Isolation and characterization of six microsatellites in the Mexican beaded lizard Heloderma horridum

Six polymorphic microsatellite loci were isolated and characterized in the threatened Mexican beaded lizard, Heloderma horridum. The number of alleles per locus ranged from three to 12, with observed heterozygosity estimates ranging from 0.00 to 0.77, and expected heterozygosity estimates ranging from 0.00 to 0.73. These microsatellites will provide a valuable tool for the investigation of the genetic variation and structure of both wild and captive H. horridum populations.     

Polymorphic microsatellite markers for the Iberian rock lizard species,Iberolacerta cyreni,and cross‐species priming in other Iberolacerta species

Eight polymorphic microsatellite loci are described for the Iberian rock lizard species, Iberolacerta cyreni. Loci were isolated from a partial genomic library that had been enriched for AAAG repeat sequence. Number of alleles per locus ranged from two to 13 in a sample of 24 individuals from the Sierra de Guadarrama (central Spain). Observed heterozygosity ranged from 0.13 to 0.96. At least three loci were amplified and polymorphic in four other Iberian rock lizard species: Iberolacerta monticola, Iberolacerta bonnali, Iberolacerta aranica and Iberolacerta aurelioi. These markers will be used to study mating strategies and reproductive success in I. cyreni.     

<Emphasis Type="Italic">Tiliqua rugosa</Emphasis> microsatellites: isolation via enrichment and characterisation of loci for multiplex PCR in <Emphasis Type="Italic">T. rugosa</Emphasis> and the endangered <Emphasis Type="Italic">T. adelaidensis</Emphasis>

We used an enrichment technique to isolate 18 novel di and tri microsatellites for the socially monogamous lizard Tiliqua rugosa. These loci were amplified in conjunction with previously described loci in two and three PCR multiplexes for T. rugosa and the endangered T. adelaidensis, respectively. The loci were highly polymorphic in both species, exhibiting between 2 and 32 alleles with observed heterozygosity ranging from 0.43 to 0.96. These markers will be useful for population-level analyses and can contribute to a genetic foundation for conservation strategies for the endangered T. adelaidensis.     

Isolation and characterization of 12 tetra‐repeated microsatellite loci from the Formosan grass lizard (Takydromus formosanus)

Twelve tetra‐repeated microsatellite loci were developed for the island‐endemic grass lizard Takydromus formosanus (Squamata: Lacertidae). We characterized these loci by genotyping 51 individuals sampled from six localities. The number of alleles per locus ranged between 12 and 23, whereas the observed heterozygosity ranged between 0.686 and 0.941. None of these loci showed gametic disequilibrium or sex linkage. With such high polymorphism, we believe that these loci should be suitable for fine‐scaled analysis of population structure, hybridizations or individual identification of grass lizards.     

Thirty-three microsatellite loci for noninvasive genetic studies of the giant panda (Ailuropoda melanoleuca)

Limited microsatellite markers useable in noninvasive genetic methods have hampered the studies of dispersal patterns and mating systems of giant pandas. Therefore, we describe in this paper the characterization of 15 novel microsatellite loci from genomic DNA-enriched libraries and 18 redesigned microsatellite loci from published papers on the giant panda. The number of alleles per locus in 60 individuals ranged from 2 to 13, the average observed heterozygosity per locus from 0.168 to 0.800, and the average expected heterozygosity per locus from 0.152 to 0.882. All loci followed Hardy-Weinberg expectations. Four pairs of significant linkage association were found among all these loci. Moreover, the 33 microsatellite loci showed high amplification successes rate in noninvasive samples, which indicated that these loci will be of use in studying dispersal patterns and mating systems of giant pandas using noninvasive genetic methods.     

