PERMANENT GENETIC RESOURCES: Isolation and characterization of nine microsatellite loci from the chub mackerel, Scomber japonicus (Perciformes, Scombridae)

The stock abundance of the chub mackerel Scomber japonicus - a very important species for fisheries, particularly in Japan - in the Pacific Ocean off Japan has remained at a low level. For studying the population genetics of the chub mackerel, we isolated nine polymorphic microsatellite loci (12-31 alleles/locus; expected heterozygosity, 0.762-0.983) from this species. Cross-species amplification indicated that eight of the nine microsatellite loci in the blue mackerel S. australasicus were polymorphic and functional.     

Molecular cytogenetic analysis of the Appenine endemic cyprinid fish Squalius lucumonis and three other Italian leuciscines using chromosome banding and FISH with rDNA probes

Karyotype and other chromosomal characteristics of the Appenine endemic cyprinid fish, Toscana stream chub Squalius lucumonis, were analysed using conventional banding and FISH with 45S and 5S rDNA probes. The diploid chromosome number (2n = 50) and karyotype characteristics including pericentromeric heterochromatic blocks and GC-rich CMA₃-positive sites corresponding to both positive Ag-NORs and 45S rDNA loci on the short arms of a single medium-sized submetacentric chromosome pair were consistent with those found in most European leuciscine cyprinids. On other hand, 5S rDNA FISH in the Toscana stream chub and three other Italian leuciscines, S. squalus, Rutilus rubilio and Telestes muticellus, revealed a species-specific hybridization pattern, i.e. signals on four (S. lucumonis), three (S. squalus and R. rubilio) and two (T. muticellus) chromosome pairs. Whereas all the species shared the 5S rDNA loci on the largest subtelocentric chromosome pair, a “leuciscine” cytotaxonomic marker, S. lucumonis showed both classes of rDNA loci tandem aligned on the short arms of chromosome pair No. 12. The present findings suggest that the observed high variability of 5S rDNA loci provides a powerful tool for investigation of karyotype differentiation in karyologically conservative leuciscine fishes.     

Characterization of 16 highly variable tetranucleotide microsatellite loci for Oregon chub (Oregonichthys crameri) and cross amplification in Umpqua chub (O. kalawatseti)

We describe the isolation and development of 16 polymorphic tetranucleotide microsatellite loci for the endangered Oregon chub (Oregonichthys crameri). Two loci appear to be duplicated. For the remaining 14 loci, we observed between three and 19 alleles per locus in a sample of 42 fish. Thirteen of these loci were also polymorphic in the closely related Umpqua chub (O. kalawatseti). These loci will aid in our understanding of the molecular ecology and conservation of these two species.     

Range-wide comparisons of northern leatherside chub populations reveal historical and contemporary patterns of genetic variation

Patchily distributed species are those taxa whose populations occupy geographically insular habitats and their conservation often depends on an understanding of the relationship among disjunct populations. The objective of our study was to use molecular data and analytical techniques to separate the effects of historical and contemporary processes influencing the distribution of a high-desert minnow, the northern leatherside chub (Lepidomeda copei). Individuals from 23 populations were sequenced for 1,140 base pairs of the cytochrome B gene of the mitochondrial genome and genotyped at seven nuclear microsatellite loci. We estimated gene flow and examined population structure using both microsatellite and mtDNA data. Low sequence divergence and the distribution of shared haplotypes in multiple watersheds suggest historical connectivity between populations over a large geographic area. In contrast, patterns of microsatellite diversity indicate that populations of leatherside chub are isolated from one another with low levels of contemporary gene flow between populations. Our results suggest that populations of leatherside chub were historically more widely inter-connected and have recently been isolated, likely through a combination of natural and anthropogenic habitat fragmentation. As populations become increasingly isolated, they are more vulnerable to extirpation as a result of stochastic events. For northern leatherside chub, recent isolation and lack of gene flow among populations may affect their long-term survival in the arid landscapes of the Great Basin and surrounding watersheds because of widespread and increasing habitat alteration and fragmentation.     

Characterization of microsatellite loci for five members of the minnow family Cyprinidae found in the Sacramento–San Joaquin Delta and its tributaries

Two microsatellite‐enriched libraries [(CAGA)_n, (TAGA)_n] were constructed using pooled DNA from three cyprinid species native to the Sacramento–San Joaquin Delta of California: Sacramento splittail (Pogonichthys macrolepidotus); Sacramento pikeminnow (Ptychocheilus grandis); and tui chub (Siphateles bicolor). Primers were designed for 105 loci and tested for levels of polymorphism in five cyprinid species found in the Delta: Sacramento splittail, Sacramento pikeminnow, tui chub, hitch (Lavinia exilicauda), and Sacramento blackfish (Orthodon microlepidotus). Fifty‐one loci were polymorphic for at least one species and 31 loci were polymorphic for multiple species. The number of polymorphic loci per species ranged from 16 to 26.     

