Characterization of six microsatellite loci in the African wild silk moth (Gonometa postica,Lasiocampidae)

Six microsatellite markers were developed for the African wild silk moth, Gonometa postica (Lasiocampidae), using an enrichment protocol. The total number of alleles ranged from three to 17 for a sample of 130 individuals across the distribution of the species. Observed levels of heterozygosity ranged from 0.17 to 0.78. Deviation from Hardy–Weinberg equilibrium detected in some loci is probably the result of large interannual population size fluctuations characteristic of this species. No evidence of linkage disequilibrium was detected among loci. These loci will be useful for the inference of demographic processes in a moth species that is of potential economic importance.     

Characterization of 31 EST-derived microsatellite markers for the pearl oyster Pinctada martensii (Dunker)

We developed and characterized 31 microsatellite markers from expressed sequence tags of Pinctada martensii (Dunker). The number of alleles per locus ranged from 4 to 18 as determined in 44 individuals from a wild population. The expected heterozygosity ranged from 0.4121 to 0.9436, while the observed heterozygosity ranged from 0.4054 to 0.7273. Most of the loci are in Hardy-Weinberg equilibrium. These markers should be useful for population genetics studies, parentage and genome mapping in this species.     

基于SSR分子标记分析云南月季种质资源亲缘关系(英文)

利用简单重复序列SSR(simple sequence repeat)标记技术对48份月季种质资源(包括14份野生种、20份古老月季品种和14份栽培品种)的遗传多样性和亲缘关系进行了分析.结果表明,(1)18对SSR引物在18个SSR位点上共检测到160个等位基因,每一位点的等位基因变幅为6～13个,平均8.9个,材料间遗传相似系数变化范围为0.157 8～0.754 9,表明在分子水平上云南月季种质资源具有丰富的遗传多样性.(2)聚类分析结果显示,在相似系数为0.44处,可将 14个野生种明显分为6个组,这与植物形态学分类结果上基本一致;在遗传相似系数为0.40水平上将48份供试材料分为八大组群.(3)亲缘关系分析结果显示,5个野生种和所有古老月季品种与大多数栽培品种的亲缘关系较近.     

Cross-species amplification of cassava (Manihot esculenta) (Euphorbiaceae) microsatellites: allelic polymorphism and degree of relationship

Microsatellite amplification was performed on cassava (Manihot esculenta) and six other different species (all wild) of the Manihot genus. We used ten pairs of microsatellite primers previously developed from cassava, detecting 124 alleles in a sample of 121 accessions of the seven species. The number of alleles per locus ranged from four to 21 alleles, and allelic diversity was greater in the wild species than in cassava. Seventy-nine alleles, including unique ones, were detected in the wild species but were not found in the crop. The lower level of heterozygosity in some wild species probably resulted from a combination of fine-scale differentiation within the species and the presence of null alleles. Overall, microsatellite primers worked across the genus, but, with increasing genetic distance, success in amplifying loci tended to decrease. No accession of M. aesculifolia, M. carthaginensis, and M. brachyloba presented a banding pattern at locus Ga-140; neither did one appear for M. aesculifolia at locus Ga-13. Previous work with amplified fragment length polymorphism (AFLP) markers and this microsatellite analysis show that these three wild taxa are the most distant relatives of the crop, whereas the wild forms M. esculenta subsp. flabellifolia and M. esculenta subsp. peruviana appear to be the closest.     

Microsatellite loci from red sea urchins (Strongylocentrotus franciscanus)

Strongylocentrotus franciscanus is a keystone marine herbivore in the Pacific North West of North America, that is commercially harvested for its roe in wild fisheries and is under development as an aquaculture species. We report the development of 11 novel, highly polymorphic di‐ and trinucleotide loci for use in population studies of this species. In a survey of over 200 individuals from three coastal sites in British Columbia, observed heterozygosities of the loci ranged from 0.39 to 0.85 and an average of 26 alleles per locus were resolved. Seven of the 11 microsatellite loci were in Hardy–Weinburg equiulibruim (HWE).     

Microsatellite loci for Gossypium davidsonii (Malvaceae) and other D-genome, Sonoran Desert endemic cotton species

? Premise of the study: Microsatellite primers previously developed for domesticated cotton (Gossypium hirsutum; tetraploid) were screened for their utility in investigating genetic structure and gene flow within G. davidsonii and five other wild, Mexican, D-genome cotton species (all diploid). ? Methods and Results: We screened 50 microsatellite primer pairs from the Cotton Marker Database, identifying 10 loci as polymorphic within G. davidsonii. In genotyping approximately 200 individuals from four populations, we found that the number of alleles per locus ranged from seven to 17, and mean observed and expected heterozygosity ranged from 0.145 to 0.492 and from 0.436 to 0.734, respectively. We genotyped six to 20 individuals from each of the remaining species, finding these 10 loci to cross-amplify in all cases and in most cases to be polymorphic. ? Conclusions: These markers may be useful for further investigation of population genetics of G. davidsonii and other wild D-genome cotton species.     

