Variability of microsatellite loci of Greenland cod <Emphasis Type="Italic">Gadus ogac</Emphasis> Richardson 1836: Comparison with other species of <Emphasis Type="Italic">Gadus</Emphasis> genus (Gadidae)

Comparative analysis of variability of seven microsatellite loci—Gmo3, Gmo-G12, Gmo-G18, Gmo19, Gmo34, Gmo35 and Pgmo32—was performed for the Greenland cod Gadus ogac, Pacific cod G. macrocephalus, Atlantic cod G. morhua, and White Sea cod G. morhua marisalbi. High genetic identity was observed between the Greenland cod and Pacific cod (I = 0.9520). Pair analysis of genetic differentiation was performed on the studied microsatellite loci according to θ (analogue of F _ST). The Greenland cod differed significantly from the Pacific, Atlantic, and the White Sea cod; however, the differentiation level varied. The lowest value was observed for the pair Greenland cod-Pacific cod (0.123), and the highest levels were registered for the pairs Greenland cod-Atlantic cod (0.605) and Greenland cod-White Sea cod (0.535).     

Microsatellite Variability of Pacific Herring <Emphasis Type="Italic">Clupea pallasii</Emphasis> Valenciennes, 1847 from the Sea of Okhotsk and Bering Sea

The genetic variability of ten microsatellite loci was examined in samples of the herring from the Sea of Okhotsk and the Bering Sea. All loci were polymorphic; the expected heterozygosity estimates varied in the range of 0.3–94.3% (mean 66.7%). The degree of genetic differentiation of the herring was statistically significant (θ = 1.38%). The level of pairwise genetic differentiation F_ST was–0.002–0.046; R_ST was–0.003–0.166. Genetic differentiation of the herring from the Sea of Okhotsk and the Bering Sea correlated with the spatial-geographic structure of the species in the studied range on the basis of F_ST (P = 0.001).     

Population genetic analysis of Euro-Arctic polar cod <Emphasis Type="Italic">Boreogadus saida</Emphasis> suggests fjord and oceanic structuring

Polar cod, Boreogadus saida, is a key species in Arctic marine ecosystems; however, its genetic population structure is largely undescribed. The population genetic structure of 472 B. saida specimens among nine locations in the north-east Atlantic was revealed using 12 microsatellite loci. Pairwise F _ST comparisons showed significant population differentiation between B. saida sampled inside fjords in Svalbard and north-east Greenland, as compared to B. saida from the shelf. The observed genetic variation was not a function of isolation by distance, and it is speculated that B. saida populations inhabiting fjords may have become reproductively isolated from shelf-dwelling B. saida during the last post-glacial recolonization.     

Population Structure of Pacific Cod <Emphasis Type="Italic">Gadus macrocephalus</Emphasis> in the Southern Part of the Range Based on the Microsatellite Analyses

The population structure of Pacific cod Gadus macrocephalus in the southern part of the range and adjacent regions is studied on the basis of the results of microsatellite analyses. Collected data indicate heterogeneity of this species population within the studied area. According to the obtained F_ST values, Pacific cod from the waters of the Republic of Korea (Yellow Sea side) and northwestern part of the Sea of Okhotsk significantly differ from all other studied regions (Table 4). Significant differentiation was also revealed between samples from the waters of the Tatar Strait and all other regions except for South Kurils Pacific cod (both Sea of Okhotsk and Pacific Ocean sides). These two latter sample collections were similar to each other as well. A low level of differentiation was also shown for the Peter the Great Bay and the East Sea/Sea of Japan waters of the Republic of Korea.     

Polymorphism of pigmentation genes (<Emphasis Type="Italic">OCA2</Emphasis> and <Emphasis Type="Italic">ASIP</Emphasis>) in some populations of Russia

In Russian populations, polymorphism of two pigmentation system genes, OCA2 (loci 305, 355, and 419, tested in Russians, Buryats, Chukchi, Koryaks, and Evens) and ASIP (locus 8818, tested in Russians and Buryats) was examined. Pairwise comparisons of the F _ST distances between the populations showed that only the populations from Northeast Asia (Chukchi, Koryaks, and Evens) were statistically significantly different from all other populations, at least relative to one of the OCA2 locus. In Russians from Pskov oblast and Novgorod oblast, increased frequency (up to 6%) of the OCA2 allele 419A was revealed. In earlier studies, as association of this allele with green eye color was demonstrated. The data obtained in terms of their application for ethnic population genetics.     