Habitat patchiness affects distribution and microhabitat use of endangered Mira chub Squalius torgalensis (Actinopterygii,Cypriniformes)

We hypothesized that distribution and microhabitat use by imperilled chub Squalius torgalensis in the Torgal stream, Portugal, during low flows, were related to spatial patchiness in physical resources and shifts in ontogenetic preferences. We mapped fish abundance and sampled microhabitat use and availability via snorkelling. We used the coefficient of dispersion in abundance, and spatial autocorrelation analyses to characterize chub distribution, and Hurdle models to relate chub presence and abundance to habitat characteristics. We showed that chub displayed an aggregated distribution, apparently in association with patchily distributed and autocorrelated physical resources, such as debris, roots and aquatic vegetation. Microhabitat use generally was unrelated to velocity, and ontogenetic differences in microhabitat use were not substantial. Nevertheless, sometimes small chub preferred low-velocity patches with coarse substrata, debris and roots, whereas large chub preferred deeper patches with roots and aquatic vegetation. Results suggest that, in low flow conditions, chub respond to resource patchiness by congregating in favourable microhabitats, and that maintenance of mosaics of patches with variable substrata, cover and depth may be important for the persistence of fish in Mediterranean streams.     

Development of microsatellite markers in bonytail (Gila elegans) with cross‐species amplification in humpback chub (Gila cypha)

Bonytail, Gila elegans, is an endangered species of fish native to the Colorado River. Primers are presented for 17 microsatellites cloned from bonytail as well as the results of test amplifications in bonytail and humpback chub, G. cypha. Bonytail exhibited three to 18 alleles per locus across the 17 microsatellites and a mean expected heterozygosity of 0.58 among 10 loci used to screen 160 broodstock. Humpback chub exhibited one to six alleles and a mean expected heterozygosity of 0.69 among 10 loci that were successfully amplified in that species.     

Genetic characterization and management of the endangered Mohave tui chub

The Mohave tui chub (Siphateles bicolor mohavensis) is the only fish native to the Mojave River, California. The fish were displaced by introduced arroyo chubs (Gila orcutti) throughout most of their range, starting in the 1930s. Two potentially relictual populations and two transplanted populations were genetically characterized using 12 microsatellite DNA loci, along with contemporary cyprinid populations in the Mojave River. We found only un-hybridized Mohave tui chubs in the refuge populations, and only un-hybridized arroyo chubs in the Mojave River. The two largest Mohave tui chub populations (Lake Tuendae and China Lake) exhibit similar, comparatively high genetic variation. Another large population (Camp Cady) with low genetic diversity shows the effect of a bottleneck of ten individuals during the historic founding event. The fourth population (MC Spring) has the fewest alleles, lowest heterozygosity, and is the most divergent, suggesting that genetic drift from a persistently low effective population size has reduced genetic diversity since its apparent isolation in 1934. We recommend instituting artificial gene flow to rebuild genetic variation in Camp Cady from both Lake Tuendae and China Lake, and the establishment of new populations with founders from both Lake Tuendae and China Lake. Additionally, we comment on the infeasibility of restoring populations of Mohave tui chub in their historic habitats.     

Enzyme polymorphisms in the mullet,Liza dussumeiri from Shatt Al-Arab river Khor Al-Zubair and the Arabian gulf

Samples of the muscles and other tissues of Lisa dussumeiri were collected from the Shatt al-Arab river, Khor al-Zubair and Kuwait (Arabian Gulf) and subjected to starch gel electrophoretic analysis of two enzymes-coding loci. All loci at all localities were in Hardy-Weinberg equilibrium. Allele frequencies at the two loci were remarkably constant throughout the area studied. The data provide no evidence of subpopulation differentiation and there is one stock of L. dussumeiri in the area studied.     

Observations on the age, growth, reproduction and food of the chub Squalius cephalus (L.) in the River Stour, Dorset

Opercular bones from 399 chub from the River Stour, Dorset were used for age and backcalculated growth measurements. Scales were only used to aid the interpretation of difficult operculars. Annuli were laid down through the period mid-April to mid-June. Growth in length was minimal between October and March. Growth rates were similar to those published for chub in other European waters, but the Stour chub were longer-lived and attained a greater ultimate size. Female chub grew faster than the males. Spawning occurred from late May into June and elaboration of the gonads took place between September and May. Immature chub had an annual cycle of condition; the 0 group having a maximum in August and older immature fish reaching their maximum in June. Both categories had a minimum condition in early spring. The cycle of gonad development affected the condition of mature fish. The numbers of eggs in chub of lengths 359–467 mm ranged from 27 000–65 000. Some females attained sexual maturity at age V or VI, but most by age VII. The majority of males matured at age V, though some at ages III or IV. Growth rates and year-class strengths varied from year to year but independently of one another. Thirty-one per cent of chub aged II and over belonged to the 1959 year class. Young chub ate insect larvae and small crustacea, but the occurrence of fish and macrophytes was greater in the diet of older fish.     

