Knowledge of the life history traits, reproductive biology and demography of rare species is fundamental for their conservation, yet plant population monitoring is uncommon. The restricted ranges of the Limonium species endemic to the Mediterranean area, combined with the vulnerability of their specialized littoral habitats, indicate the need for appropriate conservation measures. We evaluate the conservation status and estimate the future extinction risk of three Limonium species endemic to Zakynthos Island in the Ionian Islands, Greece (Limonium korakonisicum, Limonium phitosianum and Limonium zacynthium) using 5 years of monitoring data. We compile information on their geographical distribution, population dynamics, reproductive biology and genetic diversity. Population sizes and survival rates of seedlings exhibited marked annual fluctuations, although fecundity and relative reproductive success remained high throughout the monitoring period. We observed a dominance of mature individuals in all three species, indicating their increased tolerance to salinity. Three subpopulations each of L. phitosianum and L. zacynthium were genotyped using five microsatellite loci. The observed number of alleles and the low gene flow value potentially indicate reduced genetic diversity, inbreeding, and limited gene flow within and among subpopulations of both species. Based on the IUCN categories and criteria, we assess L. korakonisicum as Critically Endangered, L. phitosianum as Near Threatened and L. zacynthium as Endangered. Population viability analyses predict that, among the three species, L. zacynthium will face the highest risk of extinction within the next 50 years. Knowledge of the biology of these species provides data essential for identifying critical factors for their survival and for proposing targeted conservation measures.

Salvia miltiorrhiza is an outcrossing and perennial herb native to China. Although well-known for its medicinal value, there is a lack of knowledge regarding its natural population genetics. Here, we used 12 microsatellite markers to investigated population genetic diversity and structure of 215 samples from populations naturally distributed in central eastern China. A moderate level of genetic diversity was detected probably due to the over-mining of its roots. The allelic richness (AR) ranged from 3.034 to 4.889 with an average of 3.891. Moreover, pairwise estimates of FST among the populations of S. miltiorrhiza varied from 0.036 to 0.312 and two clusters were obtained by STRUCTURE and discriminant analysis of principal components. It is likely that the genetic differentiation of these two clusters was formed during glacial periods. Our result provides insights into the conservation of this valuable medicinal plant.

There is a need for a low-cost strategy that accelerates the genetic change in local chicken breeds. Therefore, the objective of this study is to develop a strategy for marker-assisted selection for growth in local chickens in the developing countries. The studied lines were a homozygous normally feathered selected line (CE1), homozygous naked-neck selected line (CE3) and their corresponding control lines (CE2 and CE4). Lines CE1 and CE3 have been selected for high 6-week body weight (BW) for five generations. Three generations were obtained for this study. In the current study, we consider the 6th generation as the base generation for marker-assisted selection study. Two subsequent selected generations were obtained from the base generation. The selected lines were significantly heavier at 6 weeks of age than their corresponding control lines by 24.15 and 27.07% in the base generation and by 51.4 and 34.5% in the second selected generation. A total of 34 polymorphic alleles in 19 loci were flown over generations and the flow trends were different in different lines and families within lines. Principal component analysis was applied to the data of both lines and two main components were found and three canonical correlations were obtained. Four and five alleles were persistently concomitant with the highly performed families in lines CE1 and CE3, respectively. These alleles would have brought 6-week BW into higher levels of performance by 25.1 and 16.6% in the second generation if they would have been considered in the selection for 6-week BW.

Dibothriocephalus latus is the most frequent causative agent of fish-borne zoonosis (diphyllobothriosis) in Europe, where it is currently circulating mainly in the Alpine lakes region (ALR) and Russia. Three mitochondrial genes (cox1, cob and nad3) and 6 microsatellite loci were analysed to determine how is the recently detected triploidy/parthenogenesis in tapeworms from ALR displayed at the DNA level. A geographically distant population from the Krasnoyarsk Reservoir in Russia (RU-KR) was analysed as a comparative population. One or 2 alleles of each microsatellite locus was detected in plerocercoids from RU-KR, corresponding to the microsatellite pattern of a diploid organism. In contrast, 1–3 alleles were observed in tapeworms from ALR, in accordance with their triploidy. The high diversity of mitochondrial haplotypes in D. latus from RU-KR implied an original and relatively stable population, but the identical structure of mitochondrial genes of tapeworms from ALR was probably a consequence of a bottleneck typical of introduced populations. These results indicated that the diploid/sexually reproducing population from RU-KR was ancestral, located within the centre of the distribution of the species, and the triploid/parthenogenetically reproducing subalpine population was at the margin of the distribution. The current study revealed the allelic structure of the microsatellite loci in the triploid tapeworm for the first time.

To facilitate population-genetic studies, we developed simple sequence repeat (SSR) markers and a molecular species identification assay for Peltigera membranacea (Ascomycota, Peltigerales), a common ground-dwelling lichen of forest and tundra ecosystems. Additional markers were developed for its Nostoc photobiont. Twenty-one fungal markers for P. membranacea were found to be polymorphic, with the number of alleles ranging from 3–21. Nei's unbiased gene diversity ranged from 0.588 to 0.640 in four significantly structured (FST = 0.059) mycobiont populations. For the Nostoc photobiont, 14 polymorphic SSR were developed, yielding 4–14 alleles each, with gene diversity ranging from 0.062 to 0.771 in four populations showing substantial population structure (FST = 0.278). The new markers developed are suitable for population genetic studies of Peltigera membranacea and of its cyanobiont, and at the same time allowed us to distinguish 98.5% of P. membranacea specimens from morphologically similar species of Peltigera.

