Integrative Transformation System for the Metabolic Engineering of the Sphingoid Base-Producing Yeast Pichia ciferrii

We have developed an integrative transformation system for metabolic engineering of the tetraacetyl phytosphingosine (TAPS)-secreting yeast Pichia ciferrii. The system uses (i) a mutagenized ribosomal protein L41 gene of P. ciferrii as a dominant selection marker that confer resistance to the antibiotic cycloheximide and (ii) a ribosomal DNA (rDNA) fragment of P. ciferrii as a target for multicopy gene integration into the chromosome. A locus within the nontranscribed region located between 5S and 26S rDNAs was selected as the integration site. A maximum frequency of integrative transformation of approximately 1,350 transformants/μg of DNA was observed. To improve the de novo synthesis of sphingolipid, the LCB2 gene, encoding a subunit of serine palmitoyltransferase, which catalyzes the first committed step of sphingolipid synthesis, was cloned from P. ciferrii and overexpressed under the control of the P. ciferrii glyceraldehyde-3-phosphate dehydrogenase promoter. After transformation of an LCB2 gene expression cassette, several transformants that contained approximately five to seven copies of transforming DNA in the chromosome and exhibited about 50-fold increase in LCB2 mRNA relative to the wild type were identified. These transformants were observed to produce approximately two times more TAPS than the wild type.     

Prevalence of Yeasts in English Full Blood Mares

The aim of the study was a quantitative and qualitative analysis of microflora, presentation of current data about prevalence of the microflora on the skin and mucous membranes, and determination of its possible effect on reproduction of English full blood horses bred in Poland. The material for analyses was sampled from the skin and mucous membranes (385 samples) of 55 English full blood mares. Taking into account reproduction traits, the mares were classified into three groups. Six yeast-like species, including five species from the genus Candida (C. albicans, C. guiliermondii, C. lusitaniae, C. sp., and C. tropicalis) and Trichomonascus ciferrii, were detected on the skin and mucous membranes in the English full blood mares. Growth of yeasts was observed in more than half of the samples taken from mares that had foaled and approximately 46 % of non-conceiving and barren mares. The high prevalence of various yeast strains in the mouth, nostrils, and collateral groove may suggest widespread occurrence of the microflora in the breeding environment. The results obtained indicate that the yeasts isolated in this study may be components of the normal microflora of the skin and mucous membranes in horses. The analysis results do not indicate unambiguously that the isolated microflora affects reproduction in mares, although this cannot be excluded.     

The effect of growth conditions on production and excretion of extracellular antigens by three ascomycetous yeasts

Ascomycetous yeasts produce extracellular antigens that are almost specific for the species. The antigen production by Hansenula wickerhamii and Stephanoascus ciferrii was independent of the carbon source and was proportional to the final cell density of the cultures. The same was true of chemostat cultures of Stephanoascus ciferrii, irrespective of the dilution rate and whether glucose or ammonia was the limiting nutrient. In cultures of Saccharomyces cerevisiae, however, antigen excretion mainly took place in the late exponential growth phase. Large amounts of antigen were extracted from the cell wall of Saccharomyces cerevisiae. A small amount was detected in the cytoplasm.     

Cronisia and its two species,C. fimbriata and C. weddellii

The systematic position of Riccia fimbriata Nees, a species formerly known only in sterile condition and tentatively placed in the genus Exormotheca, is reexamined in the light of the recently discovered reproductive structures. The species proved to be a member of the genus Cronisia, identical to Cronisia mexicana Hicks, described in 1993. A key is provided to the two known species of Cronisia, C. fimbriata and C. weddellii, and they are described and illustrated.     

Scale insects (Homoptera, Coccinea) of the volga area: III. Nudicauda gen. n. and other mealybugs from astrakhan

The present paper is the third part of a revision of the fauna of the Volga Area. Five species new for the Volga Area, including four new for Russia are recorded. Nudicauda gen. n. is erected with type species Nudicauda nigra (Matesova, 1957) (comb. n.). The new genus differs from other genera of mealybugs in a complex of characters and mainly in the presence of very specific oral rim tubular ducts with a broad flat ring and in an unusual ovisac with the opening over posterior tergites of the female body. In addition to the type species, N. orientalis (Borchsenius, 1949) and N. salina (Matesova, 1981) (comb. n.) are also included in the new genus. N. nigra and N. orientalis are considered as separate species, contrary to Danzig (1980). All the three species of the new genus are redescribed and illustrated. The structure of the tubular ducts of the genus Humococcus Ferris, 1953 is briefly discussed and illustrated.     

