The use of microsatellite polymorphism in genetic mapping of the ostrich (<Emphasis Type="Italic">Struthio camelus</Emphasis>)

The aim of this study was to determine microsatellite polymorphism in ostriches and using it in creation the genetic map of the ostrich. The polymorphism analysis covered 30 microsatellite markers characteristic of ostrich, for the CAU (China Agricultural University) group. The material consisted of 150 ostriches (Struthio camelus). The 30 microsatellite loci was examined and a total of 343 alleles was identified. The number of alleles at a single locus ranged from 5 at locus CAU78 to 34 at locus CAU85. The values for the observed heterozygosity H_o ranged from 0.467 (locus CAU78) to 0.993 (locus CAU16), whereas for the expected heterozygosity H_e - from 0.510 (locus CAU78) to 0.953 (locus CAU85). Analyzing the individual loci, the highest PIC value, more than 0.7 was observed for: loci CAU85 (0.932), CAU64 (0.861) and CAU32, 75 (0.852), respectively. It should be noted, that the microsatellite markers used in our study were very polymorphic as evidenced by the large number of detected alleles and high rates of heterozygosity, PIC and PE as well. The analysed microsatellite markers may be used in genetic linkage mapping of ostrich, the construction of a comparative genetic map with other ratites, such as emu and rhea, and population genetics studies or phylogenetic studies of these birds.     

Genetic diversity of the endangered Chinese endemic herb Dayaoshania cotinifolia (Gesneriaceae) revealed by simple sequence repeat (SSR) markers

Dayaoshania cotinifolia W.T. Wang is a critically endangered perennial herb endemic to southern China. Simple sequence repeat (SSR) markers were employed to assess genetic diversity in two populations of D. cotinifolia. Eight primer pairs generated a total of 36 alleles, with a mean of 4.5 alleles per locus. The expected heterozygosity (H_e = 0.416) and observed heterozygosity (H_o = 0.508) indicated a moderate level of genetic diversity, though the genetic differentiation between the populations was low (F_st = 0.014), a result that was supported by a higher gene flow (N_m = 18.000). No severe bottleneck effect was detected in the two populations. Thus, the endangered status of this species is most likely due to anthropologic effects rather than a lack of genetic diversity. In situ conservation strategies should be promoted, and the sizes of the populations should be increased through artificial breeding.     

Low levels of realized seed and pollen gene flow and strong spatial genetic structure in a small,isolated and fragmented population of the tropical tree Copaifera langsdorffii Desf

Over the past century, the Brazilian Atlantic forest has been reduced to small, isolated fragments of forest. Reproductive isolation theories predict a loss of genetic diversity and increases in inbreeding and spatial genetic structure (SGS) in such populations. We analysed eight microsatellite loci to investigate the pollen and seed dispersal patterns, genetic diversity, inbreeding and SGS of the tropical tree Copaifera langsdorffii in a small (4.8 ha), isolated population. All 112 adult trees and 128 seedlings found in the stand were sampled, mapped and genotyped. Seedlings had significantly lower levels of genetic diversity (A=16.5±0.45, mean±95% s.e.; H_e=0.838±0.006) than did adult trees (A=23.2±0.81; H_e=0.893±0.030). Parentage analysis did not indicate any seed immigration (m_seeds=0) and the pollen immigration rate was very low (m_pollen=0.047). The average distance of realized pollen dispersal within the stand was 94 m, with 81% of the pollen travelling <150 m. A significant negative correlation was found between the frequency and distance of pollen dispersal (r=−0.79, P<0.01), indicating that short-distance pollinations were more frequent. A significant SGS for both adults (∼50 m) and seedlings (∼20 m) was also found, indicating that most of the seeds were dispersed over short distances. The results suggested that the spatial isolation of populations by habitat fragmentation can restrict seed and pollen gene flow, increase SGS and affect the genetic diversity of future generations.     

Characterization of eight microsatellite loci for the sea urchin Meoma ventricosa (Spatangoida,Brissidae) through Next Generation Sequencing

Eight microsatellite loci were characterized for Meoma ventricosa (Lamarck, 1816), a burrowing sea urchin that can be afflicted by a bacterial disease causing localized mass mortality. For the analyzed population (29 individuals from St. Croix, US Virgin Islands), we observed 8.125 mean number of alleles, 0.640 mean observed heterozygosity (H_o) and 0.747 mean expected heterozygosity (H_e). Two loci showed significant deviations from Hardy–Weinberg equilibrium. Overall, the described loci were characterized by a moderately high level of polymorphism suggesting that these markers are useful for a population genetic study in the Caribbean Sea.     

