Genetic connectivity between sympatric populations of closely related char species,Dolly Varden <Emphasis Type="Italic">Salvelinus malma</Emphasis> and white char <Emphasis Type="Italic">Salvelinus albus</Emphasis>

The closely related chars Salvelinus malma and Salvelinus albus, which sympatrically inhabit the Kamchatka River basin and Kronotsky Lake (Kamchatka), attract the attention of the researchers because of their debated origin and taxonomic status. Previous studies of sympatric populations of these chars revealed small but statistically significant genetic differences between these species at a number of molecular markers, suggesting the presence of the genetic exchange and hybridization. In this study, based on genotypic characterization of nine microsatellite loci, a considerable level of historical and contemporary genetic migration between sympatric populations of these chars was demonstrated. At the individual level, a high degree of hybridization was observed, mainly among the Dolly Varden individuals from the studied populations. The obtained evidences on the genetic connectivity between sympatric S. malma and S. albus do not support the separate species status of S. albus.     

Offshore Dolly Varden charr (<Emphasis Type="Italic">Salvelinus malma</Emphasis>) in the North Pacific

Many studies have investigated the ecology of charrs in freshwater, however, little is known about charrs in the ocean. This study examined the distribution, seasonal abundance, and some biological features of Dolly Varden (Salvelinus malma) in the Pacific Ocean. An analysis of by-catch data of Japanese offshore salmon monitoring showed that Dolly Varden were distributed across a wide range in the offshore waters of the Pacific Ocean, including the Japan Sea, Bering Sea, and Okhotsk Sea. The catch per unit effort showed a sharp increase from May to August, followed by a sharp decrease in September. Offshore areas served as an important summer habitat for anadromous Dolly Varden.     

Diversity of nosed charrs <Emphasis Type="Italic">Salvelinus malma</Emphasis> (Salmonidae) of Lake Kronotskoe (Kamchatka)

The littoral benthivorous charrs Salvelinus malma with subterminal mouth distributed in Lake Kronotskoe basin are separated into three distinct morphs. These morphs significantly differ in head proportions, jaws' length, skull structure, and degree of gill rakers' reduction. The morphs spawn in distant lake tributaries at the spawning grounds of various types. Joint spawning has never been observed for these morphs in any type of spawning grounds. The differences in the external morphology and growth rate of the morphs become evident during the riverine life stage. The adult charrs inhabit the same sites of the littoral zone. No reliable difference is found in the diet among morphs, since all nosed charrs dominantly feed on amphipods with the exception of a few individuals that fed on chironomid larvae.     

Genetic divergence of mitochondrial DNA in white char <Emphasis Type="Italic">Salvelinus albus</Emphasis> and northern Dolly Varden char <Emphasis Type="Italic">Salvelinus malma malma</Emphasis>

Comparative analysis of mitochondrial DNA variation was performed in white char Salvelinus albus Glubokovsky and in its putative ancestor species, northern Dolly Varden char Salvelinus malma malma Walbaum. Highly statistically significant differentiation of S. albus and S. m. malma in the areas of sympatric (Kamchatka River basin) and allopatric (Kronotskoe Lake and Kronotskaya River) residence was demonstrated. The mtDNA divergence between S. albus and S. m. malma did not exceed the range of intraspecific variation in the populations of northern Dolly Varden char. At the same time, clusterization pattern of the Salvelinus chars provides hypothesis on the common origin of two allopatric populations of white char. Genealogical analysis of haplotypes indicates that S. albus and S. m. malma currently demonstrate incomplete divergence of mitochondrial lineages. The low nucleotide divergence estimates between S. albus and S. m. malma reflect the short time period since the beginning of the divergence of ancestral lineages. These estimates are determined by ancestral polymorphism and haplotype exchange between the diverged phylogenetic groups as a result of introgressive hybridization.     