The prevalence rates of trypanosomes, including those that require cyclical transmission by tsetse flies, are widely distributed in Africa. Trypanosoma brucei and Trypanosoma congolense are actively maintained in regions where there are no tsetse flies although at low frequencies. Whether this could be due to an independent evolutionary origin or multiple introduction of trypanosomes due to continuous movement of livestock between tsetse-free and -infested areas is not known. Thus, the aim of the study was to carry out microsatellite genotyping to explore intra-specific genetic diversity between T. (Trypanozoon), T. congolense and Trypanosoma vivax from the two regions: tsetse infested and tsetse free. Microsatellite genotyping showed geographical origin-based structuring among T. (Trypanozoon) isolates. There was a clear separation between isolates from the two regions signalling the potential of microsatellite markers as diagnostic markers for T. brucei and Trypanosoma evansi isolates. Trypanosoma vivax isolates also clustered largely based on the sampling location with a significant differentiation between the two locations. However, our results revealed that T. congolense isolates from Northern Kenya are not genetically separated from those from Coastal Kenya. Therefore, these isolates are likely introduced in the region through animal movement. Our results demonstrate the occurrence of both genetic connectivity as well as independent evolutionary origin, depending on the trypanosome species between the two ecologies.

Tall fleabane [Conyza sumatrensis (Retz.) E. Walker] is commonly invasive in agricultural fields, reducing yield in various infested crops. The current study investigates the genetic diversity within and between a significant number of invasive C. sumatrensis biotypes in soybean fields in southern, southeastern, and midwestern Brazil, using microsatellites as molecular markers. High and low observed and expected heterozygosity estimated in microsatellite loci supported our hypothesis that different levels of genetic diversity may be detected within biotypes from different invaded fields. Analysis of a significant number of biotypes in several fields showed high and low genetic diversity not associated with geographic distribution, bottleneck effect, or susceptibility to glyphosate. A deficit of heterozygous plants, high genetic divergence, and moderate allelic transference were also observed. Allelic fixation was different in the different biotypes. The bottleneck effect was seen in biotypes with reduced genetic diversity and in biotypes with the highest genetic diversity. Data on genetic diversity, bottleneck effect, and glyphosate resistance showed contrasts in biotypes from nearby invaded fields. Our study showed different genetic diversity levels in biotypes from invaded areas under the same climatic conditions.

Nipa (Nypa fruticans Wurmb.), aside from being part of the mangrove forest ecosystem that has been under assault at an alarming rate, lacks information on its genetic variation at the population level for conservation planning purposes. Microsatellites, or simple sequence repeats (SSRs), are molecular markers employed to detect genetic variations in population studies. Here, the primers for this palm were identified through whole-genome sequencing using the Illumina® Sequencing Technology platform. Over half (52.4%) of SSRs consisted of mononucleotide repeats, whereas the remainder mostly consisted of dinucleotide (22.7%) and trinucleotide (22.4%) repeats. The frequencies of longer repeats (tetranucleotides, pentanucleotides and hexanucleotides) were exceptionally rare (2.5%). At least 18 out of 75 genomic loci were polymorphic, based on 37 randomly selected samples assayed for screening. Polymorphic loci (P) reached a mean of 72% across the Indo-West Pacific region. Estimates of genetic diversity showed a lower mean observed heterozygosity (Ho) (0.195) than expected (He) (0.316), suggesting evolutionary forces were acting across nipa populations. The fixation (F) index (0.388) supported this result, which indicates considerable genetic differentiation among populations in the region. An average of 2.111 alleles (Na) was obtained. In conclusion, these microsatellite markers are promising tools for genetic evaluation of nipa populations to facilitate characterization of this species in its distribution range.

Genetic data on threatened plant populations can facilitate the development of adequate conservation strategies to reduce extinction risk. Such data are particularly important for species affected by habitat fragmentation such as Magnolia cubensis subsp. acunae, a Critically Endangered magnolia subspecies endemic to Cuba. Using genetic data from 67 individuals, we aimed to evaluate the effect of habitat fragmentation on two subpopulations in the Guamuhaya mountain range, in Topes de Collantes Protected Natural Landscape and Lomas de Banao Ecological Reserve. We characterize the structure and genetic diversity of these subpopulations, with the objective of managing their conservation more effectively. We used Landsat satellite images to determine land-cover types at the two locations and calculated indices of habitat fragmentation. For genetic analyses, we extracted DNA from the leaf tissue of individuals from the two subpopulations and used 11 microsatellite markers to genotype them. We calculated heterozygosity, allelic richness and the F-statistics, to evaluate genetic variability. The montane rainforest in Topes de Collantes was most affected by habitat fragmentation, with smaller patches of more irregular shapes, compared to submontane forest at this location and both montane and submontane forests in Lomas de Banao. Genetic diversity was higher in Topes de Collantes, but we found no genetic differentiation between subpopulations. Our findings suggest the two subpopulations can be considered a single evolutionary unit and conservation entity. We propose to use individuals from both subpopulations for reinforcement to increase the overall genetic diversity of the subspecies.

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.