Case study of microsatellite polymorphism of European perch in selected commercially important lakes of Latvia

Polymorphism of selected microsatellite markers was described for European perch populations of inland Lake Kala and costal Lake Babites in Latvia. The data consisted of ten microsatellite loci analysed for 90 individuals from the Lake Babites (n?=?45) and the Lake Kala (n?=?45). Both lakes differ in water area, connection with the Gulf of Riga (Baltic Sea), lake type, and fish species. In the Lake Kala population, the average number of alleles per locus varied from 4 to 15 and in the population of Lake Babites from 4 to 12. Low but significant genetic differentiation was detected between populations (F_ST?=?0.046). Observed and expected heterozygosity in both lakes was similar (Kala: H_o?=?0.680 and H_e?=?0.816; Babites: H_o?=?0.693 and H_e?=?0.815). The number of unique alleles per locus was 2.3 in Lake Kala and 0.6 in Lake Babites. The present study showed relatively high polymorphism of analysed microsatellite markers.     

Genetic diversity of wild population of Pyropia haitanensis based on SSR analysis

Genetic diversity analysis was conducted on 80 individuals of 4 populations of non-bred Pyropia haitanensis by simple sequence repeat (SSR) method. Using 15 pairs of microsatellite primers, 37 polymorphic loci were amplified, representing 94.9% of all loci. At the population level, the percentage of polymorphic bands (P) and polymorphism information content (PIC) were 66.67–84.62% and 0.481–0.488, with average value at 73.72% and 0.483, respectively. Expected heterozygosity (H_e) and Shannon's information index (I) were 0.279 and 0.434, respectively, at the species level, and 0.233 and 0.356 at population level. According to the coefficient of gene differentiation (G_ST), a large proportion of genetic variance (83.6%) of P. haitanensis was among individuals within populations, only 16.4% genetic variance was among populations, which was identified with the moderate gene flow value (N_m = 2.542). UPGMA clustered the 4 populations into 3 groups, and no significant correlation was found between the genetic distance and the corresponding geographic distance among the populations.     

Genetic diversity and fine-scale genetic structure in Hancornia speciosa Gomes (Apocynaceae)

We studied the genetic diversity and spatial genetic structure (SGS) of adult and juvenile individuals in a population of Hancornia speciosa in Central-West Brazil. For this, we sampled and mapped 113 adults and 100 juveniles in an area of 2.5 ha. Genomic DNA was obtained from leaves and seven microsatellite loci were used to genotype all individuals. The studied population showed high genetic diversity (H_e) but with significant inbreeding (f) for both life stages most likely due to biparental inbreeding. Spatial genetic structure was weak for both life stages and the values of S_P were low and neighborhoods (Nb) was high for both generations showing a potential long-distance gene dispersal.     

Genetic Structure of Oryza rufipogon Griff. Natural Populations in Malaysia: Implications for Conservation and Genetic Introgression of Cultivated Rice

Thirty polymorphic Oryza sativa microsatellite loci (SSRs) were used to study population genetic structure of O. rufipogon Griff. natural populations in Malaysia. A total of 445 alleles were detected with an average of 14.8 alleles per locus in 176 individuals of O. rufipogon sampled from the states of Penang, Kedah, Kelantan and Terengganu where the natural populations are still found. The Kelantan population in the northeast of Peninsular Malaysia had the highest level of genetic diversity as measured by the mean number of alleles per locus, A_a?=?7.67, average number of effective alleles, A_e?=?5.50, percentage of polymorphic loci, P?=?100%, observed heterozygosity, H_o?=?0.631 and expected heterozygosity, H_e?=?0.798. In contrast, the Terengganu population in the east showed the lowest level of genetic diversity measured by the same criteria (A_a?=?4.23, A_e?=?2.10, P?=?100%, H_o?=?0.549 and H_e?=?0.449). Model–based clustering analysis using the STRUCTURE 2.2 program placed all the individuals into 12 clusters that corresponded to the geographic sampling locations. Neighbour-joining tree was constructed based on Nei’s genetic distance to further assess the genetic structure of the O. rufipogon individuals, showed good agreement (93.8%) with the model-based cluster analysis. However, the neighbour-joining tree identified sub-populations that STRUCTURE could not identify. The classification of individuals from the same populations under the same cluster supported the population differentiation. These two analyses seemed to indicate expansion of populations from the northeast of Peninsular Malaysia (Tumpat, Pasir Mas and Kota Bahru, Kelantan) not only to the immediate south of the region i.e. Terengganu but also into the northwest (i.e. Penang and Kedah) with the former being more recent. Oryza rufipogon accession IRGC105491 and O. sativa ssp. indica cultivar MR219, which were included in this study for comparisons with the local wild rice accessions, indicated that introgression of cultivated rice could change genetic composition and affect the population genetic structure of wild rice. This possibility should be carefully considered in plans to conserve this wild rice.     