Effects of forestry on the thermal habitat of Dolly Varden (<Emphasis Type="Italic">Salvelinus malma</Emphasis>)

Forestry activities in riparian areas are known to affect stream communities considerably. Not only do riparian deforestation resulting from agriculture or urbanization developments affect stream communities but extensive commercial plantation and forestry practices can alter stream environments adjacent to remaining, intact or secondary forests. Because forestry often includes the construction of logging roads through the riparian zone, this can directly degrade stream environments. Twelve streams in the Shiretoko Peninsula, Hokkaido were investigated so as to determine the effects of forestry practices on stream temperature, periphyton biomass, grazer (benthic invertebrates) biomass and Dolly Varden (Salvelinus malma Walbaum) biomass. The greater the proportion of planted area in the catchment, the higher the stream temperature. Stream temperature directly affects periphyton biomass and Dolly Varden biomass negatively. Neither stream temperature nor periphyton biomass predicted grazer biomass, whereas a positive correlation was found between grazer biomass and Dolly Varden biomass that forage on invertebrates. The overall results indicated that Dolly Varden in the Shiretoko streams were negatively affected by forestry practices and the resultant stream temperature increases. Without effective future riparian forest management, the complex effects of both riparian disturbance and ongoing global warming could further reduce Dolly Varden populations in the region.     

Population genetic structure of northern Dolly Varden char <Emphasis Type="Italic">Salvelinus malma malma</Emphasis> in Asia and North America

The level of genetic differentiation of northern Dolly Varden char Salvelinus malma malma from Asia and North America was evaluated using the data on mtDNA variation (regions ND1/ND2, ND5/ND6, and Cytb/D-loop) obtained by means of PCR-RFLP analysis. For S. m. malma, the mean values of haplo-type and nucleotide diversity were 0.5261 ± 0.00388 and 0.001558, respectively. The mean estimate of the population nucleotide divergence constituted 0.055%. It was demonstrated that S. m. malma on the most part of the species range examined (drainages of the Beaufort Sea, Chukotka Sea, Bering Sea, and the Sea of Okhotsk) was characterized by the population genetic structure with the low level of genetic differentiation and divergence. At the same time, populations from the Pacific Ocean Gulf of Alaska demonstrated marked genetic differentiation, supported by the high pairwise G4ST values (from 0.4198 to 0.5211) and nucleotide divergence estimates (mean divergence, 0.129%), from Asian and North American populations. Analysis of molecular variance (AMOVA) showed that most of the mtDNA variation in S. m. malma fell in the intrapopulation component (72.5%). At the same time, the differences between the populations (21.1%) and between the regions (6.4%) made lower contribution to the total variation.     

Flexible seaward migration of Dolly Varden <Emphasis Type="Italic">Salvelinus malma</Emphasis> in the Shiretoko Peninsula,Hokkaido, Japan

The migratory history of Dolly Varden Salvelinus malma was investigated in 30 streams in the Shiretoko Peninsula, Hokkaido, Japan, which is in the southern part of their distribution. Captured Dolly Varden were classified into two categories by their body coloration: silvery fish were thought to be an anadromous form, and non-silvery fish a fluvial form that remains in freshwater throughout its life. Forty-seven of 1,739 Dolly Varden collected in the spring and ten of 2,126 Dolly Varden collected in the autumn were silvery fish. Twenty silvery fish and 120 non-silvery fish were examined by analysis of otolith Sr:Ca ratio. Three silvery fish from autumn and 15 non-silvery fish had migrated to salt water or brackish water. The other specimens of silvery fish were regarded as smolt or pre-smolt before seaward migration. The present study suggests that Dolly Varden in the Shiretoko Peninsula have a flexible migratory history, where some of the non-silvery fish utilize marine environments while some individuals follow an anadromous life history.     