Highly polymorphic tetranucleotide microsatellite loci for the eastern Canary Island lizard,Gallotia atlantica

We describe 10 polymorphic tetranucleotide microsatellite loci from the eastern Canary Island lacertid lizard, Gallotia atlantica. Loci were isolated from a partial genomic library that had been enriched for AAAG repeat sequence. All loci were highly polymorphic (eight alleles or more) with observed heterozygosities from 0.75 to 1.00. At least four loci were successfully amplified and polymorphic in the Gran Canarian lacertid, Gallotia stehlini. These loci will be used to examine correlations between patterns of gene flow and recent volcanism on the island of Lanzarote.     

Isolation and characterization of tetramicrosatellite DNA markers in the Eurasian otter (Lutra lutra)

Eight new tetramicrosatellite loci for Eurasian otter (Lutra lutra) were designed. Polymorphism of these eight loci in 29 otter individuals was tested. The results indicated that the allele numbers of each of loci ranged from three to five and the observed heterozygosity from 0.483 to 0.828. These new loci can be useful for population genetic research on otters and help improve the resolution of individual identification using noninvasive method.     

Isolation and characterization of nine microsatellite loci in Podarcis bocagei (Squamata: Lacertidae)

Nine dinucleotide microsatellite loci were developed through an enrichment protocol for Bocage's wall lizard, Podarcis bocagei Seoane 1884, a lacertid endemic to the Iberian Peninsula. Nineteen primer pairs were designed and tested. From these, nine loci yielded satisfactory results and were screened on 15–19 individuals. These loci revealed a high level of polymorphism (8–15 alleles) and heterozygosity (0.611–0.947) and will certainly be useful in the study of population structure and evolutionary history of this species.     

Clone identification and clinal allozyme variation in populations ofAbies alba from the Eastern Alps (Austria)

Allelic frequencies at the polymorphic enzyme loci IDH-1, IDH-2, LAP-1, and 6-PGDH-1 were determined in population samples ofAbies alba from 5 different areas in Austria. The results reveal clear genetic differentiation between the eastern and the western provenances. Gradual frequency clines are observed with all four loci. The amount of variation, measured as average heterozygosity and genotypic diversity, is significantly higher in the west. The multilocus genotypes proved to be useful markers for the identification of clones. Functional differences with respect to substrate specifity were found for the two allozymes coded by the coexisting alleles of the IDH-2 locus.     

Eleven polymorphic microsatellite loci in Lindera benzoin,Lauraceae

Microsatellite markers were developed for studies of the genetic diversity and population substructure of Lindera benzoin, Lauraceae (spicebush). Nuclear microsatellite sequences were obtained from DNA libraries that were enriched for (CA), (GA), (AAG) and (ATG) repeat motifs. From 69 microsatellite sequences, 20 primer sets were developed. Of these, 11 primer pairs resulted in amplified polymorphic loci. In 29 samples of eastern Pennsylvania spicebush plants, the number of microsatellite alleles ranged from two to 16 per locus with observed heterozygosity values ranging from 0.10 to 0.82.     

Development and characterization of microsatellite loci in the eastern chipmunk (Tamias striatus)

We isolated and characterized 16 microsatellite loci in the eastern chipmunk, Tamias striatus. The loci were screened across 25 individuals from one population and shown to be polymorphic with the number of alleles per locus ranging from two to 17. Polymorphic information content ranged from 0.212 to 0.887 and observed heterozygosity ranged from 0.200 to 0.960. Only two loci (Chip 10 and Chip 25) deviated from Hardy–Weinberg equilibrium.     

Isolation and characterization of nuclear microsatellite loci for the common green darner dragonfly Anax junius (Odonata: Aeshnidae) to constrain patterns of phenotypic and spatial diversity

Fourteen polymorphic microsatellite loci were developed from an enriched genomic library of the widely distributed migratory North American dragonfly species, the common green darner (Anax junius). For a group of 22 larvae, these loci averaged 16 alleles, with individual loci ranging from nine to 29 alleles. Observed heterozygosity averaged 0.784 per locus.     

