Evidence Of Linkage Disequilibrium Research Articles

The Toll-like Receptor 4 Polymorphism Asp299Gly Is Associated with an Increased Risk of Ovarian Cancer.

Ovarian cancer (OC) is one of the most common cancers threatening women’s lives around the world. Epithelial ovarian tumors represent the most common ovarian neoplasms. Most OC patients are diagnosed at the advanced stage, and there is an urgent need to identify novel biomarkers of the disease. Single-nucleotide polymorphisms (SNPs) in TLR genes may serve as crucial markers of cancer susceptibility. We investigated the frequency of TLR polymorphisms in a group of 200 women, including 70 with OC. Four SNPs, two each in TLR4 (rs4986790 and rs4986791) and TLR9 (rs187084 and rs5743836), were analyzed using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP). The digested fragments were separated and identified by multicapillary electrophoresis. The load quantification of human papillomavirus (HPV) types 16/18 was determined using a digital droplet PCR method. We found an increased frequency of heterozygous genotype and minor allele of the TLR4 rs4986790 SNP in women with OC compared with healthy controls, and this result remained highly significant after Bonferroni’s correction for multiple testing (p < 0.0001). No evidence of linkage disequilibrium was found with any of the examined TLR SNPs. The findings suggest that the TLR4 Asp299Gly polymorphism could be a genetic risk factor for the development of OC.

Microsatellite Markers from Peronospora tabacina, the Cause of Blue Mold of Tobacco, Reveal Species Origin, Population Structure, and High Gene Flow.

Peronospora tabacina is an obligate parasite that causes blue mold of tobacco. The pathogen reproduces primarily by sporangia, whereas the sexual oospores are rarely observed. A collection of 122 isolates of P. tabacina was genotyped using nine microsatellites to assess the population structure of individuals from subpopulations collected from central, southern, and western Europe; the Middle East; Central America; North America; and Australia. Genetic variations among the six subpopulations accounted for ∼8% of the total variation, including moderate levels of genetic differentiation, high gene flow among these subpopulations, and a positive correlation between geographic and genetic distance (r = 0.225; P < 0.001). Evidence of linkage disequilibrium (P < 0.001) showed that populations contained partially clonal subpopulations but that subpopulations from Australia and Mediterranean Europe did not. High genetic variation and population structure among samples could be explained by continuous gene flow across continents via infected transplant exchange and/or long-distance dispersal of sporangia via wind currents. This study analyzed the most numerous P. tabacina collection and allowed conclusions regarding the migration, mutation, and evolutionary history of this obligate biotrophic oomycete. The evidence pointed to the species origin in Australia and identified intracontinental and intercontinental migration patterns of this important pathogen.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Characterization of a New Set of Microsatellite Markers Suggests Polygyny and Polyandry in Atta cephalotes (Hymenoptera: Formicidae).

The leaf-cutting ant, Atta cephalotes L. (1758), is a major herbivore with great economic impact in the Neotropics. Because of its broad range and human-mediated dissemination, the ecology of this ant has received considerable attention; however, questions concerning its population genetics, dispersal, and social structure remain unexplored. Here, we aimed to identify and provide information on molecular and statistical performance of a suite of polymorphic microsatellite markers for A. cephalotes while demonstrating their utility for further genetic studies. We designed primer sequences targeting thousands of microsatellite loci and then screened 30 of these for amplification and polymorphism. Fifteen of these loci were selected and used to evaluate their polymorphism using 74 ants sampled from 15 different nests of the same location. This set of 15 loci exhibited variation of 2-20 alleles, with a mean heterozygosity of 0.57. All loci followed Hardy-Weinberg expectations with no evidence of linkage disequilibrium, while two loci showed evidence of null alleles. Our preliminary analysis suggested substantial nest differentiation with no population viscosity within the sampled location, as well as colonies with multiple queens (polygyny) and paternity (polyandry). Our newly identified microsatellites have proven to be highly informative to investigate gene flow, social structure and reproduction of this key agricultural pest.

