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The species of Anisakis constitute one of the most widespread groups of ascaridoid nematodes in the marine ecosystem. Three closely related taxa are recognised in the A. simplex (s. l.) complex, i.e. A. pegreffii, A. simplex (s. s.) and A. berlandi. They are distributed in populations of their intermediate/paratenic (fish and squids) and definitive (cetaceans) hosts. A panel of seven microsatellite loci (Anisl 05784, Anisl 08059, Anisl 00875, Anisl 07132, Anisl 00314, Anisl 10535 and Anisl 00185), were developed and validated on a total of N = 943 specimens of A. pegreffii and A. simplex (s. s.), collected in fish and cetacean hosts from allopatric areas within the range of distribution of these parasite species. In addition, the locus Anisl 7, previously detected in those Anisakis spp., was investigated. The parasites were first identified by sequence analysis of the EF1 α-1 nDNA. The panel of the microsatellites loci here developed have allowed to: (i) detect diagnostic microsatellite loci between the two species; (ii) identify specimens of the two species A. pegreffii, A. simplex (s. s.) in a multi-marker nuclear genotyping approach; (iii) discover two sex-linked loci in both Anisakis species and (iv) estimate levels of genetic differentiation at both the inter- and intra-specific level.
When a promising natural enemy of a key pest exists locally, it is a common practice in biological control (BC) to rear and release it for supplementary control in the targeted agroecosystem even though significant knowledge gaps concerning pre/post release may still exist. Incorporating genetic information into BC research fills some of these gaps. Habrobracon hebetor, a parasitoid of many economically important moths that infest stored and field crops worldwide is commonly used, particularly against the millet head miner (MHM), a key pest of millet in Sahelian countries. To advance our knowledge on how H. hebetor that occurs naturally in open-field cropping systems and grain stores as well as being mass-produced and released for MHM control, performs in millet agroecosystems in Niger we evaluated its population genetics using two mitochondrial and 21 microsatellite markers. The field samples were genetically more diverse and displayed heterozygote excess. Very few field samples had faced significant recent demographic bottlenecks. The mating system (i.e. nonrandom mating with complementary sex determination) of this species may be the major driver of these findings rather than bottlenecks caused by the small number of individuals released and the scarcity of hosts during the longlasting dry season in Niger. H. hebetor population structure was represented by several small patches and genetically distinct individuals. Gene flow occurred at local and regional scales through human-mediated and natural short-distance dispersal. These findings highlight the importance of the mating system in the genetic diversity and structure of H. hebetor populations, and contribute to our understanding of its reported efficacy against MHM in pearl millet fields.
The genus Rhagoletis (Diptera: Tephritidae) comprises more than 65 species distributed throughout Europe, Asia and America, including many species of high economic importance. Currently, there are three Rhagoletis species that infest fruits and nuts in Europe. The European cherry fruit fly, Rhagoletis cerasi (may have invaded Europe a long time ago from the Caucasian area of West Asia), and two invasive species (recently introduced from North America): the eastern American cherry fruit fly, R. cingulata, and the walnut husk fly, R. completa. The presence of different Rhagoletis species may enhance population dynamics and establish an unpredictable economic risk for several fruit and nut crops in Europe. Despite their excessive economic importance, little is known on population dynamics, genetics and symbiotic associations for making sound pest control decisions in terms of species-specific, environmental friendly pest control methods. To this end, the current paper (a) summarizes recently accumulated genetic and population data for the European Rhagoletis species and their association with the endosymbiont Wolbachia pipientis, and (b) explores the possibility of using the current knowledge for implementing the innovative biological control methods of sterile insect technique and incompatible insect technique.
The range-wide genetic structure of the highly productive and valuable timber species Sequoia sempervirens (D. Don) Endl. is still insufficiently studied, although published data based on different genetic markers (nuclear and chloroplast microsatellites, AFLP, RFLP and isozymes) demonstrated relatively low population structure. However, more genetic markers are needed to increase the efficiency of population genetic studies in coast redwood. Therefore, we developed seven nuclear and five chloroplast microsatellite or simple sequence repeat (SSR) markers based on expressed sequence tags (ESTs) and the complete chloroplast genome sequence, respectively. All selected markers were tested in a range-wide sample representing trees from 16 locations. They are highly polymorphic microsatellite loci with the number of alleles ranging from 3 to 17, and the number of effective alleles from 1.1 to 2.48. Coast redwood is a hexaploid species, and its chloroplasts are paternally inherited. Therefore, the chloroplast SSR markers are especially useful for this species, because their genotyping is not affected by nuclear genome ploidy. Moreover, they showed high gene diversity for each locus within and across all populations and can be used to study range-wide population genetic structure, pollen-based gene flow and long-distance gene transfer. Coast redwood can propagate clonally, and nuclear polymorphic EST-SSRs can be used for clonal identification. They are linked with expressed genes and their variation can reflect variation in genes under selection, including those that could be potentially important for local adaptation of coast redwood considering the threat of climate change.
