-diaminopimelic acid in whole-organism hydrolysates, complex mixtures of straight- and branched chain fatty acids, di- and tetrahydrogenated menaquinones as predominant isoprenologues, and complex polar lipid patterns containing diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides and uncharacterised components. The chemical and numerical data support the taxonomic integrity of the validly described species ofStreptosporangium and suggest that the genus is markedly underspeciated.     

Streptomyces yatensis sp. nov., a novel bioactive streptomycete isolated from a New-Caledonian ultramafic soil

The taxonomic position of an actinomycete isolated from an ultramafic soil in New Caledonia was examined using a polyphasic approach. The organism, which was designated SFOCin 76, was found to have chemical and morphological properties typical of streptomycetes and formed a distinct phyletic line in the Streptomyces violaceusniger clade of the 16S rDNA tree. It also showed a unique pattern of phenotypic properties that distinguished it from representatives of all of the validly described species classified in this clade. It is, therefore, proposed that strain SFOCin 76 be classified in the genus Streptomyces as Streptomyces yatensis sp. nov. This revised version was published online in June 2006 with corrections to the Cover Date.     

Phenetic studies in Nuphar Sm. (Nymphaeaceae):variation in sect. Nuphar

Previous taxonomic assessments of the hydrophytic genus Nuphar Sm. (Nymphaeaceae) have inadequately considered the range of morphological variation existing in the genus. A comprehensive statistical analysis of morphological features among the recognized taxa of Nuphar is presented here, first using 10 fruit characters to delimit the separation of two infrageneric groups of taxa, and secondly using 31 characters to quantify the morphological variation found within sect. Nuphar. Results from combined phenetic analyses provided empirical support for recognition of two infrageneric groups of species in the genus (largely by fruit, stigmatic disk margin, neck, and stigmatic disk size) and for separation of taxa within sect. Nuphar (by leaf size, and fruit disk and neck size). Statistical analyses demonstrate that extensive variation in ranges of character states exists among sect. Nuphar taxa. However, a suite of qualitative and quantitative characters can be used to recognize five species and three subspecies within this predominantly Old World assemblage.     

<Emphasis Type="Italic">Nonomuraea rhizosphaerae</Emphasis> sp. nov., an actinomycete isolated from the rhizosphere soil of a rubber tree (<Emphasis Type="Italic">Hevea brasiliensis</Emphasis> Muell. Arg)

A novel actinomycete, designated strain NEAU-mq18^T, was isolated from the rhizosphere soil of a rubber tree (Hevea brasiliensis Muell. Arg) collected from Xianglu Mountain in Heilongjiang Province, northeast China, and subjected to a polyphasic taxonomic study. The 16S rRNA gene sequence analysis showed that the isolate belongs to the genus Nonomuraea with high sequence similarity to Nonomuraea guangzhouensis NEAU-ZJ3^T (98.5%). Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain clusters phylogenetically with N. guangzhouensis NEAU-ZJ3^T and Nonomuraea glycinis NEAU-BB2C19^T. Moreover, key chemotaxonomic properties including the major menaquinones, fatty acid composition and phospholipid profile also confirmed the affiliation of the strain to the genus Nonomuraea. However, some physiological, morphological and biochemical properties, and low DNA–DNA relatedness values, enabled the strain to be differentiated from closely related species of the genus Nonomuraea. Thus, strain NEAU-mq18^T is concluded to represent a novel species of the genus Nonomuraea, for which the name Nonomuraea rhizosphaerae sp. nov. is proposed. The type strain is NEAU-mq18^T (=CGMCC 4.7431^T=DSM 105761^T).     

Gordonia sinesedis sp. nov., a novel soil isolate

The taxonomic position of an actinomycete isolated from soil was evaluated using a polyphasic approach. The organism, strain J72, was found to have chemical and morphological properties consistent with its assignment to the genus Gordonia. A nearly complete 16S rDNA sequence of the strain was determined by direct sequencing of the amplified gene. The tested strain formed a distinct phylogenetic line within the evolutionary radiation occupied by the genus Gordonia and was most closely related to G. polyisoprenivorans DSM 44302^T. The phenotypic profile of strain 372 readily distinguishes it from representatives of the validly described species of Gordonia. The combined genotypic and phenotypic data show that strain J72 merits recognition as a new species of Gordonia. The name proposed for the new species is Gordonia sinesedis; the type strain is J72^T (= DSM 44455^T = NCIMB 13802^T). This revised version was published online in June 2006 with corrections to the Cover Date.     

