Isolation and characterization of 14 polymorphic microsatellite loci in the ark shell Scapharca subcrenata (Bivalvia: Arcidae)

The ark shell Scapharca subcrenata is an important fishery resource, but has been suffering from severe population decline due to the deterioration of the coastal environment and over-exploitation. To investigate the genetic variation and structure among populations of S. subcrenata, 14 polymorphic microsatellite loci were developed. The number of alleles per polymorphic locus ranged from 11 to 29 and the observed and expected heterozygosity varied from 0.400 to 0.923 and from 0.705 to 0.965, respectively. These microsatellites will help advance the investigation of the genetic population structure and genetic diversity in this species.     

Isolation and characterization of microsatellite loci in endangered Cycas changjiangensis (Cycadaceae)

Eight microsatellite loci were isolated from repetitive DNA enriched libraries for Cycas changjiangensis, an endangered endemic species in Hainan Island, China. The number of allele ranged from three to seven. The observed (H _O) and expected (H _E) heterozygosities ranged from 0.0000 to 0.8750 and from 0.2359 to 0.7582, respectively. These microsatellite loci will enrich our scientific understanding for C. changjiangensis conservation.     

Isolation and characterization of microsatellite DNA markers from mangrove crab,Scylla paramamosain

The first five variable microsatellite DNA loci for mangrove crab, Scylla paramanosain, were developed. Allelic variation and other characteristics at these loci were examined in this species captured at the Urado Bay, Japan. The number of alleles per locus ranged from 24 to 44. The expected heterozygosity across loci ranged from 0.900 to 0.999 and probability of identity (P_I) ranged from 2.8 × 10^?3 to 1.7 × 10^?2. Therefore, these microsatellite markers could be useful for estimating effect of stock enhancement release and population genetic structure of mangrove crab.     

Development of microsatellite loci for Pelophylax plancyi and cross-amplification in other ranid species

Thirteen polymorphic microsatellite loci were obtained from the gold-striped pond frog, Pelophylax plancyi. The number of alleles per locus ranged from 5 to 13 with an average of 8.38. The observed and expected heterozygosities (H _O and H _E) ranged from 0.533 to 0.890 and from 0.620 to 0.897, with averages of 0.686 and 0.751, respectively. In order to assess interspecific amplification, all primer pairs were tested with the same PCR conditions on three other Pelophylax species: Pelophylax hubeiensis, Pelophylax fukienensis, Pelophylax nigromaculata and two species from the genus Hylarana and Hoplobatrachus, respectively. All of these loci can be amplified successfully in the Pelophylax species, with eight loci amplified in all species tested.     

Characterization of microsatellite markers in the mosquito Ochlerotatus caspius (Diptera: Culicidae)

Seven polymorphic microsatellite loci were developed for the mosquito species Ochlerotatus caspius, using an enriched genomic library. The number of alleles per locus varied between two and 11; the expected heterozygosity (H_E) ranged from 0.18 to 0.77. These microsatellite primers should prove useful for population genetic studies of this mosquito species.     

Isolation and characterization of 13 microsatellite loci from Incarvillea mairei (Bignoniaceae), an endemic species to the Himalaya-Hengduan mountains region

Incarvillea mairei (H. Léveillé) Grierson (Bignoniaceae) is an endemic species to the Himalaya-Hengduan Mountains region. Here, we developed 13 microsatellite markers from I. mairei using a modified biotin-streptavidin capture method. Number of alleles per locus (NA) ranged from 3 to 7 with an average of 4.615. The observed (H _O) and expected (H _E) heterozygosities were from 0.050 to 0.800 and from 0.249 to 0.815, respectively. Additionally, among the 13 identified microsatellite markers, 12 of them were successfully amplified in other three congeneric species, and most of them showed polymorphic. Obtained evidences suggest that these markers provide a useful tool for further study of the population genetic structure and the breeding system in this species or/and infra-generic species.     

Characterization of microsatellite loci in Coenonympha hero (Lepidoptera: Nymphalidae)

Microsatellite markers are suitable tools for studying dispersal pattern among local populations. I report on the characterization of seven polymorphic microsatellite loci in the scarce heath butterfly (Coenonympha hero), from one unenriched and one enriched genomic library. Number of alleles ranged from two to 20 when 108 individuals from seven populations were screened. H_O ranged from 0.140 to 0.889. Primers were also tested on three other butterfly species. Amplification was successful for all loci in Erebia triaria, while only one gave products in Maculinea alcon and Maculinea rebeli. To my knowledge, these are the first microsatellite markers published in the Nymphalidae subfamily Satyrinae.     

Isolation and characterization of microsatellite loci in Lesquerella fendleri (Brassicaceae) and cross‐species amplification

Fifteen novel microsatellite primer pairs are presented for Lesquerella fendleri, which were developed from seven dinucleotide, five trinucleotide and three tetranucleotide microsatellite DNA loci. These loci were characterized for 40 individuals from 24 localities throughout the species range. The number of alleles observed per locus ranged from three to 16, the observed heterozygosity ranged from 0.175 to 0.750, and the polymorphic information content ranged from 0.218 to 0.889. Cross‐species transferability tested on nine species of Lesquerella and one species of the related genus Physaria indicates that these primer pairs may be useful for population genetic studies of other species in Lesquerella and possibly other closely related genera.     