Analysis of DNM3 and VAMP4 as genetic modifiers of LRRK2 Parkinson’s disease

The LRRK2 gene has rare (p.G2019S) and common risk variants for Parkinson’s disease (PD). DNM3 has previously been reported as a genetic modifier of the age at onset in PD patients carrying the LRRK2 p.G2019S mutation. We analyzed this effect in a new cohort of LRRK2 p.G2019S heterozygotes (n = 724) and meta-analyzed our data with previously published data (n = 754). VAMP4 is in close proximity to DNM3, and was associated with PD in a recent study, so it is possible that variants in this gene may be important. We also analyzed the effect of VAMP4 rs11578699 on LRRK2 penetrance. Our analysis of DNM3 in previously unpublished data does not show an effect on age at onset in LRRK2 p.G2019S carriers; however, the inter-study heterogeneity may indicate ethnic or population-specific effects of DNM3. There was no evidence for linkage disequilibrium between DNM3 and VAMP4. Analysis of sporadic patients stratified by the risk variant LRRK2 rs10878226 indicates a possible interaction between common variation in LRRK2 and VAMP4 in disease risk.

Genetic diversity evidence a mixed reproduction mode in Venturia oleaginea populations in Uruguay

Olive scab caused by Venturia oleaginea is one of the most important diseases of olive worldwide. The fungus infects young leaves and fruits by asexually produced conidia. The sexual stage and its importance in the epidemiology and genetics of the pathogen population is unknown. A collection of 52 fungal isolates was obtained from scabbed leaves collected in 16 orchards from three major regions in Uruguay. All isolates were identified as V. oleaginea by PCR with specific primers and sequence analysis of the TEF-1α gene region. Five colony morphotypes and different growing rates were observed on potato dextrose agar (PDA) medium. Universally primed polymerase chain reaction (UP-PCR) was successfully used for assessing the genetic diversity and population structure of the V. oleaginea isolates, whereas ISSR was unable to generate reproducible and polymorphic bands. Two populations genetically different were identified: [1] composed by 21 isolates mainly from Southern and South-Western regions; [2] composed by 31 isolates mainly from South-Eastern and Southern regions. Both populations showed a moderate gene diversity expressed as hNei = 0.163 and 0.212 for population [1] and [2], respectively. There was evidence of linkage disequilibrium in both populations, rejecting the null hypothesis of random mating as the predominant reproduction mode, a typical result of clonal populations. However, all isolates were genotypically different, an indirect evidence of recombination. We conclude that V. oleaginea populations in Uruguay present a mixed mode of reproduction, a characteristic of many pathogens that are difficult to control.

Isolation and Characterization of Microsatellite Markers for Soybean Looper (Lepidoptera: Noctuidae).

We constructed 13 microsatellite markers for Chrysodeixis includens (Walker), a serious crop pest in the Americas. All SSR (Simple Sequence Repeat) markers were polymorphic, with no evidence of linkage disequilibrium between any loci in any population. The total number of alleles per locus ranged from 5 for L3, L9, and L11 to 26 for L6; the mean number of alleles per locus in three populations of C. includens ranged from 2.33 for L3 to 14.67 for L6. Hardy–Weinberg equilibrium (HWE) deviation was not observed in four loci for at least one population (L3, L5, L9, L10). Markers L6, L7, L8, L10, L11, L12, and L13 showed a frequency of null alleles > 0.2 for at least one population. STRUCTURE and F-statistics revealed low population structure among the populations (FST = 0.013) and a high degree of inbreeding (FIS = 0.658). The SSR markers developed here will be useful in future studies on the ecology, demography, host dynamics, and gene flow of C. includens. This information is essential to understand the recent status of C. includens as a key pest in South America.

Isolation and characterization of 12 microsatellite loci in Epipedobates anthonyi (Amphibia: Anura: Dendrobatidae) for population genetic analysis.

Tropical anurans are among the most diverse and vulnerable organisms on Earth, yet the evolutionary mechanisms behind their diversity remain relatively unexplored. Epipedobates anthonyi is a poison frog that inhabits southern Ecuador and northern Peru along a broad elevational range (0-1800m). Throughout its range, this species exhibits variation in phenotypic traits, such as color, advertisement calls, and alkaloid composition. The aim of this study is to isolate and characterize microsatellite loci to investigate patterns of genetic variation within the species. Using a next-generation sequencing approach to screen an enriched genomic library, we report twelve polymorphic microsatellite loci. The number of alleles per locus ranged from 7 to 15 per population. For the two populations tested, mean observed heterozygosity was 0.69 and 0.79, and mean expected heterozigosity was 0.84 and 0.85 respectively. Only locus EAN002 showed significant departure of HWE in both populations. None of the loci showed consistent null alleles in both populations. Also, no evidence of linkage disequilibrium was found across loci. In this paper, we report for the first time 12 microsatellite loci for E. anthonyi. These markers will be used to further elucidate evolutionary mechanisms underlying genetic and phenotypic variation across the species' range.

