Cabernet Sauvignon grapes and wine produced in the Helan Mountains were evaluated. Three different representative soil types—aeolian, sierozem and irrigation silting soil were studied. The compositions of grapes and wines were measured, and in addition, the weights of 100-berry samples were determined. The grapes that grown on the aeolian and sierozem soils matured sooner than those grown on the irrigation silting soil. The highest sugar content, total soluble solids content, sugar to acid ratio and anthocyanin content were found in the grapes that grown on the aeolian soil. The wine produced from this soil had improved chroma and tone and higher-quality phenols. The grapes grown on the sierozem soil had the highest total phenol and tannin contents, which affected the wine composition. The grapes grown on the irrigation silting soil had higher acidities, but the remaining indices were lower. In addition, the grapes grown on the aeolian soil resulted in wines with better chroma and aroma. The sierozem soil was beneficial for the formation of wine tannins and phenols and significantly affected the wine composition. The quality of the grapes from the irrigation silting soil was relatively low, resulting in lower-quality wine.Different soil types can significantly affect the composition of wine grapes and the final wine product. In this study, the effects of soil types on the composition of  Soil provides the foundation for grape vine growth, providing the necessary water and nutrition . The phyThe effects of regional characteristics such as soil type, climate and topography on grape phenolic compounds have been widely investigated ,11. All The composition of wine mainly depends on the composition of the grapes, followed by the wine-making technique . The sugDifferent soils types have different soil physical structures and mineral nutrient contents. Because aeolian soil has high sand contents, it has highly permeable, low nutrient and plant available water contents (PAWC). Irrigation silting soil has equivalent percentages of clay, silt and sand, which makes it less permeable, with medium levels of PAWC. The structure of sierozem soil is loose and highly permeable, which results in moderate PAWC and fertility with low organic matter and mineral nutrient contents and high Ca concentrations ,15. GeneThis study aimed to analyze the differences in wine grapes and wine composition among three soil types in the Helan Mountains. The objective of this research was to evaluate the relationships between soil types and wine grape composition and to provide practical information for adjusting vineyard growing practices and optimizing wine flavor.The study was approved by the Grape and Wine Engineering Research Centre of Education Ministry and was conducted at their experimental station in Ningxia (NW China).Cabernet Sauvignon grape variety was planted in 2006 in rows orientated north to south and with an individual vine density of 0.5 × 3.0 m.The vineyards are located on the Helan Mountain alluvial plain in Ningxia (NW China)  at an altitude of approximately 1035 m with a cool, semi-arid climate and large diurnal temperature differences. The annual accumulated temperature varies from 3298–3351°C, with abundant sunshine of annual average sunshine 2800~3000 hours and low annual rainfall of 158–200 mm. Early frosts are common on the slopes of the Helan Mountains, and winter temperatures are very low (<-10). Therefore, the grapevines must be covered with soil to prevent injury. Any harvesting delay will reduce the available time for pruning and soil covering. In 2013, three wine-growing areas with distinct soil types were selected: an aeolian soil area , a sierozem area  and an irrigation silting soil area . The three regions showing a triangular distribution and direct distance of them are less than 20 km, the altitude, light, rainfall and other climatic conditions are similar. All test points are our school experimental base, in order to understand the difference in quality wines under different soil types, all cultivation and management are the same, unified pruning, fertilization, irrigation and disease prevention. The basic soil physical and chemical parameters from these areas are shown in 0Brix was determined using a Brix refractometer . Reducing sugars were determined using titration with Fehling reagent, and titratable acidity was determined using standardized 0.1 N NaOH (end-point pH 8.2). The pH of the juice was recorded using a Metmorph 702SM automatic neutralizer  [In 2013, sixty plants were selected from each soil type in the vineyards as the study subjects. At the commencement of veraison (~mid-August), samples were collected weekly over a 8-week period until final harvest, which usually occurred during the first week of October. During the final harvest of each replicate, a random sub-sample of 100 grapes was weighed to determine the average grape weight. The samples were frozen and stored for later analyses, including tannin, anthocyanin, and iron-reactive phenolic content analyses. zerland) . Whole gzerland) ,30. The zerland) . In thisp values of less than 5% (p<0.05) were considered statistically significant.Significant differences among soil types were random determined by methods of one-way ANOVA and two-way analysis of variance (ANOVA) followed by