Licorice (Glycyrrhiza glabra) is an important medicinal herb and has long been used in traditional medicine for the treatment of several diseases worldwide. Understanding the genetic diversity within Glycyrrhiza species is important for the efficient conservation of these medicinal herbs. In this study, we have developed 20 polymorphic chloroplast microsatellite (cpSSR) markers using the chloroplast genome of G. lepidota. The cpSSR markers were tested on a total of 27 Glycyrrhiza individual plants. The number of alleles per locus ranged from two to eight among the Glycyrrhiza accessions. Overall, the Shannon index (I) for each cpSSR ranged from 0.315 to 1.694, the diversity indices (h) were 0.140–0.793 and the unbiased diversity indices (uh) were 0.145–0.825. In addition, the cpSSR markers were successfully divided and classified the 27 Glycyrrhiza individuals into four groups. The cpSSR markers developed in this study could be used in the assessment of genetic diversity and rapid identification of Glycyrrhiza species.
The Ceratitis FAR complex (Diptera, Tephritidae) includes four economically important frugivorous flies (Ceratitis anonae, Ceratitis fasciventris, Ceratitis quilicii, Ceratitis rosa) whose immature stages and adult females cannot be properly resolved through morphological identification. In order to develop a simplified molecular tool for the identification of two of these species (C. rosa, C. quilicii), we selected a subset of six microsatellite markers out of a panel of 16 loci that were previously developed for the molecular differentiation of the taxa within the complex. These six markers were first tested in silico and then used for the actual genotyping of C. quilicii and C. rosa, resulting in the correct identification of all male reference specimens. Here, we propose an integrated morphological and molecular setup for the identification of the four species of the FAR complex. The decision map relies on preliminary DNA barcoding or morphological identification (when possible) to exclude species not belonging to the complex followed by (a) morphological identification of all adult male specimens and female C. anonae, (b) molecular identification via a panel of 16 microsatellite markers for immature stages, damaged vouchers and samples potentially including adult female C. fasciventris/C. quilicii/C. rosa and (c) molecular identification via a reduced panel of six microsatellite markers for samples including only C. quilicii and C. rosa. This simplified diagnostic setup was profitably implemented in the framework of the ERAfrica fruit fly project and will help correctly identify species within the FAR complex for their early detection and monitoring.
Understanding demographic processes over multiple spatial scales is vital for the optimization of conservation/management strategies, particularly for commercially harvested taxa such as the brown crab (Cancer pagurus L). Brown crab population genetic structure was investigated at (i) a local scale within the Irish Sea, which included comparisons with the Lundy No Take Zone (NTZ) and (ii) across the NE Atlantic. The results indicate that the brown crab does not exhibit strong spatial structure either within the Irish Sea or at the regional level, suggesting high gene flow within and among the Irish Sea, English Channel and North Sea. Comparisons between the Lundy NTZ and harvested areas revealed similarly high levels of genetic diversity. An intriguing result was that the Lundy NTZ sample exhibited a degree of genetic patchiness (ephemeral geographically unpatterned differentiation) which may indicate elevated recruitment skews within the NTZ. Overall, the results support the view that brown crabs within the sampled area belong to a single genetically panmictic stock and that if breeding stock sizes are maintained genetic drift will not be strong enough to reduce neutral genetic diversity. The highly connected nature of this species requires international cooperation for sustainable management, an important component of which will be the application of more powerful population genomic approaches to assess finer scale aspects of stock structure as well drivers of genetic patchiness reported for the species. This is a timely consideration in light of potential future misalignments between biological and geopolitical stock boundaries in the Irish Sea following Brexit.
