Taxonomic implications of the morphological and genetic variation of cultivated and domesticated populations of the <Emphasis Type="Italic">Agave angustifolia</Emphasis> complex (Agavoideae,Asparagaceae) in Oaxaca,Mexico

The Agave angustifolia complex, distributed from Mexico to Costa Rica, comprises four species and five varieties, including three species used for mescal production. The complex is represented in the Mexican state of Oaxaca by two wild taxa, A. angustifolia var. angustifolia and A. angustifolia var. rubescens, the cultivated form A. angustifolia “Espadín” and the partially cultivated species A. rodacantha. The aims of this study were to investigate the morphological and genetic variation of the A. angustifolia complex in the state of Oaxaca and to identify traits useful for taxonomic delimitation. Four wild and three cultivated populations of A. angustifolia from Oaxaca, one population of A. tequilana from Guanajuato and one population of A. angustifolia from Sonora were sampled for morphological, genetic and cytometric analyses. We showed that cultivated populations of A. angustifolia “Espadin,” A. rhodacantha and A. tequilana could be clearly differentiated from wild populations. Furthermore, the domesticated populations of A. angustifolia, known locally as “Espadin,” had a higher ploidy level and lower genetic variation than their related wild populations. The population of A. angustifolia from Sonora could be recognized as a different entity. Populations of A. rhodacantha need to be studied throughout their entire distribution area to further evaluate their taxonomic delimitation.     

Arbuscular mycorrhizal fungi associated with<Emphasis Type="Italic"> Araucaria angustifolia</Emphasis> (Bert.) O. Ktze

Specimens of Araucaria angustifolia from a native forest reserve and a reforested area in the State Park of Campos do Jordão were studied to determine the number and diversity of spore populations of arbuscular mycorrhizal fungi (AMF) and root colonization. Six randomly chosen plots (planted with 8- to 12-year-old plants) were delimited, four in the native forest and two in the reforested area. Rhizosphere and root samples were collected during two periods of the year corresponding to the rainy and dry seasons. A greenhouse experiment was set up for multiplication of field propagules (from the native forest and reforested area) for two consecutive generations. Araucaria leaves from the experimental plots were collected during the first sampling for nutrient analysis. Twenty-four AMF taxa were found and percent AM colonization was determined in all plots. Not all AMF species observed in the field were re-isolated through the recovery pot cultures, even after a second cultivation cycle. The foliar nutrient analysis showed higher nutrient levels in plants from the native forest than the reforested area. Generally, spore richness and diversity were highest during the warmer and more humid period and in the native forest plants.     

Isolation and Characterization of Microsatellite Markers for <Emphasis Type="Italic">Viola websteri</Emphasis> (Violaceae) and Cross-Species Amplification within the Genus <Emphasis Type="Italic">Viola</Emphasis>

We isolated and characterized microsatellite loci in Viola websteri (Violaceae), an endangered species from Korea and endemic to Northeast Asia. A total of 27 microsatellite loci were developed and tested in Korean and Chinese populations. The number of alleles per locus varied from two to eight. The observed and expected heterozygosities within two populations were 0.000 to 1.000 and 0.080 to 0.816, respectively. Korean and Chinese populations were clearly distinguished by the private alleles from 16 loci. A total of 21 loci out of the 27 developed loci were successfully cross-amplified in 39 other Viola species. We believe that these microsatellite loci will be useful for future studies on genetic diversity and population structure of V. websteri, as well as other Viola species.     

