Development of polymorphic microsatellite markers for the tree pathogen and sapstain agent,Ophiostoma ips

Twelve pairs of simple sequence repeat markers (SSR) were developed using a single ascospore isolate of Ophiostoma ips, isolated from the bark beetle, Orthotomicus erosus, infesting Pinus elliottii in South Africa. All markers were found to be polymorphic when tested on seven isolates of O. ips collected from Austria, Chile, Israel, Mexico, South Africa, Sweden, and the USA. These will now be useful in population and phylogenetic studies on O. ips.     

Development and characterization of microsatellite markers for the fungus Ceratocystis fimbriata

Ceratocystis fimbriata is a serious fungal pathogen on a wide range of plants, but many cryptic species within C. fimbriata are apparently host‐specialized. Anchor polymerase chain reaction (PCR) and simple sequence repeat (SSR) enriched libraries were used to develop 16 microsatellite markers for C. fimbriata. All markers were polymorphic when tested against isolates from four host‐specialized lineages of the pathogen. These markers will be valuable for phylogenetic and population genetic studies, as well as for tracking accidental introductions of host‐specialized forms of the pathogen.     

Development and characterization of nine polymorphic microsatellite markers in the sand goby Pomatoschistus minutus (Gobiidae)

A microsatellite‐enriched genomic library was constructed for the sand goby, Pomatoschistus minutus (Pallas 1770), and nine polymorphic DNA microsatellite markers of high quality were successfully optimized. Characterization of 96 individuals from the Vaccarès lagoon (France) showed moderate to high levels of polymorphism (two to 54 alleles). All the markers conformed to Hardy–Weinberg equilibrium and showed no evidence of null alleles, large allele dropout, stuttering and linkage disequilibrium between pairs of loci. These markers successfully amplify in three closely related species and can be employed to investigate population genetic structure and to clarify paternity in Pomatoschistus species.     

Isolation and characterization of eight polymorphic microsatellite markers in Aphanius fasciatus (Teleostei: Cyprinodontidae)

The first set of eight polymorphic microsatellites markers was successfully optimized from a partial genomic library enriched for an AC motif of the killifish Aphanius fasciatus Nardo, 1827 (Teleostei, Cyprinodontidae), a brackish‐water fish widely distributed along the coasts of the Mediterranean Sea. The developed loci proved to be polymorphic, displaying from two to six alleles per locus with expected heterozygosity ranging from 0.277 to 0.752. All loci were under Hardy–Weinberg equilibrium, except Af8 that showed significant heterozygotes deficiency, and there was no evidence of linkage associations between any pair of loci. Markers reported here will be potentially useful in monitoring of genetic population structure in this species.     

Isolation and characterization of 15 polymorphic microsatellite markers for the devastating dry rot fungus,Serpula lacrymans

Fifteen polymorphic microsatellite markers were developed from microsatellite‐enriched DNA libraries of the devastating dry rot fungus, Serpula lacrymans. The loci exhibited two to four alleles per locus and the expected heterozygosity ranged from 0.13 to 0.47. The codominant markers, described here for this fungus, will permit further studies in population genetics and phylogeography of this economically highly important species.     

Isolation and characterization of polymorphic microsatellite markers for Alternaria alternata

Alternaria alternata is of major significance as a food and feed contaminant and is able to produce a range of mycotoxins that may elicit adverse effects in both animals and humans. We describe the isolation and characterization of five microsatellite markers for studying A. alternata. Marker polymorphism was screened in 64 isolates of A. alternata. The number of alleles per locus ranged from eight to 24, and allelic diversity ranged from 0.425 to 0.882. These markers will be useful in the study of relationships and population genetics amongst isolates of A. alternata.     

Development and characterization of 11 microsatellite markers in the rock sparrow,Petronia petronia

Eleven polymorphic microsatellite loci were isolated in the passeriform Petronia petronia using the enrichment protocol FIASCO (fast isolation by AFLP of sequences containing repeats). We detected three to 13 alleles per locus in 25 specimens collected from an Italian population. The level of expected heterozygosity ranged from 0.439 to 0.856. One locus is sex linked to the Z chromosome. The total exclusionary probabilities using these loci for the first and the second parent were 0.978 and 0.999, respectively. These are the first microsatellite loci characterized from the rock sparrow that can be used for estimating population structure and for large‐scale parentage analysis.     