Conservation genetics of the short-beaked common dolphin (<Emphasis Type="Italic">Delphinus delphis</Emphasis>) in the Mediterranean Sea and in the eastern North Atlantic Ocean

Mediterranean Sea common dolphins have recently been listed as ‘endangered’ in the IUCN Red list, due to their reported decline since the middle of the 20th century. However, little is know about the number or distribution of populations in this region. We analysed 118 samples from the Black Sea, Mediterranean Sea and eastern North Atlantic at nine microsatellite nuclear loci and for 428 bps of the mtDNA control region. We found small but significant population differentiation across the basin between the eastern and the western Mediterranean populations at both nuclear and mtDNA markers (microsatellite F _ST = 0.052, mtDNA F _ST = 0.107, P values ≤ 0.001). This matched the differential distribution and habitat use patterns exhibited by this species in the eastern and the western parts of the Mediterranean Sea. The assignment test of a small number of samples from the central Mediterranean could not exclude further population structure in the central area of the basin. No significant genetic differentiation at either marker was observed among the eastern north Atlantic populations, though the Alboran population (inhabiting the Mediterranean waters immediately adjacent the Atlantic ocean) showed significant mtDNA genetic differentiation compared to the Atlantic populations. Directional estimates of gene flow suggested movement of females out of the Mediterranean, which may be relevant to the population decline. Phylogenetic analysis suggested that the observed population structure evolved recently.     

High genetic diversity in an endangered medicinal plant, <Emphasis Type="Italic">Saussurea involucrata</Emphasis> (<Emphasis Type="Italic">Saussurea</Emphasis>, Asteraceae), in western Tianshan Mountains,China

Saussurea involucrata (Asteraceae) is a medicinal and second-degree national priority endangered plant that is mainly distributed in the high latitude region of the western Tianshan Mountains. The population is fragmented and isolated, and extensive human impact merits a suitable and specific conservation strategy, which can be compiled based on the genetic diversity, population structure, and demographic history. Phylogeographic studies were conducted on a total of five natural populations and 150 individuals were sampled. Data from three cpDNA intergenic spacer regions (trnL-F, matK, and ndhF-rpl32) and nrDNA ITS sequences showed that twelve haplotypes in cpDNA and five haplotypes in nrDNA indicated high genetic diversity among populations sampled (H _T?=?0.820 and 0.756) and within populations sampled (H _S?=?0.792 and 0.721). Additionally, the high genetic diversity did not mirror genetic structure in either cpDNA (F _ST?=?0.03153, G _ST?>?N _ST, p?<?0.05) or nrDNA (F _ST?=?0.03666, meaningless G _ST and N _ST). Two groups (north and south) were determined for a SAMOVA analysis. Based on this analysis, the demographic history was conducted with a Bayesian Skyline Plot and Isolation with Migration analysis, which showed sustainable and stable extension without a marked bottleneck. Divergence time was indicated at c. 6.25 Mya (90%HPD: 15.30–0.22 Mya) in the Miocene, which is consistent with the formation of the Kelasu section of Tianshan. The southern populations in the Bayanbulak and Gonglu regions require additional attention and transplanting would be an effective way to restore rare cpDNA haplotypes, increase effective population size, and migration rate. Our results suggested that in situ conservation of S. involucrata in western Tianshan should be the main strategy for protection and recovery of the species.     

Genetics and fine mapping of a yellow-green leaf gene (<Emphasis Type="Italic">ygl</Emphasis>-<Emphasis Type="Italic">1</Emphasis>) in cabbage (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">capitata</Emphasis> L.)

Cabbage (Brassica oleracea var. capitata L.) is one of the most popular cultivated vegetables worldwide. Cabbage has rich phenotypic diversity, including plant height, head shape, head color, leaf shape and leaf color. Leaf color plays an important role in cabbage growth and development. At present, there are few reports on fine mapping of leaf color mutants in B. oleracea. In this study, a naturally occurring yellow-green leaf cabbage mutant (YL-1), derived from the self-pollinated progenies of the hybrid ‘Hosom’, was used for inheritance analysis and gene mapping. Segregation populations including F₂ and BC₁ were generated from the cross of two inbred lines, YL-1 and 01–20. Genetic analysis with the F₂ and BC₁ populations demonstrated that the yellow-green leaf color was controlled by a single recessive nuclear gene, ygl-1. Insertion–deletion (InDel) markers, designed based on the parental re-sequencing data, were used for the preliminary mapping with BSA (bulked segregant analysis) method. A genetic map constructed with 15 InDels indicated that ygl-1 was located on chromosome C01. The ygl-1 gene is flanked by InDel markers ID2 and M8, with genetic distances of 0.4 cM and 0.35 cM, respectively. The interval distance between two markers is 167 kb. Thus, it enables us to locate the ygl-1 gene for the first time in B. oleracea. This study lays the foundation for candidate gene prediction and ygl-1gene cloning.     