Structural Polymorphism of <Emphasis Type="Italic">Sinorhizobium meliloti</Emphasis> Genes Related to Virulence and Salt Tolerance

Analysis of the structural polymorphism of eight genes in Sinorhizobium meliloti (nodA, nodB, nodC, and nodH, as well as betA, betB, betC, and betB2) involved in virulence control and salt tolerance, respectively, was carried out in native populations from two geographically distant areas of alfalfa diversity. These areas are located in the North Caucasian gene center of cultivated plants (NCG) and in the modern center of introgressive hybridization of alfalfa located next to the Aral Sea area (PAG) subjected to salinization. RFLP types (alleles) of the nod and bet genes, similar to those in the reference strain Rm1021 (A-type) and different from them (divergent, or D-type alleles) were revealed. The combinations for A- and D-type alleles of the aforementioned genes (analysis of the linkage disequilibrium, LD) were studied in both populations. It was shown that D-type alleles of the nod genes were two times more frequent in the NCG population, while D-type alleles of the bet genes were predominantly identified in the PAG population. At the same time, different combinations of D-type alleles of both the nod and bet genes prevailed in populations. For instance, in the case of the glycine betaine metabolism pathway, these were the betC and betB2 genes in NCG population and betB and betA genes in PAG population. The state of linkage disequilibrium was shown for 60.7% of combinations of alleles of the nod and bet genes in the S. meliloti strains from NCG and more than twice less in strains from the PAG population. It was concluded that clonal lines prevailed in NCG, while the PAG population of S. meliloti had a panmictic structure with revealed single clonal lines.     

Effects of different silvicultural systems on the genetic diversity of <Emphasis Type="Italic">Shorea parvifolia</Emphasis> populations in the tropical rainforest of Southeast Asia

Selective logging systems have been used to prevent the rapid decline of forest resources in Southeast Asia, but little is known about the impacts of selective logging on the genetic diversity of Southeast Asian rainforests. We evaluated the effects of silvicultural systems with differing cutting rotations and enrichment planting regimes on the genetic diversity of Shorea parvifolia, an abundant and ecologically important tree in Southeast Asian rainforests. Our result showed that in most respects the genetic diversity is not significantly different between primary forest and the other silvicultural systems; however, the proportion of private alleles is significantly different between them. Intensive second-rotation (L3) harvesting of individuals >40 cm in diameter at breast height (dbh) resulted in a sizable reduction in the number of reproductive trees and a dramatic decrease in the numbers of rare and private alleles, suggesting a negative impact on the genetic diversity of the remaining tree population. Enrichment planting with S. parvifolia in the logged forest improved some genetic parameters, significantly increasing the number of rare alleles in L3 in particular. We conclude that the genetic diversity of logged tropical forests gradually decreases depending on logging rotation times, especially with respect to sensitive genetic parameters such as the numbers of rare and private alleles, and that enrichment planting with native dipterocarps can maintain or even increase the genetic diversity of logged tropical forests in Southeast Asia.     

Environmental transition zone and rivers shape intraspecific population structure and genetic diversity of an Amazonian rain forest tree frog

Diversification processes acting across geographically continuous populations have been rarely documented in Amazonia, because of the lack of fine-scale sampling over extensive areas. We aimed to determine the geographic effects of an environmental transition zone and large rivers on the intraspecific population structure of the Manaus slender-legged tree frog (Osteocephalus taurinus) along a ~ 900 km transect of tropical rain forest. Using one mitochondrial (16S), two nuclear genes (TYR, POMC) and three microsatellites, we estimated the population structure, phylogenetic relationships and geographic variation of 262 O. taurinus and 5 O. oophagus (a close relative) along the Purus–Madeira interfluve (PMI) and opposite banks of the central Amazon and upper Madeira rivers, at central-southern Amazonia, Brazil. Six genetic clusters were identified: two corresponding to sympatric populations of O. taurinus and O. oophagus from their type locality, north of Amazon river. Within PMI, there were three distinct O. taurinus genetic clusters distributed along the geographic gradient with one main phylogeographic break found (concordant between 16S and TYR), that corresponds to a transition zone (ecotone) between dense and open rain forest ecotypes. The sixth cluster was an O. taurinus population isolated at the east bank of the upper Madeira river. In addition, restricted haplotype sharing was identified from the west to east banks at upper Madeira river. Within PMI, parapatric genetic structure is explained by a potential association of the genetic clusters to the different forest ecotypes they inhabit coupled with isolation by distance, thus supporting the gradient hypothesis for diversification. Differentiation of populations that are external to the PMI is most likely explained by the barrier effect of the Madeira and Amazon rivers. Our findings provide new evidence on diversification processes across continuous Amazonian landscapes; however, the specific mechanisms underlying the origin and maintenance of the identified phylogeographic break need to be further studied.     