the Student-Newman-Keuls test using the SAS 8.1 software  followed by the least significant difference (LSD) test for multiple comparisons among groups. Differences with During the maturation period, the 100-berry weight varied significantly among the different soil types . It incrGenerally, the total soluble solids increased during the maturation period of the grapes . During During the first 3 weeks of maturation, the grape reducing sugar content increased steadily in all soil types. However, the accumulation rates increased during the subsequent 2 weeks for the grapes that were grown on the aeolian and sierozem soils . The redThe titratable acidity increased steadily during maturation and decreased rapidly during the first 3 weeks . The titDuring the first 5 weeks of maturation, the pH of the grape juice generally increased by 0.10 pH units each week . AlthougSugar to acid ratios in the grapes from the aeolian and sierozem soils were generally higher than those in the grapes from the irrigation silting soil. Until September 5th, the values were similar, but they tended to diverge in the final 4 weeks . At harvThe anthocyanin contents in the grape during the first 5 weeks of maturation increased before decreasing over the final 2 weeks  in all t-1, which was significantly greater than that of the grapes grown on the aeolian soil. The total phenol contents of the grapes from the irrigation silting soil were the lowest among the treatments and were significantly lower than in the grapes grown on the aeolian soil.During the early stages of the mature period, the total phenol contents in the wine grapes from the sierozem soil were lower than those grown on the aeolian soil . After S-1, followed by 42.65 mg g-1 from the aeolian soil. However, the difference between the sierozem and aeolian soils was not significant. The tannin content of the grapes from the irrigation silting soil was only 30.35 mg g-1, which was significantly lower than that of the grapes from the other two soil types.Before August 29th, the tannin contents of the wine grapes from the three soil types were similar. However, after August 29th, the tannin contents of the wine grapes from the sierozem and aeolian soils rapidly increased, reaching a stable value by September 26th. However, the tannin contents of the wine grapes from the irrigation silting soil continued to increase gradually . At harvAccording to Soil types played an important role in the tannin and total phenol contents of the grapes. The wine made from grapes from the sierozem soil had the highest tannin and total phenol contents, followed by the aeolian soil and the irrigation silting soil, the latter of which was significantly lower than the other two soil types.Similar to the phenolic compound results, the chromas of the wines made from the grapes grown on the aeolian and sierozem soils were not significantly different. The wines from the aeolian and sierozem soils both had chromas of approximately 8.20, which was significantly greater than that of the wine from the irrigation silting soil . The winSoil composition as a determining factor for the sensory profile of a wine, the types and texture will influence the root systems and the soil water-holding capacity and mineral composition ,32. In aSugar, organic acid, phenolic compounds, anthocyanin and aroma substances are all important for wine composition, with sugar being the most important substance. Grapes with higher sugar contents can produce full-bodied wines ,35. ThusSome differences in the various phenolic compounds were observed in the regional wines, which suggested that the accumulation of phenolic compounds in grape berries is strongly affected by soil type ,37–39. TSoil texture had the most wide-reaching influence on the wines from the various sites. However, consistency among vintages was not observed ,41. ZoneAt the same grape-ripening level, soil may affect several wine characteristics. Wines from poorer soils with higher coarse fractions exhibit higher total phenolic contents and color intensity but lower resveratrol concentrations ,41. The Different soil types resulted in significant differences in the total sugar and total acid contents of the grapes and, to some extent, the anthocyanin, tannin, phenolic and aroma contents in the berry skin. The grapes from the aeolian soil have small berry size and loose fruit clusters. The skin color was nicer and the skin fruit ratio was higher for the grapes from the sierozem soil. Therefore, the sugar and anthocyanin contents and the color density were greater, which resulted in better tasting and looking wine with a high tannin content. However, the composition of the wine made from the grapes grown on the irrigation silting soil was comparatively lower.The grapes grown on the sierozem soil that matured early had high sugar and anthocyanin contents. Consequently, these grapes produced wine with better chroma. Furthermore, the grapes grown on the sierozem soil that matured during a moderate period had medium tannin contents and low acidity. In general, the sierozem soil was favorable for the formation of tannins and phenolic compounds, which influenced the mouthfeel and composition of the wine. The grapes grown on the irrigation silting soil had high acidity and lowest contents of other investigated substances.