Revision of the genus <Emphasis Type="Italic">Melamphaes</Emphasis> (Melamphaidae). 5. Oligo-raker species: <Emphasis Type="Italic">M. indicus</Emphasis>, <Emphasis Type="Italic">M. eurous</Emphasis>, and <Emphasis Type="Italic">M. typhlops</Emphasis>

Three oligo-raker species (?19 rakers on the first gill arch) of the genus Melamphaes out of the “M. typhlops” group are considered. The validity of M. indicus Ebeling is restored. This species inhabits equatorial and tropical waters of the Indian Ocean and the western part of the Pacific Ocean. M. eurous sp. n., which is related to M. indicus, is described from equatorial waters of the eastern part of the Pacific Ocean. M. typhlops (Lowe) inhabiting the northern part of the Atlantic Ocean, from the equatorial zone about to 45° N, is redescribed.     

Development and Characterization of Microsatellite Markers for Genetic Analysis of the Swimming Crab, <Emphasis Type="Italic">Portunus trituberculatus</Emphasis>

This study isolated and characterized 11 novel microsatellite markers for the commercially important swimming crab species, Portunus trituberculatus. Genetic diversity and population structure of two populations of P. trituberculatus in the East China Sea were assessed using these loci. The microsatellite markers produced 242 alleles, varying from 17 to 26 alleles per locus. In all the samples, the range of heterozygosity was 0.6324–0.9403 (observed) and 0.8998–0.9547 (expected). An F-statistic analysis revealed low genetic differentiation between the populations (mean F _ST = 0.0197), with 98% of the variation resulting from the within-population component. In addition, cross-amplification was tested in two other portunid species, and we found that many loci yielded useful information. The high degree of polymorphism exhibited by the 11 microsatellites suggests that these markers will be useful for both aquaculture and studies of natural populations of the genus.     

The <Emphasis Type="Italic">Adh</Emphasis> locus and adaptation of <Emphasis Type="Italic">cn</Emphasis> and <Emphasis Type="Italic">vg</Emphasis> mutants in experimental populations of <Emphasis Type="Italic">Drosophila melanogaster MEIG</Emphasis>

A complex study on the adaptation of cn and vn mutants and the allozymes of alcoholdehydrogenase (ADH) was carried out in initially pure lines, and their panmixia populations during exchange of the mutant genotype with that of wild-type flies (C-S) and D) through saturating crossings. The relative adaptation of the genotypes was estimated by their effect on reproductive efficiency in the experimentally obtained population. Fecundity, lifespan, and the resistance of the studied genotypes to hyperthermia were investigated individually. It was shown that the high level of adaptation of the cn mutants and the low level of adaptation of the vg mutants was correlated with the presence of different ADH allozymes. In the studied population, the F-allozyme of ADH accompanied the vg mutation, while the S-allozyme of the enzyme was detected in cn mutants. Saturating crossings of C-S(Adh ^S)×vg(Adh ^F) and D(Adh ^F) × cn(Adh ^S), along with the parallel determination of the allele composition of the Adh locus, demonstrated that the complete substitution of the F-allozyme of ADH in the vg mutants by the S-allozyme in D flies, as well as the substitution of the S-allozyme of ADH in the cn mutants by the F-allozyme in D flies was realized only after the 15th–20th backcrosses. These results favor the coadaptation of cn and vg marker genes with alleles of the Adh locus and indicate the important role of the latter in the adaptation of genotypes. In the studied population, selection acted primarily against the vg mutants, which were inferior to the cn mutants, and heterozygote genotypes in indices of the main adaptation components.     

Population structure of North Atlantic and North Pacific sei whales (<Emphasis Type="Italic">Balaenoptera borealis</Emphasis>) inferred from mitochondrial control region DNA sequences and microsatellite genotypes

