Genetic signature of amphimixis allows for the detection and fine scale localization of sexual reproduction events in a mainly parthenogenetic nematode

Asexuality is an important mode of reproduction in eukaryotic taxa and has a theoretical advantage over sexual reproduction because of the increased ability to propagate genes. Despite this advantage, hidden signs of cryptic sex have been discovered in the genomes of asexual organisms. This has provided an interesting way to address the evolutionary impact of sex in plant and animal populations. However, the identification of rare sexual reproduction events in mainly asexual species has remained a challenging task. We examined the reproductive history in populations of the plant parasitic nematode Xiphinema index by genotyping individuals collected from six grapevine fields using seven microsatellite markers. A high level of linkage disequilibrium and heterozygous excess suggested a clonality rate of 95–100%. However, we also detected rare sexual reproduction events within these highly clonal populations. By combining highly polymorphic markers with an appropriate hierarchical sampling, and using both Bayesian and multivariate analysis with phylogenetic reconstructions, we were able to identify a small number of sexually produced individuals at the overlapping zones between different genetic clusters. This suggested that sexual reproduction was favoured when and where two nematode patches came into contact. Among fields, a high degree of genetic differentiation indicated a low level of gene flow between populations. Rare genotypes that were shared by several populations suggested passive dispersal by human activities, possibly through the introduction of infected plants from nurseries. We conclude that our method can be used to detect and locate sexual events in various predominantly asexual species.     

Molecular biodiversity in the moss <Emphasis Type="Italic">Leptodon smithii</Emphasis> (Neckeraceae) in relation to habitat disturbance and fragmentation

Bryophytes seem particularly suitable to investigate genetic diversity in relation to habitat disturbance due to their large employment as bioindicators and to the recent application of molecular markers to moss population studies. Genetic variation and structure were analysed in seven urban, extraurban and remote populations of Leptodon smithii, an epiphytic moss of Quercus ilex, a phanerogamic species of Mediterranean climax vegetation. A total of 210 individual shoots were DNA extracted and amplified with internal simple sequence repeat (ISSR) primers, and 54 haplotypes were identified. An uneven distribution of haplotype number and frequencies was observed among sites, with a higher number of haplotypes and more homogeneous haplotype frequencies in the extraurban/remote populations. Molecular diversity indices were overall higher in the extraurban sites than in the urban ones. Multilocus linkage disequilibrium values were in line with the occurrence of sexual/asexual reproduction in the seven populations. The isolation-by-distance model was not supported by Mantel test among sites; however, within-population fixation index (F_ST) highlighted a clear relation between genetic and physic distances among trees, suggesting a limited dispersal range for L. smithii’s spores. The genetic structure was mainly affected by population size, wood structure and extent, and genetic drift consequent to habitat fragmentation and human-induced disturbance.     

Freshwater protists do not go with the flow: population structure in Gonyostomum semen independent of connectivity among lakes

Many recent studies have found genetically differentiated populations in microorganisms despite potentially high dispersal. We designed a study to specifically examine the importance of physical dispersal barriers, i.e. geographic distance and lack of hydrological connectivity, in restricting gene flow and enhancing divergence in limnic microorganisms. We focused on the nuisance microalga Gonyostomum semen, which has recently expanded in Northern Europe and differentiated into genetically distinct populations. G. semen was sampled from six lakes distributed in two adjacent watersheds, which thereby comprised, both connected and non‐connected lakes. The individual isolates were genotyped by amplified fragment length polymorphism. Several lake populations were differentiated from each other, but connectivity within watersheds could not explain the observed population genetic pattern. However, isolation by distance was moderate and might limit the gene flow among distant populations. In addition, we found low, but significant linkage disequilibrium, which indicates regular sexual recombination in this species, despite its high degree of asexual reproduction. Therefore, we conclude that the genetic properties of microalgae with occasional sexual reproduction essentially mirror regularly recombining species. Furthermore, the data indicated bottlenecks supporting the hypothesized recent range expansion of this species.     