Isolation and characterization of microsatellite loci in wild and domestic turkeys (Meleagris gallopavo)

We describe the isolation, development and application of seven microsatellite loci in the eastern wild turkey, Meleagris gallopavo silvestris, as well as their amplification and levels of polymorphism in the domestic turkey. The number of alleles per locus ranged from 5 to 15 and average heterozygosity was high for almost all loci. Domestic turkeys showed significantly reduced numbers of alleles per locus and overall heterozygosities when compared to eastern wild turkeys. The high variability in these markers should provide the level of resolution required to continue studies of wild turkey population genetics.     

基于粪便DNA 的藏狐微卫星位点筛选及个体识别

藏狐是我国青藏高原东部多房棘球绦虫和石渠棘球绦虫最主要的野生动物终末宿主。棘球绦虫会导致一类称为棘球绦虫病的致死性人兽共患疾病,青藏高原东部牧区是该病重要的流行区。因此作为终末宿主,评估藏狐种群的棘球绦虫感染率对于该病的流行病学研究意义明显。而要获取这方面信息,首先必须了解藏狐的种群数量。为此,我们基于非损伤取样的原则,使用藏狐新鲜粪便作为研究材料,从已发布的藏狐及近缘种的48个微卫星位点中筛选了11 个用于藏狐粪便DNA 多态性分析。对2011 -2012 年7 -8 月间收集的128 份有效藏狐粪便样品(2011 年68 份,2012 年60 份)进行特异性PCR 扩增,并用琼脂糖凝胶电泳和荧光引物标记法进行基因分型,根据各位点的等位基因频率计算出各位点的基因型数(N),期望杂合度(He )、观测杂合度(Ho)、多态信息含量(PIC)以及不同个体基因型相同概率值(PI)。结果发现,各位点N 介于4 - 7,H e为0.66 - 0. 80,H o为0.17 -0.68,PIC 为0.5496 - 0.7623。11 个位点的累积PI 值满足个体识别的需要(PIbiased = 1. 283 × 10 ^{- 11} ;PIsi bs =7.572 × 10 ^{- 5} )。但是,由于粪便DNA 质量差异较大,不同位点的扩增成功率差异较大(0.176 - 0. 926)。我们发现,按照扩增成功率由高到低排列,前6 个微卫星位点(P03,CXX172,CPH6,CPH8,P01i,P08)的扩增成功率均超过0.6,且累积PI 值小于0.004 (PIbiased =2.775 × 10 ^{- 7} ;PIsibs = 3. 606 ×10 ^{- 3} ),表明这6 个位点可以对藏狐进行个体识别。因此,针对本研究的数据,制定了如下的个体识别原则: (1)只有粪便DNA 至少成功扩增出前6 个微卫星位点的样品可以进入下一步分析; (2)所有位点的信息均相同的两个样品被认为是来自同一个体;(3)保险起见,如果仅有一对位点信息不相等,此两个样品依然被判定来自同一个体。在此基础上,我们从2011 年样品中识别出30 个藏狐个体,从2012 年样品中识别出21 个个体。     

Allozyme genotype-environment relationships in natural populations of Drosophila buzzatii

Allozyme frequency data from natural populations of Drosophila buzzatii were analyzed for genotype-environment relationships. Allele frequency and heterozygosity at six loci polymorphic throughout eastern Australia and a number of environmental factors (both means and variabilities) were examined by a variety of multivariate techniques. Significant genotype-environment associations were found for five of the six loci, and after correcting for geographic location significant associations remained for Est-2 and Adh-1 gene frequencies and heterozygosities and for Pgm gene frequencies. The results are discussed in relation to selection and gene flow and provide the basis for laboratory studies to disentangle confounded effects of (1) environmental means and environmental variabilities and (2) allele frequency and heterozygosity, and thus to further test for and determine the nature of any natural selection at particular allozyme loci.This work was supported by a grant from the Australian Research Grants Committee to J. S. F. B.     

