Molecular identification and the features of genetic diversity in interspecific hybrids of Amur sturgeon ( Acipenser schrenckii × A. baerii, A. baerii × A. schrenckii, A. schrenckii × A. ruthenus , and A. ruthenus × A. schrenckii ) based on variability of multilocus RAPD markers

The method of polymerase chain reaction with random primers (RAPD PCR) was used to identify the progeny of the crosses between three sturgeon species, Amur sturgeon (Acipenser schrenckii Brandt, 1869), Siberian sturgeon (A. baerii Brandt, 1869), and sterlet (A. ruthenus Linnaeus, 1758). Using ten primers, genetic variation in 70 yearlings, produced in seven individual crosses: Acipenser schrenckii × A. schrenckii, A. baerii × A. baerii, A. ruthenus × A. ruthenus, A. schrenckii × A. baerii, A. baerii × A. schrenckii, A. schrenckii × A. ruthenus, and A. ruthenus × A. schrenckii was described and evaluated. It was demonstrated that the samples composed of hybrids from individual crosses were more variable than the samples of parental species. On the other hand, pooled samples of hybrids from two cross directions were genetically less variable than the pooled samples of their parents. The three main features of the hybrid RAPD profiles identified included: (1) preservation of marker DNA fragments of both parents in one genome; (2) presence of specific DNA fragments, absent from both parents; and (3) dependence of the frequency of some DNA fragments from the cross direction. Multidimensional scaling clearly distinguishes in the space of three coordinates the individuals of original species and the hybrid progeny with differentiation in the groups of direct and backcross hybrids. Analysis of relationships (UPGMA and NJ) pointed to substantial differentiation between the species, as well as between the species and hybrid progeny. Close genetic relationships between direct and backcross hybrids were demonstrated. Multilocus RAPD markers in association with statistical methods are considered to be the useful tool for discrimination of interspecific hybrids of sturgeon. Possible reasons for the differences in the hybrid RAPD profiles are discussed. Original Russian Text ? K.V. Rozhkovan, G.N. Chelomina, E.I. Rachek, 2008, published in Genetika, 2008, Vol. 44, No. 11, pp. 1453–1460.     

Neotropical Monogenoidea. 54. Proposal of Aetheolabes n. g. (Dactylogyrinea: Diplectanidae), with the description of A. goeldiensis n. sp. from the gills of ‘pescada’ Plagioscion sp. (Teleostei: Sciaenidae) in Brazil

Aetheolabes goeldiensis n. g., n. sp. (Diplectanidae) is described from the gills of ‘pescada’ Plagioscion sp. (Sciaenidae) collected from the Baía de Marajó, about 30 km north of Belém, Pará, Brazil. The monotypic Aetheolabes n. g. is characterised, in part, by its type-species having the haptor and haptoral sclerites modified as a clasp for attachment to the gill tissue of its host, the copulatory complex situated far posterior to the intestinal bifurcation near the mid-length of the trunk, the vaginal pore apparently within the genital atrium, the tegument lacking scales, anchors atypical for diplectanids, and by lacking peduncular spines and squamodiscs. A. goeldiensis n. sp. closely resembles Diplectanum umbrinum Tripathi, 1957 from India and China by the haptoral sclerites forming a clasp, but differs from it primarily by the orientation of the reproductive organs and absence of squamodiscs.     

Plant regeneration from suspension cells induced from hypocotyls derived from interspecific cross Alstroemeria pelegrina × A. magenta and transformation with Agrobacterium tumefaciens

Embryogenic cell suspension cultures were established using the ovule culture of an interspecific cross, Alstroemeria pelegrina var. rosea × A. magenta. Ovules harvested 14 days after pollination were cultured on Murashige and Skoog (MS) medium without plant growth regulators (PGRs); calli were produced on the hypocotyl surface in germinating zygotic embryos. Suspension cells were induced from the calli by using liquid MS media containing 2,4-dichlorophenoxyacetic acid or 4-amino-3,5,6-trichloropyridine-2-carboxylic acid (picloram). Adventitious embryos developed from the suspension cells on half-strength MS medium supplemented with 0.5 mg l⁻¹ of both α-naphthaleneacetic acid and N⁶-benzylaminopurine; they grew into plantlets on the same medium. The plantlets formed rhizomes following transfer to half-strength MS medium without PGRs, and acclimatized plants were easily established. Subsequently, Agrobacterium-mediated transformation system was applied. The suspension cells were co-cultivated with A. tumefaciens strain EHA101/pIG121Hm or LBA4404/pTOK233, both of which contain neomycin phosphotransferase II, hygromycin phosphotransferase and intron-containing ?-glucuronidase (intron-GUS) genes. Seven days after co-cultivation, the cells were subjected to GUS assay; staining was most pronounced in the cells subcultured in a picloram-containing liquid medium and co-cultivated with EHA101/pIG121Hm. The co-cultivated cells were transferred to the MS medium containing picloram and 20 mg l⁻¹ hygromycin; 1 month later, several hygromycin-resistant callus lines showing GUS activity were obtained. Transgenic plants were obtained through our plant regeneration system, and foreign gene insertion into the regenerated plants was confirmed by polymerase chain reaction.     