Population genetic consequences of the reproductive system in the liverwort <Emphasis Type="Italic">Mannia fragrans</Emphasis>

Ecological factors affecting reproduction and dispersal are particularly important in determining genetic structure of plant populations. Polyoicous reproductive system is not rare in bryophytes; however, to date, nothing is known about its functioning and possible population genetic effects. Using the liverwort Mannia fragrans as a model species, the main aims of this study were to separate the relative importance of the components of the polyoicous reproductive system and to assess its consequences on the genetic structure of populations. High sex expression rates increasing with patch size and strongly female-biased sex ratios were detected. Additional input into clonal growth after production of sex organs was found in males compared to females. Similar clonal traits of the rare bisexual and asexual plants and preference toward newly colonized patches suggest that selection prefers colonizers that first develop organs of both sexes, hence ensuring sexual reproduction when no partner is present. Despite frequent spore production, ISSR markers revealed low genetic diversity, probably resulting from the effective clonal propagation of the species and frequent crossing between genetically identical plants. The presence of numerous rare alleles and unique recombinant haplotypes indicates occasional recombination and mutation. Effective spreading of new haplotypes is probably hampered however by large spore size. Since populations are small and isolated, such haplotypes are probably continuously eliminated by genetic drift. These results suggest that although both sexual and asexual reproductions seem to be effective, asexual components of the reproductive system play a greater role in shaping the genetic composition of the populations.     

Biogeography and population structure of the Neotropical endemic yeast species <Emphasis Type="Italic">Metschnikowia lochheadii</Emphasis>

The genetic structure of populations of the out-crossing haplontic yeast species Metschnikowia lochheadii was investigated. The species is associated with floricolous beetles in Central America and Hawaii. The objective was to determine whether sexual reproduction is prevalent and to what extent the geographic distribution of genotypes can be viewed as historical. The genetic markers examined include the mating type (h (+) or h (-)) and nine polymorphic DNA loci. The data were used to assess population structuring based on F (ST) and linkage disequilibrium and the distribution of alleles using parsimony haplotype networks. In Central America, M. lochheadii is subdivided into sexually active demes between which gene flow is limited. Isolates from five Hawaiian islands had identical haplotypes, confirming that the species has undergone a founder effect concomitant with the recent import of a nitidulid beetle into the archipelago.     

The phylogeographical and population genetic approach to the investigation of the genetic diversity patterns in self‐incompatible clonal and polyploid Linnaea borealis subsp. borealis

Surveys of genetic diversity patterns of self‐incompatible clonal polyploid plant species are still scarcer than those of diploid plant species. Therefore, I studied the phylogeographical history of Linnaea borealis subsp. borealis to shed light on the colonization history of this clonal self‐incompatible polyploid plant in Eurasia using selected regions of plastid DNA and genetic diversity patterns of 22 populations of this species employing AFLP markers. I also addressed the question of whether the genetic diversity patterns in L. borealis subsp. borealis in Eurasia are similar to those of earlier published studies of clonal self‐incompatible diploid or polyploid plants. This survey revealed that the shallow phylogeographical history (six plastid haplotypes forming one haplogroup, 100% bootstrap support) and moderate genome‐wide diversity estimated using AFLP markers (Frag_poly = 10.8–38.9%, I = 0.060–0.180, F_ST = 0.289) were general characteristics of L. borealis subsp. borealis in its Eurasian range. The sampling strategy, in most cases at 1–2‐m or even 3–5‐m intervals, showed that a balance between vegetative and sexual reproduction and limited pollen dispersal among compatible mates can be important for genetic diversity patterns in populations of this taxon. Despite the fact that one‐half of the investigated populations were strongly isolated, they still preserved similar levels of genetic diversity across the geographical range. I found no support for the hypothesis that a bottleneck and/or inbreeding had accompanied habitat fragmentation as factors shaping genetic diversity. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 64–76.     

Transmission of the apomictic mode of reproduction in Pennisetum: co-inheritance of the trait and molecular markers

Summary Apomixis, asexual reproduction through seed, is an obligate mode of reproduction in several species from the genus Pennisetum. Transfer of apomixis to sexual, cultivated pearl millet (P. glaucum) from a wild species P. squamulatum has resulted in an obligate apomictic backcross line with a low, but unknown number, of chromosomes from the wild species. Molecular markers (restriction fragment length polymorphisms and random amplified polymorphic DNAs) have been identified that unequivocally demonstrate the presence of P. squamulatum DNA in BC₃. Three of the informative RFLP clones have been sequenced and converted to sequence-tagged sites that can be amplified by the polymerase chain reaction. Molecular genetic analysis of more advanced back-cross individuals, using the two types of polymerase chain reaction-based molecular markers, has demonstrated co-inheritance of apomictic reproduction and two of the molecular markers. The remaining five molecular markers generally co-segregate with each other but are not linked with the mode of reproduction. These results suggest that genes for apomixis apparently can be transmitted by a single chromosome. Chromosome-specific markers will provide a starting point for the mapping of this genetically intractable reproductive trait.     