Polymorphic Microsatellite Markers Selection by Cross-Species Amplification in Black Francolin (Francolinus francolinus asiae)

The cross-species amplification is an effective method in obtaining microsatellite loci for genetic studies of closely related species. In this study, the cross-species amplified markers were characterized and selected for black francolin (Francolinus francolinus asiae). It is a ground dwelling game bird. In this study total eight markers were cross-species amplified. These eight markers were selected from chicken. The number of alleles per locus ranged from 4 to 21. The observed heterozygosity ranged from 0.022 to 0.98 and the expected heterozygosity from 0.23 to 0.79. There was no evidence of linkage disequilibrium among the pair of loci. These loci were polymorphic and offered valuable tool for genetic diversity and population genetic studies of black francolin.

Identification and characterization of microsatellite loci in West Atlantic sea cucumber Holothuria grisea (Selenka 1867)

The sea cucumber Holothuria grisea has become the subject of intense and unregulated fishing in northeastern Brazil due to their growing demand in Asian market. However, there is little knowledge about the dynamics and genetics of H. grisea wild populations on the South American coast. In this study, we present the first set of H. grisea microsatellite markers, identified and characterized using Illumina paired-end reads of whole genome shotgun sequencing. From 50 strictly selected candidates, eight novel microsatellite markers were successfully developed. We then genotyped 30 individuals to evaluate the degree of polymorphism and validate the markers. The number of alleles ranged from three to 14, while observed and expected heterozygotes ranged from 0.156 to 0.906 and from 0.283 to 0.774, respectively. After correcting for multiple tests,we found no evidence of linkage disequilibrium in all pairwise combinations between the loci. One locus (Hgr15607) revealed deviation from the Hardy-Weinberg equilibrium, as well as the presence of null alleles. However, we observed significant differences in frequency distribution between males and females at locus Hgr15607. We believe that the markers describedhere will be useful for conservation efforts and management of H. grisea fisheries and for prospective aquaculture of these organisms.

Genetic variation and population homogeneity of the sea star Coscinasterias tenuispina (Forcipulatida: Asteroidea) on the coast of Rio de Janeiro, Brazil

Genetic variation and population structure of the sea star Coscinasterias tenuispina were investigated using allozyme electrophoresis (seven polymorphic loci out of nine sampled) at seven sites on the coast of Rio de Janeiro State. Observed values of heterozygosity’s were high (HOBS = 0.31 - 0.53), although similar to those found for others marine invertebrates. Departures from Hardy-Weinberg expectations were found only for one locus in three populations. No evidence of linkage disequilibrium was found among any loci for any population. The genetic identities vary from 0.882 to 0.988. The average  was low and insignificant statistically ( = 0.017). Furthermore, Bayesian analysis of population partitioning showed that the highest LnP(D) value was achieved for K = 1. These results do not corroborate any effect by upwelling limiting gene flow among the analyzed populations or the hypothesis that the strong sex ratio imbalance previously recorded to Itaipu population has been maintained by asexual reproduction.

Isolation and Characterization of Novel Polymorphic Microsatellite Markers for Cimex hemipterus F. (Hemiptera: Cimicidae).

Due to the growing public health and tourism awareness, Cimex hemipterus Fabricius (Hemiptera: Cimicidae) has gained a great interest in increasing reported infestation cases in tropical regions of the world, including Malaysia. Since the information on the molecular ecology and population biology of this species are tremendously lacking, the isolation and development of molecular markers can be used to determine its genetic structure. In this study, novel microsatellite primers isolated from enriched genomic libraries of C. hemipterus were developed using 454 Roche shotgun sequencing. Seven validated polymorphic microsatellite primers were consistently amplified and characterized from 70 tropical bed bugs collected from seven locations throughout Malaysia. The number of alleles per locus identified ranged from 6 to 14. Comparison of loci for overall and between population were done with mean observed and expected heterozygosity were determined at 0.320 and 0.814, 0.320 and 0.727, respectively. Polymorphic information criteria (PIC) valued the markers as highly informative as PIC >0.5. Overall population, they are possibly in Hardy-Weinberg equilibrium with loci Ch_09ttn, Ch_01dn, and Ch_13dn showing signs of a null allele. There were no scoring errors caused by stutter peaks, no large allele dropout was detected for all loci and showed no evidence of linkage disequilibrium. In conclusion, all seven molecular microsatellite markers identified can be beneficially used to gain more information on the population genetic structure and breeding patterns of C. hemipterus as well as the relationship of dispersal and infestation.