Morphophysiological patterns of life history strategy adoption in Dolly Varden <Emphasis Type=Italic>Salvelinus malma</Emphasis> in Kamchatka

Growth, age composition, gonad maturity and patterns of life history strategy adoption in the population of Dolly Varden from the Kol River (West Kamchatka) are studied. Major factors controlling differentiation of juveniles and trajectories of ontogenetic developments, growth rate and gametogenesis in generations in particular years. The differentiation in a generation into anadromous and resident groupings is completed mainly at the age of 4 years. The process of formation of resident and migratory life history strategy in Dolly Varden is epigenetic. Bifurcation in the life of one specimen is observed once in life, and that in a generation is observed four times: at the first, the second, the third, or the fourth year. The life cycle of females from the Kol River is invariant until they reach the age of 1 year, and that in males is during the 4 years of the freshwater phase. In Kamchatka, all spawners, including precocious males, may reproduce several times during life.     

Differentiation of Dolly Varden Char <Emphasis Type="Italic">Salvelinus malma</Emphasis> from Asia and North America Inferred from PCR-RFLP Analysis of Mitochondrial DNA

Genetic differentiation of Dolly Varden char Salvelinus malma Walbaum from the Asian and North American Pacific coasts was studied. We examined restriction fragment length polymorphism of three mitochondrial DNA (mtDNA) fragments amplified in polymerase chain reaction, which encoded four NADH dehydrogenase subunits, the cytochrome b gene, and a D-loop segment. The mtDNA haplotypes were shown to form three phylogenetic groups, whose geographic distribution corresponded to three Dolly Varden subspecies: S. malma malma, S. malma krascheninnikovi, and S. malma lordi. The nucleotide sequence divergence between S. malma malma and S. malma krascheninnikovi was 3.8%; between S. malma malma and S. malma lordi, 3.1%; and between S. malma krascheninnikovi and S. malma lordi, 2.5%. The northern Dolly Varden S. malma malma from Asia was shown to be genetically identical to that from North America.__________Translated from Genetika, Vol. 41, No. 5, 2005, pp. 626–634.Original Russian Text Copyright © 2005 by Oleinik, Skurikhina, Brykov, Crane, Wenburg.     

Trophic differentiation of the nosed charr <Emphasis Type="Italic">Salvelinus schmidti</Emphasis> Viktorovsky, 1978 in Lake Kronotskoe (Kamchatka)

The nosed charr Salvelinus schmidti that inhabits the littoral zone of Lake Kronotskoe is divided into two groups according to food preferences and parasite fauna. Fish of the first group (G) predominantly feed on gammarids and are characterized by a high infestation with Cystidicola farionis, Cyathocephalus truncatus, and Crepidostomum spр. Fish of the second group (A) do not consume gammarids but feed mostly on chironomid larvae and pupae and on mollusks; the predominant parasites are Phyllodistomum umblae and Proteocephalus longicollis. The significant difference in the abundance of C. farionis, which remains in fish at least for 2 years, is indicative of a long-term and persistent trophic diversification between the charr groups. Moreover, significantly higher growth rates, a larger body size, and a longer lifespan are observed for fish of group G. The feeding strategy of the nosed charr is maintained throughout the life and does not depend on the sex of fish and their distribution within the littoral zone of the lake.     

A comparison of different structures and methods for estimating age of northern-form Dolly Varden <Emphasis Type="Italic">Salvelinus malma malma</Emphasis> from the Canadian Arctic

Assessment of anadromous northern-form Dolly Varden Salvelinus malma malma in the Western Canadian Arctic requires reliable methods for estimating ages. Additionally, conservation efforts warrant determining whether fin rays provide a non-lethal alternative to otoliths. Precision and bias of whole and sectioned otoliths, and sectioned pectoral and pelvic fin rays were examined. Two age readers with different levels of experience ageing this species read each structure three times. Coefficient of variation (CV) was calculated to measure precision, and age bias plots were created for each method of preparation/structure within and between readers. The experienced reader demonstrated the highest precision with sectioned otoliths (CV = 1.6 %) followed by whole otoliths (CV = 4.2 %) while pectoral and pelvic fins were the lowest, CV = 7.7 % and 7.5 %, respectively. The age bias plot showed little difference between whole and sectioned otoliths, although greater imprecision/bias was evident for whole otoliths at age ≥9. Compared to otoliths, fin rays produced younger age estimates starting at 5 years; however, pelvic fins were more biased towards younger estimates than pectoral fins. The less experienced reader had greater inconsistencies, tending to overage younger and underage older samples for all methods compared to the more experienced reader, underscoring the importance of experience when estimating age for this species. We conclude that both types of fin rays are a poor non-lethal alternative to otoliths for fish ≥5 years and recommend an experienced ager could use whole otoliths up to age 8 and sectioned otoliths for fish ≥9 years (>500 mm fork length).     