AFLP and RAPD Characterization of Phaeoacremonium aleophilum Associated with Vitis vinifera Decline in Spain

The genetic diversity among Spanish isolates of the fungus Phaeoacremonium aleophilum, one of the major causes of grapevine decline, was determined using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) techniques. Using RAPD, a large genetic variation was observed among 36 Pm. aleophilum single‐spore cultures, with 76 (82.6%) polymorphic bands generated by 12 RAPD primers. A neighbour‐joining dendrogram showing the RAPD patterns of diversity revealed four groups of haplotypes. The Bayesian and principal components clustering analysis revealed three groups of haplotypes. When more than one isolate of Pm. aleophilum was obtained from a single vine, different haplotypes were found. Seventeen single‐spore isolates were used for AFLP analysis. Five primer combinations produced 358 scorable markers, of which 309 (86.3%) were polymorphic. The analysis based on genetic distance as well as clustering analysis confirmed three main groups largely in agreement with those returned by the RAPD results. The Mantel correlation between the RAPD and AFLP distance matrices ranged from R = 0.5931 to R = 0.6294. The high level of haplotype diversity among the RAPD and AFLP markers suggests that sexual reproduction and genetic recombination may occur between Pm. aleophilum haplotypes in Spain. The AFLP approach revealed a greater number of polymorphic markers. A relationship between the genetic profile of the infecting isolate of Pm. aleophilum and the age or decline symptoms of the grapevines may exist.     

Notes on African Mosses. I. Andreaea camerunensis sp.nov. and other Mosses mostly from Nigeria and the British Cameroons

Abstract

Reproductive ecological traits such as success of fertilization, partitioning of sexes, the relative success of sexual versus asexual reproduction and dispersal distances are likely to considerably influence genetic structure within and among plant populations. In the liverwort Mannia fragrans both sexual and asexual reproduction can be frequently observed: sporophytes are produced abundantly every year and asexual propagation by fragmentation of thalli is also common. The aim of this study was to use ecological and molecular methods (ISSR markers) to separate the role of sexual and asexual components in shaping the partitioning of genetic variability within and among populations. In addition to genetic analyses conducted seasonally, sex expression and fertilization rates, sex ratios, regeneration from vegetative fragments and outcrossing was estimated in 3 populations of the species. Sex expression rates were high and, in spite of strongly female biased sex ratios, high fertilization rates were detected. However, capacity for regeneration from fragments was also high. Despite frequent spore production genetic diversity was low within populations. This is probably the result of the predominance of asexual reproduction s. 1. including crossing between genetically identical plants. Although recombination and mutation occasionally generates new haplotypes, these have little chance to spread because of the large spores mainly falling into their own patch, where chances for germination are low. Due to small size and isolation of the populations, genetic drift is likely to eliminate these haplotypes. Remote populations differed significantly, each being dominated by a few clones, reflecting negligible gene flow among them. Differences among individual populations can partly be related to differences in their reproductive behaviour and degree of isolation.     

Expanding the Species and Chemical Diversity of Penicillium Section Cinnamopurpurea

A set of isolates very similar to or potentially conspecific with an unidentified Penicillium isolate NRRL 735, was assembled using a BLAST search of ITS similarity among described (GenBank) and undescribed Penicillium isolates in our laboratories. DNA was amplified from six loci of the assembled isolates and sequenced. Two species in section Cinnamopurpurea are self-compatible sexual species, but the asexual species had polymorphic loci suggestive of sexual reproduction and variation in conidium size suggestive of ploidy level differences typical of heterothallism. Accordingly we use genealogical concordance analysis, a technique valid only in heterothallic organisms, for putatively asexual species. Seven new species were revealed in the analysis and are described here. Extrolite analysis showed that two of the new species, P. colei and P. monsserratidens produce the mycotoxin citreoviridin that has demonstrated pharmacological activity against human lung tumors. These isolates could provide leads in pharmaceutical research.     