Allele and haplotype diversity of 12 X-STRs in Sardinia

The analysis of clusters of tightly linked X-chromosome short tandem repeat (STR) markers can assist the interpretation of complex kinship cases. However, when linkage disequilibrium (LD) is present in the population of origin of tested individuals, haplotype rather than allele frequencies should be used in likelihood calculations. The diversity of twelve X-STRs arranged in four linkage groups (I: DXS10148-DXS10135-DXS8378; II: DXS7132-DXS10079-DXS10074; III: DXS10103-HPRTB-DXS10101; IV: DXS10146-DXS10134-DXS7423) was tested in a Sardinian population sample (n=516) including three open populations from the Northern, Central and Southern part of the island, and three isolates (Benetutti, Desulo, Carloforte). Evidence of LD was detected in Sardinia within each linkage group. Significant differences in haplotype and allele frequency distribution of X-STR markers was seen between isolates and open populations, which on the contrary appeared highly homogeneous.The percentage of Sardinian haplotypes previously unobserved in a similar dataset compiled for the Italian population was: 76.3% (linkage group I), 61.3% (linkage group II), 54.1% (linkage group III), 58.9% (linkage group IV). Significant pairwise genetic differences were seen between mainland Italy, the three Sardinian isolates, and the open population of Southern Sardinia.The study confirms the presence of high levels and complex patterns of LD along the X chromosome in Sardinia, and provides population-specific haplotype data for biostatistical evaluation in kinship testing.

Development and characterization of novel microsatellite loci for the amphipod Cymadusa filosa

Microsatellites are molecular tools for answering a wide variety of questions in population genetics. Here, we developed a novel set of microsatellite loci for the amphipod Cymadusa filosa using a microsatellite-enriched library. In total, 22 loci were described, and ten polymorphic markers were characterized for 30 individuals from southeastern Brazil. The number of alleles ranged from two to 26 and the observed heterozygosity from 0.033 to 0.933. No significant deviation from Hardy-Weinberg equilibrium was found; there was no evidence for linkage disequilibrium, and null alleles were detected only in three loci at low frequencies. These markers will be useful tools for ecological and evolutionary studies involving C. filosa.

Development and evaluation of microsatellite loci for Gynochthodes boninensis (Rubiaceae), a woody climbing plant endemic to the Bonin (Ogasawara) Islands, Japan

Microsatellite loci were developed for Gynochthodes boninensis, an endemic climbing plant in the Bonin Islands. Using a Roche 454 GS Junior next-generation sequencer, 158 microsatellite loci were designed. Of the 48 microsatellite loci tested, 37 were successfully amplified and 25 were polymorphic in two populations of G. boninensis. For the 25 polymorphic loci, the mean expected heterozygosities per locus were 0.303 in the Chichijima Island population and 0.310 in the Hahajima Island population, respectively. There was no evidence of linkage disequilibrium in either population, but one locus showed significant deviation from Hardy–Weinberg equilibrium in one population. The microsatellite loci developed in this study will be useful for future studies of population genetics of G. boninensis. In particular, because this species is androdioecious (males and hermaphrodites coexist), characterizing the species gene flow is crucial to understanding the evolution and maintenance of this rare sexual system.