Driving a taxicab remains one of the most dangerous occupations in the United States, with leading homicide rates. Although safety equipment designed to reduce robberies exists, it is not clear what effect it has on reducing taxicab driver homicides.Taxicab driver homicide crime reports for 1996 through 2010 were collected from 20 of the largest cities  in the United States: 7 cities with cameras installed in cabs, 6 cities with partitions installed, and 7 cities with neither cameras nor partitions. Poisson regression modeling using generalized estimating equations provided city taxicab driver homicide rates while accounting for serial correlation and clustering of data within cities. Two separate models were constructed to compare (1) cities with cameras installed in taxicabs versus cities with neither cameras nor partitions and (2) cities with partitions installed in taxicabs versus cities with neither cameras nor partitions. Cities with cameras installed in cabs experienced a significant reduction in homicides after cameras were installed  and compared to cities with neither cameras nor partitions . Cities with partitions installed in taxicabs experienced a reduction in homicides  compared to cities with neither cameras nor partitions, but it was not statistically significant.The findings suggest cameras installed in taxicabs are highly effective in reducing homicides among taxicab drivers. Although not statistically significant, the findings suggest partitions installed in taxicabs may be effective. Taxicab drivers work in one of the most violent occupations in the United States . Two covariates were included in the statistical analysis: the concurrent decline in homicide rates in the United States since 1990  and the number of licensed taxicabs from municipal transportation regulators (denominator). A standardized search strategy tailored for municipal police crime departments was used to locate taxicab driver homicides using any one of the following criteria: (1) crime premise designated as ‘vehicle’, (2) name of licensed taxicab companies, (3) keywords ‘cab’, ‘taxi’ or ‘driver’ in the crime report or (4) news clippings reporting taxicab driver homicides. Each crime report provided by each municipal police department was reviewed for relevance by the first author. Information on installation year of camera or partition (or neither) was obtained directly from city transportation regulators. A city was designated annually as a “camera city” or “partition city” if >70% of the cabs had cameras or partitions during the study period; a cut point of 70% was decided Generalized estimating equations accounted for serial correlation and clustering of data within cities. Annual taxicab driver homicide rates were modeled on installation status (camera or partition) compared to control cities in two separate models adjusting for covariates. A separate model restricted to camera cities only compared post-installation versus pre-installation homicide rates. The Wald test statistic determined the significance of installation status. The natural logarithm of the number of licensed taxicabs by city per year was used as an offset variable. The taxicab driver homicide counts were assumed to follow a Poisson distribution; the offset variable provided the denominator used to calculate the homicide rates. The data were tested for dispersion and found to be slightly under-dispersed . Therefore, all confidence intervals were considered conservative in their range.Police crime report data were analyzed for 7 cities where cameras were installed in taxicabs, 6 cities where partitions were installed in taxicabs and 7 cities where neither partitions nor cameras were installed in taxicabs. From 1996 through 2010, 95 taxicab driver homicides were investigated by law enforcement authorities   after controlling for pre-existing annual changes in city taxicab driver homicide rates (“year”) and the background city homicide rate. A sub-analysis exploring changes in taxicab driver homicide rates among cities with only a company policy  found a statistically significant reduction in rate  that persisted  after covariate adjustment. Modeling annual citywide taxicab driver homicide rates on camera installation in camera cities compared with control cities post-installation versus pre-installation revealed a statistically significant reduction in homicide rate  (Model 2) that persisted  after controlling for covariates. A sub-analysis exploring changes in taxicab driver homicide rates in cities where camera installation is not citywide but due to a company policy found a reduced rate compared to control cities in both unadjusted and adjusted models . Modeling annual citywide taxicab driver homicide rates on partition installation in partition cities compared with control cities revealed a slightly higher rate  which, after controlling for declining homicide rates and background crime rates, presented a reduction in taxicab driver homicide rate that was not statistically significant .Modeling annual citywide taxicab driver homicide rates among camera cities post-installation compared with pre-installation revealed a statistically significantly reduced homicide rate  taxicab company implemented a policy of camera-equipped taxicabs. Findings suggest that cities with partition-equipped