Pollen morphology of selected Central European species from subgenera Vignea and Carex (Carex,Cyperaceae) and its relation to taxonomy

This study analysed the taxonomic utility of selected features of pollen grains of 45 Central European species of subgenera Vignea and Carex of the genus Carex. Seven quantitative features were analysed: [length of polar axis (P), length of equatorial axis (E), exine thickness on the pole (Exp), exine thickness on the equator plane (Exe), P/E ratio and the relative thickness of exine (Exp/P and Exe/E ratio)] and outline, shape and exine sculpture. Among the biometrical features, the most important for Carex taxonomy (particularly at the level of subgenus) is the thickness of exine (Exp). Marked differences in the morphological structure of inflorescences at the subgenus level are reflected in the differences of pollen features. Results of our studies clearly show an explicit (and not previously described in the palynological literature) taxonomic division of the genus Carex on the basis of pollen biometrical features into two units corresponding exactly to the subgenera (Vignea and Carex). However, the features of pollen grains analysed did not contribute any essential taxonomic criteria at levels lower than subgenus (e.g. sections). An exception was for C. disticha, where it was observed that the poroid areas occurred in pairs, a new feature for sedges. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 164 , 422–439.     

Proposal of Monorhizochytrium globosum gen. nov., comb. nov. (Stramenopiles,Labyrinthulomycetes) for former Thraustochytrium globosum based on morphological features and phylogenetic relationships

Thraustochytrium is the type genus of the family Thraustochytriaceae in the class Labyrinthulomycetes. This genus is characterized by zoospore formation, namely, shape of the cell wall of sporangia and presence or absence of a proliferous body. However, there are several issues associated with the taxonomy of this genus, and these include polyphyletic taxa and overlapping of taxonomic features among species. In particular, the first and second species, T. proliferum and T. globosum, were described based on observations of the morphological features of natural samples in the absence of culture conditions. Before addressing the taxonomic issues associated with this genus, it is important to consider the taxonomic features of each species, i.e., the life history under culture conditions and the phylogenetic position. The objective of the present study was to isolate T. globosum, the second described species in the genus Thraustochytrium, from the type locality. We successfully isolated strain NBRC 112723, which exhibited characteristic features of T. globosum. Under culture conditions, strain NBRC 112723 exhibited taxonomic features observed in other thraustochytrid species. Our molecular phylogeny indicated that this strain isolated from the type locality was located in an unidentified thraustochytrid group; moreover, some strains located in this group exhibited characteristic features of strain NBRC 112723. We clearly distinguished T. globosum based on the taxonomic criteria used to classify the T. proliferum type species. Therefore, we propose the establishment of a new genus, Monorhizochytrium, for the species T. globosum in the family Thraustochytriaceae.     

Phylogenetic Relationships Within the Genus Potorous (Marsupialia: Potoroidae) Based on Allozyme Electrophoresis and Sequence Analysis of the Cytochrome b Gene

Controversy over the taxonomic designations within the genus Potorous has highlighted problems associated with using only morphological data. The recent rediscovery of Gilbert's Potoroo has allowed us to reexamine the relationship of this species to other extant potoroos. Phylogenetic relationships and estimated divergence times are presented, based on electrophoretic and cytochrome b sequence data. There was considerable concordance between the data sets. We conclude that Gilbert's Potoroo is a separate species and should be referred to by its original name, Potorous gilbertii (Gould 1841). Estimates for potoroid divergences are in line with those of Flannery (1989).     

Is Oryza malampuzhaensis Krish. et Chand. (Poaceae) a valid species? Evidence from morphological and molecular analyses

Phylogenetic relationship between O. malampuzhaensis Krish. et Chand. (2n = 4x = 48; Poaceae, Oryzeae), a South Indian endemic wild rice with a disputed taxonomic identity, and eight other species belonging to the O. officinalis complex of the genus Oryza was examined using 62 morphological characters and 445 random amplified polymorphic DNA (RAPD) markers. Multivariate and cluster analyses using both the data sets clearly separated all accessions of O. malampuzhaensis into a distinct group. Genetic distances between O. malampuzhaensis and other species in O. officinalis complex were comparable with the distance between any other two taxa with species rank in this complex. Case-by-case taxonomic treatment of O. malampuzhaensis in relation to other species examined is presented. A taxonomic key for the discrimination of O. malampuzhaensis from other species in the O. officinalis complex has been constructed. Based on the present results, we strongly argue to restore the species rank to O. malampuzhaensis, as originally proposed by Krishnaswamy and Chandrasekharan (1958).     