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.     

Transferability and characterization of microsatellite markers from Byrsonima cydoniifolia A. Juss. (MALPIGHIACEAE) in seven related taxa from Cerrado biome reveal genetic relationships

Byrsonima Rich. is one of the largest genera of the Malpighiaceae family with 97 species occurrence in Brazil and multiple potentialities, including pharmaceutical and food industries. In this study, 17 microsatellite markers characterized in Byrsonima cydoniifolia were tested for seven related taxa, all species are native to Brazil and four are endemic. Genomic DNA was extracted from leaves tissues and 17 microsatellite markers were used to cross-amplification of microsatellite regions. Polymorphism and genetic diversity were evaluated for B. intermedia, B. verbascifolia, B. laxiflora, B. subterranea, B. umbellata, B. linearifolia. from 16 individuals and for B. viminifolia from 14 individuals. Transferred microsatellite markers panels ranged from 11 (64.8%) in B. viminifolia to 6 (35.2%) in B. umbellata. The total number of alleles per locus ranged from 5 (B. linearifolia) to 8 (B. subterranea) alleles. B. umbellata showed lower values of observed and expected heterozygosity (H_O?=?0.312; H_E?=?0.436) and B. subterranea presented the highest values (H_O?=?0.687; H_E?=?0.778). A greater number of microsatellite markers should be developed for B. umbellata. The microsatellite marker panels transferred to the species B. intermedia, B. verbascifolia, B. laxiflora, B. subterranea, B. viminifolia and B. linearifolia are very informative, with a high combined probability of exclusion of paternity (Q?≥?0.976) and the low combined probability of identity (I?≤?9.91?×?10^–6), potentially suitable for future genetic-population studies, supporting strategies for maintaining the genetic diversity and for exploration of Byrsonima species as genetic resources.

     

Isolation of microsatellite DNA and analysis on genetic diversity of endangered fish,Hucho taimen (Pallas)

Hucho taimen (Pallas) is an endangered species in China. To evaluate the species’ genetic diversity and population genetic structure, we isolated six polymorphic microsatellite markers from its genomic libraries by (ACA)₁₆ enriched, and characterized using 61 wild individuals. The number of alleles per locus ranged from two to seven, expected heterozygosity ranged from 0.3200 to 0.7410, polymorphism information content ranged from 0.3047 to 0.6896. These markers will be useful for the genetic variation assessment of taimen.     

Isolation of polymorphic microsatellite loci from the endangered mudflat crab Chasmagnathus convexus (Crustacea: Decapoda: Varunidae)

Chasmagnathus convexus is a mudflat crab that is distributed throughout East Asia. However, most populations of this species in Korea are endangered due to continuing seashore developments and habitat fragmentations. In this study, we isolated and characterized nine microsatellite loci from C. convexus. The number of alleles per locus ranged from 3 to 23. The expected heterozygosities ranged from 0.145 to 0.974 and the observed heterozygosities ranged from 0.150 to 0.947. Cross-species amplifications for 10 other Varunid crab species were performed. These microsatellite markers could be useful for planning conservation strategies for C. convexus and related species.     

Isolation and characterization of polymorphic microsatellite markers in Anthyllis vulneraria

To study the reproductive system of the putative autogamous species Anthyllis vulneraria in Belgian calcareous grasslands, eight polymorphic microsatellite loci were developed using the sequence‐tagged microsatellite profiling technique and 35 plants were typed for all loci. No linkage disequilibrium was found between any pair of loci. Over the loci, the number of alleles ranged from two to five. Observed and expected heterozygosities ranged from 0.053 to 0.688 and from 0.053 to 0.631, respectively. The estimated F_IS values (F_IS = 0.031 and 0.122) are not in accordance with a predominantly selfing reproductive system.     

Isolation and characterization of eight polymorphic microsatellite loci from Ludwigia polycarpa (Onagraceae), a threaten herb in North America

Ludwigia polycarpa is the only species of section Microcarpium occurring north of 37°N in the central midwest of the United States. Recently, the loss of wetlands in North America reduced the population number and size of L. polycarpa dramatically. In this study, for the purpose of the conservation of the endangered species, we described eight microsatellite DNA loci. High variabilities enable these molecular markers to assess the population structure. The number of alleles per locus ranged from 8 to 23. The expected (H _E) and observed (H _O) heterozygosities ranged from 0.86 to 0.96 and 0.00 to 1.00, respectively. Seven of the eight microsatellite loci displayed significant departures from Hardy–Weinberg expectations, likely due to the loss of habitats and the small population size. No linkage disequilibrium was observed in the pairwise comparisons of loci. The application of these microsatellite loci in L. polycarpa may provide a tool for understanding its demography and population structure.     