Genetic divergence and morphological difference between the spotted and common mackerel

Three types of mackerel,Scomber species, the typical spotted, non-spotted and their intermediate types collected from three locations in the adjacent waters of Japan were analyzed by starch gel electrophoresis and examined for some morphometric and meristic characters. Clear genetic differences between the typical spotted and non-spotted were observed at six loci, i.e.αGpd-B, Idh-A, Ldh-A, Sod, Hem-1 andHem-2, out of the 23 loci examined. The intermediate individuals genetically belong to either the spotted or non-spotted type and a genetical hybrid possessing both kinds of allele from the two typical types was not observed at all. These three types could be divided into two groups. Average genetic distances for intra- or inter groups were estimated as 0.006 and 0.414, respectively. Two morphological characters, the length of separated anal spine and the number of interneurai spines associated with the first dorsal fin spine, were found to be significant character separating the two groups. From the above results, the two groups of mackerel were clearly divided into two separate species,Scomber australasicus andS. japonicus.     

Natural variation in the genes responsible for maturity loci E1, E2, E3 and E4 in soybean

Background and Aims

The timing of flowering has a direct impact on successful seed production in plants. Flowering of soybean (Glycine max) is controlled by several E loci, and previous studies identified the genes responsible for the flowering loci E1, E2, E3 and E4. However, natural variation in these genes has not been fully elucidated. The aims of this study were the identification of new alleles, establishment of allele diagnoses, examination of allelic combinations for adaptability, and analysis of the integrated effect of these loci on flowering.

Methods

The sequences of these genes and their flanking regions were determined for 39 accessions by primer walking. Systematic discrimination among alleles was performed using DNA markers. Genotypes at the E1–E4 loci were determined for 63 accessions covering several ecological types using DNA markers and sequencing, and flowering times of these accessions at three sowing times were recorded.

Key Results

A new allele with an insertion of a long interspersed nuclear element (LINE) at the promoter of the E1 locus (e1-re) was identified. Insertion and deletion of 36 bases in the eighth intron (E2-in and E2-dl) were observed at the E2 locus. Systematic discrimination among the alleles at the E1–E3 loci was achieved using PCR-based markers. Allelic combinations at the E1–E4 loci were found to be associated with ecological types, and about 62–66 % of variation of flowering time could be attributed to these loci.

Conclusions

The study advances understanding of the combined roles of the E1–E4 loci in flowering and geographic adaptation, and suggests the existence of unidentified genes for flowering in soybean,     

Factors affecting the establishment success of the invasive piscivorous chub in small irrigation ditches in northern Kyushu,Japan

Habitat matching is an important factor for the establishment of alien fishes in recipient environments. The piscivorous chub, Opsariichthys uncirostris uncirostris, originally exclusively inhabited large water areas; however, the species has been introduced and established in small irrigation ditches on Kyushu Island. The evaluation of the habitat use of the piscivorous chub suggested that this alien fish uses the lentic-type irrigation ditches as a nursery ground and the lotic-type ditches as a spawning ground. This finding implies that the establishment of the piscivorous chub may require connectivity between lentic and lotic habitats.     

Spawning migrations of the chub in the River Spree, Germany

Out of 52 chub Leuciscus cephalus tagged with internal radio‐transmitters, 46 migrated to spawning grounds in the River Spree, Germany. All chub started their first spawning migration on 21–22 May in both years of study, 1995 and 1996. They arrived at three different spawning grounds within a day and left after 1 to 6 days to return to their original sites. On 17 and 18 June 1995 and 1996 chub started their second spawning migration with the same pattern as the first. About 60% of all migrating chub moved between 1 and 13 km upstream to the nearest spawning ground. Some exceptions were observed: a few chub did not use the nearest spawning ground, some chub used different spawning grounds at first and second spawning or moved downstream to a spawning ground. Spawning grounds were characterized by 0·4 m s⁻¹ current velocity, shallow depth (0·1–0·8 m) and a stony bottom, where most eggs adhered.     