Currently, three stocks of sei whales (Balaenoptera borealis) are defined in the North Atlantic; the Nova Scotian, Iceland-Denmark Strait and Eastern North Atlantic stocks, which are mainly based upon historical catch and sighting data. We analyzed mitochondrial control region DNA (mtDNA) sequences and genotypes from 7 to 11 microsatellite loci in 87 samples from three sites in the North Atlantic; Iceland, the Gulf of Maine and the Azores, and compared against the North Pacific using 489 previously published samples. No statistically significant deviations from homogeneity were detected among the North Atlantic samples at mtDNA or microsatellite loci. The genealogy estimated from the mtDNA sequences revealed a clear division of the haplotypes into a North Atlantic and a North Pacific clade, with the exception of one haplotype detected in a single sample from the Azores, which was included in the North Pacific clade. Significant genetic divergence between the North Atlantic and North Pacific Oceans was detected (mtDNA Φ_ST?=?0.72, microsatellite Weir and Cockerham’s ? = 0.20; p?<?0.001). The coalescent-based estimate of the population divergence time between the North Atlantic and North Pacific populations from the sequence variation among the mtDNA sequences was at 163,000 years ago. However, the inference was limited by an absence of samples from the Southern Hemisphere and uncertainty regarding mutation rates and generation times. The estimates of inter-oceanic migration rates were low (Nm at 0.007 into the North Pacific and at 0.248 in the opposite direction). Although estimates of genetic divergence among the current North Atlantic stocks were low and consistent with the extensive range of movement observed in satellite tagged sei whales, the high uncertainty of the genetic divergence estimates precludes rejection of multiple stocks in the North Atlantic.     

Novel polymorphic microsatellite loci for distinguishing rock bass (<Emphasis Type="Italic">Ambloplites rupestris</Emphasis>), Roanoke bass (<Emphasis Type="Italic">Ambloplites cavifrons</Emphasis>), and their hybrids

The rock bass (Ambloplites rupestris) is a popular sport-fish native to the Mississippi and Great Lakes basins of North America. The species has been widely introduced outside its native range, including into Atlantic-slope streams of Virginia where it may hybridize with an imperiled, similar-looking congener, the Roanoke bass (Ambloplites cavifrons). In this study, we identified and evaluated novel molecular markers to facilitate identification of these species and study the extent of hybridization. Using molecular libraries developed from A. rupestris, we identified a suite of candidate nuclear microsatellite loci, synthesized primer sets, and tested these markers for amplification and polymorphism in populations of both species. We then calculated standard diversity statistics within and differentiation statistics between species, the latter providing an indication of marker power for distinguishing the species and their hybrids. Additionally, we evaluated our efficiency for identifying hybrids by classifying simulated genotypes of known ancestry. Eleven loci were polymorphic (2–22 alleles per locus) and reliably amplified in both species. Multilocus genetic differentiation between A. cavifrons and A. rupestris was quite high (F _ST  = 0.66; D _LR  = 19.3), indicating the high statistical power of this marker set for species and hybrid identification. Analyses of simulated data suggested these markers reliably distinguish between hybrids and non-hybrids, as well as between F1 hybrids and backcrossed individuals. This panel of 11 loci should prove useful for understanding patterns of hybridization between A. rupestris and A. cavifrons. As the first microsatellite markers developed for Ambloplites, these markers also should prove broadly useful for population genetic studies of this genus.     

Identification of <Emphasis Type="Italic">Pm58</Emphasis> from <Emphasis Type="Italic">Aegilops tauschii</Emphasis>

Key message

A novel powdery mildew-resistance gene, designated Pm58, was introgressed directly from Aegilops tauschii to hexaploid wheat, mapped to chromosome 2DS, and confirmed to be effective under field conditions. Selectable KASP? markers were developed for MAS.

Abstract

Powdery mildew caused by Blumeria graminis (DC.) f. sp. tritici (Bgt) remains a significant threat to wheat (Triticum aestivum L.) production. The rapid breakdown of race-specific resistance to Bgt reinforces the need to identify novel sources of resistance. The d-genome species, Aegilops tauschii, is an excellent source of disease resistance that is transferrable to T. aestivum. The powdery mildew-resistant Ae. tauschii accession TA1662 (2n?=?2x?=?DD) was crossed directly with the susceptible hard white wheat line KS05HW14 (2n?=?6x?=?AABBDD) followed by backcrossing to develop a population of 96 BC₂F₄ introgression lines (ILs). Genotyping-by-sequencing was used to develop a genome-wide genetic map that was anchored to the Ae. tauschii reference genome. A detached-leaf Bgt assay was used to screen BC₂F_4:6 ILs, and resistance was found to segregate as a single locus (χ?=?2.0, P value?=?0.157). The resistance gene, referred to as Pm58, mapped to chromosome 2DS. Pm58 was evaluated under field conditions in replicated trials in 2015 and 2016. In both years, a single QTL spanning the Pm58 locus was identified that reduced powdery mildew severity and explained 21% of field variation (P value?<?0.01). KASP? assays were developed from closely linked GBS-SNP markers, a refined genetic map was developed, and four markers that cosegregate with Pm58 were identified. This novel source of powdery mildew-resistance and closely linked genetic markers will support efforts to develop wheat varieties with powdery mildew resistance.