A new suite of twenty-two polymorphic microsatellite loci in the parasitic wasp, Habrobracon hebetor (Hymenoptera: Braconidae): Promising molecular tools for studying the population genetics of several beneficial braconid species

Combining a biotin-enrichment protocol and 454GS-FLX titanium pyrosequencing technology, we characterised 22 polymorphic microsatellite loci from the parasitic wasp, Habrobracon hebetor (Say) (Hymenoptera: Braconidae), a cosmopolitan species commonly used in biological control against a wide range of both major lepidopterous pests of stored products and field crops in different parts of the world. Three multiplex PCR sets were optimised and characterised across 46 H. hebetor specimens from two samples collected from millet fields in Niger. Two to 11 alleles were found per locus and observed heterozygosity ranged from 0.289 to 0.826. Polymorphism was detected in both samples with a similar level of observed heterozygosity (0.482 vs. 0.502) and number of alleles (4.1 vs. 3.6). Deviation from Hardy-Weinberg equilibrium was detected at the same five loci in both samples and five or seven more loci in each sample but was not associated with heterozygote deficiencies. Even though evidence for linkage disequilibrium was found between a few alleles, these new loci segregated independently. The variability of the 22 loci will enable estimates of genetic diversity and structure patterns, as well as gene flow between H. hebetor populations at different spatial scales. Cross-species amplifications were successful among the six Bracon spp. tested and nine loci will be particularly appropriate for population genetic studies in B. brevicornis.

Characterisation of 12 microsatellite loci in the Vietnamese commercial clam Lutraria rhynchaena Jonas 1844 (Heterodonta: Bivalvia: Mactridae) through next-generation sequencing

The marine clam Lutraria rhynchaena is gaining popularity as an aquaculture species in Asia. Lutraria populations are present in the wild throughout Vietnam and several stocks have been established and translocated for breeding and aquaculture grow-out purposes. In this study, we demonstrate the feasibility of utilising Illumina next-generation sequencing technology to streamline the identification and genotyping of microsatellite loci from this clam species. Based on an initial partial genome scan, 48 microsatellite markers with similar melting temperatures were identified and characterised. The 12 most suitable polymorphic loci were then genotyped using 51 individuals from a population in Quang Ninh Province, North Vietnam. Genetic variation was low (mean number of alleles per locus = 2.6; mean expected heterozygosity = 0.41). Two loci showed significant deviation from Hardy-Weinberg equilibrium (HWE) and the presence of null alleles, but there was no evidence of linkage disequilibrium among loci. Three additional populations were screened (n = 7-36) to test the geographic utility of the 12 loci, which revealed 100 % successful genotyping in two populations from central Vietnam (Nha Trang). However, a second population from north Vietnam (Co To) could not be successfully genotyped and morphological evidence and mitochondrial variation suggests that this population represents a cryptic species of Lutraria. Comparisons of the Qang Ninh and Nha Trang populations, excluding the 2 loci out of HWE, revealed statistically significant allelic variation at 4 loci. We reported the first microsatellite loci set for the marine clam Lutraria rhynchaena and demonstrated its potential in differentiating clam populations. Additionally, a cryptic species population of Lutraria rhynchaena was identified during initial loci development, underscoring the overlooked diversity of marine clam species in Vietnam and the need to genetically characterise population representatives prior to microsatellite development. The rapid identification and validation of microsatellite loci using next-generation sequencing technology warrant its integration into future microsatellite loci development for key aquaculture species in Vietnam and more generally, aquaculture countries in the South East Asia region.

Development and characterization of 10 microsatellite markers in the Cape horseshoe bat, Rhinolophus capensis (Chiroptera, Rhinolophidae) and cross-amplification in southern African Rhinolophus species.

BackgroundThe Cape horseshoe bat, Rhinolophus capensis, is endemic to the Cape region of South Africa. Coalescent analysis of mitochondrial DNA sequence data suggests extensive historical gene flow between populations despite strong geographic variation of their echolocation call phenotype. Nevertheless the fine-scale genetic structure and evolutionary ecology of R. capensis remains poorly understood. Here we describe the development of 10 novel polymorphic microsatellite loci to investigate of the dispersal ecology of R. capensis and to facilitate taxonomic studies of Rhinolophus species in southern Africa.FindingsWe report 10 microsatellite primer pairs that consistently amplify scorable and polymorphic loci across 12 African rhinolophid species. Initial analysis of two populations of R. capensis from South Africa revealed moderate to high levels of allelic variation with 4–14 alleles per locus and observed heterozygosities of 0.450–0.900. No evidence of linkage disequilibrium was observed and eight of the loci showed no departure from Hardy–Weinberg equilibrium. Cross-species utility of these markers revealed consistently amplifiable polymorphic loci in eleven additional rhinolophid species.ConclusionsThe cross-amplification success of the microsatellites developed here provides a cost-effective set of population genetic marker for the study of rhinolophid evolutionary ecology and conservation in southern Africa.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-015-1465-5) contains supplementary material, which is available to authorized users.