New endemic deepwater charr morphs of the genus <Emphasis Type="Italic">Salvelinus</Emphasis> (Salmoniformes: Salmonidae) from Lake Kronotskoe,Kamchatka

Two previously unknown profundal dwelling charr morphs of the genus Salvelinus from Lake Kronotskoe are described in this article. According to their lifestyle peculiarities, these morphs were named the “bigmouth charr” and “smallmouth charr.” The former group is near-bottom benthivorous, while the latter one inhabits the water column and occupies the omnivorous niche. Bigmouth and smallmouth charrs are distinguished from the rest of the sympatric charr morphs by their smaller size, shorter snout, larger eyes, as well as by several craniological traits and lower parasite abundance. Spawning of both morphs takes place directly in the profundal zone and lasts from late October to February. Additional studies are required to determine the systematic status of the new morphs.     

Phylogeography of southern Asian Dolly Varden char <Emphasis Type="Italic">Salvelinus malma krascheninnikovi</Emphasis>: Genealogical analysis of mitochondrial DNA

Phylogeography of southern Asian Dolly Varden char was studied using the data on mtDNA variation (regions ND1/ND2, ND5/ND6, and Cytb/D loop) obtained using PCR-RFLP analysis. Analysis of contemporary population genetic structure showed that Salvelinus malma krascheninnikovi throughout the whole species range was characterized by high population differentiation in combination with rather small differences between the populations from remote regions. The genealogy of mtDNA haplotypes was reconstructed and nested clade analysis of geographical distances was performed. Geographical distribution of mtDNA haplotypes of S. m. krascheninnikovi was explained by population genetic processes (restricted gene flow), as well as by historical demographic events (range expansion and fragmentation). It was demonstrated that the main demographic events were associated with cyclic processes of the geological formation of the Sea of Japan and adjacent territories. Furthermore, genealogical tree of S. m. krascheninnikovi contained the traces of secondary contact between isolated phylogeographical lineages.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.     

Genetic relationships of Chukchi charr <Emphasis Type="Italic">Salvelinus andriashevi</Emphasis> and Taranetz charr <Emphasis Type="Italic">Salvelinus taranetzi</Emphasis>

On the basis of sequence analysis of the mitochondrial DNA (mtDNA) control region (CR), cytochrome b (Cytb), and cytochrome oxidase-1 (CoI) genes, the relationships of endemic species Salvelinus andriashevi Berg, 1948, represented by the only population from Lake Estikhed (Chukotka), were estimated. The data on the genealogical analysis of mtDNA haplotypes supported phylogenetic closeness of S. andriashevi and S. taranetzi. It was also demonstrated that the specimens of Chukchi charr, along with Salvelinus sp. 4 (Lake Nachikinskoe), S. krogiusae (Lake Dal’nee), S. boganidae and S. elgyticus (Lake El’gygytgyn), and S. a. erythrinus from Canada’s Northwest Territories (NWT) belonged to the Arctic group of Taranetz charr. The problem of coincidence of taxonomic differentiation of charrs of the genus Salvelinus based on morphological and genetic analyses is discussed.     