Anonymous nuclear markers for the eastern fence lizard,Sceloporus undulatus

We present results from a screen for de novo variable nuclear loci using a genomic library approach in Sceloporus undulatus, the eastern fence lizard. We tested amplification success for 77 primer pairs in S. undulatus, Sceloporus occidentalis and Sceloporus grammicus. Many loci amplified in all three species suggesting that our primers will be useful for developing sequencing or single nucleotide polymorphism (SNP) genotyping markers in other sceloporine lizards. We also sequenced 19 loci, containing 158 variable sites, for 91 S. undulatus individuals. We report high levels of nucleotide variation in this species with an average of 38 SNPs per kilobase.     

Enzyme function and polymorphism: A test in two Anolis lizard species

Electrophoretic surveys of two lizard species were used to test hypotheses that relate levels of enzyme variability to enzyme function (single-substrate versus multiple-substrate enzymes, regulatory versus nonregulatory enzymes). Anolis roquet behaviorally regulates its body temperature, but its congener A. gundlachi is passive to variations in the thermal environment. As a result, populations of A. gundlachi probably experience the thermal environment as temporally coarse-grained, whereas populations of A. roquet do not. We therefore predicted that A. gundlachi would exhibit greater enzyme heterozygosity than A. roquet and that different enzyme classes would contribute disproportionately to this interspecific difference. The data show (1) that A. gundlachi does have a higher heterozygosity and (2) that this difference appears to result from high levels of heterozygosity at loci coding for multiple-substrate enzymes. The dichotomy between regulatory and nonregulatory enzymes offers no explanation for the variability in heterozygosity among enzyme loci in these species.E.Z. was supported by a grant from the Natural Sciences and Engineering Council of Canada. The study was accomplished while P.E.H. held a postdoctoral fellowship from the Killam Trust of Dalhousie University and a grant from the Research Development Fund of Dalhousie University. The collection of material was made possible by grants (to P.E.H.) from National Science Foundation (DEB 75-16334), the Explorers Club of New York, Sigma Xi, and the Richmond and Anderson Funds of Harvard University. We thank Dr. D. W. Foltz for his help with the calculations.     

Age-dependent allozymic variation in a natural population of lizards

An analysis of allozymic variation at 17 loci in a population of the sagebrush lizard (Sceloporus graciosus) in southern Utah yielded an estimate of genic heterozygosity of 0.028. Seven of the loci were variable, but only one, Est-1, was strongly polymorphic. The observation that the frequency of the common genotype (MM) at the Est-1 locus declined monotonically in successive age classes from 0.74 in hatchlings to 0.58 in adults 4 years old or older suggests that allele frequencies at this locus are not independent of selective influences.This research was supported by NSF grant GB-29141 to D.W.T. and NIH grant GM-15769 to R.K.S.     

Isolation and characterization of 14 polymorphic microsatellite markers for the blue and red shrimp, Aristeus antennatus (Crustacea, Decapoda)

Eighteen microsatellite loci were isolated and characterized from the blue and red shrimp, Aristeus antennatus Risso 1816, a commercially exploited marine crustacean widely distributed throughout the Mediterranean Sea and in the eastern Atlantic. Polymorphism was assessed in a population (n = 20) from the southwestern Sardinian seas; 14 loci resulted polymorphic and showed from three to 13 alleles. The observed heterozygosity ranged from 0.2 to 0.85. These microsatellites will be potentially useful for the study of A. antennatus population genetic structure.     

Development of microsatellite loci for population studies of the pink ling,Genypterus blacodes (Teleostei: Ophidiidae)

Fifteen microsatellite loci were developed for pink ling (Genypterus blacodes), a fish of significant commercial importance to Australasia. Nine loci were examined in samples from five regions of Australia’s South East Fishery. All nine were highly polymorphic; numbers of alleles per locus ranged from 11 to 52 in total samples of 270–306 individuals. The average observed heterozygosity per locus per sample (0.823) was a little lower than the average Hardy–Weinberg expected heterozygosity per sample (0.895), perhaps reflecting the possible presence of null alleles at two loci. There was no significant evidence of genetic stock heterogeneity.