Assessing genetic diversity for the USA endemic carnivorous plant <Emphasis Type="Italic">Pinguicula ionantha</Emphasis> R.K. Godfrey (Lentibulariaceae)

Understanding patterns of genetic diversity and population structure for rare, narrowly endemic plant species, such as Pinguicula ionantha (Godfrey’s butterwort; Lentibulariaceae), informs conservation goals and can directly affect management decisions. Pinguicula ionantha is a federally listed species endemic to the Florida Panhandle in the southeastern USA. The main goal of our study was to assess patterns of genetic diversity and structure in 17 P. ionantha populations, and to determine if diversity is associated with geographic location or population characteristics. We scored 240 individuals at a total of 899 AFLP markers (893 polymorphic markers). We found no relationship between the estimated population size with either of two measures of diversity (proportion of loci polymorphic, P = 0.37; Nei’s gene diversity, P = 0.50). We also found low levels of population genetic structure; there was no clear relationship of genetic isolation by distance (P = 0.23) and only a small (but significant) proportion of genetic variation was partitioned amongst regions (2.4 %, P = 0.02) or populations (20.8 %, P < 0.001). STRUCTURE analysis found that the model with two inferred clusters (K = 2) best described the AFLP data; the dominant cluster at each site corresponded to the results from PCoA and Nei’s genetic distance analyses. The observed patterns of genetic diversity suggest that although P. ionantha populations are isolated spatially by distance and both natural and anthropogenic barriers, some gene flow occurs among them or isolation has been too recent to leave a genetic signature. The relatively low level of genetic diversity associated with this species is a concern as it may impair fitness and evolutionary capability in a changing environment. The results of this study provide the foundation for the development of management practices that will assist in the protection of this rare carnivorous plant.     

Genetic diversity and genetic structure of Persian walnut (<Emphasis Type="Italic">Juglans regia)</Emphasis> accessions from 14 European,African, and Asian countries using SSR markers

Persian walnut (Juglans regia L.) is the world’s most widely grown nut crop, but large-scale assessments and comparisons of the genetic diversity of the crop are notably lacking. To guide the conservation and utilization of Persian walnut genetic resources, genotypes (n = 189) from 25 different regions in 14 countries on three continents were sampled to investigate their genetic relationships and diversity using ten microsatellite (SSR) loci. The SSRs amplified from 3 to 25 alleles per locus, with a mean value of 11.5 alleles per locus. The mean values of observed and expected heterozygosity were 0.62 and 0.73, respectively. Based on Nei’s genetic identity, accessions from Bratislava (Slovakia) and Antalya (Turkey) showed the lowest similarity (0.36), while accessions from Algeria and Tunisia as well as accessions from Debrecen (Hungary) and Trnava (Slovakia) had the highest similarity (0.97). Two populations from Iran (Alborz and Ardabil) had the highest number of private alleles (7 and 5), but they were quite different as they also had the lowest genetic identity when compared to the remaining populations as well as to each other. Although overall differentiation among regions was relatively low (F _st  = 0.07), cluster analysis grouped accessions generally but not completely according to geography. STRUCTURE software confirmed these results and divided the accessions into two main groups, separating accessions collected from Europe and North Africa from those from Greece and the Near East. Results indicate the presence of a likely center of diversity for Persian walnut in Eastern and Southeastern Europe. They also provide information that can be used to devise conservation actions. Notably, the genetic diversity of threatened populations from two regions in Iran should be conserved.     

