Microsatellite loci for the New Zealand trematode Maritrema novaezealandensis

Seven polymorphic microsatellite loci were isolated and characterized from the coastal New Zealand trematode Maritrema novaezealandensis, an important driving force in the population dynamics of its snail and crustacean intermediate hosts. Observed heterozygosities ranged from 0.33 to 0.82, and three to 17 alleles were detected in 22 trematode sporocysts. These loci will be integral for identifying parasite genetic clones within hosts and addressing a range of questions concerning the molecular ecology of this species.     

Microsatellite DNA markers for Lysiphlebus testaceipes

Microsatellite loci were isolated from the aphid parasitoid Lysiphlebus testaceipes (Hymenoptera: Braconidae). Ten loci were obtained from an enriched partial genomic library. Genetic diversity was analysed at seven of these loci and two natural populations, one on oleander and one on citrus. The observed number of alleles ranged from one to 17, and the observed heterozygosity ranged from 0.37 to 0.82. In both populations, no departure from Hardy–Weinberg equilibrium was detected except for one locus. The differentiation between the two populations was characterized by an F_ST of 0.09.     

藏酋猴微卫星富集文库的构建及微卫星分子标记的筛选

通过磁珠富集法构建了藏酋猴AC重复和AAAG重复的微卫星富集文库,分离微卫星序列并对其进行分析。将藏酋猴基因组DNA经Sau3AI酶切后纯化回收,连接特定接头。用生物素标记的探针与酶切片段杂交,捕获300～1000bp片段,随后将获得的片段连接到pMD-19T载体上,转化至JM109中,成功构建藏酋猴微卫星富集文库。(AC)n富集文库和(AAAG)n富集文库的阳性克隆率分别为50%和10%左右。根据测序得到的48个微卫星序列成功设计了24对引物,最终筛选出6个微卫星标记,这些标记将为藏酋猴的遗传多样性研究、圈养种群结构的分析和遗传图谱的构建等奠定一定的基础。     

Microsatellite loci for the progenetic trematode,Coitocaecum parvum (Opecoelidae)

Nine polymorphic microsatellite loci were characterized from the freshwater trematode Coitocaecum parvum. This parasite can either reproduce sexually in the definitive host or produce eggs by selfing inside its second intermediate host. Two to 11 alleles per locus were detected in 24 trematode sporocysts and observed heterozygosities ranged from 0.04 to 0.96. These loci will be useful for identifying parasite genetic clones within hosts and testing for effects of relatedness on parasite life history strategy.     

Microsatellite DNA markers for a grasshopper: Prionotropis hystrix rhodanica (Orthoptera,Pamphagidae)

Microsatellite loci were isolated from Prionotropis hystrix rhodanica, an endangered grasshopper species, endemic to a steppic area southeast of France. Six polymorphic loci were obtained from an enriched partial genomic libray. A high genetic diversity was observed at all loci, with an observed number of alleles ranging from seven to 29, and an observed heterozygosity between 0.71 and 0.90. Test of cross‐amplifications on two closely related taxa are presented.     

Microsatellite DNA markers for the pea aphid Acyrthosiphon pisum

Microsatellite loci were isolated from enriched partial genomic libraries of Acyrthosiphon loti and Acyrthosiphon pisum. Twenty of those loci were characterized in A. pisum. Fifteen of those loci were polymorphic. Genetic diversity varied across loci, allele repeat number ranging from two to 15, and observed heterozygosity from 0.1 and 0.96. An additional eight microsatellite loci originally isolated from other aphids but cross‐priming with A. pisum showed polymorphism as well. Allele size ranged from three to 9 and observed heterozygosity from 0.43 to 0.84. Overall, we present 23 microsatellite loci that can be used to reveal polymorphism in pea aphids.     

PCR primers for polymorphic microsatellite loci in the desert locust,Schistocerca gregaria (Orthoptera: Acrididae)

Nine pairs of polymerase chain reaction (PCR) primers that amplify polymorphic microsatellite loci in the desert locust, Schistocerca gregaria (Forskal), were developed using a magnetic bead‐based enrichment protocol. A sample of 48 locusts collected during the 1993 and 1995 upsurge periods in Eritrea, East Africa, were genotyped. The number of alleles per locus ranged from six to 20; the average was 12.67. Allelic distributions were significantly different between samples from different localities.     