Comparative study (AFLP and morphology) of three species of Prosopis of the Section Algarobia: P. juliflora, P. pallida, and P. limensis. Evidence for resolution of the “P. pallida–P. juliflora complex”

The problems of delimitation of species of Prosopis originate from the few morphological discontinuities which exist among some of them; some, however, originated as a result of wide distribution of germplasm without proper knowledge of the species, in particular, much material catalogued as P. juliflora, but being of other species, was distributed for reforestation projects worldwide. This work tests the morphological results obtained for P. pallida and P. limensis of the Peruvian–Ecuadorian coast and for P. juliflora of the Caribbean Basin of Colombia and Venezuela utilizing a study of AFLPs and a study of the morphology of plantlets developed in a conventional garden study. The phenogram obtained for the AFLPs demonstrates each of the three species to be a well differentiated cluster and the molecular variance between them is significantly greater than the variance within each species. Study of the plantlets also indicates statistically significant differences for four morphological characters between P. juliflora and the other two species (P. pallida and P. limensis). These results, in addition to the morphological differentiation evident between adult plants of P. pallida and P. limensis and the clear separation of these two species from P. juliflora, corroborate the genetic identity of the three taxa analyzed.     

Galactosylation of antibiotics using the β-galactosidase from Aspergillus oryzae

The β-galactosidase from Aspergillus oryzae has been shown to catalyze the synthesis of β-galactosides of antibiotics such as chlorphenisin and chloramphenicol using β-lactose as the galactosyl donor. Among the water-miscible organic solvents tested, 20% (v/v) acetonitrile in the reaction mixture gave the highest yield in galactoside synthesis. The products obtained were purified by preparative TLC and liquid chromatography and analyzed by ¹H-and ¹³C-NMR, and MS (FAB). Chlorphenisin and chloramphenicol were galactosylated exclusively at their primary hydroxy groups. The pH optimum for the transgalactosylation reaction was between pH 4–5. Increasing concentrations of galactosyl donor and aglycon caused increasing yields of galactosides. When the resulting galactosylated antibiotic was withdrawn from the sample, further synthesis was observed. This could be accelerated either by withdrawing the resulting monosaccharides (glucose and galactose) or exchanging them for mannose.     

Molecular evidence of a reed warbler × great reed warbler hybrid ( Acrocephalus scirpaceus × A. arundinaceus) in Belgium

We report on the second case of a reed warbler × great reed warbler hybrid (Acrocephalus scirpaceus and A. arundinaceus). The bird was captured during a standardised ringing session in Belgium in autumn 1999, and fell between the parental species in all measurements. Molecular analyses of two microsatellite loci verified that the bird was a female that had a reed warbler father and a great reed warbler mother.     

Androgenic response of genotypes selected from Capsicum annuum L.?×?C. chinense Jacq. hybrids

In research on androhaploids in the progeny of interspecific hybrids within the Capsicum genus, three genetically stable lines of F₇ generation, selected from the C. annuum L. × C. chinense Jacq. hybrid, were used. In the first line, only callus tissue was formed as a reaction to the conditions of culture. Cytometric analysis of this tissue revealed the presence of cells with DNA content in nuclei at the level of 1C to 16C. The tissue was mixoploid and non-embryogenic. Anthers of the other line did not respond. In the third one, nine embryos were obtained, and they developed into plants. By means of cytometric analysis, the 1C DNA level was found in eight of them and these were androgenic plants. The origin of one of the diploid plant was not established due to the homozygous character of the donor plants. The experiment results confirm the already known diversity of genotype reaction to the conditions of culture. It moreover point to the possibility of selection of the forms with an androgenic potency from interspecific hybrids.     