Development of ten microsatellite markers from the keystone mistletoe <Emphasis Type="Italic">Tristerix corymbosus</Emphasis> (Loranthaceae) using 454 next generation sequencing and their applicability to population genetic structure studies

Tristerix corymbosus (Loranthaceae) is a keystone mistletoe from the South American temperate rainforests. As most mistletoes, T. corymbosus relies on biotic pollination and seed dispersal, which may cause population structure. For a better understanding of its ecology, we isolated and characterized ten polymorphic microsatellite loci for this species. We used 454 Next Generation Sequencing to build a microsatellite library from a high quality DNA sample. We tested 90 sequences from which we obtained ten polymorphic markers. In order to assess their variability, the novel markers were tested in 48 individuals from three locations of the Valdivian Coastal Reserve in Chile. We also estimated genetic differences between pairs of populations using the F_ST statistic. The mean number of alleles per locus in the 48 individuals studied was 7.1 (2–17 alleles per locus). The observed and expected heterozygosity ranged from 0.298 to 0.634 and from 0.310 to 0.881, respectively. There were genetic differences among the three populations assessed, according to the F_ST values (ranging from 0.048 to 0.100, all significant) and, the number of alleles per locus ranged from 3.9 to 5.1. These are the first microsatellite markers developed for T. corymbosus, and they arise as a powerful tool for studying population structure, genetic diversity and gene flow at the landscape scale, along its distribution.     

A fifth major genetic group among honeybees revealed in Syria

Background

Apiculture has been practiced in North Africa and the Middle-East from antiquity. Several thousand years of selective breeding have left a mosaic of Apis mellifera subspecies in the Middle-East, many uniquely adapted and survived to local environmental conditions. In this study we explore the genetic diversity of A. mellifera from Syria (n?=?1258), Lebanon (n?=?169) and Iraq (n?=?35) based on 14 short tandem repeat (STR) loci in the context of reference populations from throughout the Old World (n?=?732).

Results

Our data suggest that the Syrian honeybee Apis mellifera syriaca occurs in both Syrian and Lebanese territories, with no significant genetic variability between respective populations from Syria and Lebanon. All studied populations clustered within a new fifth independent nuclear cluster, congruent with an mtDNA Z haplotype identified in a previous study. Syrian honeybee populations are not associated with Oriental lineage O, except for sporadic introgression into some populations close to the Turkish and Iraqi borders. Southern Syrian and Lebanese populations demonstrated high levels of genetic diversity compared to the northern populations.

Conclusion

This study revealed the effects of foreign queen importations on Syrian bee populations, especially for the region of Tartus, where extensive introgression of A. m. anatolica and/or A. m. caucasica alleles were identified. The policy of creating genetic conservation centers for the Syrian subspecies should take into consideration the influence of the oriental lineage O from the northern Syrian border and the large population of genetically divergent indigenous honeybees located in southern Syria.

     

Patterns of population genetic structure and diversity across bumble bee communities in the Pacific Northwest

Patterns of genetic structure and diversity are largely mediated by a species’ ecological niche and sensitivity to climate variation. Some species with narrow ecological niches have been found to exhibit increased population differentiation, limited gene flow across populations, and reduced population genetic diversity. In this study, we examine patterns of population genetic structure and diversity of four bumble bee species that are broadly sympatric, but do not necessarily inhabit the same ecological niche in the Pacific Northwest of the United States. Testing for the effect of isolation by geographic distance (IBD) with linearized F _st and D _est found that Bombus sylvicola and B. mixtus exhibited significant IBD across populations. In contrast, both B. melanopygus and B. flavifrons, two species that are distributed across a broad elevation gradient, exhibited no IBD, a result further corroborated by Bayesian a priori population assignment tests. Furthermore, we discovered that B. sylvicola populations distributed on the Olympic Peninsula have significantly less average allelic diversity than populations distributed in the Cascade Mountains. Our results suggest that populations distributed in the Olympic Mountains represent a distinct genetic cluster relative to the Cascade Mountains, with B. sylvicola and B. mixtus likely experiencing the greatest degree of population genetic differentiation relative to B. flavifrons and B. melanopygus. While bumble bees are known to co-exist across a diversity of habitats, our results demonstrate that underlying population genetic structure and diversity may not necessarily be similar across species, and are largely governed by their respective niches.     

