Cross-amplification of microsatellites from the codling moth Cydia pomonella to three other species of the tribe Grapholitini (Lepidoptera: Tortricidae)

This study examined cross-species amplification of 33 microsatellite markers, previously developed for Cydia pomonella, in three related fruit moth species of the same tribe (Grapholitini), namely Grapholita molesta, Grapholita funebrana and Grapholita lobarzewskii. Eight microsatellite loci yielded polymorphic products for G. molesta, nine for G. funebrana and 11 for G. lobarzewskii. At all these loci, the number of alleles ranged between four and 11 in G. molesta, and between four and nine in G. funebrana and G. lobarzewskii each. The successful cross-amplified loci can be used for research on population genetics and gene flow of the three target species.     

Eleven novel microsatellite markers developed from the living fossil Ginkgo biloba (Ginkgoaceae)

Ginkgo biloba L. (Ginkgoaceae), a famous and widely cultivated plant, is often referred to as a living fossil. Here we isolated and characterized 11 polymorphic microsatellite loci from G. biloba using a modified biotin-capture method. These loci showed high levels of allelic diversity, alleles sizes ranged from 4 to 16 per locus with polymorphism information content ranging from 0.432 to 0.909 (average = 0.752). The observed and expected heterozygosities ranged from 0.208 to 0.708 (average = 0.484) and from 0.501 to 0.934 (average = 0.795), respectively. These microsatellite markers can be used as genetic markers for further studies on genetic diversity, genetic structure and the gene flow among populations of this species.     

Genetic analysis of male fertility restoration in wild cytoplasmic male sterility<Emphasis Type="Italic"> G</Emphasis> of beet

Cytoplasmic male sterility (CMS) has been used in the breeding of sugar beet for decades but is also more generally an important feature of the reproductive system in its wild relative, Beta vulgaris ssp. maritima. Among the several CMSs found in wild populations, the G CMS is a mitochondrial variant of the respiratory chain. The segregants derived from a cross between a restored plant and a female (male-sterile) plant on G cytoplasm exhibited three sexual phenotypic classes: female, hermaphrodite and intermediate. The pattern of segregation suggests a genetic inheritance with two loci in epistatic interaction. Nevertheless, it was possible to apply a bulk segregant analysis approach to search for AFLP and microsatellite markers linked to the restorer locus (RfG₁) which controls the capacity to produce pollen [female versus non female (i.e. intermediates and hermaphrodites)] in the segregating population. A linkage group was constructed with four AFLP markers and nine microsatellites, and a total size of 40 cM (Kosambi). The closest marker, a microsatellite, was totally linked to RfG1, which was also flanked by two AFLP markers delimiting a 5 cM window. This linkage group was identified as being chromosome VIII where neither of the restorer loci of the Owen CMS are located.Communicated by H.C. Becker     

Isolation and characterization of polymorphic microsatellite loci in the field vole,Microtus agrestis,and their cross‐utility in the common vole,Microtus arvalis

We developed nine polymorphic microsatellite loci for the field vole, Microtus agrestis. The number of alleles ranged from five to 15 and observed heterozygosities ranged from 0.40 to 1.00. We also tested the microsatellite loci for amplification and polymorphism in the congeneric species Microtus arvalis. Five of the nine loci were successfully analysed in this species. The microsatellite markers will be employed in studies of reproductive success and fine‐scale spatial genetic structure.     

Isolation and characterization of 12 microsatellite loci for cross-species amplification in the cyprinid gudgeons Squalidus chankaensis and S. japonicus

Twelve dinucleotide markers were successfully isolated and characterized from a microsatellite‐enriched genomic library obtained for the gudgeon Squalidus chankaensis biwae. These markers were also available for the congeners S. c. tsuchigae and S. japonicus from Japan, which had five to 46 alleles and an expected heterozygosity ranging from zero to 0.946. Linkage equilibrium was observed at all loci, and most loci did not show significant deviation from Hardy–Weinberg equilibrium. The isolated microsatellite markers will be useful for genetic diversity studies of Squalidus populations.     