Microsatellite DNA markers for Plasmopara viticola,the causal agent of downy mildew of grapes

Microsatellite loci were isolated from Plasmopara viticola (Oomycetes), the causal agent of downy mildew of grape, one of the most damaging fungal diseases of grapevine worldwide. Seven polymorphic loci were obtained from an enriched partial genomic library. A low genetic diversity was observed at all loci, with a mean observed allele number of 3.75 and an observed heterozygosity ranging from 0.074 to 0.547. Cross‐amplification tests on three closely related taxa indicated that two loci could be used in other Oomycetes species. These microsatellite loci were proved to be useful for population genetic analysis.     

Microsatellite markers in peach [Prunus persica (L.) Batsch] derived from an enriched genomic and cDNA libraries

Twenty‐four and 12 microsatellite loci were developed in peach [Prunus persica (L) Batsch cv. Akatsuki] by using an enriched genomic and fruit cDNA libraries, respectively. The microsatellite loci obtained from an enriched library produced 1–9 alleles per locus, 24 in total, of which 22 showed polymorphisms. The average values of observed and expected heterozygosities among the 24 loci were 0.15 and 0.68, respectively. The microsatellite loci derived from cDNA showed 1–7 alleles per locus. Eight sequences showed significant homology to the registered genes in a database.     

A Modified Enrichment Method to Construct Microsatellite Library from Plateau Pika Genome （Ochotona curzoniae）

A microsatellite-enriched library of plateau pika(Ochotona curzoniae)was constructed according to the strong affinity between biotin and streptavidin.Firstly,genomic DNA was fragmented by ultrasonication,which is a major improvement over traditional methods.Linker-ligated DNA fragments were hybridized with biotinylated microsatellite probes,and then were subjected to streptavidin-coated magnetic beads.PCR amplification was performed to obtain double-stranded DNA fragments containing microsatellites.Ligation and transformation were carried out by using the pGEM-T Vector System Ⅰ and Escherichia coli DH10B competent cells.Sequencing results showed that 80.2% of clones contained microsatellite repeat motif.Several modifications make this protocol time-efficient and technically easier than the traditional ones; particularly,composition and relative abundance of microsatellite repeats in plateau pika genome were truly represented through the optimized PCR conditions.This method has also been successfully applied to construct microsatellite-enriched genomic libraries of Chinese hamster(Cricetulus griseus)and small abalone[Haliotis diversicolor(Reeve)]with high rates of positive clones,demonstrating its feasibility and stability.     

Development of Rainbow Trout Microsatellite Markers from Repeat Enriched Libraries

The efficiency of developing polymorphic microsatellite markers from 2 repeat enriched libraries was evaluated. Thirty-six polymorphic microsatellite markers were developed for rainbow trout, 27 of which were informative in a mapping family. The ability of each marker to amplify genomic DNA from other salmonids was also observed. Received March 1, 2001; accepted May 1, 2001.     

Isolation and characterization of microsatellite markers in two basidiomycete species: Pleurotus eryngii and P. ferulae

Six polymorphic microsatellite loci were developed for the closed basidiomycete species Pleurotus eryngii and P. ferulae using an enriched genomic library. The number of alleles per locus varied between two and eight and the expected heterozygosity ranged from 0.30 to 0.72 in P. eryngii and from 0.20 to 0.73 in P. ferulae. This enrichment technique is useful in those species where microsatellite loci are rare in the genome.     

Polymorphic dinucleotide microsatellites for the Mediterranean angler species (Lophiidae)

A microsatellite‐enriched genomic library was obtained from black‐bellied angler Lophius budegassa and six polymorphic dinucleotide markers were successfully optimized. These markers showed levels of polymorphism ranging from 0.363 to 0.662 and allele numbers ranging from three to seven. These markers were also found to successfully amplify in the closely related Lophius piscatorius (angler) and thus appear to be useful for genetic stock structure analysis in both species.     

Microsatellite loci for paternity analysis in the fathead minnow,Pimephales promelas (Teleostei: Cyprinidae)

Many recent studies have employed molecular markers to uncover important aspects of mating systems in teleost fishes. The fathead minnow (Pimephales promelas) is a nest‐building North American cyprinid that spawns multiply and exhibits exclusive male parental care. A battery of microsatellite markers was developed to analyse paternity in this species. The seven characterized loci possess four to 31 alleles and expected heterozygosities of 0.455–0.974. In combination, they elicit an exclusion probability of 0.999, a desirable level for paternity analysis. In addition, cross‐amplifications were conducted to test primer efficacy in 13 other taxa, including two congeners.     