Zygotorulaspora cornina sp. nov. and Zygotorulaspora smilacis sp. nov., Two Novel Ascomycetous Yeast Species Isolated from Plant Flowers and Fruits

Three isolates belonging to the ascomycetous genus Zygotorulaspora were obtained from the fruits of Cornus officinalis and Smilax china, and flowers of Dendranthema zawadskii var. latilobum in Gongju-si, Korea. Phylogenetic Analyses of the LSU D1/D2 domain and ITS region sequences supported the recognition of two new species: Zygotorulaspora cornina sp. nov. (type strain NIBRFGC000500475 = KACC93346P^PP) and Zygotorulaspora smilacis sp. nov. (type strain NIBRFGC000500476 = KACC93347P^PP). The two novel species revealed no growth on D-Galactose, unlike the other six species in the genus Zygotorulaspora. They are distinguished from each other by their phylogenetic differences and phenotypic characteristics such as assimilation of xylitol, 5-keto-D-gluconate, and ethanol. All species in the genus Zygotorulaspora including the two novel species have phenotypic traits of genus Zygotorulaspora: asci are persistent, sucrose and raffinose are assimilated, and m-inositol is not required for growth, and they are mainly associated with plants.     

Pancreatonema torriensis gen. nov., sp. nov. (Nematoda: Rhabdochonidae) from the pancreatic duct of Raja naevus

A new genus and species of nematode, Pancreatonema torriensis, from the pancreatic duct of Raja naevus caught off Aberdeen and north-west Scotland is described and aspects of its biology are discussed. According to the classification of Skrjabin, Sobolev & Ivashkin (1967) [Spirurata of animals and man and the diseases caused by them (4). Essentials of Nematodology, Vol. 16 (I.P.S.T. Translation No. 5904, Jerusalem, 1971)]it belongs to the family Rhabdochonidae, sub-family Prosungulonematinae, and is closely related to the genus Johnstonmawsonia. The taxonomy of the Prosungulonematinae is confused but it is suggested that Johnstonmawsonia should at present be restricted to those species of the genus described from marine teleosts, and that the species from freshwater teleosts should be included in the genus Prosungulonema.     

Pachyseris inattesa sp. n. (Cnidaria,Anthozoa, Scleractinia): a new reef coral species from the Red Sea and its phylogenetic relationships

A new scleractinian coral species, Pachyseris inattesa sp. n., is described from the Red Sea. Despite a superficial resemblance with some species in the agariciid genus Leptoseris with which it has been previously confused, P. inattesa sp. n. has micro-morphological characters typical of the genus Pachyseris. This genus, once part of the Agariciidae, is comprised of five extant species and is widely distributed throughout the tropical Indo-Pacific. It is currently incertae sedis as a result of recent molecular analysis and appears to be closely related to the Euphylliidae. A molecular phylogenetic reconstruction including P. inattesa sp. n., the genus type species P. rugosa, and P. speciosa, all present in the Red Sea, was performed using the mitochondrial intergenic spacer between COI and 16S-rRNA. The results confirm that P. inattesa sp. n. is a monophyletic lineage closely related to the other Pachyseris species examined.     

Sumatrella chelonica gen. n., sp. n., a new remarkable genus and species from Indonesia,Sumatra (Acari,Uropodina, Oplitidae)

A new genus Sumatrella gen. n. is described and illustrated based on the new species Sumatrella chelonica sp. n. collected in Sumatra, Indonesia. The new genus belongs to the family Oplitidae based on its hypertrichous internal malae and the absence of strongly sclerotized structures on the dorsal shield. The new genus is closely related to the genus Chelonuropoda Sellnick, 1954 but the transverse furrow on ventral idiosoma close to coxae IV and the strongly sclerotized C-shaped dorsal line are missing in the new genus. These characters can be found in species of Chelonuropoda.     