Isolation and characterization of microsatellite loci in the highland papaya Vasconcellea × heilbornii V. Badillo (Caricaceae)

Nine polymorphic microsatellite loci were identified in the tropical plant Vasconcellea ×heilbornii and used to estimate allelic diversity in two populations of southern Ecuador. Allelic richness ranged from two to five alleles, observed heterozygosity ranged from 0.150 to 0.947 and expected heterozygosity ranged from 0.186 to 0.701. Most of these markers also amplified microsatellite loci in two other Vasconcellea species (Vasconcellea stipulata and Vasconcellea cundinamarcensis). Hence, these markers will be useful for population genetic analysis and the evaluation of genetic diversity and gene flow in these species.     

Development, characterization and transferability of microsatellite markers for Cullen australasicum (Leguminosae)

The development of eight polymorphic microsatellite markers for use in the Australian native legume Cullen australasicum is described. The number of alleles per locus ranged from 3 to 8. Observed heterozygosity ranged from 0.208 to 0.667. The cross-species amplification of these eight markers and a ninth marker, which is monomorphic in the populations examined, but may be used to distinguish between species, was also tested in five other species. These microsatellite markers will be useful for investigating the population structure of natural populations of C. australasicum and other Cullen species which may be susceptible to genetic contamination via pollen mediated gene flow from planted pastures of C. australasicum.     

A simple method for isolation of microsatellites from the mudskipper (Boleophthalmus pectinirostris), without constructing a genomic library

The mudskipper Boleophthalmus pectinirostris is an important aquaculture species in China. It is endangered due to the loss and fragmentation of natural habitat, seawater pollution and overfishing. In this study, ten polymorphic microsatellite loci were isolated by the 5′-anchored polymerase chain reaction technique. Population genetic parameters were estimated on 60 wild individuals collected from Jiuduansha wetland in Yangtze River estuary. The number of alleles ranged from 3 to 14. The observed(H _O) and expected(H _E) heterozygosities ranged from 0.2500 to 0.8500 and from 0.3679 to 0.8692, respectively. This suite of microsatellite loci will facilitate future studies of the genetic status of wild and hatchery bred populations of B. pectinirostris.     

生态因子对红螯相手蟹捕食毛蚶苗种的影响

敌害生物的捕食在控制海洋底栖生物群落的丰度和组成中起着关键性的作用。以红螯相手蟹(Sesarma haematocheir)和毛蚶(Scapharca subcrenata)为试验对象,研究了红螯相手蟹的密度、规格、性别,以及毛蚶的密度、规格、海水温度和底质条件对毛蚶苗种存活的影响。结果表明,蟹表现出了第二种类型的功能反应,高密度底播毛蚶苗种可以显著提高成活率;当毛蚶苗种壳长达到20mm以上时,蟹的摄食速率显著下降;随着蟹个体的增大,其摄食速率显著增加,毛蚶的存活率下降;当蟹的密度逐渐增加的时候,同种个体之间的干扰竞争显著提高了毛蚶存活率;雄蟹凭借强有力的螯导致了更多毛蚶苗种的死亡;海水温度较低的春季和秋季底播毛蚶苗种可以显著提高成活率;底质条件的复杂性和异质性为毛蚶的存活提供了"庇护空间",从而减少了敌害生物捕食所带来的损失。     

Isolation and cross-amplification of microsatellites in pink abalone (Haliotis corrugata)

Ten novel microsatellite loci were isolated in pink abalone, Haliotis corrugata, using (GT)₁₅ and (CT)₁₅ enriched genomic libraries. Two previously reported Haliotis kamtschatkana microsatellites cross‐amplified in H. corrugata. A set of 12 polymorphic microsatellites were evaluated in a wild population sample (N = 49). The number of alleles ranged from two to 55, and the observed and expected heterozygosities ranged from 0.104 to 0.939 and from 0.213 to 0.982, respectively. Significant deviations from Hardy–Weinberg equilibrium at three loci and no linkage disequilibrium were observed. Haliotis corrugata microsatellites cross‐amplified in other abalone species, two in H. fulgens, and seven in H. rufescens.     

Isolation and characterization of novel polymorphic nuclear microsatellite markers from Ophrys fusca (Orchidaceae) and cross-species amplification

The present study reports the isolation and characterization of eight new polymorphic microsatellite loci from the sexually deceptive orchid Ophrys fusca. Microsatellites were isolated from two partially enriched genomic libraries using FIASCO (Fast Isolation by AFLP of Sequences COntaining repeats). Seventy-three loci were screened for primer design and primer pairs corresponding to eight different loci were selected for microsatellite characterization of two Portuguese populations. Total number of alleles per locus ranged from 10 to 32. All loci showed a high level of observed heterozygosity (H₀) ranging from 0.33 to 1 and were possible to amplify in 16 other species of Ophrys using the same primers. H. C. Cotrim and F. A. Monteiro have contributed equally to this work.