     

Post-release fishing mortality of blue (<Emphasis Type="Italic">Prionace glauca</Emphasis>) and silky shark (<Emphasis Type="Italic">Carcharhinus falciformes</Emphasis>) from a Palauan-based commercial longline fishery

Accounting for components of fishing mortality, including post-release mortality (F_r), is necessary for robust assessments of the effects of fishing. Forty-eight blue (Prionace glauca) and 35 silky sharks (Carcharhinus falciformes) were tagged with pop-up satellite archival tags to monitor F_r rates from pelagic longline vessels in the western tropical Pacific Ocean. There is a paucity of F_r studies at low latitudes and identifying factors that significantly explain F_r is critical for understanding fishing mortality. Mean F_r rates were 0.17 [95% CI 0.09–0.30] for blue shark and 0.20 [95% CI 0.10–0.36] for silky shark. When it occurred, F_r was acute with 87% of mortalities within 2 days of release. Several prognostic operational, environmental, biological and handling variables were evaluated to assess their influence on survival outcomes. Using Kaplan–Meier survival curves, logistic regression, accelerated failure time and Cox proportional hazards models to screen variables, the only significant prognostic or risk variable was health condition at haulback. There was close correspondence (~?83% accuracy) between condition at capture and survival outcomes. Reliable methods to classify at-vessel condition represent an inexpensive and simple metric for estimating both F_r and at-vessel (F_c) mortality rates. Examining F_c rates in detail in longline fisheries using capture information on depth, temperature and dissolved oxygen that may act in synergy with condition code and hooking duration is a research priority. Results suggest that a large proportion of shark survive following release and that F_r rates can be increased by improving the haulback condition of captured sharks.     

Distribution and parameters of genetic polymorphism in northern red-backed vole (<Emphasis Type="Italic">Clethrionomys rutilus</Emphasis>) and bank vole (<Emphasis Type="Italic">Clethrionomys glareolus</Emphasis>) in West Siberia

This article presents data on the genetic variability of the northern red-backed vole and the bank vole that live sympatrically in West Siberia. The two species of voles have comparable, relatively high indices of genetic variability of inter simple sequences repeats DNA. The proportion of polymorphic DNA markers is 95–98%, and the Nei’s genetic diversity index is 0.33–0.35. A total of 47–58% of allozyme loci in the voles are polymorphic, and the average heterozygosity per locus is 0.058 in the northern red-backed vole and 0.054 in the bank vole. Interpopulation differentiation is less pronounced in the red-backed vole (F _ST 0.293) compared to the bank vole (F _ST 0.475). Individuals of the hybrid line of the bank vole with the mitochondrial haplotype of the red-backed vole have been found by PCR typing of cytochrome b gene fragment of mtDNA. The distribution boundary of the hybrid line of bank voles goes farther to the northeast than was shown in earlier works. The proportion of hybrid specimens range from 2 to 34%. The indices of genetic variability in the hybrid line of the bank vole are lower than those of the parental species.     

Comparison of molecular genetic utilities of TD,AFLP, and MSAP among the accessions of <Emphasis Type="Italic">japonica,indica</Emphasis>, and <Emphasis Type="Italic">Tongil</Emphasis> of <Emphasis Type="Italic">Oryza sativa</Emphasis> L.

Rice is one of the most important food crops in the world. Genetic diversity is essential for cultivar improvement programs. We compared genetic diversity derived from insertion–deletion (in–del) or base substitutions by amplified fragment length polymorphism (AFLP), from transposon transposition mutations by transposon display (TD), and from cytosine methylation by methylation-sensitive amplified polymorphism (MSAP) in japonica, indica, and Tongil type varieties of Oryza sativa L. Polymorphic profiles from the three marker systems allowed us to clearly distinguish the three types of varieties. The indica type varieties showed the highest genetic diversity followed by the Tongil and japonica type varieties. Of the three marker systems, TD produced the highest marker indices, and AFLP and MSAP produced similar marker indices. Pair-wise comparisons of the three marker systems showed that the correlation between the two genetic markers systems (AFLP and TD, r = 0.959) was higher than the correlations between the genetic and epigenetic marker systems (AFLP and MSAP, r = 0.52; TD and MSAP, r = 0.505). Both genetic marker systems had similar levels of gene differentiation (G _ST ) and gene flow (N _m ), which differed in the epigenetic marker system. Although the G _ST of the epigenetic marker system was lower than the genetic marker systems, the N _m of the epigenetic marker system was higher than in the genetic marker systems, indicating that epigenetic variations have a greater influence than genetic variations among the O. sativa L. types.