Genetic evidence for the recognition of two fiddler crabs, Uca iranica and U.?albimana (Crustacea: Brachyura: Ocypodidae), from the northwestern Indian Ocean, with notes on the U.?lactea species-complex

The status of two poorly known fiddler crabs, Uca iranica Pretzmann, 1971, from the Persian Gulf, and U. albimana (Kossmann, 1877), from the Red Sea, was studied using two mitochondrial genes: the large subunit (16S) ribosomal (r)RNA and cytochrome oxidase subunit I (COI). A molecular phylogeny shows both U. iranica and U. albimana to be members of a monophyletic U. lactea species-complex containing six taxa, with three highly supported internal clades. Uca iranica and U. albimana are the closest genetically, but are different enough to be considered valid species (16S rRNA nucleotide divergence > 7.0%, and COI > 11.9%), and form a highly supported “western” clade with U. annulipes (in line with the original morphological concept). A West Pacific “eastern” clade includes U. lactea in the north and the more widely ranging U. perplexa. An Australian endemic species, U. mjoebergi, forms a third monotypic clade.     

Effects of β-lactam antibiotics, auxins, and cytokinins on shoot regeneration from callus cultures of two hybrid aspens, Populustremuloides?×?P. tremula and P. xcanescens?×?P. gradidentata

The effects of β-lactam antibiotics (penicillin, carbenicillin and cefotaxime), cytokinins, and auxins including phenylacetic acid, a β-lactam breakdown product, were evaluated during in vitro shoot morphogenesis in two hybrid aspens; P. tremuloides × P. tremula (XTTa) and P. x canescens × P. grandidentata (XCaG). Although different callus and shoot induction media were used for both hybrids, the β-lactams often engendered similar responses. At concentrations of 1,000 mg l⁻¹, carbenicillin adversely impacted shoot elongation and, to a lesser degree, shoot regeneration. Cefotaxime enhanced caulogenesis for all of the concentrations evaluated (125–500 mg l⁻¹) especially when the cytokinin thidiazuron was used for shoot induction. The shoots formed faster and in greater numbers; and the improvements were significant (α = 0.05) for both hybrids. However, hyperhydricity was more problematic when cefotaxime was included in the media. The incidence of shoot hyperhydricity for the XCaG hybrid was more than twice as great for the highest cefotaxime concentration evaluated (500 mg l⁻¹) than for the control (>90% vs. ~40%). Penicillin had an opposite effect. Hyperhydricity frequencies for the XCaG hybrid were lower when the media were supplemented with penicllin and the reductions were statistically significant at concentrations of 500–1,000 mg l⁻¹. The effects of the antibiotics were generally not reproduced by the auxins (0.1–100 μM), including phenylacetic acid, or the other potential β-lactam degradation products evaluated (e.g. phenylmalonic acid, aminopenicillanic acid). The antibiotics may have affected shoot hyperhydicity indirectly via changes in the time course of shoot regeneration.     

Somatic embryogenesis and plant regeneration in diploid Allium fistulosum × A. cepa F1 hybrid onions

Procedures were developed for disinfestation of non-dormant basal plate tissue excised from field grown basal plate tissue of diploid Allium fistulosum × A. cepa F₁ hybrid onions. Contamination levels varied with the season and vegetative development of plant material. Callus initiated from basal plate tissue and immature inflorescences of the F₁ hybrids was maintained on a BDS-based medium containing 0.75 mg/l picloram and 2.0 mg/l BA. When this medium was supplemented with vitamins and glycine, and with proline at 2.5 gm/1, somatic embryos began to form. Their development continued on a BDS-based shoot promotion medium containing 0.03 mg/l picloram and 0.32 mg/l 2iP supplemented with vitamins, glycine and proline. Genotypes differed significantly in the numbers of structures regenerated. Plantlets from somatic embryos were rooted into BDS or half-strength BDS medium without growth substances and were successfully transferred to sterilized potting mix in plastic commercial corsage boxes.Abbreviations BA benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - picloram 4-amino-3,5,6-trichloropicolinic acid - 2iP N6-(2-isopentenyl)adenine - NAA 1-naphthylacetic acid - BDS Gamborg's B5 medium modified by Dunstan and Short (1977a)     

A comparative description and distribution of the flying fishes—Cypselurus poecilopterus, C. simus, and C. callopterus, sorted out into the species group of spotwing species of the subgenus Poecilocypselurus