Genetic diversity of populations of a rare species of Dictamnus gymnostylis Stev. in Bashkir Cis-Urals

We examined genetic diversity in populations of Dictamnus gymnostylis Stev., a rare species growing in the Bashkir Cis-Urals, based on the analysis of 8 gene-enzyme systems and detected a fairly high level of intraspecific genetic diversity and population differentiation. We determined the average number of alleles per locus (A) as 1.57; the portion of polymorphic loci (P ₉₅) as 0.508; the observed heterozygosity (H _o) as 0.139; and the expected heterozygosity (H _e) as 0.169. Of the total genetic diversity, 88.3% stems from variability within populations, and 11.7% is due to variation among populations. The average value for the Nei’s genetic distance (D) constituted 0.028.     

Microsatellite development for a tetrodotoxin-containing sea slug (Pleurobranchaea maculata)

Using 454 pyrosequencing data, 24 polymorphic microsatellite markers were identified for the grey side-gilled sea slug, Pleurobranchaea maculata. The grey side-gilled sea slug is found throughout the western and south Pacific and is known to contain high concentrations of tetrodotoxin. Polymorphism was assessed in 20 individuals obtained from geographically distinct locations within New Zealand. Between 2 and 15 alleles were identified at each locus. The observed heterozygosity (H_o) and expected heterozygosity (H_e) ranged from 0.10 to 1 and 0.10–0.94, respectively. No significant linkage disequilibrium between pairs of loci or deviations from the Hardy–Weinberg proportions were observed. The markers are central to understanding the population biology and genetic structure of P. maculata.     

Genetic Structure of Wild Rice Oryza Glumaepatula Populations in Three Brazilian Biomes Using Microsatellite Markers

The existence of Oryza glumaepatula is threatened by devastation and, thus, the implementation of conservation strategies is extremely relevant. This study aimed to characterize the genetic variability and estimate population parameters of 30 O. glumaepatula populations from three Brazilian biomes using 10 microsatellite markers. The levels of allelic variability for the SSR loci presented a mean of 10.3 alleles per locus and a value of 0.10 for the average allelic frequency value. The expected total heterozygosity (H_e) ranged from 0.63 to 0.86. For the 30 populations tested, the mean observed (H_o) and expected heterozygosities (H_e) were 0.03 and 0.11within population, respectively, indicating an excess of homozygotes resulting from the preferentially self-pollinating reproduction habit. The estimated fixation index ( _IS ) was 0.79 that differed significantly from zero, indicating high inbreeding within each O. glumaepatula population. The total inbreeding of the species (_IT ) was 0.98 and the genetic diversity indexes among populations, _ST and _ST, were 0.85 and 0.90, respectively, indicating high genetic variability among them. Thus, especially for populations located in regions threatened with devastation, it is urgent that in situ preservation conditions should be created or that collections be made for ex situ preservation to prevent loss of the species genetic variability.     

Polymorphic microsatellites identified by cross-species amplifications in the European Coot Fulica atra

We tested the cross-amplification of 26 microsatellites developed for passerines and an additional three developed for Gallinula species in eight European Coots from two populations. Sixteen microsatellite markers successfully amplified, of which nine were polymorphic with 2–6 alleles (mean 3.7 alleles) and an expected heterozygosity (H _e) ranging from 0.375 to 0.805 (mean H _e = 0.589). On average, we found 2.22 alleles/locus and a mean H _e of 0.440 in one nest, and 2.56 alleles/locus and a mean H _e of 0.494 in the other one. These nine polymorphic markers could be of potential use in studies of genetic variability, population structure and reproductive strategy of European Coots.     