Genetic markers of adaptive processes in the Far Eastern pink salmon <Emphasis Type="Italic">Oncorhynchus gorbuscha</Emphasis>: Allelic diversity at the locus of major histocompatibility complex MHC I-<Emphasis Type="Italic">A1</Emphasis>

To clarify allelic diversity at the locus of major histocompatibility complex MHC class I-A1 in the Far Eastern pink salmon Oncorhynchus gorbuscha, sequencing of the electrophoretic alleles isolated from the gel (DGGE alleles) was performed. In 47 individuals, the genotypes of which consisted of ten DGGE alleles, 18 MHC I-A1 nucleotide sequences were revealed, and thus, eight cryptic alleles not detected by electrophoresis were identified. Eleven of these alleles were identified earlier in pink salmon from Hokkaido, Alaska, and British Columbia, and seven, possibly, were unique to the populations from some Far Eastern regions. Six of the previously determined DGGE alleles corresponded to more than one nucleotide sequence. However, the sequences attributed to the same DGGE allele differed on average by less than 1 nucleotide. These findings point to sufficient sensitivity of the DGGE method, although the genetic diversity and differentiation estimates obtained with it will obviously be somewhat underestimated. Considerable predominance of nonsynonymous substitutions over the synonymous ones in the codons of the MHC I-A1 antigen-binding site confirms the presence of positive selection aimed at providing the population resistance to local spectrum of pathogens. Refinement of the allelic composition of the adaptively important MHC genetic marker will contribute to more complete understanding of the adaptive genetic structure of pink salmon as an important element of the overall population structure of the species.     

Sporulation and diversity of arbuscular mycorrhizal fungi in Brazil Pine in the field and in the greenhouse

The aim of this work was to assess the sporulation and diversity of arbuscular mycorrhizal fungi (AMF) at different forest sites with Araucaria angustifolia (Bert.) O. Ktze. (Brazil Pine). In addition, a greenhouse experiment was carried out to test the use of traditional trap plants (maize + peanut) or A. angustifolia to estimate the diversity of AMF at each site. Soil samples were taken in two State Parks at southwestern Brazil: Campos do Jordão (Parque Estadual de Campos do Jordão [PECJ]) and Apiaí (Parque Estadual Turístico do Alto Ribeira [PETAR]), São Paulo State, in sites of either native or replanted forest. In PECJ, an extra site of replanted forest that was impacted by accidental fire and is now in a state of recuperation was also sampled. The spore densities and their morphological identification were compiled at each site. In the greenhouse, soil samples from each site were used as inoculum to promote spore multiplication on maize + peanut or A. angustifolia grown on a sandy, low-fertility substrate. Plants were harvested, respectively, after 4 months or 1 year of growth and assessed for mycorrhizal root colonization. Spore counts and identification were also performed in the substrate, after the harvest of plants. Twenty-five taxa were identified considering all sites. Species richness and diversity were greater in native forest areas, being Acaulospora, the genus with the most species. Differences in number of spores, diversity, and richness were found at the different sites of each State Park. Differences were also found when maize + peanut or A. angustifolia were used as trap plants. The traditional methodology using trap plants seems to underestimate the diversity of the AMF. The use of A. angustifolia as trap plant showed similar species richness to the field in PECJ, but the identified species were not necessarily the same. Nevertheless, for PETAR, both A. angustifolia and maize + peanut underestimated the species richness. Because the AMF sporulation can be affected by many conditions, it is impossible to draw detailed conclusions from this kind of survey. More precise experiments have to be set up to isolate the different factors that modulate the ecophysiological interactions between host plant and endophyte.     