Polymorphic MHC loci in an asexual fish,the amazon molly (Poecilia formosa; Poeciliidae)

Genes of the major histocompatibility complex (MHC) encode molecules that control immune recognition and are highly polymorphic in most vertebrates. The remarkable polymorphisms at MHC loci may be maintained by selection from parasites, sexual selection, or both. If asexual species show equal (or higher) levels of polymorphisms at MHC loci as sexual ones, this would mean that sexual selection is not necessary to explain the high levels of diversity at MHC loci. In this study, we surveyed the MHC diversity of the asexual amazon molly (Poecilia formosa) and one of its sexual ancestors, the sailfin molly (P. latipinna), which lives in the same habitat. We found that the asexual molly has polymorphic MHC loci despite its clonal reproduction, yet not as polymorphic as the sexual species. Although the nucleotide diversity was similar between the asexual and sexual species, the sexual species exhibited a greater genotypic diversity compared to the asexual one from the same habitats. Within‐genome diversity was similar for MHC class I loci, but for class IIB, the sexual species had higher diversity compared to the asexual — despite the hybrid origins and higher levels of heterozygosity at microsatellite loci in the asexual species. The level of positive selection appears to be similar between the two species, which suggests that these polymorphisms are maintained by selection. Thus, our findings do not allow us to rule out the sexual selection hypothesis for the evolution of MHC diversity, and although the sexual fish has higher levels of MHC‐diversity compared to the asexual species, this may be due to differences in demography, parasites, or other factors, rather than sexual selection.     

Genetic structure of South Australian Pyrenophora teres populations as revealed by microsatellite analyses

The aim of this study was to determine the genetic structure of South Australian field populations of the barley net blotch pathogens, Pyrenophora teres f. sp. teres (PTT) and P. teres f. sp. maculata (PTM), using microsatellite DNA markers. Three PTT populations (76 isolates total) and two PTM populations (43 isolates total) were sampled from separate fields during a single growing season. The results showed that of the 20 microsatellite loci examined, 17 (85 %) were polymorphic within the PTT and PTM populations. In total, 120 distinct alleles were identified of which only 11 (9 %) were shared between the two population types. Nei’s measure of gene diversity across the PTT and PTM populations was similar at 0.38 and 0.40, respectively, and also much higher than previously reported from studies in which other types of molecular markers were used. The coefficient of genetic differentiation among both populations was the same (G_ST = 0.03) and the low and insignificant estimates of F_ST, as indicated by θ, between populations of the same type (PTT: θ < 0.008, PTM: θ = 0.014) indicated that isolates sampled from different areas within the same field were genetically similar. In contrast, high and significant genetic differentiation was observed among and between populations of different type (G_ST = 0.42, θ > 0.567). The high number of unique multilocus haplotypes observed within the PTT (84 %) and PTM (100 %) populations, combined with a 1:1 distribution of both mating types, suggested that sexual reproduction was predominant among these populations. However, tests for multilocus associations showed that both PTT and PTM populations were in significant linkage disequilibrium. Although the levels of disequilibrium were low, an asexual reproductive component could not be excluded.     

Sequence Diversity and Linkage Disequilibrium within the Merozoite Surface Protein-1 (Msp-1) Locus of Plasmodium falciparum: A Longitudinal Study in Brazil

ABSTRACT. The merozoite surface protein‐1 (MSP‐1) is a major vaccine candidate for the asexual blood stage of malaria. We examined both the extent of sequence diversity in block 17, the 3′end of Msp‐1 gene coding for a 19‐kDa polypeptide (MSP‐1₁₉) putatively involved in red blood cell binding, and the patterns of linkage disequilibrium between polymorphic sites throughout the Msp‐1 locus. The parasite population sample consisted of Plasmodium falciparum isolates collected between 1985 and 1998 in Rondônia. an area of hypoendemic malaria transmission in the southwestern Brazilian Amazon. Results were summarized as follows. (I) Seven block‐17 sequence variants or haplotypes were found among 130 isolates, including two new haplotypes (novel combinations of previously reported amino acid replacements), here named Brazil‐1 (E‐TSR‐F) and Brazil‐2 (Q‐TSR‐F). (2) As previously shown for other Msp‐1 polymorphisms, frequencies of block‐17 haplotypes displayed significant temporal variation. (3) Extensive linkage disequilibrium was demonstrated between neighboring dimorphic sites within block 17, as well as between polymorphisms at the 5′and 3′ends of Msp‐1 (map distance range: 3.83–4.99 kb). (4) The overall patterns of linkage disequilibrium within Msp‐1 remained stable over a period of nearly one decade, and examples of possible ‘epidemic’ expansion of parasites carrying particular Msp‐1 alleles were found in the 1980s and 1990s. These results are discussed in relation to the population biology of P. falciparum and the development of malaria vaccines based on MSP‐1.     