taxicabs may experience reduced taxicab driver homicide rates compared to nonpartition-equipped other cities.The observance of a significant reduction in taxicab driver homicides in cities where taxicabs are equipped with cameras, and the more pronounced effect in cities where taxicabs are equipped with cameras as mandated by a city ordinance, was an encouraging one. Based on anecdotal evidence from transportation regulators in cities already mandating cameras by ordinance, transportation regulators are currently considering promulgating city ordinances for their cities that will mandate the use of security cameras in taxicabs. Our findings suggest cities where taxicabs are equipped with cameras could result in fewer taxicab driver homicides particularly in cities with an ordinance mandating taxicabs are equipped with cameras. Furthermore, these findings are consistent with a previous analysis examining news clippings to identify homicides  we could not estimate individual risk of homicide for taxicab drivers across safety equipment types and (2) the lack of uniform data in the crime reports on homicide circumstances precluded more detailed analyses. Significant strengths of this study are the 15-year time span, systematic collection of data in 20 major US cities, pre-post design that includes use of comparison cities with neither camera-equipped nor partition-equipped taxicabs, and statistical analysis that accounts for serial correlation of data that adjusts for two crucial covariates. Additionally, in working with city regulators and counting only licensed drivers, we were able to assume the drivers were in taxicabs installed with required safety equipment. The study and its findings make a significant contribution to understanding the possible effect of taxicab safety equipment on taxicab driver homicides.Taxicab driver personal safety in Seattle and King County, Final report and recommendations. The report of the Taxicab Advisory Group Committee on Driver Safety to the Director of the Department of Executive Administration for the city of Seattle. June 18, 2004.


Vespula pensylvanica in its native range of North America and its introduced range in the archipelago of Hawaii. Our goal was to gain a better understanding of the invasion dynamics of social species and the processes affecting biological invasions. We found that V. pensylvanica showed no significant genetic isolation by distance and little genetic structure over a span of 2000 km in its native range. This result suggests that V. pensylvanica can successfully disperse across large distances either through natural‐ or human‐mediated mechanisms. In contrast to the genetic patterns observed in the native range, we found substantial genetic structure in the invasive V. pensylvanica range in Hawaii. The strong patterns of genetic differentiation within and between the Hawaiian Islands may reflect the effects of geographic barriers and invasion history on gene flow. We also found some evidence for gene flow between the different islands of Hawaii which was likely mediated through human activity. Overall, this study provides insight on how geographic barriers, invasion history, and human activity can shape population genetic structure of invasive species.Invasive species cause extensive damage to their introduced ranges. Ocean archipelagos are particularly vulnerable to invasive taxa. In this study, we used polymorphic microsatellite markers to investigate the genetic structure of the social wasp  Invasive species are recognized as one of the top threats to the environment . Native samples were collected from 170 traps within the states of California, Colorado, Oregon, Wyoming, and New Mexico. Invasive samples were collected from 178 traps from the Hawaiian Islands of Kauai, Oahu, Molokai, Lanai, Maui, and Hawaii , and expected heterozygosity (He), were calculated with GenAlEx v 6.5  v 1.1 in order to assess levels of genetic differentiation  present in the native range and invasive range, we performed different simulations, each under the assumption of a different K value (1 to 44), representing the total number of transects. For each simulation, we used an admixture model with uncorrelated allele frequencies to account for wasps with mixed ancestry and the LOCPRIOR model to use sampling location to inform clustering. Each simulation was run 10 times with 10,000 steps of burn‐in and 50,000 Markov chain Monte Carlo (MCMC) repetitions. The most likely number for K was selected based on log likelihood and the ΔK statistic developed by Evanno et al.  which is a Bayesian approach that compares the posterior probabilities of a large number of simulated datasets under given models to those calculated from observed data  trees for individuals within transects and regions  and effective number of alleles (P = 0.001) than wasps from the invasive range (Table P = 0.0219) from the number of private alleles in the invasive range . Finally, the native range had a significantly higher level of expected heterozygosity (P = 0.001) and observed heterozygosity (P = 0.023) when compared to the invasive range. Overall, these results suggest that there is a slightly, but significantly, higher level of genetic diversity in the native range than the invasive range of V. pensylvanica.We investigated whether levels of