Description of Oculogryphus shuensis sp. n. (Coleoptera,Lampyridae), the first species of the genus in the Sino-Japanese realm,with a modified key to the subfamily Ototretinae

A new species of the lampyrid genus Oculogryphus Jeng, Engel, and Yang, O. shuensis sp. n. from China (Sichuan Province) is described and figured. The genus previously was known only from Vietnam, and the new species is the first representative of the genus in the Sino-Japanese zoogeographic realm. Some morphological variations of Oculogryphus and the allied genus Stenocladius are discussed and a modification to the most recent key to ototretine genera is proposed to accommodate Oculogryphus.     

Afroprinus cavicola gen. et sp. n. from the Afrotropical region with notes on cave-dwelling Saprininae (Coleoptera,Histeridae)

A new genus and species from Kenya, Afroprinus cavicola is herein described and illustrated and its systematic position is discussed. By the prosternal pre-apical foveae connected by marginal prosternal stria it resembles most of the Afrotropical species of the genus Chalcionellus Reichardt, 1932, or some species of the genus Pholioxenus Reichardt, 1932 from South Africa and Namibia. Afroprinus can be distinguished from Chalcionellus chiefly by the lack of pronotal depressions and a coarsely sculptured, non-metallic dorsum; from Afrotropical species of Pholioxenus it can be most easily distinguished by the asetose pronotal hypomeron. The new taxon was discovered in a cave, but lacks obvious troglophilic adaptations. Notes on other Saprininae taxa found in caves are given. An identification key to the genera of Afrotropical Saprininae is provided.     

A revision of digger wasps of the genus Oxybelomorpha (hymenoptera, crabronidae, oxybelini): 6. The O. persa and O. steckii species-groups

The Palaearctic species-groups of the digger wasp genus Oxybelomorpha Brauns 1897, O. persa and O. steckii, are revised. Three species were investigated. Oxybelomorpha steckii (Kohl 1923) and O. maroccana (de Beaumont, 1957) are considered separate species. The male of O. maroccana is described for the first time. The lectotype of Belomicrus (Pseudoxybelus) persa Gussakovskij, 1933 is designated. A key to 24 revised species of the genus Oxybelomorpha is given.     

Taxonomic revision of European Apium L. s.l.: Helosciadium W.D.J.Koch restored

European representatives of Apium sensu lato (Apiaceae), and Apium prostratum and Naufraga balearica, were studied with morphological, fruit anatomical, and palynological methods. Morphometric data were compared with phylogenetic results from previous molecular studies. This confirms that most of the European Apium species belong to a separate group corresponding to the previously named genus Helosciadium. All these species had previously been formally named as Helosciadium species, except for the new combination Helosciadium bermejoi, which is formally described here. Molecular studies place Apium prostratum and Naufraga balearica close to Apium graveolens, the type species of Apium. Our morphometric results show similarities of Naufraga with H. bermejoi, but fruit anatomy distinguishes it both from Helosciadium and from A. graveolens/prostratum. The placement of Cyclospermum leptophyllum in a separate genus is confirmed. Diagnostic keys to the genera and Helosciadium species, and an annotated checklist are given.     

Gall midges of the supertribe stomatosematidi (Diptera, Cecidomyiidae) in the Palaearctic region, with description of new taxa from the Late Eocene Rovno amber: 3. Groveromyia gen. n. analysis of biometric parameters of the recent and late eocene genera and species

The new genus Groveromyia gen. n. was described from the Late Eocene Rovno amber. The genus includes 4 species: G. occlusa sp. n., G. digna sp. n., G. astrosa sp. n., and G. concinna (Fedotova, 2005) comb. n. (Bryocrypta). Some changes in the biometric parameters of the genera and species were revealed relative to those in the recent and the Late Eocene Stomatosematidi. Keys to the recent and fossil genera of the tribe Stomatosematidi and the species of the genus Groveromyia are given.     

History and current taxonomic status of genus Agrobacterium

The genus Agrobacterium was created a century ago by Conn who included it in the family Rhizobiaceae together with the genus Rhizobium. Initially, the genus Agrobacterium contained the non-pathogenic species Agrobacterium radiobacter and the plant pathogenic species Agrobacterium tumefaciens and Agrobacterium rhizogenes. At the end of the past century two new pathogenic species, Agrobacterium rubi and Agrobacterium vitis, were added to the genus. Already in the present century these species plus Agrobacterium larrymoorei were reclassified into genus Rhizobium. This reclassification was controversial and for a time both genus names were used when new species were described. Few years ago, after a taxonomic revision based on genomic data, the old species A. rhizogenes was maintained in the genus Rhizobium, the old species A. vitis was transferred to the genus Allorhizobium and several Rhizobium species were transferred to the genus Agrobacterium, which currently contains 14 species including the old species A. radiobacter, A. tumefaciens, A. rubi and A. larrymoorei. Most of these species are able to produce tumours in different plants, nevertheless the genus Agrobacterium also encompasses non-pathogenic species, one species able to nodulate legumes and one human pathogenic species. Taking into account that the species affiliations to five Agrobacterium genomospecies have not been determined yet, an increase in the number of species within this genus is expected in the near future.     