Multiplex PCR system applied for analysing microsatellite loci of Schlegel's black rockfish,Sebastes schlegeli

Two multiplex PCR amplifications were performed to analyse six microsatellite loci of Schlegel's black rockfish, Sebastes schlegeli, an important commercial fish in the northern part of Japan and an important species for the stock enhancement program in this area. We analysed 67 wild samples from Yamada Bay, Iwate Prefecture, Japan. The observed genotype frequencies agreed with the Hardy–Weinberg expectations at all loci, and the observed heterozygosities ranged from 0.072 to 0.897.     

Isolation and characterization of polymorphic microsatellites in the groundnut seed beetle,Caryedon serratus (Coleoptera,Bruchidae)

Caryedon serratus feeds on the seeds of four genera of wild Caesalpinioideae. It recently became a major pest of groundnut in Africa. To determine the extent of gene flow and isolation between wild and groundnut‐feeding populations, we isolated and characterized 10 microsatellite loci in this species. Polymorphism ranged from three to 15 alleles in the 80 individuals collected in four localities in Senegal on five host plants.     

Isolation and characterization of polymorphic microsatellite markers for the endangered Korean freshwater fish Hemibarbus mylodon

Korean barbel, Hemibarbus mylodon, is an endangered freshwater species endemic to Korea. In order to undertake a conservation program for this species in Korea, it is essential to evaluate its genetic diversity and population structure. For this purpose, we developed 10 polymorphic microsatellite markers and examined their allelic variation using a total of 60 wild individuals collected from three different localities. The number of alleles per locus ranged from three to 26. The expected and observed heterozygosities per locus ranged from 0.19 to 0.94 and from 0.20 to 0.98, respectively. Significant departures from Hardy–Weinberg equilibrium expectations occurred in four loci. These markers will be useful for the assessment of genetic diversity for this species.     

Seventeen novel microsatellite markers from an enriched library of the pest species Bactrocera dorsalis sensu stricto

Seventeen novel microsatellite markers were isolated and characterized from an enriched DNA library of the pest species Bactrocera dorsalis sensu stricto. The polymorphism of these loci was tested in individual fruit flies from a Thai mass‐rearing strain and three wild Asian populations: two from Thailand and one from Myanmar. Allele numbers per locus ranged from two to 15. In the natural populations, a high level of polymorphism was detected in all loci suggesting the usefulness of these markers to quantify the genetic variation in this highly invasive insect.     

Development and characterization of 30 polymorphic microsatellite markers for the Atlantic surfclam, Spisula solidissima (Dillwyn, 1817)

Thirty polymorphic microsatellite markers were developed for the Atlantic surfclam, Spisula solidissima, from an enriched library and characterized in 24 clams from a wild population. The number of alleles ranged from 3 to 16 per locus. The expected and observed heterozygosities ranged from 0.1942 to 0.9238 and 0.0833 to 0.875 respectively. Six loci showed significant (P < 0.05 after Bonferroni correction) deviation from Hardy–Weinberg equilibrium, probably because of the presence of null alleles. Three primer pairs amplified duplicated loci with two involving tandem mini‐satellite repeats. Most of the microsatellite markers developed here should be useful for genetic studies in this species.     

A simple method for isolation of microsatellites from the mudskipper (Boleophthalmus pectinirostris), without constructing a genomic library

The mudskipper Boleophthalmus pectinirostris is an important aquaculture species in China. It is endangered due to the loss and fragmentation of natural habitat, seawater pollution and overfishing. In this study, ten polymorphic microsatellite loci were isolated by the 5′-anchored polymerase chain reaction technique. Population genetic parameters were estimated on 60 wild individuals collected from Jiuduansha wetland in Yangtze River estuary. The number of alleles ranged from 3 to 14. The observed(H _O) and expected(H _E) heterozygosities ranged from 0.2500 to 0.8500 and from 0.3679 to 0.8692, respectively. This suite of microsatellite loci will facilitate future studies of the genetic status of wild and hatchery bred populations of B. pectinirostris.     

High variability and disomic segregation of microsatellites in the octoploid Fragaria virginiana Mill. (Rosaceae)

The objectives of the present study were to develop microsatellite markers for the wild strawberry, Fragaria virginiana, to evaluate segregation patterns of microsatellite alleles in this octoploid species, and assess genetic variability at microsatellite loci in a wild population. A genomic library was screened for microsatellite repeats and several PCR primers were designed and tested. We also tested the use of heterologous primers and found that F. virginiana primers amplified products in cultivated strawberry, Fragaria × ananassa Duch. and Fragaria chiloensis. Similarly, microsatellite loci developed from cultivated strawberry also successfully amplified F. virginiana loci. We investigated four microsatellite loci in detail, three developed from F. virginiana and one from cultivated strawberry. A survey of 100 individuals from a population of F. virginiana in Pennsylvania demonstrated high heterozygosities (H_e or gene diversity ranged from 0.80 to 0.88 per locus) and allelic diversity (12–17 alleles per locus), but individual plants had no more than two alleles per locus. Segregation patterns in parents and progeny of two controlled crosses at these four loci were consistent with disomic Mendelian inheritance. Together these findings suggest that the genome of F. virginiana is "highly diploidized" and at least a subset of microsatellite loci can be treated as codominant, diploid markers. Significant heterozygote deficiencies were found at three of the four loci for hermaphroditic individuals but for only one locus among females in this gynodioecious species.Communicated by J. Dvorak     