Polymorphic microsatellites in a mangrove species,Rhizophora mangle L. (Rhizophoraceae)

Twenty‐six microsatellite loci were isolated and characterized from the mangrove species Rhizophora mangle using (GT)_n and (CT)_n repeats. Eighty‐four per cent of the clones contained microsatellite sequences; the most common dinucleotides were the (GA/CT) and (CA/GT) repeats. Ten primers were selected to investigate the polymorphism among individuals of R. mangle from two natural populations of the Colombian Pacific Coast. The observed heterozygosity per locus varied from 0.20 to 0.80, the power of discrimination was 0.32–0.84 and the power of exclusion was 0.03–0.75. This set of microsatellites offers an efficient tool for population genetics studies on this species.     

Microsatellite primers for Australian and New Guinean pythons isolated with an efficient marker development method for related species

Microsatellites are powerful molecular genetic markers for many evolutionary and biotechnological investigations, however, development of sufficient microsatellite markers is time‐consuming and expensive especially considering the vast numbers of species for which they could be used. In light of the conservative nature of microsatellite loci between related species we describe an alternative approach to microsatellite development. A single round of microsatellite isolation enabled the characterization of sufficient loci for a large number of related python species. From 21 loci isolated in the focal species, an average of 86.2% were conserved within the other species while an average of 60.5% were polymorphic in all 13 python species analysed. Our approach will decrease significantly the expense and time required for microsatellite development for large numbers of related species.     

Microsatellite primers for Sorbus torminalis and related species

This study reports the cloning and characterization of nine microsatellite primer pairs in a scattered woody species (Sorbus torminalis), and shows their potential for further use in 36 species of the Maloideae, a Rosaceae subfamily containing important fruit crop and ornamental species. These primers were designed from a microsatellite library constructed from genomic DNA of S. torminalis and enriched for CA and GA repeats. Genotyping 48 S. torminalis of a natural population with the six best markers yielded a mean of 10.7 alleles per locus, and an expectation of exclusion probability for paternity analysis greater than 0.993.     

Microsatellite primers for the Atlantic coastal killifish,Fundulus heteroclitus,with applicability to related Fundulus species

The mummichog (Fundulus heteroclitus), a common Atlantic coastal killifish, is a model vertebrate species for the study of molecular genetic variation in natural populations and of environmental toxicology. We report the development of a set of 20 microsatellite loci in this species. Average expected heterozygosity across all loci was 0.84 (range: 0.60–0.97), revealing a high level of variability at most loci. A survey of seven additional Fundulus species yielded one or two robust amplification products in over half (63%) of the species–primer combinations tested. Therefore, many of these loci will also prove useful in studies of other members of the genus Fundulus.     

Microsatellite primers for the African mole‐rat genus Cryptomys and cross‐species amplification within the family Bathyergidae

We report 11 microsatellite loci for the African mole‐rat genus Cryptomys (Rodentia: Bathyergidae), isolated from the Damaraland mole‐rat (Cryptomys damarensis) and the Common mole‐rat (C. hottentotus hottentotus). All loci are highly polymorphic in the source species, with between six and 13 alleles and observed heterozygosity ranging from 0.54 to 0.86. Two C. damarensis loci (DMR1 and 6) may be located on the X‐chromosome. Cross‐species amplification was investigated in 10 Bathyergid species, representative of the five genera. The majority of loci amplified polymorphic product in all Cryptomys species tested. Amplification was also widespread in the other genera, except in Heterocephalus.     

Microsatellite primers in the Chinese dove tree, Davidia involucrata (Cornaceae), a relic species of the Tertiary

? Premise of the study: The first microsatellite primers were developed for Davidia involucrata, an endangered relic species of the Tertiary in China, to further describe its genetic variability and population structure. ? Methods and Results: Using the Fast Isolation by AFLP of Sequences Containing Repeats (FIASCO) protocol, 15 polymorphic microsatellite loci were isolated and characterized in 20 individuals from the germplasm collections of D. involucrata at the Hunan Forest Botanical Garden. High levels of polymorphism were revealed, with the total number of alleles per locus and the number of alleles per locus per individual ranging from two to 13 and from one to six, respectively. ? Conclusions: The multibanded patterns of microsatellite loci obtained in the current study confirmed that D. involucrata might be a polyploid species. The primers will be useful for studies of genetic diversity and for guiding conservation strategies for D. involucrata.