Goats have played a key role as source of nourishment for humans in their expansion all over the world in long land and sea trips. This has guaranteed a place for this species in the important and rapid episode of livestock expansion triggered by Columbus’ arrival in the Americas in the late 1400s. The aims of this study are to provide a comprehensive perspective on genetic diversity in American goat populations and to assess their origins and evolutionary trajectories. This was achieved by combining data from autosomal neutral genetic markers obtained in more than two thousand samples that encompass a wide range of Iberian, African and Creole goat breeds. In general, even though Creole populations differ clearly from each other, they lack a strong geographical pattern of differentiation, such that populations of different admixed ancestry share relatively close locations throughout the large geographical range included in this study. Important Iberian signatures were detected in most Creole populations studied, and many of them, particularly the Cuban Creole, also revealed an important contribution of African breeds. On the other hand, the Brazilian breeds showed a particular genetic structure and were clearly separated from the other Creole populations, with some influence from Cape Verde goats. These results provide a comprehensive characterisation of the present structure of goat genetic diversity, and a dissection of the Iberian and African influences that gave origin to different Creole caprine breeds, disentangling an important part of their evolutionary history. Creole breeds constitute an important reservoir of genetic diversity that justifies the development of appropriate management systems aimed at improving performance without loss of genomic diversity.
Western flower thrips (WFT) (Frankliniella occidentalis) is an introduced pest that harms French bean production in Kenya and other countries. Since new WFT management approaches are being developed, a closer look at the genetic makeup of WFT populations can give new insights into source habitats, crop colonization patterns or host plant preferences, which are prerequisites for integrated pest management (IPM) strategies. For this purpose, we used six microsatellite loci to analyse the genetic structure, diversity and gene flow of WFT sampled on French beans, intercrops and weeds in Kenyan French bean production areas. The results of this preliminary study indicate that the available microsatellites are sufficiently polymorphic for more detailed analyses on local dispersal patterns of WFT in Kenya. Even with the limited data set, the results reveal that F. occidentalis populations show considerable genetic differentiation between host plant species but not between regions, which suggests reduced gene flow and a possible development of biotypes. Possible consequences of the results on IPM are discussed.
The giant reed, Arundo donax is one of the worst invasive alien species globally, including South Africa, where it invades riparian areas across the country. Biological control is being considered to address the invasive potential and negative impacts of the weed. This study investigated the phylogeography of A. donax to guide the biological control program. To determine plant haplotype and genetic diversity, three regions of the chloroplast were sequenced and three microsatellite markers were analyzed in 40 samples from across the plant’s distribution in South Africa. It was determined that all populations of A. donax in South Africa were haplotype M1, which is the most widely distributed haplotype worldwide, believed to originate from the Indus Valley, Asia. In addition, no genetic diversity was found, indicating that all the A. donax populations in South Africa are essentially one clone. The results indicate that suitable biological control agents are likely to be found in the ancient native range of haplotype M1. This research has contributed to the global understanding of the phylogeography of A. donax and will guide the biological control program in South Africa.
The study of colonizing and of dominant grass species is essential for
prairie conservation efforts. We sought to answer how naturalized Kentucky
bluegrass in the northern Great Plains has become successful in the last 20
yr despite its long history in the northern Great Plains. We tested for
evidence of geographical differentiation using flow cytometry and
microsatellite markers to ascertain the population genetics of Kentucky
bluegrass. Across all tested wild populations, high levels of genetic
diversity were detected along with moderate levels of structure. Mantel
tests of geographical patterns were not significant. Using clonal
assignment, we found two major clones that made up the majority of the
tested wild populations. When we compared the wild individuals to pedigree
cultivars, we found virtually no genetic overlap across all tests, which did
not support our hypothesis of developed cultivars contributing to high
genetic diversity in natural populations. Furthermore, DNA content tests
indicated a narrow range in ploidy in wild populations compared with lawn
cultivars, further supporting a hypothesis of divergence between wild and
pedigree cultivars. These results indicate the recent invasion of Kentucky
bluegrass in the northern Great Plains is not because of adaptation or
propagule pressure, but rather likely an environmental or land use
Knowledge of linkage disequilibrium (LD) patterns is considered a prerequisite for effective association mapping studies. However, no LD analysis in the Argentine public temperate maize collection has been reported to date. In this study, a panel of 111 temperate maize inbreds genotyped at 74 single sequence repeats (SSRs) loci was used to assess LD, genetic diversity and population structure to evaluate the suitability of the panel for association mapping. Mini-core sets were also designed for in-depth phenotyping and allele mining purposes. The panel consisted of: (1) locally developed orange flint germplasm; (2) temperate inbred lines with Iowa Stiff Stalk Synthetic background; and (3) eight historic flint lines, some of them from the Cuarentín race. As a result, four subpopulations were defined. Joint analysis of population structure and combining ability allowed identifying two main heterotic patterns. High molecular diversity, a low extent of LD and a high ratio of linked to unlinked SSR loci pairs in significant LD were detected indicating the suitability of the entire collection for association mapping. The fact that the LD extent in the mini-core sets was similar to that observed in the entire collection and that only a small percentage of allelic richness was reduced suggests that these mini-core sets are suitable to capture diversity, exploit phenotypic variance and discover useful variants representative of the entire collection.