Isolation and characterization of microsatellite markers for Pinellia ternata and cross-species amplification

In this study, we isolated and characterized 12 microsatellite loci for Pinellia ternata. Polymorphism of these 12 loci was assessed in 46 individuals collected from two wild populations. All the loci were polymorphic with four to 13 alleles per locus and the observed and expected heterozygosities ranged from 0.312 to 0.680 and from 0.506 to 0.734, respectively. None of the loci showed significant deviation from Hardy–Weinberg equilibrium (P < 0.05). No significant linkage disequilibrium was observed between pairs of studied loci. In addition, most markers amplified successfully in three closely related taxa that are Pinellia cordata, P. peltata and P. pedatisecta. These microsatellite markers could provide a useful tool for genetic structure studies of the Pinellia species.     

Isolation and characterization of microsatellite loci in <Emphasis Type="Italic">Cryptocarya chinensis</Emphasis> in lower subtropical China

We report on the isolation and characterization of eight microsatellite markers from repetitive DNA enriched libraries for Cryptocarya chinensis from lower subtropical China. These are the first microsatellite loci reported for Cryptocarya species. The number of alleles ranged from three to nine. Observed and expected heterozygosities ranged from 0.0518 to 0.9910 and 0.5241 to 0.7935 for polymorphic loci, respectively. These markers will allow analyses of the baseline genetic variability and population structure of C. chinensis to enrich our scientific understanding of forest fragmentation on genetic health of this species and provide strategies for effective conservation and management in this area.     

Characterization of new microsatellite loci from Vitis vinifera and their conservation in some Vitis species and hybrids

We present here characterization data for seven new microsatellite markers designed from new microsatellite loci isolated from a microsatellite‐enriched DNA library from Vitis vinifera. The observed heterozygosity varied from 0.73 up to 0.93 and the number of alleles per locus ranged from 12 to 26. This high polymorphism makes these new markers interesting for use in genotyping studies and completing the set of microsatellite markers already available for V. vinifera. Additionally these seven new markers appear to be conserved in four other Vitis species and 15 Vitis hybrids used as rootstocks for V. vinifera cultivation.     

Isolation and characterization of microsatellite loci in the Japanese pipistrelle (Pipistrellus abramus)

We describe the first set of ten microsatellite markers isolated in Pipistrellus abramus. The number of alleles per locus ranged from 7 to 13. The observed and expected heterozygosities values ranged from 0.486 to 0.971 and from 0.752 to 0.876, respectively. Three loci revealed significant departure from Hardy–Weinberg equilibrium and no linkage disequilibrium was found between loci pairs. These informative microsatellite markers will be a powerful molecular tool for studying the population genetic structure of P. abramus, as well as other species of this genus.     

Isolation and characterization of the first microsatellite markers for the endangered swan mussel <Emphasis Type="Italic">Anodonta cygnea</Emphasis> L. (Bivalvia: Unionoidea)

Freshwater molluscs are globally critically threatened, yet little is known about their genetic diversity. We describe the isolation and characterization of the first microsatellite markers for the European swan mussel, Anodonta cygnea. Nine loci were polymorphic in a panel of 110 individuals from seven German and Estonian populations. Number of alleles ranged from 2 to 14 and heterozygosity levels ranged from 0.021 to 0.498 for H _o and from 0.069 to 0.634 for H _e. Deficiency of heterozygous genotypes was observed in six loci. The length of the microsatellite repeat motives was positively correlated with the allele numbers in the respective loci.     

Characterization of six polymorphic microsatellites for the polychaete tubeworm Hydroides elegans and cross‐species amplification in the congener Hydroides hexagonus

We isolated and characterized six polymorphic microsatellite loci for the polychaete tubeworm, Hydroides elegans. Two additional loci were not reliably scorable and estimates of heterozygosity were obtained for the other six. In addition, cross‐species amplification was successful for two loci using the congener H. hexagonus. Given that few microsatellite loci are available for polychaetes, these markers will be useful in assessing dispersal and gene flow in H. elegans and probably also other polychaetes.     

