New microsatellite markers for the bush rat,Rattus fuscipes greyii: characterization and cross‐species amplification

We describe here 16 new microsatellite markers for the bush rat, Rattus fuscipes greyii, and characterize their cross‐species amplification within the Australian Rattus and at a greater level of divergence in Rattus rattus and Rattus norvegicus. Within R. f. greyii, all of the loci are highly polymorphic, with six to 24 alleles per locus across the species range and expected heterozygosity ranging from 0.48 to 0.90 per locus within a sample of 24 rats from a large population on Kangaroo Island. Cross‐species amplification rates were approximately 87% within the Australian Rattus and approximately 50% within R. rattus and R. norvegicus. These loci are highly polymorphic with a high success rate of cross‐species amplification, making them potentially useful for a wide range of genetic studies.     

A trial re-introduction of the Greater Stick-nest Rat (Leporillus conditor) in arid South Australia

Summary Eight Greater Stick‐nest Rats (Leporillus conditor S.) were re‐introduced into a 8 ha release pen within the Arid Recovery Reserve, South Australia, in September 1998. Their weight, reproductive condition, activity, habitat use and grazing impacts were investigated during the 8 month experimental trial. Greater Stick‐nest Rats increased in weight and bred successfully after the end of summer (April 1999). They consumed native vegetation including Bladder Saltbush (Atriplex vesicaria), Ruby Saltbush (Enchylaena tomentosa) and Sturt's Pigface (Gunniopsis quadrifida) but Sturt's Pigface was most heavily browsed and no monitored plants died from over‐browsing during the trial. Rats preferred dune habitat and the two largest female rats maintained discrete activity areas and shelter sites. Nocturnal activity of adult male rats was centred around adult females with both sexes generally staying within 150 m of daytime shelter sites. Shelter sites were commonly within dead Umbrella Wattle (Acacia ligulata) or Narrow‐leafed Hopbush (Dodonaea viscosa) shrubs covered in low vegetation, and rats also adapted burrows of other species to use as below‐ground components. Although supplementary water was provided and may have ensured survival over the hot summer months, the trial release was considered a success and a full‐scale re‐introduction of the Greater Stick‐nest Rat was conducted in 1999.     

Development and characterization of microsatellite markers for Prosopis chilensis and Prosopis flexuosa and cross‐species amplification

Prosopis chilensis and Prosopis flexuosa (Fabaceae) are closely related hardwood arboreal species that are widely distributed in the arid regions of Argentina. The development of highly polymorphic markers, such as microsatellites, is desirable for genetic studies of these species. Here, we present the development and characterization of six polymorphic microsatellite markers in P. chilensis and P. flexuosa. These markers showed a polymorphism information content between 0.14 and 0.85 and the number of alleles varied from two to 13 considering both species. All markers revealed a broad cross‐species affinity when tested in seven other Prosopis species. All primers amplified in at least five species.     

Development and cross‐species amplification of 18 microsatellite markers in the Sumatran tiger (Panthera tigris sumatrae)

Eighteen polymorphic microsatellite loci from the highly endangered Sumatran tiger (Panthera tigris sumatrae) were isolated and characterized. Upon polymerase chain reaction amplification, 16 of these markers produced a single, sharp band in all three tiger and 10 non‐tiger felid species examined. Of the two remaining loci, 6HDZ057 and 6HDZ635 failed to amplify genomic DNA from puma (Felis concolor) and cheetah (Acinonyx jubatus), respectively. The amplification of these markers across four genera is an indication of their usefulness for population genetics studies and conservation work in a wide range of felid species.     

Isolation and characterization of microsatellite loci in Pseudoplatystoma corruscans (Siluriformes: Pimelodidae) and cross‐species amplification

A total of five polymorphic microsatellites loci from Pseudoplatystoma corruscans were isolated and characterized. A population survey involving 43 specimens resolved a large number of alleles (range seven to eight among loci) and high observed heterozygosity (0.500–0.615), indicating its usefulness in population genetics studies. Cross‐species amplification was successful in four other Pimelodidae species.     

Ten polymorphic microsatellite markers in the invasive round goby (Neogobius melanostomus) and cross‐species amplification

Ten tetranucleotide, dinucleotide and compound microsatellite loci were isolated and characterized for the round goby, Neogobius melanostomus. Analysis of 64 gobies from one nonindigenous population in Lake Erie, Ontario, Canada, indicated that allele number varied from three to 12 per locus, while observed heterozygosity ranged between 0.33 and 0.86. Eight of these primers showed some amplification in other species in four genera. These newly developed microsatellite markers are a powerful tool that will provide insights into population structure and dispersal of the round goby in their novel environment.     