The three species of flying fishes—Indo-Pacific Cypselurus poecilopterus, Central Pacific C. simus, and Eastern Pacific C. callopterus form a particular species group within the subgenus Poecilocypselurus. These species rather weakly differ in the body height, number of predorsal scales and number of gill rakers. The sculls of these three species are very much alike; there are just very small differences in the proportions of some bones and the number of skull lateral line canals and pores. It could be imagined that C. poecilopterus has appeared in the Indo-Malayan Archipelago area and thence extended in all directions (up to the coasts of the Eastern Africa, Northern and Eastern Australia, New Guinea and adjacent islands, the western islands of Polynesia and the waters of Japan). C. callopterus inhabiting warm waters of the Eastern Pacific and C. simus whose area is situated in the central part of the Pacific Ocean are seemingly derived from this species.     

Three new species, Lamellodiscus tubulicornis n. sp., L. magnicornis n. sp. and L. parvicornis n. sp. (Monogenea: Diplectanidae) from Gymnocranius spp. (Lethrinidae: Monotaxinae) off New Caledonia, with the proposal of the new morphological group ‘tubulicornis’ within Lamellodiscus Johnston & Tiegs, 1922

Three new species of Lamellodiscus are described from four (including two undescribed) species of Gymnocranius off New Caledonia, South Pacific. All three species have a similar body anatomy and morphology of the haptoral hard parts and are distinguished on the basis of the male copulatory organ (MCO). Lamellodiscus tubulicornis n. sp. (type-host: G. grandoculis; other host: Gymnocranius sp. B) has an MCO with a tube and horn; L. magnicornis n. sp. (type-host: G. grandoculis; other hosts: G. euanus, Gymnocranius sp. A and Gymnocranius sp. B) has an MCO with a long horn and a membrane; L. parvicornis n. sp. (type-host: G. euanus; other hosts: G. grandoculis and Gymnocranius sp. B) has an MCO with a small horn and a membrane. Lamellodiscus epsilon Yamaguti, 1968 is redescribed based on the type-specimens (from Monotaxis grandoculis off Hawaii). Lamellodiscus sp. is recorded from Gnathodentex aureolineatus off New Caledonia. All these five species have lamellodiscs that exhibit a unique characteristic: the second lamella forms an almost closed circle, in contrast to all other described species of Lamellodiscus in which the second lamella has the form of one or two crescents. Lamellodiscus spp. are usually classified in two groups, the ‘ignoratus’ and ‘elegans’ groups, according to the structure of the lamellodisc; we propose a new ‘tubulicornis’ group for these five species. In addition to their lamellodisc structure, species of the tubulicornis group are also characterised by their egg (elongate in contrast to tetrahedral in the two other groups) and their hosts (monotaxine lethrinids as opposed to mainly sparids). The generic diagnosis of Lamellodiscus given by Amine & Euzet (2005) is amended to include species with elongate eggs.     

Acanthocephalus?amini n. sp. (Acanthocephala: Echinorhynchidae) from the freshwater fish Cichlasoma?urophthalmus (Günther) (Cichlidae) in Mexico

Acanthocephalus amini n. sp. (Palaeacanthocephala: Echinorhynchidae) is described from the intestine of Cichlasoma urophthalmus (Günther) (Pisces: Cichlidae) collected in the Río Champotón, a river in Campeche State, Mexico. It is the fourth species of Acanthocephalus Koelreuther, 1771 described from North American freshwater fishes, although two other species are known from South America. The new species is distinguished from other members of Acanthocephalus by features of its trunk, which is small, clavate, slightly expanded medially and bluntly pointed posteriorly. It is further distinguished by having a cylindrical proboscis armed with 13–14 longitudinal rows of 11–12 stout hooks; the apical and medial proboscis hooks are almost uniform in size and shape, decreasing in size towards the base; the posteriormost hooks are smaller, straighter and more slender than the anterior and middle hooks; and the lateral rows of hooks are more widely spaced, forming a conspicuous longitudinal area devoid of hooks. Furthermore, the lemnisci are saccate and shorter than the proboscis receptacle; and the neck is very short with a thick collar of trunk tegument, which encircles the base of proboscis. In males, the testes are in the middle third of trunk, diagonal, spherical and small relative to the body size, and there are six clavate cement glands. In females, the uterus forms a conspicuous, elongate, cylindrical egg reservoir. The new species is most similar to A. alabamensis Amin & Williams, 1983, but can be distinguished by its swollen, clavate trunk, the largest proboscis hooks being present apically and medially, smaller testes, a shorter male reproductive system relative to body size and females with a prominent uterus. They have different hosts and geographical distribution. The new species can be differentiated from Brasacanthus sphoeroides Thatcher, 2001, a similar species in a monotypic echinorhynchid genus, because the latter is larger, has smaller proboscis hooks and its lemnisci are longer than the receptacle.     