Genetic assessment of <Emphasis Type="Italic">Abies koreana</Emphasis> (Pinaceae), the endangered Korean fir,and conservation implications

To establish a management plan for endangered and rare species, genetic assessment must first be conducted. The genetic characteristics of plant species are affected by demographic history, reproductive strategy, and distributional range as well as anthropological effects. Abies koreana E. H. Wilson (Pinaceae), Korean fir, is endemic to Korea and found only in sub-alpine areas of the southern Korean Peninsula and Jejudo Island. This species has been designated as critically endangered by the International Union for Conservation of Nature due to a continuous decline in its range and population fragmentation. We genotyped 176 individuals from seven natural populations and two afforested populations on the Korean Peninsula using 19 microsatellite loci. STRUCTURE analysis revealed two genetic clusters in natural populations (F _st  = 0.040 and R _st  = 0.040) despite low differentiation. We did not detect a significant reduction in genetic diversity or the signature of a genetic bottleneck despite population fragmentation and small population size. We deduced that this species exhibits a metapopulation structure, with the population on Jirisan Mountain acting as a source of genetic diversity for other local small populations on the Korean Peninsula, through contemporary asymmetric gene flow. However, the majority of afforested individuals on the Korean Peninsula originated from a different gene cluster. Thus, we recommend a conservation strategy that maintains two genetically unique clusters.     

Hordein locus polymorphism of cultivated barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) in Turkey

Starch gel electrophoresis has been used to study the polymorphism of hordeins encoded by the Hrd A, Hrd B, and Hrd F loci in 93 landrace specimens of barley assigned to 17 ancient provinces located in modern Turkey. Forty-five alleles of Hrd A with frequencies of 0.11–29.34%, 51 alleles of Hrd B with frequencies of 0.11–8.07%, and 5 alleles of Hrd F with frequencies of 0.75–41.29% have been detected. Cluster analysis of the matrix of allele frequencies has demonstrated that barley populations from different old provinces of Turkey are similar to one another. Cluster structure of local barley populations has been found, most populations (82%) falling into three clusters. The first cluster comprises barley populations from six provinces (Thracia, Bithynia, Pontus, Lydia, Cappadocia, and Armenia); the second cluster, populations from five provinces (Paphlagonia, Galatia, Lycaonia, Cilicia, and Mesopotamia); and the third one, populations from three provinces (Phrygia, Karia, and Lycia). Barley populations from Mysia, Pamphlya, and Syria do not fall in any cluster.     

Development and Characterization of Microsatellite Markers for Genetic Analysis of the Swimming Crab, <Emphasis Type="Italic">Portunus trituberculatus</Emphasis>

This study isolated and characterized 11 novel microsatellite markers for the commercially important swimming crab species, Portunus trituberculatus. Genetic diversity and population structure of two populations of P. trituberculatus in the East China Sea were assessed using these loci. The microsatellite markers produced 242 alleles, varying from 17 to 26 alleles per locus. In all the samples, the range of heterozygosity was 0.6324–0.9403 (observed) and 0.8998–0.9547 (expected). An F-statistic analysis revealed low genetic differentiation between the populations (mean F _ST = 0.0197), with 98% of the variation resulting from the within-population component. In addition, cross-amplification was tested in two other portunid species, and we found that many loci yielded useful information. The high degree of polymorphism exhibited by the 11 microsatellites suggests that these markers will be useful for both aquaculture and studies of natural populations of the genus.     