Development of microsatellite markers for southern catfish (Silurus meriaionalis) and cross‐species amplification in northern sheatfish (Silurus soldatovi)

Forty microsatellite markers were developed from a (CA)_n enrichment library created from the DNA of southern catfish (Silurus meriaionalis). Also, the population structure of northern sheatfish (Silurus soldatovi) was examined by 24 microsatellite loci. They are polymorphic in at least one of the two geographically distant populations sampled from the Heilongjiang River and the Songhuajiang River in North China, respectively. Unbiased expected heterozygosity levels varied from 0.435 to 0.946 and number of alleles per locus varied from three to 20. Results indicated that these microsatellite loci were highly polymorphic and could be used as genetic markers.     

Di‐ and tetranucleotide microsatellite markers for the Alpine newt (Triturus alpestris): characterization and cross‐priming in five congeners

We developed a set of di‐ and tetranucleotide microsatellite markers for the Alpine newt, Triturus alpestris. Polymorphism as detected in 39 individuals ranged from 3 to 32 alleles at a locus. Cross‐priming with samples of five other Triturus species showed extremely poor levels of cross‐species utility. Still, these markers are suitable for studies of inter‐ and intrapopulation genetic diversity in the focal species.     

Isolation,characterization and cross‐species amplification of microsatellite DNA loci in the tropical American yam Dioscorea trifida

We developed microsatellite markers in American yam (Dioscorea trifida). A microsatellite sequence‐enriched genomic library was screened, and after sequencing, primers were designed for 20 microsatellites. Among these, eight primer pairs yielded amplification products that were both interpretable and polymorphic in 24 yam cultivars. The number of alleles per locus ranged from two to 13 and the overall expected heterozygosity was around 0.5. Six of the eight Dioscorea trifida microsatellite loci gave amplification products in other Dioscorea species.     

Characterization of microsatellite DNA markers for a mahseer species,Tor tambroides (Cyprinidae) and cross‐amplification in four congeners

This study reports the isolation and characterization of microsatellite DNA markers in a mahseer species, Tor tambroides (Pisces, Cyprinidae). Of a total of 14 loci evaluated, 10 were polymorphic in T. tambroides samples, with an average of 2.86 alleles per locus. Deviations from Hardy–Weinberg equilibrium were observed at one locus and there was no indication of linkage disequilibrium among loci. A high level of cross‐amplification among four congeners was achieved, with 12 loci successfully amplifying and 11 loci showing polymorphism in at least one other species. These markers will be a useful resource for population genetic studies and broodstock management of closely related mahseer species.     

Development of microsatellite markers in Abies nordmanniana (Stev.) Spach and cross‐species amplification in the Abies genus

Abies nordmanniana (Stev.) Spach is a widely grown Christmas tree in Denmark, from where it is exported to most of Europe. An ongoing breeding programme is taking place, and as a tool for that, we report the development of five polymorphic nuclear microsatellite markers. To investigate the potential for transferring the markers to other species in the Abies genus, polymerase chain reaction amplification was tested in 19 other species. In general, amplification occurred in a very high proportion of the tested species.     

Isolation and characterization of microsatellite loci in Lampsilis abrupta (Bivalvia: Unionidae) and cross‐species amplification within the genus

We have documented the first microsatellites isolated from a unionid and demonstrated that these markers can be useful for surveys of neutral genetic variation in several Lampsilis species. We describe the isolation and characterization of 15 polymorphic microsatellite DNA loci for the endangered unionid Lampsilis abrupta. Among individuals from five collections, allelic diversity ranged from six to 17 alleles and averaged 10.4 alleles per locus. Individual heterozygosity was observed to range from 20.0% to 86.7% and averaged 46.9%. Cross‐species amplification was investigated in nine additional Lampsilis species. A high level of flanking sequence similarity was inferred as 13 of 15 loci amplified in at least seven species.     

Isolation and characterization of novel microsatellite loci for the short‐beaked common dolphin (Delphinus delphis) and cross‐amplification in other cetacean species

Seven tetranucleotide and three dinucleotide microsatellite loci were isolated from the short‐beaked common dolphin (Delphinus delphis). Number of alleles and observed heterozygosity ranged from three to twelve (mean 8.4) and from 0.087 to 0.978 (mean 0.741), respectively. All loci were tested in four other odontocete species and showed polymorphisms in most locus–species combinations. This panel of nuclear markers will be useful for the investigation of population ecology and social interactions in common dolphins and other cetacean species.     