Isolation and characterization of microsatellite markers for the endangered <Emphasis Type="Italic">Taxus yunnanensis</Emphasis>

Eleven polymorphic microsatellite loci were characterized for the endangered conifer Taxus yunnanensis. Eight loci were isolated through SSR-anchored PCR, one locus was developed by cross-species amplification tests, while the last two loci were obtained from cross-species microsatellite sequences available in GenBank. Variability of these markers was tested in 48 individuals collected from its main distribution range in Southwest China. The number of alleles per locus ranged from 2 to 9, and the observed and expected heterozygosity varied from 0.000 to 0.625 and from 0.062 to 0.853, respectively. Ten of these eleven loci were significantly deviated from Hardy–Weinberg expectation, except TS07 which showed a distinct heterozygote excess. The availability of these new polymorphic microsatellite markers will provide an ideal marker system for detailed population genetics studies in T. yunnanensis and potentially also for closely related species.     

Development of twelve novel microsatellite markers in the medicinal mushroom Cordyceps militaris (Ascomycota: Clavicipitaceae) for its strain identification and genetic analysis

Cordyceps militaris (a caterpillar fungus), which belongs to the class Ascomycetes, has extensively been used for medicinal purposes in East Asia. Here, we isolated and characterized 12 microsatellite loci from the medicinal mushroom, C. militaris. Twenty‐nine individual samples were taken from a single locality in southwestern Korea and used to characterize the developed markers. The number of alleles of these loci ranged from two to 13 (mean allelic richness = 6.44). Observed heterozygosity and expected heterozygosity in the population ranged from 0.035 to 0.880 and from 0.035 to 0.886, respectively. Five of 12 loci significantly deviated from Hardy–Weinberg Equilibrium, most likely due to heterozygote deficiency caused by inbreeding. Tests of genotypic linkage disequilibrium between the 12 loci showed no significant association of alleles. These microsatellite markers will provide valuable tools for genetic analyses for the strain identification of C. militaris as well as for its resource conservation.     

Genetic Variability Assessed by Microsatellites in a Breeding Program of Pacific White Shrimp (<Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>)

Genetic diversity in a shrimp-breeding program was monitored for 2 generations by microsatellite DNA markers (Pvan1578 and Pvan1815) to establish levels of variation and proceed with a selection program. An increase in the number and frequencies of some alleles in both microsatellite loci from G₀ to G₂ was induced by foreign sire contributions. Most common alleles and high heterozygosities (around 70% in both loci) were maintained through the generations, indicating that there had not been a significant loss of genetic variability in the breeding program. However, when compared with variability in other wild and cultured stocks, the presence of 4 main alleles at both loci may be an indication that a certain reduction in variability already was present in the line used as founder stock (G₀). Therefore, it is recommended that additional genetic variability be introduced to the breeding stock by crossing it with a different line.     

Characterization of novel microsatellite loci in the great leaf-nosed bat, Hipposideros armiger and cross-amplification in other related species

Eight microsatellite loci were isolated from an enriched genomic library of the great leaf-nosed bat, Hipposideros armiger. The polymorphism of these loci was tested on a population of 48 individuals from Anhui Province, China. All loci revealed the polymorphism ranging from three to 12 alleles. The observed heterozygosity values were from 0.213 to 0.875 and expected heterozygosity from 0.232 to 0.820. No significant linkage disequilibrium was detected. Two loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction. In addition, successful cross-amplification also suggested that these microsatellite markers will facilitate research on the population genetics and gene flow of H. armiger and other related species.     

Development of STMS markers from the medicinal plant Madagascar periwinkle [Catharanthus roseus (L.) G. Don.]

We report the isolation and characterization of the first set of sequence‐tagged microsatellites sites (STMS) markers in Catharanthus roseus, a plant with a vast range of medicinal uses. The microsatellite loci were cloned from an enriched library constructed using degenerate primers. Based on the microsatellite motifs, seven STMS primer pairs were designed. They were used to amplify 32 accessions of C. roseus and one accession of Catharanthus trichophyllus. The primers amplified an average of 3.86 alleles per locus. The observed heterozygosity ranged from 0.2903 to 0.9688 with an average of 0.7511. The STMS markers of C. roseus also amplified corresponding loci in a related species (C. trichophyllus) suggesting conservation of the loci across the genus. These markers will prove useful for genetic diversity analysis and linkage map construction in C. roseus.     