genetic diversity differed between native and invasive ge Table . We alsoP = 0.0023), expected heterozygosity (P < 0.001), effective number of alleles (P < 0.001), and number of private alleles (P < 0.001) among the islands. Interestingly, the island of Hawaii had the highest effective number of alleles, observed heterozygosity, expected heterozygosity, and allelic richness. Maui had highest number of private alleles. In contrast, Kauai had the lowest observed heterozygosity, expected heterozygosity, allelic richness, effective number of alleles, and number of private alleles.We next investigated differences in genetic diversity between different islands in the invasive Hawaiian range . There were significant differences in observed heterozygosity  and New Mexico (P < 0.001). Additionally, there were marginally significant excesses of heterozygosity present in the islands of Hawaii (P = 0.052), Lanai (P = 0.052), and Molokai (P = 0.086). In contrast, Oahu (P = 0.852) and Kauai (P = 0.380) displayed no signs of bottlenecks. Overall, there is some evidence for population bottlenecks in the eastern islands of Hawaii but not the western islands.Recently bottlenecked populations may display an excess of heterozygosity compared to expected heterozygosity calculated from observed allele number  and geographic distance (km) for individuals sampled from different traps in the native range   , Oahu , Molokai , or Lanai . However, we did find significant isolation‐by‐distance relationships in Hawaii  and Maui .We tested for the presence of genetic isolation by distance. Our analysis revealed no significant correlation between genetic distance , as the ΔK metric was similar for K = 2 and 3. Nevertheless, K = 3 had the highest ΔK. Interestingly, almost all individuals in the native range were partially assigned to all three putative populations . Overall, the mean assignment scores for the western regions were higher than those found for the central regions (Table P > 0.05), suggesting that we cannot exclude either the western or the central regions as the source population for invasive V. pensylvanica .We assigned individuals from the invasive range to regions in the native range to determine the potential origins of invasive ns Table a. This sIn contrast to the assignment tests, approximate Bayesian computation suggested that the central regions of the native range were the most likely source of the invasive populations Table b. SpecifWe visualized the relationships between individuals sampled from different transects with neighbor‐joining trees. Transects from the native regions formed a starlike structure, indicating a lack of strong genetic relationships in the native range Fig. A. In conIntroduced species tend to experience drops in genetic diversity due to population bottlenecks derived from founder events .Table S2. Measures of genetic diversity at microsatellite loci for V. pensylvanica from invasive regions, including number of samples (N), number of alleles (Na), effective number of alleles (Ne), observed heterozygosity (Ho), expected heterozygosity (He), allelic richness (A), and number of private alleles (Np).Table S3. Assignment and Exclusion test of invasive Hawaiian populations to mainland populations in the United States.Table S4. Locations of, total numbers of, and genotypes of V. pensylvanica wasps collected from, each trap in the sampled transects, ranges, and regions (NA = location not determined) at 15 loci.Click here for additional data file.


Melanaphis sacchari is a pest of sugarcane and sorghum which originated in the old world, was introduced into the Americas, and is now distributed worldwide. Our purpose was to assess the genetic diversity and structuring of populations of this species according to host and locality. We used 10 microsatellite markers to genotype 1333 individuals  collected mainly on sugarcane or sorghum. Five multilocus lineages (MLL) were defined, grouping multilocus genotypes (MLG) differing by only a few mutations or scoring errors. Analysis of a 658 bp sequence of mitochondrial COI gene on 96 individuals revealed five haplotypes, with a mean divergence of only 0.19 %. The distribution of MLL appeared to be strongly influenced by geography but not by host plant. Each of the five MLL grouped individuals from (A) Africa, (B) Australia, (C) South America, the Caribbean and the Indian Ocean including East Africa, (D) USA, and (E) China. The MLL A and C, with a wide geographic distribution, matched the definition of superclone. Among aphids, M. sacchari has one of the lowest known rates of genetic diversity for such a wide geographical distribution.Numerous studies have examined the genetic diversity and genetic structure of invading species, with contrasting results concerning the relative roles of genetic diversity and phenotypic plasticity in the success of introduced populations. Increasing evidence shows that asexual lineages of aphids are able to occupy a wide geographical and ecological range of habitats despite low genetic diversity. The anholocyclic aphid  Range expansion of exotic species can result from either evolutionary adaptation or generalism and plasticity often associated with a change in niche Melanaphis van der Groot 1917 comprises around 20 species mainly associated with Poaceae, most of which originate