Exotic tree monocultures play a limited role in the conservation of Atlantic Forest epiphytes

The Brazilian Atlantic Forest suffered a severe geographic contraction along the last five centuries that reduced drastically most vascular epiphyte populations. Among the range of man-made matrixes, tree monocultures have the potential to contribute positively to the maintenance of the regional epiphyte diversity. Here, we test the similarity in abundance, richness, and species composition between vascular epiphytic communities established in managed monocultures of exotic and native species with natural communities occurring in neighboring native Araucaria Forest patches. In the São Francisco de Paula National Forest (Rio Grande do Sul state, Brazil), we recorded 62 epiphyte species from 300 phorophytes occurring in 12, one-hectare plots of Araucaria Forest and managed plantations of Pinus, Eucalyptus and Araucaria. Species richness, rarefied richness and abundance were significantly higher in Araucaria Forest in comparison to the exotic stands. Species composition was also substantially differentiated as Araucaria Forest patches harbored a greater number of zoochorous species than those of the exotic stands. Additionally, plantations of Araucaria angustifolia, a native species, sustained more individuals and more species than the exotic plantations. Neither tree height nor DBH explained epiphyte richness; however, both phorophyte diversity and stand age together accounted for 92% of the among-site variation in epiphytic species richness. We conclude that substrate heterogeneity in combination with time available for colonization contribute significantly to beta-diversity of epiphytes in Araucaria forests. However, demographic experimental studies are necessary in order to disentangle the role of substrate quality from metapopulation processes, such as dispersal limitation, at both temporal and spatial scales.     

Microsatellite analysis of the genetic relationships between wild and cultivated giant grouper in the South China Sea

The giant grouper (Epinephelus lanceolatus) is a coral fish with high commercial value in Southeast Asia. In the present study, we isolated 11 microsatellite DNA markers, and analysed the genetic diversity and differentiation between cultured stocks and wild populations of the giant grouper originating from the South China Sea. A total of 390 alleles at 11 microsatellite loci were detected in 130 individuals from five different populations. The expected heterozygosity varied from 0.131 to 0.855 with a mean value of 0.623 and the observed heterozygosity varied from 0.145 to 0.869 with a mean value of 0.379. The allelic richness and heterozygosity studies revealed that the genetic diversity of the cultured population was significantly reduced when compared with that of the wild population. The F_is, pairwise F_st values, analysis of molecular variance (AMOVA), three-dimensional factorial correspondence analysis and structure analysis revealed significant population differentiation between the cultured stocks and the wild populations, among the three cultured populations and between the two wild populations. These differences may be caused by random genetic drift, the effects of artificial selection and founder effects. Our results will be useful in the management of cultured stocks and conservation of wild populations of the giant grouper.     

Microsatellite markers for the Ussuri white-toothed shrew (Soricidae: <Emphasis Type="Italic">Crocidura lasiura</Emphasis>) developed by Ion Torrent sequencing and their application to the shrew populations in disturbed forests

Understanding the impacts of forest management practices on genetic diversity is essential for effective animal management and conservation. We characterized novel microsatellite loci in the Ussuri white-toothed shrew (Crocidura lasiura Dobson, 1890) to test the impacts of anthropogenic thinning of forest trees on the shrew populations and their genetic diversity. Using Ion Torrent sequencing technology, we characterized 611 potential microsatellite markers with complete di- to tetra-nucleotide motifs, identifying nine polymorphic loci. The observed and expected heterozygosities across the nine loci were 0.526 and 0.527, respectively. Mean allelic diversity was 5.2 alleles per locus, with the mean polymorphism information content at 0.498. In comparison among shrew populations, which inhabited in the forests thinned in 2004 (CLA; n = 10), 2008 (CLB; n = 9) and 2014 (CLC; n = 3), the observed heterozygosities are similar among the three populations (0.525 at CLA, 0.532 at CLB and 0.519 at CLC), whereas the expected heterozygosities were much lower in population of CLC (0.377) than that of CLA (0.509) and CLB (0.533). The small sample size at CLC limited effective comparison and evaluation of the impact of forest thinning on genetic diversity in this shrew population. Future application of the species-specific microsatellite markers described here to a larger sample size would be valuable in estimating the ecological parameters of shrew populations associated with existing forest management practices.