Identification and Evaluation of 21 Novel Microsatellite Markers from the Autumnal Moth (Epirrita autumnata) (Lepidoptera: Geometridae).

The autumnal moth (Epirrita autumnata) is a cyclically outbreaking forest Lepidoptera with circumpolar distribution and substantial impact on Northern ecosystems. We have isolated 21 microsatellites from the species to facilitate population genetic studies of population cycles, outbreaks, and crashes. First, PCR primers and PCR conditions were developed to amplify 19 trinucleotide loci and two tetranucleotide loci in six multiplex PCR approaches and then analyzed for species specificity, sensitivity and precision. Twelve of the loci showed simple tandem repeat array structures while nine loci showed imperfect repeat structures, and repeat numbers varied in our material between six and 15. The application in population genetics for all the 21 microsatellites were further validated in 48 autumnal moths sampled from Northern Norway, and allelic variation was detected in 19 loci. The detected numbers of alleles per locus ranged from two to 13, and the observed and expected heterozygosities varied from 0.04 to 0.69 and 0.04 to 0.79, respectively. Evidence for linkage disequilibrium was found for six loci as well as indication of one null allele. We find that these novel microsatellites and their multiplex-PCR assays are suitable for further research on fine- and large-scale population-genetic studies of Epirrita autumnata.

Lack of concordance and linkage disequilibrium among brothers for androgenetic alopecia and CAG/GGC haplotypes of the androgen receptor gene in Mexican families.

Androgenetic alopecia (AGA) or common baldness is the most prevalent form of hair loss in males. Familial predisposition has been recognized, and heritability estimated in monozygotic twins suggests an important genetic predisposition. Several studies indicate that the numbers of CAG/GGC repeats in exon 1 of the androgen receptor gene (AR) maybe associated with AGA susceptibility. To investigate a possible correlation between AR CAG/GGC haplotypes and the presence or not of alopecia in sibships with two or more brothers among them at least one of them has AGA. Thirty-two trios including an alopecic man, one brother alopecic or not, and their mother were enrolled. Sanger sequencing of the exon 1 of the AR gene was conducted to ascertain the number of CAG/GGC repeats in each individual. Heterozygous mother for the CAG/GGC haplotypes was an inclusion criterion to analyze the segregation haplotype patterns in the family. Concordance for the number of repeats and AGA among brothers was evaluated using kappa coefficient and the probability of association in the presence of genetic linkage between CAG and GGC repeats and AGA estimated by means of the family-based association test (FBAT). The median for the CAG and GGC repeats in the AR is similar to that reported in other populations. The CAG/GGC haplotypes were less polymorphic than that reported in other studies, especially due to the GGC number of repeats found. Kappa coefficient resulted in a concordance of 37.3% (IC 95%, 5.0-69.0%) for the AGA phenotype and identical CAG/GGC haplotypes. There was no evidence of linkage disequilibrium. Our results do not confirm a possible correlation or linkage disequilibrium between the CAG/GGC haplotypes of the AR gene and androgenetic alopecia in Mexican brothers.

Isolation and characterization of microsatellite loci in the northern grass lizard, Takydromus septentrionalis (Squamata: Lacertidae)

The northern grass lizard (Takydromus septentrionalis) is an oviparous lacertid lizard that is endemic to China and has a wide distributional range covering the central and south-eastern parts of the country. It is listed on the ‘‘lists of terrestrial wildlife under state protection, which are beneficial or of important economic or scientific value’’ promulgated by China’s State Forestry Administration. Here, nine polymorphic microsatellite loci were isolated from T. septentrionalis genomic libraries. These markers revealed a high degree of genetic diversity (2–22 alleles per locus) and heterozygosity (H O ranged from 0.063 to 0.938, and H E ranged from 0.06 to 0.928). No locus exhibited deviations from Hardy–Weinberg equilibrium. There was no evidence of linkage disequilibrium among pairs of loci. These microsatellite markers will be useful for future study of population genetic analysis and parentage assignment of T. septentrionalis.