Development of polymorphic simple sequence repeat markers for Pisolithus microcarpus

Six polymorphic simple sequence repeat (SSR) markers were developed for the ectomycorrhizal fungus Pisolithus microcarpus. A polymerase chain reaction (PCR)‐based technique was used in which random amplified polymorphic DNA (RAPD) fingerprints were probed with labelled SSR oligonucleotides by southern hybridization. The number of alleles per locus ranged from two to nine with expected heterozygosity values from 0.33 to 0.76. These loci will be potentially useful for genetic structure and gene flow studies of P. microcarpus populations. Cross‐species amplification with Pisolithus albus isolates at all loci was also observed.     

Isolation and characterization of polymorphic simple sequence repeat loci in Armillaria ostoyae

A modified hybridization strategy was used to construct a microsatellite enriched library from DNA of Armillaria ostoyae, a serious root pathogen on pine. Sequence characterization of 19 random clones revealed 12 distinct loci harbouring a repetitive motif. Primer design from the flanking regions allowed for their development as polymerase chain reaction based markers. Polymorphic assessment at both the population and global levels revealed levels of variation useful for genetic studies. The level of cross‐species amplification observed with closely related Armillaria species was high, raising the possible exploitation of these primers across the genus.     

Polymorphic markers for the sea cucumber Parastichopus californicus

Five polymorphic markers were isolated from a Parastichopus californicus genomic DNA library. In addition, a method was developed to extract DNA from both muscle and skin tissue. The number of alleles at these loci ranged from three to 14. Observed heterozygosity ranged from 0.2 to 0.9. These markers, the first of their type for a sea cucumber species, will prove useful for exploration of the population structure of P. californicus.     

Isolation and characterization of polymorphic tetranucleotide microsatellite loci in the pelagic perciform fish Pomatomus saltatrix (Linnaeus, 1766) from South Africa

Eight polymorphic microsatellite loci, containing simple tetranucleotide repeats, were isolated de novo from a Pomatomus saltatrix partial genomic library using the fast isolation by amplified fragment length polymorphism of sequences containing repeats protocol. These loci were further characterized in 100 individuals from two putative populations off the South African east coast. The loci are highly polymorphic with 18-37 alleles (on average 24 alleles/locus) and the observed heterozygosity in both populations was high (0.79). These loci will be used to assess population structuring in P. saltatrix along the southern African coast with consideration of implications for future management of this important linefish species.     

Development of polymorphic markers for the root pathogen Thielaviopsis basicola using ISSR‐PCR

Thielaviopsis basicola is a soil‐borne fungal pathogen affecting many important agricultural crops. Little is known regarding the population biology or origin of this pathogen. Polymorphic markers developed for Ceratocystis fimbriata, a species complex phylogenetically closely related to T. basicola, were tested and found not to be useful for T. basicola. In this study 14 primer pairs, seven of which resulted in the amplification of single polymorphic fragments in T. basicola were developed. These primers will enable further studies on this economically important pathogen, and will result in an enhanced understanding of its population structure in different parts of the world.     

Isolation and characterization of polymorphic microsatellite markers in the migratory freshwater fish Prochilodus costatus

We isolated six polymorphic microsatellite loci in the migratory freshwater fish Prochilodus costatus, which is an endemic species and important fisheries resource from São Francisco river basin, Brazil. We have evaluated the applicability of these loci to study genetic variation in wild population of this fish. Thus, based on the genotypes of 32–48 individuals, we detected two to 21 alleles per locus, observed and expected heterozygosities ranging from 0.19 to 0.89 and from 0.17 to 0.92, respectively. These polymorphic markers should provide efficient tools to study population genetic structure of this fish.     