中国特有属藤山柳属(猕猴桃科)植物的叶表皮形态及其分类学意义

由于形态特征变异和地理分布区域存在重叠,中国特有属藤山柳属(猕猴桃科)的物种划分问题长期以来存在争议,曾被分为20种或修订为含1种4个亚种的单型属。该研究选取了在形态和地理分布上有代表性的29个居群的184份标本,利用光学显微镜和扫描电镜观察了叶表皮形态和微形态特征,以探讨它们的分类学意义。结果表明:(1)藤山柳属植物叶表皮毛被的形态和微形态特征有3类,即光滑-短柱状毛、刚毛-长柱状毛/长刺毛、绒毛-单列多细胞毛,且这些特征在居群间差异明显,并各具明显的地理分布格局,支持把藤山柳属分为3类,即光滑类、刚毛类和绒毛类。(2)3类藤山柳植物在个别居群表现出部分同域分布现象,在峨眉山不同海拔高度的3个居群存在垂直地带性分布特点。(3)藤山柳属植物叶表皮的其他形态特征,如不规则型表皮细胞、6类气孔器、叶表皮初级蜡质纹饰以网状隆起为主,伴随着2～4类次级纹饰,在居群间变化多样等均没有明显的分类学意义。(4)由于具有相同的叶表皮形态特征和地理分布,建议把繁花藤山柳合并到绒毛藤山柳,故支持藤山柳属是1个正处于分化进程中的单型属,包括1个种3个亚种。     

中国金黄衣属地衣的修订

在形态和化学研究的基础上,对中国金黄衣属地衣进行了分类订正。将《中国地衣综览》中微孢衣属的3个种——黄微孢衣、枝瓣微孢衣和戈壁微孢衣正式归入金黄衣属,并分别命名为多孢金黄衣、枝瓣金黄衣和戈壁金黄衣(新拟),并增补散瓣金黄衣(新拟)。至此,中国金黄衣属包括了4个种。该文详细记述了每个种的表型特征和地理分布,并提供了分种检索表。     

CHROMOSOME NUMBERS IN PLATYSCHKUHRIA RYDBERG (COMPOSITAE) AND THEIR SYSTEMATIC SIGNIFICANCE

Diploid (2n = 12_II), tetraploid, pentaploid, and hexaploid level chromosome numbers are reported for Platyschkuhria Rydb. (Compositae). First reports of accessory chromosomes in the genus are also presented. Known polyploidy in Platyschkuhria is thus far restricted to P. integrifolia (Gray) Rydb. var. desertorum (M. E. Jones) Ellison from south-central Utah. Cytogenetic analysis, using methods and models of Jackson and Hauber (1982), suggests alloploid origins for the tetraploid levels. Diploid and polyploid populations correlate strongly with distinctively different edaphic situations, yet no consistent morphological features were identified that could justify taxonomic recognition of the polyploids.     

Cytomorphological characters supporting the taxonomic validity ofCyanothece (Cyanoprokaryota)

The fine structure of the type species of the genusCyanothece Komárek 1976,C. aeruginosa, is described and compared with the main cytological characteristics of morphologically related members of the generaCyanobium, Cyanobacterium andSynechococcus. Several morphological features, such as cell walls with thick outer layers containing a special type of vesicles, position of thylakoids, keritomy (net-like appearance of protoplast caused by arrangement of thylakoids, net-like nucleoids and/or by tendency to form intrathylakoidal spaces) and a special structure of mucilaginous envelopes were found to be characteristic of this genus, supporting its separate position among coccal cyanoprokaryotes (cyanobacteria, cyanophytes). The taxonomic significance of ultrastructural features in all mentioned genera is discussed.     