from East Asia Melanaphis sacchari  , which is considered to be mainly anholocyclic, is present in America, Australia, Asia and Africa. M. sacchari is known to be invasive in continental US Melanaphis individuals originating from sorghum or sugarcane were distinct taxa, referred to as M. sorghi and M. sacchari respectively, even though their host plant preference was not absolute. In their catalogue, Remaudière and Remaudière M. sorghi to be a synonym for M. sacchari, but both forms were still listed as separate taxa by Blackman and Eastop M. sacchari constitutes a single species or a complex of sibling taxa.The old world genus M. sacchari is a major pest of sorghum and sugarcane. On sugarcane, it is considered to be the most common and most efficient vector of the Sugarcane yellow leaf virus (ScYLV), which causes yellow leaf disease M. sacchari is one of the main control tactics suggested both for sugarcane In any case, M. sacchari; 2) molecular evidence for the existence of sibling species; and 3) its genetic diversity and structuring according to host and locality.Based on a worldwide sample of aphids covering its area of distribution (home range as well as invasive range) and using microsatellite markers and sequencing of a fragment of the mitochondrial cytochrome c oxidase I gene (COI), the purpose of this study was to evaluate 1) the mode of reproduction of Sorghum bicolor, S. halepense, S. verticilliflorum). The three sorghum species were considered as a single host plant, hereafter named ‘sorghum’.Here, an ‘individual’ refers to one individual aphid and a ‘sample’ refers a several individuals collected from the same host plant species in a given locality and date. The complete set of individuals  compriseAphids were collected from wild or cultivated plants and placed in 70% ethanol in Eppendorf tubes, kept frozen at −80°C until they were processed. Only a few aphids were collected on each plant sampled to avoid collecting several individuals from the same colony.Sampling was carried out from 2002 to 2009 by our team in Reunion Island and by colleagues in the other parts of the world (see acknowledgements). Geographic coordinates of sampling localities are provided in DNA was extracted using the “salting-out” protocol of Sunnucks and Hales M. sacchariAccording to their polymorphism, ten microsatellite loci  were selA total of 91 aphids were chosen among the worldwide sample to represent different combinations of region and host plant. COI fragments were amplified using the LCO1490 and HCO2198 primers designed by Folmer et al. M. sacchari, we assumed that the different occurrences of the same MLG in a sample were the result of local clonal reproduction. We therefore retained a single representative of each MLG in each of the 57 samples for genetic and diversity analysis.Micro-Checker software sex, the probability that the repeated MLGs originated from distinct sexual reproductive events. A psex value lower than 0.01 supported the hypothesis that MLGs originated from the same MLL MLG or MLL for RMLL), and N is the number of samples Using GENCLONE software M. sacchari individuals collected in India Melanaphis genus were also retrieved from GenBank and used as outgroups: M. donacis (referenced HQ443314), M. bambusae (referenced EU701747 and EU701746) and M. japonica (referenced GU457792). Maximum Likelihood inference performed with MEGA6 Sequence alignments of the COI gene were performed using Geneious software version 5.6.6 is). Genetic differentiation between MLLs was tested with a G test and pairwise FST were computed.We used GENEPOP We genotyped a total of 1333 aphids using the ten microsatellite markers. When we retained a single representative of each MLG in each of the 57 samples, this yielded a dataset containing 98 individuals.MLG  = 0.361. The distribution of the pairwise number of different alleles between MLG appeared multimodal, with a first minimum located at a distance of five alleles  which grouped slightly distinct MLGs resulting from step mutations or scoring errors . When the five M. sacchari sequences retrieved from GenBank were excluded, the mean divergence was 0.17% (range 0.000.30%). The mean divergence of M. sacchari sequences with the closest taxa, M. japonica, was 1.06% (range 0.92%1.39%).Within our 91 observed . One hapobserved . The secobserved . These t support . But witPlotting the results of the factorial correspondence analysis with GENETIX confirmed the grouping of the 36 MLGs in five MLLs . Factor st ranging from 0.262 to 0.694 .M. sacchari and M. sorghi. We found sequence variations peaking at 0.61%, with a mean value of 0.19%, both of which are within the range of intraspecific divergence observed in the Aphididae family by Footit et al. Sequencing the COI ‘barcoding’ region, a typical locus used for species discrimination and phylogeny, particularly in aphids Aphis gossypii, Carletto et al. A. gossypii was found in Iran A. gossypii alate spring migrants in France Brachycaudus helichrysi  sibling species H2, a sexually reproducing population was identified in India, despite almost exclusively clonal reproduction at the worldwide scale M. sacchari in Asia Population genetic parameters were consistent with populations which only reproduce by apomictic parthenogenesis, as previously described by Blackman and Eastop