Isolation and characterization of microsatellites loci in the lemon (Citrus limon)

This study reports the isolation and characterization of seven polymorphic microsatellite loci in Citrus. The loci were isolated from two libraries constructed from genomic DNA nonenriched for TC and AC repeats and enriched for AC repeats. These markers yielded four to nine alleles per locus (mean 6.14) in a survey of 32 Citrus cultivars. Average observed heterozygosity ranged from 0.43 to 0.72. The levels of polymorphism found in this study suggest that these microsatellite loci can become an important tool for genetic studies in Citrus.     

The development of eight microsatellite loci in the wild daffodil Narcissus triandrus (Amaryllidaceae)

We report microsatellite primer pairs for the wild tristylous daffodil, Narcissus triandrus (Amaryllidaceae). From enriched libraries, we identified 58 unique microsatellite loci. We designed primer pairs for 27 of these loci and screened genomic DNA from 38 to 40 adults from a single population. For eight polymorphic loci, the number of alleles per locus ranged from five to 17. As six primers also amplified loci in three other Narcissus species, including two horticultural varieties, we expect that some of these markers will be transferable to other Narcissus species.     

Development and characterization of microsatellite loci for Rosa odorata var. gigantea Rehder & E. H. Wilson (Rosaceae)

Eighteen microsatellite markers were developed from Rosa odorata var. gigantea (Rosaceae), including 11 new microsatellite markers and 7 modified microsatellite loci having been developed from other Rosa species. About 27 wild individuals from 3 populations were used to screen polymorphism of these 18 microsatellite makers. The average allele number of these markers was 3.9 per locus, ranging from 2 to 9. The expected and observed heterozygosities varied from 0.2711 to 0.8043 and from 0.0370 to 0.5556, respectively. Cross-species amplification in other eight Rosa species showed their potential use for congeneric species. These microsatellite primers will be used for population genetics studies, constructing genetic linkage maps or locating quantitative trait locus (QTL) of R. odorata var. gigantea and related species.     

High-density genetic and physical bin mapping of wheat chromosome 1D reveals that the powdery mildew resistance gene <Emphasis Type="Italic">Pm24</Emphasis> is located in a highly recombinogenic region

Genetic maps of wheat chromosome 1D consisting of 57 microsatellite marker loci were constructed using Chinese Spring (CS) × Chiyacao F₂ and the International Triticeae Mapping Initiative (ITMI) recombinant inbred lines (RILs) mapping populations. Marker order was consistent, but genetic distances of neighboring markers were different in two populations. Physical bin map of 57 microsatellite marker loci was generated by means of 10 CS 1D deletion lines. The physical bin mapping indicated that microsatellite marker loci were not randomly distributed on chromosome 1D. Nineteen of the 24 (79.2%) microsatellite markers were mapped in the distal 30% genomic region of 1DS, whereas 25 of the 33 (75.8%) markers were assigned to the distal 59% region of 1DL. The powdery mildew resistance gene Pm24, originating from the Chinese wheat landrace Chiyacao, was previously mapped in the vicinity of the centromere on the short arm of chromosome 1D. A high density genetic map of chromosome 1D was constructed, consisting of 36 markers and Pm24, with a total map length of 292.7 cM. Twelve marker loci were found to be closely linked to Pm24. Pm24 was flanked by Xgwm789 (Xgwm603) and Xbarc229 with genetic distances of 2.4 and 3.6 cM, respectively, whereas a microsatellite marker Xgwm1291 co-segregated with Pm24. The microsatellite marker Xgwm1291 was assigned to the bin 1DS5-0.70-1.00 of the chromosome arm 1DS. It could be concluded that Pm24 is located in the ‘1S0.8 gene-rich region’, a highly recombinogenic region of wheat. The results presented here would provide a start point for the map-based cloning of Pm24.