Microsatellite markers for the Eucalyptus stem canker fungal pathogen Kirramyces gauchensis

Ten microsatellite markers were developed for the fungus Kirramyces gauchensis, which causes an important stem canker disease of Eucalyptus trees in plantations. Primers for 21 microsatellite regions were designed from cloned fragments. Fourteen of the primer pairs provided single amplicons and 10 of these were polymorphic for K. gauchensis. Allelic diversity ranged from 0.21 to 0.76 with a total of 30 alleles. None of the markers was able to amplify in the phylogenetically distinct but morphologically similar species Kirramyces zuluensis. The 10 characterized polymorphic microsatellite regions will be studied to determine the population structure of K. gauchensis in plantations of different countries.     

Isolation and characterization of fourteen polymorphic microsatellite markers in the viperine snake Natrix maura

Nineteen polymorphic microsatellite loci were identified and developed for Natrix maura. Polymorphism was assessed for 120 individuals sampled across four sampling sites from the French Pyrenees Mountains. The number of alleles per locus ranged from 3 to 15, and expected heterozygosity per locus ranged from 0.227 to 0.863. We tested for deviation from Hardy–Weinberg equilibrium and linkage disequilibrium and assessed the presence of null alleles for all loci, resulting in a selection of 14 high‐quality polymorphic markers. These markers will be extremely useful in identifying fine‐scale genetic structures and providing insight into conservation management plans of this species.     

Isolation and characterization of 17 polymorphic microsatellite loci for the three-toed woodpecker (Picoides tridactylus)

We describe primers and polymerase chain reaction conditions to amplify 17 di‐, tri‐ and tetranucleotide microsatellite loci from the three‐toed woodpecker (Picoides tridactylus). The primers were tested on 26 to 30 individuals from a single population breeding in southern Finland. The developed primer pairs yielded an average of 7.6 alleles per locus (range two to 15), an average observed heterozygosity of 0.69 (range 0.07 to 0.97), and an average polymorphic information content of 0.68 (range 0.06 to 0.90).     

Development of polymorphic markers for Cirsium arvense,Canada thistle,and their amplification in closely related taxa

Suppression of invasive Canada thistle, Cirsium arvense, with biological control agents has stalled because introduced agents were not host‐specific. To aid in the development of more effective management strategies, molecular markers are needed to examine the genetic structure of Canada thistle populations. Microsatellite (simple sequence repeat) markers were developed and intersimple sequence repeat (ISSR) markers were tested for North American populations. An average of nine polymorphic alleles per microsatellite locus and 11 per ISSR locus were detected. These will be used to examine the genetic structure of C. arvense in the northern Great Plains and their transferability to endemic Cirsium spp.     

Identification and characterization of microsatellite markers in Hermann's tortoise (Testudo hermanni,Testudinidae)

The isolation and characterization of six polymorphic loci from a Testudo hermanni genomic library is reported. Enrichment was performed for AC but four of the characterized microsatellites present also an additional motif. Variability was tested on populations of the two recognized subspecies, Testudo hermanni hermanni and Testudo hermanni boettgeri. For one locus, a size range specific for the subspecies T. h. hermanni was observed. These are the first primers identified directly in the genome of T. hermanni     

Identification and characterization of 21 polymorphic microsatellite loci from the mycophagous fly Drosophila neotestacea

Drosophila neotestacea is a mushroom‐feeding fly that is common in the temperate and boreal forests of North America. Here I describe the isolation and characterization of 21 polymorphic microsatellite loci from the D. neotestacea genome. In a sample of flies from Rochester, New York, the expected and observed heterozygosities ranged from 0.344 to 1.000 and from 0.384 to 0.923, respectively. Of the 21 markers, six were likely X‐linked, seven showed a departure from Hardy–Weinberg equilibrium, and none were found to be in linkage disequilibrium. These loci will facilitate future ecological and population genetic studies of D. neotestacea.     

Isolation and characterization of microsatellite markers from the phytopathogenic fungus Alternaria dauci

Eleven polymorphic microsatellite markers were isolated from the necrotrophic phytopathogenic fungus Alternaria dauci based on enriched genomic libraries. In order to assess allelic variability, the microsatellite loci were analysed in a collection of 43 isolates. The number of detected alleles in 11 loci ranged from two to 24 (mean 10.4). Test of cross-species amplification and sequencing of the resulting amplicons showed that some of these microsatellites could be used in different species such as Alternaria solani, Alternaria bataticola and Alternaria zinniae.