Ten new microsatellite loci for the yellowhammer (Emberiza citrinella) and their cross‐species applicability among related taxa

We describe isolation and characterization of the first microsatellite loci specifically developed for a very common European passerine bird, the yellowhammer Emberiza citrinella. Nine out of our 10 loci are polymorphic within the species E. citrinella. Number of alleles ranged from two to 21 per locus and observed heterozygosity between 0.20 and 0.91. Four primer pairs also yielded reproducible results in other species of Emberizidae. These loci comprise a set of molecular markers for various applications, from moderately polymorphic loci suitable for population studies to highly polymorphic loci for pedigree analysis in Emberizidae.     

Isolation and characterization of nine microsatellite markers from Coffea arabica L., showing wide cross‐species amplifications

Genetic improvement of coffee (Coffea arabica L.) is constrained by low genetic diversity and lack of genetic markers, suitable screening tools, information on the genetic make‐up of available gene pool and long generation time. In this context, use of DNA markers such as microsatellites that provide high genetic‐resolution becomes highly desirable. Here, we report the development of nine new microsatellite markers from partial genomic library of an elite variety of Coffea arabica. The developed microsatellites revealed robust cross‐species amplifications in 17 related species of coffee, and their Polymorphic Information Content varied from 0 to 0.6, 0 to 0.78 and 0.67 to 0.90 for the arabica, robusta genotypes and species representatives, respectively. The data thus suggest their potential use as genetic markers for assessment of germplasm diversity and linkage analysis of coffee.     

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.     

Isolation and characterization of microsatellite markers in the gecko Gekko swinhonis and cross‐species amplification in other gekkonid species

Gekko swinhonis is a gekkonid lizard endemic to China, inhabiting Loess Plateau, Huabei Plain, Huanghuai Plain and areas north of the Yangtze River. We characterized 21 dinucleotide microsatellite loci from an AC/AG‐enriched genomic library of G. swinhonis. The number of alleles per locus ranged from eight to 24 and the observed and expected heterozygosities ranged from 0.160 to 0.834 and from 0.584 to 0.917, respectively. We also tested the utility of these loci in other Gekko and Hemidactylus species; many of these loci can be cross‐amplified.     

Novel microsatellite DNA markers for the threatened African endemic tree species,Milicia excelsa (Moraceae), and cross‐species amplification in Milicia regia

Eleven microsatellite primer pairs were developed for the tropical African tree Milicia excelsa. Genomic DNA was enriched for dinucleotide (TC_n and TG_n) and tretranucleotide (GATA_n), and 188 random clones were sequenced from both orientations. We designed and tested 44 oligonucleotide primer pairs, which were evaluated using genomic DNA from 30 M. excelsa mature trees collected from a natural population in Benin. Eleven of the 44 markers showed good amplification and were polymorphic. The number of putative alleles for polymorphic primer pairs varied from three to seven, with expected and observed heterozygosities ranging from 0.10 to 0.64 and from 0.10 to 0.80, respectively. All 11 loci amplified the related species Milicia regia, indicating that these primers will be useful for population and ecology genetic studies in other species of the genus Milicia.     

Isolation and characterization of 15 microsatellite loci in the poplar rust fungus,Melampsora larici‐populina,and cross‐amplification in related species

We developed 15 microsatellite loci in the poplar rust fungus, Melampsora larici‐populina, using two enrichment protocols. Polymorphism of each locus was assessed on a panel of 30 isolates, comprising three subpanels (world, regional and local scales). Thirteen loci were polymorphic with three to eight alleles detected. The 15 loci were also tested on five related Melampsora species, M. allii‐populina, M. medusae f. sp. deltoidae, M. larici‐tremulae, M. rostrupii and M. pinitorqua, and partial or global cross‐amplification events were detected.     

Isolation and characterization of 11 new microsatellite markers for Macaranga (Euphorbiaceae)

We provide primer sequences for 11 new polymorphic microsatellite markers developed in the tropical ant-plant genus Macaranga (Euphorbiaceae), after enrichment cloning of Macaranga tanarius and Macaranga hypoleuca. Allele numbers per locus ranged from two to 16 among 20 accessions of M. tanarius, and from three to 10 among 22 accessions of M. hypoleuca. Observed and expected heterozygosities ranged from 0.150 to 0.900 and from 0.375 to 0.894 in M. tanarius, and from 0.545 to 1.000 and from 0.434 to 0.870 in M. hypoleuca, respectively. Six of the 11 primer pairs successfully cross-amplified polymorphic polymerase chain reaction products in Macaranga winkleri.