Prevalence of bacteria in the spermathecae of female snow crab, Chionoecetes opilio (Brachyura: Majidae)

The spermathecal contents of primiparous, multiparous and barren female snow crabs were observed by a scanning electron microscope. Bacteria colonies were observed with a significantly higher frequency in the spermathecae of primiparous and old barren females. Bacteria infect the spermathecae and destroy the spermatophores and spermatozoa inside. These observations suggest that bacteria in the spermathecae do not exclude opportunistic microbes by modifying pH of the medium as suggested in the literature. The prevalence of bacteria in primiparous and old barren females suggests that they infect individuals with a weak anti-microbial protection. The absence of bacteria in the highly acidic seminal fluid derived from males upon copulation suggests that it may provide anti-microbial protection. Bacteria do not seem to be able to survive in an anaerobic environment for a long period.     

Evidence for a permanent establishment of the snow crab (Chionoecetes opilio) in the Barents Sea

In the Atlantic the snow crab (Chionoecetes opilio) is naturally distributed on the northwestern side, i.e. eastern Canada and west Greenland. Until recently, there have been no observations of snow crab in eastern Atlantic. However, in 1990s single and occasional reports were made of crabs captured in the eastern part of the Barents Sea, presumably introduced through ballast water. Special attention during the annual bottom-trawl surveys in the Barents Sea during February 2004–2006 were given to include recordings of snow crab to evaluate if the introduced species has succeeded to establish a self-sustaining population in this region. Recordings of snow crabs were systematically noted and biological measurements carried out. The results confirm previous Russian observations of snow crabs in the northern region of Gåsebanken. In addition, a significant number of crabs were also found in the central region of the Barents Sea, mainly in deeper waters from 180 to 350 m depth. The sizes ranged from 14 to 136 mm carapace width. All females above 70 mm were berried with fertilised eggs. A major fraction (31% in 2005; 76% in 2006) of the crabs consisted of juveniles below 50 mm CW, providing evidence for successful recruitment. The small-sized crabs were exclusively found in Gåsebanken, identifying the main recruiting area at present for snow crab in the Barents Sea. The results obtained show that the snow crab is now adapted to the northeast Atlantic.     

Isolation and characterization of polymorphic microsatellite markers in the red snow crab (Chionoecetes japonicus)

A total of 12 polymorphic microsatellite DNA loci were isolated from the red snow crab, Chionoecetes japonicus (Brachyura: Majidae), one of important fisheries resources in the Far East. The number of alleles observed at each locus ranged from two to 19, with the observed and expected heterozygosities of 0.125-0.875 and 0.156-0.949, respectively, suggesting these loci to be a useful molecular marker for population analysis in this species. Of the 12 loci, seven also were available for genotyping of the snow crab, Chionoecetes opilio, implying these loci as a useful molecular marker in the genus Chionoecetes.     

Autotomy and regeneration of limbs in the male blue king crabParalithodes platypus from the Bering Sea and the Sea of Okhotsk

Limb loss and the frequency of regeneration was investigated in the male blue king crabParalithodes platypus collected in the Sea of Okhotsk and the Bering Sea. TheP. platypus population in the Bering Sea is characterized by an anomalously high proportion of crabs with missing limbs, which is largely due to the intensive fishing. The present approach to the exploitation of the Bering Sea population, in which large intact crabs are removed and injured crabs are left to recover to the normal state, may result in a decline in the reproductive potential of this population as early as this coming year.     

Observations on the histopathology and probable progression of the disease caused by Trichomaris invadens,an invasive ascomycete,in the Tanner crab,Chionoecetes bairdi

Fifty-three Tanner crabs with varying degrees of surface encrustations and internal invasion by Trichomaris invadens were necropsied, processed, and examined to determine the extent of internal tissue invasion and histopathological effects of the infection. Sufficient data were accumulated to provide some insight on the progression, but not the rate, of tissue invasion. The epidermis and subepidermal layers are invaded and virtually replaced by proliferating hyphae. The invasion then proceeds internally, predominantly via the connective tissue. Major muscles and blood vessels are invaded, then invasion of deeper organs, including the hemopoietic tissue, gastrointestinal tract wall, and wall of the heart follows. The eyestalk is infected, usually early in the disease, and the invasion follows a similar pattern to that in the cephalothorax. Massive invasion and destruction of the retina is the major pathological effect. The gill may also be involved, with destruction of the lamellar tissues and invasion and destruction of podocytes in the stem of the gill.     