Genetic structure of Polytrichum formosum in relation to the breeding system as revealed by microsatellites

Microsatellite variation was determined for three Danish and three Dutch populations of the haploid moss species Polytrichum formosum to gain insight into the relative importance of sexual vs. asexual reproduction for the amount and structure of genetic variation. In general, low levels of microsatellite variation were observed within this species. Even when estimated for polymorphic loci only, the levels of microsatellite variability (P=90.6, A=4.3 and H_S=0.468) within populations were on average lower than those reported for most other plant species. In contrast, genotypic diversity was high within each of the examined populations, indicating that sexual reproduction is a very important determinant of the genetic structure of P. formosum within populations. In agreement with previous findings for allozyme data, no significant genetic differentiation (F_ST=0.028, R_ST=0.015) was observed neither between populations nor between regions approximately 450 km apart (Denmark vs. the Netherlands). These low levels of population differentiation observed for both types of genetic markers are probably best explained by a high level of effective spore dispersal (gene flow) between populations. Therefore, also on a large geographical scale sexual reproduction is the most important determinant of the genetic structure of P. formosum, despite the high potential to reproduce clonally.     

Evidence from comparative genomics for a complete sexual cycle in the 'asexual' pathogenic yeast <Emphasis Type="Italic">Candida glabrata</Emphasis>

Background  

Candida glabrata is a pathogenic yeast of increasing medical concern. It has been regarded as asexual since it was first described in 1917, yet phylogenetic analyses have revealed that it is more closely related to sexual yeasts than other Candida species. We show here that the C. glabrata genome contains many genes apparently involved in sexual reproduction.     

BIOGEOGRAPHY OF ASEXUAL AND SEXUAL REPRODUCTION IN CALOGLOSSA (DELESSERIACEAE,RHODOPHYTA) FROM AUSTRALIA AND NEW ZEALAND

Caloglossa species are widely distributed in mangroves and salt marshes around the world and their life history patterns are being investigated in laboratory culture. In Australia all isolates of C. monosticha, C. postiae and C. ogasawaraensis have Polysiphonia‐type (P‐type) sexual life histories. Among the 70 C. leprieurii isolates from Australia and New Zealand P‐type sexual reproduction also is dominant. However, ten isolates of C. leprieurii from the Spencer Gulf and the Gulf of St. Vincent in South Australia give rise to successive tetrasporphyte generations without gametophytes. Moreover, one isolate from Queensland is asexual. Only one South Australia isolate, obtained from Lake Alexandrina at the mouth of the Murray River, is sexual. South Australia and Pacific Mexico are two regions in which asexual reproduction is dominant. In another mangrove dwelling red alga Bostrychia moritiziana (Rhodomelaceae) non‐sexual reproduction also is frequent in Australia, New Caledonia and Bali (Indonesia). This asexual reproductive pattern of tetrasporophytic recycling appears to have arisen independently among individual populations of various red algal species in different regions. Investigations are underway on the molecular phylogeny of the Caloglossa leprieurii isolates.     