Nonnative M. vimineum has been expanding rapidly in the eastern United States, where it can negatively affect plant communities. Locally, the species is assumed to spread from roadsides into nearby forests, where it can form dense populations after disturbances, especially in light gaps. Using microsatellite markers, we quantified patterns of genetic variation and structure among populations at nine sites in West Virginia. We then examined patterns of local dispersal within each population, focusing on subpopulations along the roadside, those coalescing nearby along the forest edge, and subpopulations in the interior forest. We found that levels of genetic variation of M. vimineum were relatively low overall across populations but with genetic structure present among populations (Fst = 0.60). Within populations, subpopulations along the roadside were genetically variable, containing 4 to 22 unique, multilocus genotypes. Many of these genotypes were also identified in the adjacent forest, consistent with local, diffusive spread from the roadway. However, several genotypes in the interior forest were unique to the population, indicating that dispersal from other sites may also occur. Overall, it appears that genetic diversity and structure in M. vimineum reflects a variety of processes, including localized dispersal and long-distance migration.
In a germplasm bank collection the conservation and characterization of genetic resources is a prerequisite in order to use the material in breeding projects aiming the creation of new cultivars. In the present study, 54 Prunus salicina domestica and Prunus domestica genotypes (including seven Greek cultivars), maintained in the ex situ National Genebank Collection of Greece, were classified using microsatellite (simple sequence repeat, SSR) markers on high resolution melting (HRM) analysis. The SSR primer pairs were chosen from the published literature as originally designed on Prunus species. This combined approach was used to genotype all plum accessions of the collection highlighting the benefits of either method (HRM and SSRs) for cultivar identification. Dendrograms for P. domestica and P. salicina and a combined one with all the genotypes assayed were produced. A total of 15 from the 19 P. domestica accessions analysed, including all the Greek accessions but ‘Avgati Skopelou’, were grouped into the same clade in the combined dendrogram, whereas the remaining four were dispersed into the P. salinica clades. Bayesian structure analysis confirmed that ‘Avgati Skopelou’ differs from the rest of the Greek plum cultivars since it was not grouped into the same cluster. The combination of HRM and SSRs, provided a considerably faster, cost-effective, closed-tube microsatellite genotyping method for molecular characterization of plum cultivars.
The Virginia opossum (Didelphis virginiana) is considered highly adaptable to anthropogenic disturbances; however, the genetic effects of disturbance on this marsupial have not been studied in wild populations in Mexico. Here we evaluated the genetic diversity of D. virginiana at sites with different levels of disturbance within the Highlands and Central Depression regions of Chiapas in southern Mexico. Twelve microsatellite loci were used and the results demonstrated moderate mean heterozygosity (He = 0.60; Ho = 0.50). No significant differences in heterozygosity were found among sites with different levels of disturbance in both regions (range Ho = 0.42–0.57). We observed low but significant levels of genetic differentiation according to disturbance level. The inbreeding coefficient did not differ significantly from zero, suggesting that low genetic differentiation in these environments may be associated with sufficient random mating and gene flow, a result associated with the high dispersal and tolerance characteristics of this marsupial. Our results for D. virginiana in this particular area of Mexico provide a foundation for exploring the impact of human disturbance on the genetic diversity of a common and generalist species.
Yams (Dioscorea spp.) are one of the main root and tuber crops in the world, especially within the species complex Dioscorea cayenensis/D. rotundata. Few studies have been conducted in Brazil with these species, including genetic diversity. The objective of this study was to characterize the genetic diversity of local varieties of D. cayenensis and D. rotundata using morphological and molecular markers, and provide information on the management and use of the crop by family farmers from different regions in Brazil. Thus, yam tubers were sampled from several municipalities in the South, Southeast and Northeast regions. Eighteen morphological traits and ten microsatellite loci were used to analyse 47 yam accessions (23 D. cayenensis and 24 D. rotundata). Species identification was carried out after field morphological evaluation. Spatial genetic analysis indicated significant structure among the local varieties, mostly between regions and species. Both cluster and Bayesian analyses showed a separation of the accessions into two distinct groups: group I with accessions originated from the Southeast region and group II with accessions originated from the Northeast region, while accessions from the South region were intermediate or included in either group. The results showed a separation between D. cayenensis and D. rotundata accessions in Brazil, and that D. cayenensis occurs predominantly in the Southeast region, while D. rotundata occurs in the Northeast region. Further studies with larger sampling would be welcome in order to confirm these findings. Also, this study highlights the importance of family farmers in the genetic diversity conservation of these species in Brazil.