Expression ofHoxDGenes in Developing and Regenerating Axolotl Limbs

Hoxgenes play a critical role in the development of the vertebrate axis and limbs, and previous studies have implicated them in the specification of positional identity, the control of growth, and the timing of differentiation. Axolotl limbs offer an opportunity to distinguish these alternatives because the sequence of skeletal differentiation is reversed along the anterior–posterior axis relative to that of other tetrapods. We report that during early limb development, expression patterns ofHoxDgenes in axolotls resemble those in amniotes and anuran amphibians. At later stages, the anterior boundary ofHoxd-11expression is conserved with respect to morphological landmarks, but there is no anterior–distal expansion of the posterior domain ofHoxd-11expression similar to that observed in mice and chicks. Since axolotls do not form an expanded paddle-like handplate prior to digit differentiation, we suggest that anterior expansion of expression in higher vertebrates is linked to the formation of the handplate, but is clearly not necessary for digit differentiation. We also show that the 5′HoxDgenes are reexpressed during limb regeneration. The change in the expression pattern ofHoxd-11during the course of regeneration is consistent with the hypothesis that the distal tip of the regenerate is specified first, followed by intercalation of intermediate levels of the pattern. BothHoxd-8andHoxd-10are expressed in non-regenerating wounds, butHoxd-11is specific for regeneration. It is also expressed in the posterior half of nerve-induced supernumerary outgrowths.     

Specific Biological Traits of Males of the Blue Crab Paralithodes platypusin the Northeast Part of the Sea of Okhotsk

Data on body size, composition, and mass; peculiarities of molting; and number and seasonal migrations of males of the blue king crab Paralithodes platypusfrom the northeast part of the Sea of Okhotsk are presented. It was found that the main aggregations of juveniles (up to 65% of the total number) are located in the south of the population area. Males molt once every two years, and the relative number of groups molting in alterating years was equal. Mass molting was observed at the end of June and July. Mass migration of males to shoals occurred in June, and their return occurred in October.     

First Record of Ulca bolini (Pisces: Hemitripteridae) in the Southeastern Part of the Sea of Okhotsk

This paper presents data on the first record of Ulca bolini. The fish was caught at a depth of 280–300 m in the Sea of Okhotsk off the Kuril Islands (Prostor Bay, Iturup Island). It was 60 cm in fork length (FL) and 7 kg in weight.     

Reproduction Strategy of the Deep-sea Hermit Crabs Pagurus alatus and Pagurus excavatus of the Central-Western Mediterranean Sea

Some aspects of the reproductive biology of two hermit crabs Pagurus alatus (Fabricius, 1775) and P. excavatus (Herbst, 1791) were studied. Specimens were collected monthly from April 2000 to March 2001 on the continental shelf and the upper-middle slope off the Southwestern coasts of Sardinia, in the central-western Mediterranean Sea. Crab size (minimum and maximum shield length) was 1.7 and 9.5 mm for 1150 females of P. alatus; 4.0 and 10.4 mm for 347 females of P. excavatus, respectively. Females of P. alatus with ripe ovaries were only found in February-April and ovigerous females were observed throughout the year except in March, with the highest incidence in summer. Females of P. excavatus with fully developed ovaries were collected during all months of the year reaching a peak from April to June. Ovigerous females of P. excavatus occurred throughout the year and the main spawning period occurred in March and April. Fecundity of both species was calculated to assess seasonal variation of reproductive intensity and was positively correlated with the size of the individuals. Monthly fecundity did not reveal significant differences in P. alatus ovigerous females, whereas P. excavatus exhibited significant differences in monthly fecundity depending on the time of year. Difference in fecundity among P. alatus and P. excavatus can be related to the size of the ovigerous females and species of hermit crab, but their different reproductive strategies suggest that they are specifically related to different environmental conditions.     

The First Finding of the Actinia Synandwakia hozawai in the Sea of Okhotsk

The finding of the actinia Synandwakia hozawai in the coastal waters of northern Sakhalin (Sea of Okhotsk) suggests a wider range of this species, which was previously only known to inhabit the coastal waters of eastern Japan. Data are presented on the morphology of the S. hozawai specimen from the Sea of Okhotsk and the types of its nematocysts.     