Genetics of Vegetative Incompatibility in Cryphonectria parasitica

Vegetative incompatibility in the chestnut blight fungus, Cryphonectria parasitica, in Europe is controlled by six unlinked vic loci, each with two alleles. Four previously identified vic loci (vic1, vic2, vic3, and vic4) were polymorphic in European vegetative compatibility (vc) types. Two new loci, vic6 and vic7, also were identified among European vc types. In one cross, vic genes segregated independently at five loci, and 194 progeny were assigned to 32 vc types; none of these loci were linked. A total of 64 vc types were identified from all crosses. All 64 genotypes possible from six vic loci, each with two alleles (2⁶ = 64), were identified and assigned to vc types. Based on our model, vc types v-c 5 and v-c 10, which had been used in previous genetic studies, differ by only five vic genes. Future studies of vc types in C. parasitica can use knowledge of vic genotypes for analysis of population genetic structure based on vic allele frequencies and to determine the effect of each vic gene on virus transmission between vc types.     

Molecular and morphologic approaches to discrimination of variability patterns in chub mackerel, Scomber japonicus

The systematic status and the evolutionary biology of chub mackerel (Scomber japonicus) in the South West Atlantic Ocean is confusing with an unknown degree of genetic differentiation and reproductive isolation between units. Simultaneous genetic and morphologic analyses were made on 227 fish collected from two areas of the South West Atlantic Ocean and one from the Mediterranean Sea. The genetic analysis was based on 36 protein-coding loci, 16 of which were variable. The morphologic analyses include six morphometric length measurements and a meristic character. Correspondence between genetic and morphologic variability patterns indicates isolated Mediterranean and Southwest Atlantic subgroups of S. japonicus and, less clearly, possible additional divergence in two regional stocks within the latter group. The most conservative approach to management is to manage the stocks independently of one another.     

Genetic differentiation among eight color types of the freshwater goby,Rhinogobius brunneus,from western Japan

Genetic differentiation among eight color types of the freshwater goby,Rhinogobius brunneus, from the western part of Japan was investigated by using electrophoretic methods. Four sympatric types (Cross-band, Dark, Cobalt and Large-Dark (A) types) did not share alleles at between one and six loci out of 12 loci tested. No hybrid specimens were found among these types. The average genetic distances among these four types ranged from 0.13 to 0.72, which fall within the range of values among congeneric species of fishes. The average genetic distances among the other four types, Large-Dark (B), Orange, Shinji-Lake and Boso types, were only 0.01 to 0.03, and fall within the range of values among conspecific populations. These results suggest that the former four types are clearly discrete species and the latter four types may be considered as intraspecific variations of a fifth species.     

Gene diversity at allozyme loci in the cottonwood leaf beetle,Chrysomela scripta

Gene diversity was studied in beetles collected from a poplar plantation at Ames, Iowa. The beetles are widely distributed throughout North America wherever poplars (Populus spp) occur. Of 38 loci interpretable by simple Mendelian criteria, 22 were polymorphic (58%). Nei's mean heterozygosity per locus was 20.1±4.0%. A mean 2.29±1.35 alleles per locus was detected. The foregoing levels of diversity are typical of Coleoptera. There were significant deviations from Hardy-Weinberg expectations at six loci, and of these loci, four deviations were judged to have been caused by technical problems in resolving heterozygotes. Twelve polymorphic loci are considered suitable to carry out studies on the breeding structure of this important pest.     

Distribution and Habitat Use of Sturgeon Chubs (Macrhybopsis Gelida) and Sick-Lefin Chubs (M. Meeki) in the Missouri and Yellowstone Rivers, North Dakota

Sampling was conducted on the Missouri and Yellowstone rivers, North Dakota to obtain information on the distribution, abundance and habitat use of the sturgeon chub (Macrhybopsis gelida) and sicklefin chub (M. meeki)(Family Cyprinidae), two declining benthic fish species native to the Missouri River basin. The study area consisted of three distinct river segments, the Missouri River near Williston, the Missouri River near Bismarck (below Garrison Dam), and the Yellowstone River near its confluence with the Missouri River. Both species of chub were collected, mainly with a benthic trawl, throughout 94% of the range sampled in the Williston and Yellowstone segments. Sicklefin and sturgeon chubs were the second and third most abundant cyprinids, respectively, collected from the Williston and Yellowstone segments. Best-fit regression models indicated that the presence of sturgeon chubs increased with decreasing depth, increasing velocity and decreasing water clarity, and that the presence of sicklefin chubs increased with increasing depth, decreasing velocity and decreasing water clarity. In contrast, no chubs of either species were collected in trawls from the Bismarck segment. This segment had significantly deeper, faster, and clearer water than both the Williston and Yellowstone segments.