EVALUATION OF ELEVATED PLOIDY AND ASEXUAL REPRODUCTION AS ALTERNATIVE EXPLANATIONS FOR GEOGRAPHIC PARTHENOGENESIS IN EUCYPRIS VIRENS OSTRACODS

Transitions from sexual to asexual reproduction are often coupled with elevations in ploidy. As a consequence, the importance of ploidy per se for the maintenance and spread of asexual populations is unclear. To examine the effects of ploidy and asexual reproduction as independent determinants of the success of asexual lineages, we sampled diploid sexual, diploid asexual, and triploid asexual Eucypris virens ostracods across a European wide range. Applying nuclear and mitochondrial markers, we found that E. virens consists of genetically highly differentiated diploid sexual populations, to the extent that these sexual clades could be considered as cryptic species. All sexual populations were found in southern Europe and North Africa and we found that both diploid asexual and triploid asexual lineages have originated multiple times from several sexual lineages. Therefore, the asexual lineages show a wide variety of genetic backgrounds and very strong population genetic structure across the wide geographic range. Finally, we found that triploid, but not diploid, asexual clones dominate habitats in northern Europe. The limited distribution of diploid asexual lineages, despite their shared ancestry with triploid asexual lineages, strongly suggests that the wider geographic distribution of triploids is due to elevated ploidy rather than to asexuality.     

Sexual and asexual reproduction of the holothurian Stichopus chloronotus (Echinodermata): a comparison between La Réunion (Indian Ocean) and east Australia (Pacific Ocean)

Summary

Stichopus chloronotus (Brandt, 1835) is one among nine aspidochirotide holothurian species known to reproduce both sexually by broadcast spawning and asexually by transverse fission. New data on the sexual cycle of this species in La Réunion are presented here and information available on sexual and asexual reproduction in this species is summarised. Sexual reproduction on La Réunion shows a distinct seasonality with a main spawning period in the warm season (November-February). The spawning period the Great Barrier Reef appears to be at the same time. Some intriguing deviations from unity in sex-ratio, usually biased towards more male individuals, have been observed in both geographic regions (sex ratio at La Reunion 31:1). New data on the asexual reproduction of this species in La Réunion confirm the high rates of fission. The peak of asexual reproduction in both the Indian and Pacific Ocean was observed in winter (June-July). Thus, asexual reproduction in this species occurs outside the season for sexual reproduction. The rate of asexual reproduction appears to vary between sample locations. However, results of population genetic studies on S. chloronotus (Uthicke et al., 1999; Uthicke et al., 2001) indicated that in most populations investigated a maximum of about 60% of all individuals may be derived from sexual recruitment. Cluster analyses on genetic distances between populations grouped populations within Oceans together, with the exception of one sample from a nearshore reef of the GBR. Although genetic differences between the two regions exist, these are relatively small regarding the large geographic distance. We conclude that asexual reproduction in S. chloronotus is important to maintain local population sizes, but that larval exchange between populations mediated by sexual reproduction is important for colonisation of new areas and to provide connectivity between populations. Here, we present the first synthesis of these phenomena for a holothurian species.     

Predominance of sexual reproduction in Romanian populations of the aphid Sitobion avenae inferred from phenotypic and genetic structure

Models of coexistence of sexual and asexual lineages in aphids assume that obligate parthenogenetic lineages predominate in areas with mild winter climate because of their high reproductive output, while sexual lineages predominate in areas with severe winter because they produce eggs resistant to frost. To validate this hypothesis in natural conditions, the reproductive mode of populations of the aphid Sitobion avenae was assessed in two very contrasting climatic situations, Romania (severe winter) and Western France (mild winter). To achieve this, reproductive modes were inferred from both (1) the population composition in sexual and asexual forms in autumn, and (2) the genetic structure of Romanian and French populations of S. avenae using microsatellite markers. Romanian populations encompassed a high proportion of sexual forms and were characterised by a very high genotypic diversity and low linkage disequilibrium. In constrast, the French population showed frequent linkage disequilibria, low genetic diversity, and high level of clonal amplification with two asexual genotypes representing over 60% of the sample. In agreement with the model's predictions, these results clearly indicate that sexual reproduction in S. avenae is predominant under the continental climate of Romania, while asexual lineages prevail under the oceanic climate of Western France.     

Isolation and characterization of microsatellite DNA loci from Russell's snapper (Lutjanus russellii)

A total of 37 microsatellite loci from the Russell's snapper, Lutjanus russellii, were successfully isolated and characterized. Thirty‐four loci were polymorphic in L. russellii samples. Twenty of the 37 markers did not differ significantly from Hardy–Weinberg expected genotype proportions. Significant linkage disequilibrium was detected in one pairwise comparison. The numbers of alleles and observed heterozygosities in polymorphic loci ranged from two to 16 and from 0.41 to 0.95, respectively. These markers will be useful for studying the population genetic structure of this species.