M. sacchari, which the present study did not allow us to identify. At least two of the five MLLs covered a very wide geographic area and matched the pattern of a single asexual genotype with a high capacity for dispersal which would have spread across a large area: MLL-A was observed in West and East Africa, and MLL-C was observed in South America, the Indian Ocean, East Africa, and the Caribbean. Both MLLs match the definition of “superclone” Microsatellite analyses showed that population structuring at the worldwide scale was only influenced by geography, delimiting five MLLs corresponding to five geographic zones: Africa, China, Australia, USA, and South America – Indian Ocean (including Kenya) and the Caribbean. Variation within each of the four biggest zones  was low, with MLGs within a zone differing by a few step mutations, which we suggest is due to one or few separate introductions. These results suggest that each of the five zones was colonized separately following the introduction of one or a few clones from the region of origin of M. sacchari in the countries sampled, meant we were not able to reconstruct the invasion routes of this species. The only exception was continental USA. M. sacchari was first described in Hawaii in the late 19th century M. sacchari was introduced in continental USA from Hawaii. This finding is noteable, as one would expect an introduction into continental USA from either South America or the Caribbean, a shorter invasion route. However, Mondor et al. M. sacchari has been recorded in almost all areas where sugarcane is cultivated, the possibility for the expansion of some genotypes should be taken seriously, mainly its unknown potential impact on the epidemiology of the viral diseases it transmits.The low rate of genetic diversity observed in the whole geographic area covered by our study, and the lack of published data about the dates of introduction of Figure S1Distribution of the pairwise number of different alleles between MLGs.(PDF)Click here for additional data file.Figure S2Molecular Phylogenetic analysis by Maximum Likelihood method with bootstrap support  using 658 bp cytochrome c oxidase subunit I sequences from 100 Melanaphis spp. individuals.(PDF)Click here for additional data file.Table S1Voucher number, sampling information, GenBank accession and SSR genotyping of individual aphids.(XLSX)Click here for additional data file.Table S2ST and significance of the G test computed with GENEPOP.Genetic differentiation between MLLs: pairwise F(PDF)Click here for additional data file.Table S3Population genetics parameters for each MLL.(PDF)Click here for additional data file.


GMP) of a city and its population (pop). It has been demonstrated that GMP ∝ YYpopβ with β always greater than 1 and close to 1.2. This fundamental finding highlights a universal rule that holds across countries and cultures and might explain the very nature of cities. However, in an increasingly connected world, the hypothesis that the economy of a city solely depends on its population might be questionable. Using data for 248 cities in the European Union between 2005 and 2010, we found a double GMP/pop scaling regime. For West EU cities, β = 1 over the whole the period, while for post-communist cities β > 1 and increases from ∼1.2 to ∼1.4. The evolution of the scaling exponent describes the convergence of post-communist European cities to open and liberal economies. We propose a simple model of economic convergence in which, under stable political conditions, a linear GMP/pop scaling is expected for all cities. The results suggest that the GMP/pop super-linear scaling represents a phase of economic growth rather than a steady, universal urban feature. The results also suggest that relationships between cities are embedded in their political and economic context and cannot be neglected in explanations of cities, urbanization and urban economics.Recent advances in the urban science make broad use of the notion of scaling. We focus here on the important scaling relationship between the gross metropolitan product ( Any geographically extended social entity’s per capita gross domestic product (GDP) is expected to be independent of its population. After all, this is the mean economic output of the people living in that city, village or metropolis. A population-independent per capita GDP would imply a linear relationship between total GDP and population. However, urban historians and urban economy scholars [dynamic externalities and urbanization economies are rooted in a combination of urban density and diversity, which promotes both interactions (knowledge exchange) and economic competition that in turn promotes innovation and economic growth. These observations focused on urban vs rural economies, highlighting the primal role of cities in economic growth and fostering further understanding of urban economic output resulting from individuals’ interactions in dense urban environments. Important investigations have been proposed in this direction by Bettencourt, Lobo and colleagues [GMP is universally dependent on pop through a power law such as GMP ∝ YYpopβ with β always greater than 1 and close to 1.2 ± 0.02. Larger cities are wealthier than expected by a linear assumption when compared with smaller cities. Super-linear GMP/pop scaling (β > 