Polymorphic microsatellite loci for species of Australian freshwater cod, <Emphasis Type="Italic">Maccullochella</Emphasis>

The Australian freshwater cod genus, Maccullochella is represented by three species: Murray cod, M. peelii peelii, eastern freshwater cod, M. ikei, and trout cod, M.macquariensis. Seven novel microsatellite loci from M. ikei and six previously published loci from M. peelii peelii were tested on wild populations of Murray, eastern and trout cod. Levels of polymorphism varied between species with 13 loci polymorphic in Murray cod, 9 in trout cod and 7 in eastern cod. Observed heterozygosities ranged from 0.053 to 0.842. This suite of microsatellite loci will facilitate future studies of the genetic status of wild and hatchery bred populations of Maccullochella.     

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.     

Isolation and characterization of microsatellite DNA loci from the golden cuttlefish,Sepia esculenta Hoyle (Cephalopoda)

The first microsatellite markers were isolated from the golden cuttlefish, Sepia esculenta Hoyle. Eleven primer sets were designed to amplify the marker sequences via polymerase chain reaction. The 45–50 individuals from one wild population in the coastal waters of Ehime Prefecture, Japan were used to screen polymorphism in the 11 microsatellite loci. All the microsatellite loci were polymorphic, with the range of alleles from seven to 27 per locus. The observed and expected heterozygosities ranged from 0.380 to 0.980 and from 0.654 to 0.940, respectively. These marker loci except for one locus showing significant deviation from Hardy–Weinberg equilibrium will be useful for the assessment of genetic variation and population structure of this species.     

Isolation and characterization of ten new polymorphic microsatellite loci in the mud crab, Scylla paramamosain

We developed polymerase chain primers for ten microsatellite loci in the mud crab, Scylla paramamosain. All markers were obtained from a (CA)₁₅ and (CT)₁₅-enrichment DNA library, and characterized in 30 individuals from one wild population. The number of alleles per locus varies between 8 and 18, and the observed and expected heterozygosities ranged from 0.6207 to 0.9333 and from 0.5886 to 0.9243, respectively. These polymorphic loci provide a valuable tool for population genetic analysis and parentage determination in this species.     

Isolation and characterization of microsatellites in yellow perch (Perca flavescens)

A total of 45 microsatellite loci from yellow perch, Perca flavescens, were isolated and characterized. Among the 45 microsatellite loci, 32 had more than two alleles. A wild population of P. flavescens (n = 48) was used to examine the allele range of the microsatellite loci. Mendelian inheritance of alleles was confirmed by examining the amplified products in pair‐mated families. The number of alleles for the 32 polymorphic loci varied from two to 16, and observed heterozygosity ranged between 0.024 (YP79) and 0.979 (YP60). Cross‐species polymorphic amplification in four other Percidae species was successful for 22 loci.     

中国特有单种属血水草的微卫星分子标记开发与评价

血水草(Eomecon chionantha Hance)是中国亚热带地区特有的单种属植物。本研究采用(AG)15、(AC)15两种生物素探针及磁珠富集法从血水草基因组中开发并筛选出13个微卫星位点,且在该物种中能进行稳定的PCR扩增并表现出多态性。利用筛选出的13个SSR多态性标记对血水草4个野生居群24个个体进行PCR扩增,结果显示每个微卫星位点的等位基因数为2~7个,观测杂合度(HO)和期望杂合度(HE)分别为0.125~0.792和0.299~0.691。说明这些SSR标记可用于血水草遗传多样性的后续研究,并揭示该物种地理分布格局形成的原因,有助于保护和利用中国亚热带地区的血水草野生资源。     

Isolation and characterization of microsatellite loci in Santalum lanceolatum and Santalum leptocladum (Santalaceae)

? Premise of the study: Microsatellite primers were developed for the first time in the native Australian sandalwood species Santalum lanceolatum. ? Methods and Results: Using an enrichment cloning protocol, five novel polymorphic codominant loci were developed and characterized in S. lanceolatum and S. leptocladum. In addition to these, three existing microsatellite loci from other sandalwood species were successfully amplified and characterized for S. lanceolatum and S. leptocladum. Among the eight loci, allelic diversity ranged from 4 to 29. ? Conclusions: Primers will be useful for studies of clonality, genetic diversity and spatial genetic structure in wild populations. When coupled with other molecular techniques will help investigate the relationship between S. lanceolatum and S. leptocladum, species of commercial and conservation interest.