Genetic diversity and parentage assignment in Dojo loach,Misgurnus anguillicaudatus based on microsatellite markers

In this study, genetic diversity and structure of three Misgurnus anguillicaudatus populations from three different geographical locations in China (Hunan, Hubei and Henan province) were investigated using microsatellite markers. High level of genetic diversity of all three populations was revealed by expected heterozygosity (H_e), observed heterozygosity (H_o) and allele number. Significant genetic differentiations were found between all pairs of populations. The efficiency of eight microsatellite markers in parentage assignment of 540 progeny from twenty full-sib families was evaluated. Simulation based on allele frequency data demonstrated that probabilities of exclusion per locus range from 0.313 to 0.825 when no parent information is available and 0.504 to 0.904 when one parent is known. The assignment success rate based on the real data using eight markers was 96.85%. This study indicates that these M. anguillicaudatus resources are valuable genetic and breeding material for aquaculture and the microsatellite markers will be useful for investigation of genetic background and molecular marker-assisted selective breeding in this species.     

Genetic variation and population genetic structure of Rhizophora apiculata (Rhizophoraceae) in the greater Sunda Islands,Indonesia using microsatellite markers

Genetic variations within and among Rhizophora apiculata populations in the Greater Sunda Islands of Indonesia were studied using microsatellite markers. The study found 38 alleles on five loci in 15 populations. The observed (H _o) and expected (H _e) heterozygosity values are 0.338 and 0.378, respectively. Inbreeding effect from self-pollination might explain its heterozygote deficiency. Population genetic differentiation (F _ST = 0.381) was similar to other mangrove species. The genetic diversity of R. apiculata populations along the coastline inside the archipelago (e.g., Buleleng, Donggala, Mamuju, and Takalar) was higher than those of population along the coastline outside the archipelago, especially northern Sumatra populations (i.e., Langkat, Tapanuli Tengah, Dumai, and Padang). The isolation by distances and sea currents directions as well as their connectivity might affect the gene flow and genetic exchange. The more isolated with fewer connections by sea currents, the smaller gene flow and genetic exchange observed between populations. The higher genetic exchange, on the contrary, occurred when population location was closer to the meeting point of the sea currents. The study also showed that the patterns of sea current movement seemed to have influence genetic clustering of populations which fell into three main groups (Sunda Shelf Mangroves) and one isolated population (New Guinea Mangroves).     

Allozyme variation of the endangered endemic plant Myricaria laxiflora: Implications for conservation

Myricaria laxiflora, a riparian plant that naturally occurs in the riverbanks of the Yangtze River Valley, has become extinct across its entire geographical distribution range in the wild due to the construction of the Three Gorges Dam. The allozyme variation of M. laxiflora populations was investigated in the present study. Mean number of alleles per locus (A) was 2.35, and the observed heterozygosity (H_o) and expected heterozygosity (H_e) were 0.35 and 0.30, respectively. Six populations showed significant excesses of heterozygotes based on the examination of a multilocus fixation index (F_IS). The population genetic divergence of M. laxiflora is high (G_ST = 0.144 and ?_B = 0.131) and the analysis of molecular variance analysis shows that 19.71% of the total genetic variation is caused by the difference between populations. Based on the obtained genetic information, six management units have been identified, all of which are expected to enhance the effective management of the remaining and transplanted individuals of this endangered species in the future.     

Development of thirty new polymorphic microsatellite primers for <Emphasis Type="Italic">Paeonia suffruticosa</Emphasis>

Four new microsatellite primer pairs were developed in tree peony (Paeonia suffruticosa) based on the database mining and other twenty-six primer pairs by fast isolation by AFLP of sequences containing repeats (FIASCO) method. The polymorphism of each locus was further evaluated in 40 individuals of one population plus 5 tree peony related species. The number of alleles per locus ranged from 3 to 7 and the expected (H_e) and observed (H_o) heterozygosity at each locus ranged from 0.42 to 0.78 and 0.28 to 0.59, respectively. These microsatellite markers will be useful for investigating genetic diversity and studies of population genetic structure of tree peony.     