Hepatospora eriocheir (Wang and Chen, 2007) gen. et comb. nov. infecting invasive Chinese mitten crabs (Eriocheir sinensis) in Europe

We describe a microsporidian parasite infecting non-native Chinese mitten crabs (Eriochier sinensis) from Europe. Electron microscopy revealed merogonic and sporogonic life stages bound within a plasmalemma. The crab parasite develops polar tube precursors at the sporont stage but does not complete formation of the intact spore extrusion apparatus at the stage of the sporogonial plasmodium like Enterocytozoon bienuesi and other representatives of the Enterocytozoonidae. Its presence within an aquatic crustacean host, and a distinct molecular phylogeny based on partial small subunit ribosomal RNA (SSU rRNA) gene sequences also place it relatively close, though distinct to, existing genera within the Enterocytozoonidae. Consideration of morphological and phylogenetic characteristics of other hepatopancreas-infecting microsporidia from crustaceans suggests that certain ones (e.g. Enterospora canceri) are retained within the clade corresponding to the existing family Enterocytozoonidae, while others, including the parasite described here, may eventually be grouped in a sister taxon potentially of family rank. Based upon morphological and host similarity, it is likely that the parasite described here is the same as Endoreticulatus eriocheir (Wang and Chen, 2007), previously described from Chinese mitten crabs in Asia. However, using a combined taxonomic approach based upon morphological and phylogenetic data, we propose the formation of a new genus (Hepatospora) to replace the previous generic classification of the Asian parasite as Endoreticulatus. The microsporidian from the hepatopancreas of E. sinensis is named Hepatospora eriocheir (Wang and Chen, 2007) gen. et comb. nov. It is assumed that the parasite was introduced during initial invasions of this crab to Europe during the early 20th Century.     

Actinomycetes isolated from beehives in Thailand

Actinomycetes are one of the main microbial groups that produce bioactive compounds used as antibiotics. Although bacteria, mold and yeast have frequently been found in bees, the presence of actinomycetes in bee hives had not been previously identified or reported. The aim of our research was to focus on the diversity of actinomycetes in bee hives in Thailand. Bees, brood cells and hive materials were collected from apiaries and natural sources. Thirty-two isolates of actinomycetes were isolated and identified using morphological, physiological, chemical and molecular characterization. Most of the isolates belonged to the genus Streptomyces. Some less frequent isolates were classified in the genera Nonomuraea, Nocardiopsis and Actinomadura.     

Phylogeny in Echinocereus (Cactaceae) based on combined morphological and molecular evidence: taxonomic implications

Echinocereus is a morphologically diverse genus that includes 64 species grouped into eight taxonomic sections based on morphological traits. In previous molecular phylogenetic analyses, the relationships amongst Echinocereus species were not entirely revealed and useful characters to recognize clades were not provided. The inclusion of several sources of evidence in a phylogenetic analysis is likely to produce more supported hypotheses. Therefore, we performed a combined phylogenetic analysis with a set of 44 morphological characters and six chloroplast DNA sequences. Topologies from parsimony and Bayesian analyses were mostly congruent. However, the relationships of E. poselgeri were not consistent between analyses. A second Bayesian analysis using a long-branch extraction test resulted in a topology with the morphological position of E. poselgeri congruent with that in parsimony analysis. Parsimony and Bayesian analyses corroborated the monophyly of Echinocereus, which included eight monophyletic groups. The combined phylogeny integrated into different clades those taxa that were not determined in previous analyses and changed the relationships of some recognized clades. The clades did not recover the recent infrageneric classification. In the present study, a new sectional classification for Echinocereus is proposed based on the eight recovered clades, which is supported by a combination of morphological and molecular characters. An identification key for sections in the genus is included.     

Phylogenetics and biogeography of the broad-nosed bats, genus Platyrrhinus (Chiroptera: Phyllostomidae)

The Neotropical broad-nosed bats, genus Platyrrhinus, represent a well-defined monophyletic group of 14 recognized species. A recent study of morphological characters confirmed Platyrrhinus monophyly and species diagnosis, but offered little support to their intra-specific relationships. We conducted phylogenetic analyses of the genus, using dense taxonomic sampling in combination with four gene sequences representing both mitochondrial and nuclear DNA transmission systems. Our aim was to elucidate the phylogenetic structure among species, using the resulting 3341 bp of DNA. Maximum parsimony, maximum likelihood, and Bayesian inference analyses produced similar topologies that confirm the monophyly of the genus Platyrrhinus and strongly support many previously unrecognized groups. Paraphyly of Platyrrhinus helleri and the unclear position of P. brachycephalus in the clades were also apparent in the data. Our biogeographical analysis suggests a Brazilian Shield origin for Platyrrhinus, followed by subsequent radiations of lineages in the Amazon Basin and Andes. Secondary dispersal from Amazonian and Andean centers is responsible for the Platyrrhinus inhabiting the Guianan Shield and the Pacific lowlands and Central America, respectively.