Regeneration of Gossypium hirsutum and G. barbadense from shoot apex tissues for transformation

A method of regenerating cotton plants from the shoot apical meristem of seedlings was developed for use with particle gun and Agrobacterium-mediated transformation. This method was developed to circumvent the problems of genotype restriction and chromosomal damage frequently encountered in cotton regeneration in tissue culture through somatic embryogenesis. In this procedure, the cells of the shoot meristem are targeted for transformation. Normal and fertile plants of Gossypium barbadense Pima S-6, and 19 cultivars of G. hirsutum were regenerated using this method. Shoot regeneration from these tissues was direct and relatively rapid. A MS based, hormone-free medium could be used with all the varieties tested.This project was funded by grants from Cotton Incorporated, Nisshinbo Industries, and a grant from the Texas Agricultural Experiment Station to RHS. Texas Agricultural Experiment Station Technical Article TA-25667.     

Regeneration in Phaseolus vulgaris L. Promotive role of N6-benzylaminopurine in cultures from juvenile leaves

Plants were regenerated from cultured young leaves of Phaseolus vulgaris L. cv. Kinghorn. For inducing shoot regeneration the expiant had to consist of the petiole and a portion of the lamina, and N⁶-benzylaminopurine (BAP) had to be present in the culture medium. Furthermore, the frequency of shoot regeneration increased more than seven-fold if donor seedlings were raised on a medium containing 5 M BAP, followed by culture of the leaf explants on a medium containing 20 M BAP. Regenerated shoots developed roots on basal (hormone-free) medium and the resulting plantlets could be transplanted to soil.Abbreviations BAP N⁶-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog (1962) medium This research was supported by operating grants from the Research Board Grants Program of the University of Guelph and the Natural Sciences and Engineering Research Council of Canada to PKS. Technical and photographic assistance from Sangeeta Saxena and Jean Gerrath is gratefully acknowledged.     

Agrobacterium-Mediated Transformation and Plant Regeneration of Triticum aestivum L.

The use of two Agrobacterium tumefaciens strains for transformation of Triticum aestivum L. cv. Vesna was studied. Immature embryos, isolated 15 d after pollination, were co-cultivated with the super-binary LBA4404/pTOK233 and the binary AGL1/pDM805 vectors. While the transient GUS-intron expression was high (69.9 and 80.0 %), the number of plants regenerated on selective media containing hygromycin or phosphinotricin did not exceed 0.4 and 0.13 %, respectively. Nevertheless, the regenerated plants were fertile and produced seeds. The T₀ plants, as well as the T₁ seedlings, displayed the activity in the β-glucuronidase histochemical assay and a positive signal in PCR analysis for the presence of uidA gene sequences in their genomes. The data suggest that the transformation of wheat cv. Vesna with both Agrobacterium strains is feasible. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genus Alexandrium Halim, 1960 (Dinophyta) from the Pacific coast of Russia: Species composition, distribution, and dynamics

At present 8 species of Alexandrium genus have been found in seas and adjacent waters of Russia: A. acatenella, A. catenella, A. insuetum, A. margalefii, A. ostenfeldii, A. pseudogonyaulax, A. tamarense, and A. tamutum. The distribution and population density of Alexandrium species varied within the surveyed area of the Pacific: in the Sea of Japan and Sea of Okhotsk, 7 species were recorded; 3 species were recorded along the Pacific coast of Kamchatka; and 2 species were found in the Bering Sea. A. tamarense was the most widespread and abundant species over the area. A. insuetum was recorded only in the Sea of Japan, and A. catenella, in the Sea of Okhotsk (Terpeniya Bay). The highest concentration of Alexandrium spp. (2–7 million cells/l) was recorded along the Pacific coast of Kamchatka and in the Bering Sea; in the Sea of Okhotsk, a rather high concentration (51000 cells/l) was registered in Aniva Bay; in the Sea of Japan, the highest concentration was recorded in Peter the Great Bay (6000 cells/l). The distribution of cysts (spores) in surface sediments of the Pacific coast of Russia as a whole reflected the pattern of distribution of vegetative cells of Alexandrium. Cysts of Alexandrium cf. tamarense prevailed all over the area, with the maximum concentration along the Pacific coast of Kamchatka. Beyond that type of cysts, insignificant numbers of cysts of Alexandrium cf. minutum were recorded in Peter the Great Bay and Aniva Bay. Analysis of seasonal dynamics revealed that cells of Alexandrium spp. occurred in Peter the Great Bay from June up to September, and along the Pacific coast of Kamchatka from April to October. In the first region, the maximum density was recorded in August; it was provided by A. pseudogonyaulax (59% of the total density of Alexandrium), A. tamarense (35%), and A. insuetum (6%). In the second region, it was recorded in July, thanks only to development of A. tamarense.     