Revision of genus Melamphaes (Melamphaidae): Part 4. Multi-raker species: M. ebelingi, M. occlusus sp. nova, and M. nikolayi sp. nova

In the fourth part of the revision of the genus Melamphaes (Melamphaidae) multi-raker species (20 and more rakers at the first gill arch, rarely 19??in M. ebelingi) with eight soft rays in the ventral fin are considered. In this species, temporal spine is absent, there are 15 rays in the pectoral fin, and there are 11 to 12 abdominal vertebrae. M. ebelingi inhabits tropical and subtropical waters of the Atlantic Ocean. A new catch of this rare species in the northeastern part of the ocean is reported. The new species M. occlusus is described from one specimen from the subtropical part of the southwestern part of the Indian Ocean. Still another new species, M. nikolayi, is described from one specimen from the northwestern part of the Pacific Ocean (subtropical waters). A table for identification of 15 multi-raker species of the genus Melamphaes is provided.     

Bathycamptus eckmani gen. et spec. nov. (Copepoda,Harpacticoida) with a review of the taxonomic status of certain other deepwater harpacticoids

Bathycamptus eckmani gen. et spec. nov., which is associated with mudballs produced by the. cirratulid Tharyx luticastellus, is described from bathyal muds in San Diego Trough, off California. ?Heteropsyllus minutus Wells from the Haden Ground, Scotland is considered to be its closest relative and is placed in the same genus. The genera Bathycamptus and Psammocamptus Mielke are regarded as sister groups on the basis of the shared sexual dimorphism shown by P3-P4. Relationships with other marine Canthocamptidae are discussed, and a re-evaluation of the genus Hemimesochra Sars is made. It is concluded that this genus should encompass only the type species H. clavularis Sars. ?Leimia dubia Wells and H. nympha Por are transferred to the new genus Boreolimella, which is closely related to Bathycamptus but not to Leimia Willey. The genus Perucamptus gen. nov. is established to include H. rapiens Becker and shows no clear relationship with the other genera. H. trisetosa Coull is assigned to Caroliiaicola gen. nov., which is regarded as being an advanced member of the Paranannopidae. H. secunda Wells is recognised as belonging to Mesopsyllus Por; whilst H. nixe Por is considered the type species of a new genus Pusillargillus.     

Gastropod mollusks from the Hauterivian of Ulyanovsk (Volga Region): 2. Genera Khetella Beisel, 1977 and Cretadmete gen. nov.

The diagnosis of the genus Khetella Beisel, 1977 is emended, and a new genus, Cretadmete, is described. Two new species belonging to these genera, K. trautscholdi and C. neglecta, are established from the Upper Hauterivian of Ulyanovsk (Volga Region).     

Gall midges of the supertribe stomatosematidi (Diptera,Cecidomyiidae) in the Palaearctic region,with descriptions of new taxa from the late Eocene Rovno Amber. 2. The genera <Emphasis Type="Italic">Didactylomyia</Emphasis> and <Emphasis Type="Italic">Rovnodidactylomyia</Emphasis> gen. n.

One new genus (Rovnodidactylomyia Fedotova et Perkovsky, gen. n.) and four new species (Didactylomyia dlusskyi sp. n., Rovnodidactylomyia zosimovichi gen. n. et sp. n., R. sidorenkoi sp. n., and R. iconica sp. n.) are described from an amber sample of the late Eocene Period (Rovno District, Ukraine). Two species from a Baltic amber sample of the late Eocene Period, described earlier in the genus Bryocrypta, are attributed to the genus Rovnodidactylomyia (R. girafa (Meunier, 1904) comb. n. and R. capitosa (Meunier, 1904)). The first fossil species of the genus Didactylomyia is described for the first time; two contemporary Palaearctic species are recorded. Keys to the species of the genus Didactylomyia are given.