The commercially important chokka squid Loligo reynaudii occurring in South African waters is currently managed on a single-unit stock hypothesis. We tested this assumption through a spatial comparison of the morphology throughout the distributional range of the species. Forty-three morphometric characters were measured from 1079 chokka collected off the south coast of South Africa, the west coast of South Africa, and southern Angola. While no significant differences were found in the hard body parts, results from classification analysis showed that though all four types of morphometric attributes (soft body parts, beaks, statoliths, sucker rings) resulted in some separation, the most consistent separation of samples from the three regions was based on soft body part morphometric characters. On average, though dependent on the model, the overall correct classification rate ranged from 0.68–0.99 for males and 0.7–0.99 for females in all three regions. Previous DNA analysis had revealed some genetic differences between west coast and south coast samples, suggesting the confluence of the cold Benguela and warm Agulhas current may act as the approximate point of a phenotypic and possible genetic breakpoint. Finer scale genetic analysis of samples collected across the Benguela–Agulhas confluence reported no significant genetic structuring in this area suggesting environmental heterogeneity and not restriction of genetic flow/isolation as the primary driver of the observed phenotypic divergence.
Taraba State, Nigeria, is an important conservation site for the Endangered Nigeria–Cameroon chimpanzee Pan troglodytes ellioti. Gashaka Gumti National Park, Nigeria's largest national park and home to potentially the largest contiguous population of the Nigeria–Cameroon chimpanzee, spans a significant portion of the eastern sector of Taraba and the adjoining Adamawa State. South of the Park, Ngel Nyaki Forest Reserve comprises two forest fragments and holds a small population of chimpanzees. We investigated the existence of patterns in population structure and dispersal within this region, using microsatellite loci extracted from non-invasive sources of DNA. Our results indicate that dispersal and thus gene flow between the groups of chimpanzees at the Park and Reserve is limited, at least more so than it is within the Park, and we identified a biased sex ratio at the Reserve, forewarning of potential conservation concerns in relation to demographic and genetic stochasticity. We discuss conservation actions that may be applicable to sustaining the population within Ngel Nyaki Forest Reserve.
Our investigation was based on a molecular study of the genetic relationships among raspberry genotypes collected around selected medieval castles, Carthusian monasteries and nearby villages. We assumed that the hypothetical medieval raspberry genotypes could be traced to isolated medieval settlements that used to be highly prosperous during the feudal era but were later abandoned. Some of these genotypes could have survived in natural conditions without seed multiplication for at least three centuries. The molecular genetic analysis was based on microsatellite data. A total of 155 alleles were detected at 18 microsatellite loci. The clustering method grouped the analysed genotypes into seven main clusters. The analyses indicated that the most probable medieval genotypes had been collected around the ruins of two abandoned Carthusian monasteries: Zice and Jurkloster. They were morphologically very similar, vigorous and primitive but obviously not wild genotypes. The plants could be more than 2.3 m high, the canes were medium waxy, the lower and upper parts of the canes were covered by sparse short spines, the mid part was more or less completely smooth, the fully developed leaves were 15–25 cm long and the inflorescences were loose. In addition, the flowers were relatively small, the fruit setting was poor and the fruits were small, ovoid to conical and more aromatic than those of modern cultivars.
Salernitano (SAL) is an ancient Italian horse breed developed over the course of the ages together with Napoletano and, during the 20th century, by crossing with Thoroughbred horse lines. Excellent in hurdle jumping, this breed is currently facing a concrete risk of extinction due to the lack of appropriate management strategies. This research is the first SAL genetic characterization that aims to set up the basic knowledge for a conservation plan. A representative sample of 61 SALs was analyzed by means of a set of 16 microsatellites markers (short tandem repeats (STRs)). The sequence of hypervariable D-loop mtDNA region was also performed on a subset of 24 mares in order to study the maternal diversity and obtain a complete picture of the internal genetic variation. All the molecular data were analyzed together with those obtained from three Sicilian horse breeds investigated in a previous research (Siciliano, Sanfratellano and Sicilian Oriental Purebred). STRs markers revealed a moderate level of genetic diversity in SAL (alleles/locus 5.1, He 0.67) and confirmed the hunch of genetic erosion. Autosomal variability highlighted a very light deficit of homozygotes (FIS=−0.067). Experimental D-loop sequences were compared by multiple alignments with those retrieved from biological databases and revealed two unreported haplotypes. The phylogenetic network, which was built on mtDNA sequences, included various cosmopolitan and European horses and showed SAL haplotypes distributed among different mtDNA lineages.