Fine-scale genetic structure of the threatened rosewood Dalbergia nigra from the Atlantic Forest: comparing saplings versus adults and small fragment versus continuous forest

Assessing the population genetic structure of threatened species is important for developing successful conservation strategies. In this study, we evaluated the fine-scale spatial genetic structure (SGS) of Dalbergia nigra from a regenerating secondary forest fragment and compared it with previous data from a primary forest of a large reserve. A total of 107 adult and 111 saplings were mapped and genotyped for seven microsatellite loci. The genetic diversity was high and similar in adults (H _e?=?0.682) and saplings (H _e?=?0.680). The spatial extent of SGS was higher in adults than in saplings. Overlapping generations in the potentially reproductive individuals is the likely explanation for the higher SGS in adults (Sp?=?0.016) in relation to the saplings (Sp?=?0.010). The SGS in the adults from the secondary forest fragment was similar to that found in the primary forest. Considering the SGS found in adults, from both the secondary and primary forests, seeds for ex situ conservation should be collected from trees at least 80 m apart to reduce the genetic similarity between samples. These results highlight the importance of preserving small forest fragments to allow successful regeneration and maintenance of the genetic diversity in D. nigra.     

Genetic diversity and population structure of the Asian green mussel Perna viridis in South China Sea based on microsatellite markers

The green mussel, Perna viridis is ecologically and economically important in the coastal region of the South China Sea. Determining its population genetic structure at this fine geographic scale will help sustainable management of natural stocks. In this study, we examined the genetic diversity and population genetic structure of P. viridis from four locations in the South China Sea (n = 45–48) using nine microsatellite loci. The results showed moderate levels of genetic diversity in all four samples (mean A = 13.222–14.000, mean A_e = 7.092–7.571, mean A_r = 12.894–13.746, mean H_o = 0.596–0.656, mean H_e = 0.690–0.733) and a large effective population size estimate for the pooled sample (total N_e estimates = infinity, 95% CI = 1869.0-infinity). We did not detect any sign of recent bottleneck events in P. viridis populations in the South China Sea. The conventional and a model-based analysis reveal low, non-significant genetic divergence among the four samples (F_ST = − 0.001–0.005, P > 0.05/6). The results obtained from this study can provide valuable genetic information for the conservation and fishery management of P. viridis by retaining the high N_e estimates.     

Structure of genetic diversity in marginal populations of black poplar (Populus nigra L.)

It has been hypothesized that populations at the margins of the distributional range of a species show reduced genetic diversity and increased inter-population differentiation compared to central populations. Here, we test this hypothesis by examining the structure of genetic diversity in marginal populations of black poplar, Populus nigra L. (Salicaceae). This species occurs mainly in Europe but its range extends to central Asia. We collected 117 individuals from 10 populations at the edge of the distributional range of the species in central Asia to examine the structure of genetic diversity based on genetic polymorphisms at 20 microsatellite markers. As expected, the genetic diversity within these marginal populations is relatively low, with an average observed heterozygosity H_o of 0.337 and an average expected heterozygosity H_e of 0.466, compared to the genetic diversity of populations from central distributions. However, we recovered very low genetic differentiation between populations, with an average F_st of 0.0745, a value similar to those reported for central populations. AMOVA analyses confirmed this result, showing that only 9.2% of the total variation could be attributed to between-population variance (P < 0.001). Our findings do not fully support hypotheses about the structure of genetic diversity in marginal populations formed from observations on other species. We suggest that a high rate of outcrossing and possible postglacial colonization at the edge of the distributional range of this long-lived poplar may explain the observed structure of the genetic diversity.     

Isolation and characterization of new microsatellite markers from the Japanese scallop (<Emphasis Type="Italic">Patinopecten yessoensis</Emphasis>)

Twenty polymorphic microsatellite loci were isolated from the Japanese scallop (Patinopecten yessoensis) using a method of hybridizing twice. The genetic diversity was analyzed by these microsatellite markers in field specimens from Zhangzidao Island in Liaoning Province, China. Observed heterozygosity (H _o), expected heterozygosity (H _e), number of effective alleles (N _e), and polymorphism information contents (PIC) were calculated. The heterozygote deficiency or excess was detected by the fixation index (Fis). The Hardy–Weinberg equilibrium was tested by possibilities (P _HWE) using chi-square test. Results indicate that the average of allele numbers was 6.0. The mean values of the parameters H _o, H _e, N _e, and PIC were 0.7921, 0.7076, 4.4347, and 0.6684, respectively. Only 4 of 20 microsatellite markers showed heterozygote deficiency including loci FJ262370, FJ262381, FJ262384, and FJ262400. The P _HWE value indicates the wild population deviated somewhat from the Hardy–Weinberg equilibrium. These newly isolated markers increase the available molecular resources of the Japanese scallop.