The Role of dinB in the Appearing of Antibiotics Resistance in E.coli

The role of dinB gene in the appearing of antibiotics resistance was studied. Plasmid containing multi-copy dinB gene was transfected into E. coli to create an overexpression. The strains carrying multi-copies of dinB gene demonstrate a significant survival advantage over the wild strain. In vitro experiment, the dinB-overexpressed strain evolved resistance within 8 hours, while wild strain could not.In vivo experiment with mice model infected with dinB-overexpressed strain, resistant clones emerged significantly earlier and demonstrated significant higher level of resistance than those infected with the wild control strain. The results showed that dinB gene made a contribution in the appearing of the antibiotics resistance and has a potential as a target for prevention from the appearing of antibiotic resistance.     

A ThCAP gene from Tamarix hispida confers cold tolerance in transgenic Populus (P. davidiana?×?P. bolleana)

The ThCAP gene, which encodes a cold acclimation protein, was isolated from a Tamarix hispida NaCl-stress root cDNA library; its expression patterns were then assayed by qRT-PCR in different T. hispida tissues treated with low temperature (4°C), salt (400 mM NaCl), drought (20% PEG6000) and exogenous abscisic acid (100 μM). Induction of ThCAP gene was not only responsive to different stress conditions but was also organ specific. When transgenic Populus (P. davidiana × P. bolleana) plants were generated, expressing ThCAP under regulation of the cauliflower mosaic virus CaMV 35S promoter, they had a greater resistance to low temperature than non-transgenic seedlings, suggesting that ThCAP might play an important role in cold tolerance.     

Speciation and chromosomal evolution of the Palearctic species of chironomids of the genus Sergentia Kieffer (Diptera, Chironomidae): Divergence of the Karyotypes of S. baueri Wülker et al., 1999, S. prima Proviz et al., 1997, S. electa Proviz et al., 1999 and their relationship with the endemic species from Baikal lake

The banding structures of chromosomes of three Palearctic species of chironomids of the genus Sergentia Kieffer, 1922 from the Irkutsk reservoir were comparatively analyzed: S. baueri Wülker et al., 1999 (2n = 8), S. prima Proviz et al., 1997 (2n = 8), and S. electa Proviz et al., 1999 (2n = 6). The S. baueri karyotype was taken as a standard in chromosome mapping. It was established that in contrast to S. baueri and S. prima the S. electa karyotype was formed due to a complex translocation: transfer of chromosome IV in the median part of chromosome III. The sequences of chromosome I bands were found to be completely homologous in all Palearctic species and in S. rara Proviz et al., 1999 (2n = 8) taken as the original species in the group of Baikalian endemics. In both groups, seven homozygous inversions were detected, of which the common one is the inversion in chromosome III. It was assumed on the basis of the results of the karyological analysis that among the Palearctic species S. baueri is most similar to the Baikalian endemics and S. electa is phylogenetically closer to S. prima. Specific features of the chromosome and morphological evolutions of the Sergentia genus species are considered.     

Paraorygmatobothrium?taylori n. sp. (Tetraphyllidea: Phyllobothriidae) from the Australian weasel shark Hemigaleus?australiensis White, Last & Compagno (Carcharhiniformes: Hemigaleidae)

Paraorygmatobothrium taylori n. sp. (Tetraphyllidea: Phyllobothriidae) is described from the Australian weasel shark Hemigaleus australiensis White, Last & Compagno in Moreton Bay, off Queensland, Australia. The new species differs from 10 of the 11 described species of Paraorygmatobothrium Ruhnke, 1994 by the possession of prominent, semicircular bothridial muscle bands. From P. barberi Ruhnke, 1994, with which it shares the bothridial muscle bands, it differs in the possession of a cephalic peduncle and vitelline follicles that extend almost to the mid-line of the proglottis and are reduced, rather than completely interrupted, at the level of the ovary. P. janineae Ruhnke, Healy & Shapero, 2006 is recorded from its type–host but in a new locality, Moreton Bay, off Queensland, Australia. P. taylori is the third species of the genus recorded from the Hemigaleidae in Australian waters. Three of the eight known hemigaleid species are now recorded to harbour this genus, and three different species are now known from the two hemigaleids found in Australian waters.