Ecology of Slaty-Backed Gull (<Emphasis Type="Italic">Larus schistisagus</Emphasis>) Breeding on Kronotskoe Lake,Kamchatka Peninsula

The slaty-backed gull population of Kronotskoe Lake has not been subjected to anthropogenic impact for many decades. The number and distribution of the breeding colonies of the slaty-backed gulls on the islands of this largest freshwater reservoir of the Kamchatka Peninsula are presented. Data on the breeding phenology, the seasonal dynamics of the food composition of breeding gulls, the foraging distance, and breeding success are considered. These data are compared to the results obtained earlier for Kuril’skoe Lake. Both gull populations are unique in that they are the only freshwater colonies within the distribution range of this marine species. The breeding success of the slaty-backed gulls on Kronotskoe Lake suffers from predation, both intra- and interspecific, in the latter case from brown bears and sea eagles. The hatching period and the fledging time of chicks in this population are both shifted to the end of summer, when gulls have the most abundant food available within the lake area, which increases successful survival of the fledglings. Despite considerable differences between the ecosystems of Kronotskoe Lake and Kuril’skoe Lake, there are several common features in the diets of the gull populations living on the lakes: (1) Despite the fact that gulls perform foraging flights to the sea coast, they only take fish and almost never collect marine invertebrates there. (2) Nearly half of all food items are taken within a 40-km distance from the colony. (3) During the growth period of chicks, gulls from both lakes rely on salmonids (anadromous in Kuril’skoe Lake, resident in Kronotskoe Lake) that they hunt in the lakes near their colonies.     

Revision of the <Emphasis Type="Italic">Serrulati</Emphasis> species of <Emphasis Type="Italic">Penstemon</Emphasis> section <Emphasis Type="Italic">Saccanthera</Emphasis> subsection <Emphasis Type="Italic">Serrulati</Emphasis>

A revision of Penstemon sect. Saccanthera subsect. Serrulati includes a new species (P. salmonensis), a new variety (P. triphyllus var. infernalis), and the elevation of a subspecies to species (P. curtiflorus), bringing the total number of species to eight, which are keyed and described, complete with nomenclature and type citations.     

<Emphasis Type="Italic">Galleria</Emphasis><Emphasis Type="Italic">mellonella</Emphasis> are Resistant to <Emphasis Type="Italic">Pneumocystis</Emphasis><Emphasis Type="Italic">murina</Emphasis> Infection

Studying Pneumocystis has proven to be a challenge from the perspective of propagating a significant amount of the pathogen in a facile manner. The study of several fungal pathogens has been aided by the use of invertebrate model hosts. Our efforts to infect the invertebrate larvae Galleria mellonella with Pneumocystis proved futile since P. murina neither caused disease nor was able to proliferate within G. mellonella. It did, however, show that the pathogen could be rapidly cleared from the host.     

<Emphasis Type="Italic">Agrobacterium</Emphasis><Emphasis Type="Italic">tumefaciens</Emphasis>-mediated transformation of callus cells of <Emphasis Type="Italic">Crataegus</Emphasis><Emphasis Type="Italic">aronia</Emphasis>

A genetic transformation system has been developed for callus cells of Crataegus aronia using Agrobacterium tumefaciens. Callus culture was established from internodal stem segments incubated on Murashige and Skoog (MS) medium supplemented with 5 mg l⁻¹ Indole-3-butyric acid (IBA) and 0.5 mg l⁻¹ 6-benzyladenine (BA). In order to optimize the callus culture system with respect to callus growth and coloration, different types and concentrations of plant growth regulators were tested. Results indicated that the best average fresh weight of red colored callus was obtained on MS medium supplemented with 2 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l⁻¹ kinetin (Kin) (callus maintenance medium). Callus cells were co-cultivated with Agrobacterium harboring the binary plasmid pCAMBIA1302 carrying the mgfp5 and hygromycin phosphotransferase (hptII) genes conferring green fluorescent protein (GFP) activity and hygromycin resistance, respectively. Putative transgenic calli were obtained 4 weeks after incubation of the co-cultivated explants onto maintenance medium supplemented with 50 mg l⁻¹ hygromycin. Molecular analysis confirmed the integration of the transgenes in transformed callus. To our knowledge, this is the first time to report an Agrobacterium-mediated transformation system in Crataegus aronia.