甜椒花药培养及再生植株技术体系的研究

为建立甜椒(Capsicum annuum var.grossum)花药培养及再生植株技术体系,对影响花药胚状体诱导和分化的因素进行了研究。结果表明,培养基组成对胚状体诱导率的影响以NAA基本培养基椰乳KT,最佳胚状体诱导培养基为NTH+0.1 mg L–1 NAA+10%椰乳+1 mg L–1 KT+50μmol L–1 Ag NO3+30 g L–1蔗糖+5 g L–1琼脂+2 g L–1活性炭。花药经过24 h低温预处理和8 d高温预培养后,胚状体诱导率可达23.38%。植物生长调节剂对胚状体出芽率的影响为6-BANAAIAA,最佳胚状体分化培养基为NTH+1 mg L–1 6-BA+0.3 mg L–1 NAA+0.1 mg L–1 IAA+30 g L–1蔗糖+5 g L–1琼脂。胚芽转入1/2MS+0.5 mg L–1 IBA+30 g L–1蔗糖+5 g L–1琼脂培养基后,生根率可达92.5%。     

<Emphasis Type="Italic">Magadania skopetsi</Emphasis>, a new genus and species of Zoarcidae (Teleostei: Perciformes) from the Sea of Okhotsk

A new genus and species of zoarcid fish, Magadania skopetsi, is described on the basis of 17 specimens (83.4–115.9mm SL) from the intertidal zone of the northern coast of the Sea of Okhotsk, near Magadan, Russia. This species differs from other known zoarcid genera and species by having the following combination of characters: suborbital bones 5, the suborbital canal with 6–8 small pores; first epibranchial fan-shaped; palatopterygoid series reduced; supraoccipital broadly contacting exoccipital; posttemporal ventral ramus weak; upper lip continuous at symphysis; palatine teeth, pelvic fins and scales present; lateral line configuration mediolateral, incomplete, absent on posterior half of body; vertebrae 18–22+78–83=97–104. The closest genus cannot be determined cladistically except that the species is a member of the subfamily Gymnelinae. During the spawning season, M. skopetsi lives under stones near the outer marginal area of the intertidal zone.     

Morphological peculiarities of <Emphasis Type="Italic">Fragilidium mexicanum</Emphasis> Balech, 1988 (Dinophyta) from the Far-Eastern Seas of Russia

The vegetative cells of Fragilidium mexicanum Balech are recorded from the Far Eastern seas of Russia (Sea of Japan and Sea of Okhotsk) for the first time. Morphological study of both cultured and wild cells of F. mexicanum showed that the shape of the cell and the first (1″) and second (2″) precingular plates, the direction of the slot in plate 1″, and the shape of the anterior sulcul plate (S.a.) vary greatly. These features bear similarity to those of F. mexicanum, as well as a closely related species, F. subglobosum. The most conservative characters distinguishing the two species are the shape and size of the first and seventh postcingular plates and the shape of the posterior sulcal plate.     

Regeneration of the cell wall in protoplasts of Candida albicans

To assess the dynamics of synthesis of the wall by regenerating Candida albicans protoplasts deposition of chitin and mannoproteins were investigated ultrastructurally using wheat germ agglutinin conjugated with either horseradish peroxidase or colloidal gold, and Concanavalin A coupled to ferritin respectively.Freshly prepared protoplasts lacked wheat germ agglutinin receptor sites but after 1–2 h of regeneration, they were detected. After 4–5 h of regeneration, the cell wall showed a discrete structure which was only labelled with wheat germ agglutinin in thin sections. At this stage of regeneration the outermost layer of the wall was labelled with clusters of Concanavalin A-ferritin particles.After 8 h regeneration, the cell wall appeared compact, and homogenously marked with wheat germ agglutinin whereas only the surface layers appeared consistently labelled with Concanavalin A-ferritin.From these observations we conclude that C. albicans protoplasts are able to regenerate in liquid medium a cell wall consisting of a network of chitin fibrils and mannoproteins at least (glucan polymers were not determined in the present cytological study). The former are the fundamental component of the inner layers at early stages of regeneration, whereas the latter molecules are predominant in the outer layers of the wall.Abbreviations WGA-HRP wheat germ agglutinin conjugated with horseradish peroxidase - WGA-Au wheat germ agglutinin conjugated with colloidal gold - Con A-ferritin Concanavalin A coupled to ferritin