1) provides a strong and simple quantitative explanation for the increasing growth and expansion of cities: large urban basins offer more opportunities than disadvantages compared to smaller basins, which is why they continue to grow and attract people. Super-linear scaling is consistent with classical urban economic theories. Such theories are built upon the established fact that urbanization in developed countries was accompanied by economic growth and industrialization. These historic patterns generated expectations of a virtuous circle between economic growth and urbanization, regardless of the local conditions [GMP/pop scaling is still debated and poses some important questions about the relationships between urbanization and economic growth. For example, recent geographically sound studies show that the definition of urban boundaries is crucial to the measurement of any scaling of socio-environmental performance with population [variations on a theme?The hypothesis of urban scaling is certainly a pillar of quantitative methods for understanding cities . Far fro a theme , which a a theme . We focuscholars –6 have nlleagues , 7, 8. Illeagues , 10, thelleagues . For exanditions , 13. Howpulation , 15. As pulation , 17. ThiGMP is universally super-linear with population. European metropolitan areas do not show a uniform and super-linear behaviour in terms of GMP scaling. Former Eastern Bloc cities behave differently than other cities in Europe, suggesting that positive externalities are not sufficient for determining economic growth. We found, however, that different scaling regimes mirror the economic transitions and economic convergence patterns of post-communist and former Eastern Bloc cities. The results suggest that super-linear GMP/pop scaling represents a transition condition of growing economies rather then a universal and stable condition of all cities. Moreover, the economic growth of a town is dependent not only on its population size but indeed on higher level economic and social dynamics.We explored these questions using archival data on per capita GDP and population for 248 cities in the European Union between 2005 and 2010. We failed to verify the consensus that We used GMP and population for 243 metropolitan areas in Europe over the decade 2005-2010. The data are provided by Eurostat . These dGMP of the 248 cities is bimodal  and former members of the Eastern Bloc (EEu). The EEu countries were aligned with the USSR before its collapse in the late 1980s. EEu cities experienced transitions from totalitarian to liberal economic regimes. Theories of the dynamics of transition from totalitarian to liberal economies are widely debated . The mai bimodal , indicatGMP versus population increases, which indicates that the GMP growth rate is proportional to its population. Indeed, if a city with a larger population grows faster than a city with a smaller population, its GMP vs population curve  will become steeper over time, which this is what we observe for the emerging EEu economies. Given a city i with log population pi, let gi,t be its log GMP for year t. We assume at this stage that the population of cities does not change to the same degree as its GMP. This assumption works well with the observed data. For our analysis, we used a log scale for GMP (g) and population size (p). On a log scale, the scaling of GMP versus population size can be given a mathematical form:βt indicates whether the relationship is linear (β = 1), sub-linear (β < 1) or super-linear (β > 1). Empirically, we observe that βt is 1 for WEu cities for all years and increasingly larger than 1 for EEu cities. The growth of GMP can be modelled as a change from year to year:GMP growth have assumed a constant, population-independent GMP growth rate:We propose a simple descriptive model of the observed double scaling regime that might mirror the process of economic convergence of EEu to WEu cities incorporating the diminishing return effect. The scaling exponent of γ1 = 0. Plugging the GMP − pop scaling relationship into the GMP growth model, we find the relationship for the change in the scaling exponent:with GMP growth (where γ1 = 0) would lead to a stable scaling exponent over time, which we observe for WEu cities. For EEu cities, we observe a β that grows for cities that are poorer than the West and a β that is equal to 1 and stable for those that are as rich as cities in the West. To accommodate this latter observation, we make the GMP growth rate depend on GMP itself:Population-independent GMP such that this term is smaller when the per capita GMP of the city is larger than a threshold value of g⋆. The value of g⋆ is indeed arbitrary and can change given specific conditions. In our case, g⋆ = 11400 euros. Note that gi,t − pi is log per capita GMP. This new form of GMP growth reduces to GMP . Cities keep growing at a population- and GMP-dependent rate until their percapitaGMP crosses g⋆. When the city is rich, as is the case for all cities in the West and a handful in the East, the population- and GMP-dependent growth term drops out. Over very long periods and under desirable, stable economic and political conditions, we would expect all cities to cross the wealth threshold, and the growth term becomes constant and independent of the population. In the limit of θ → ∞, the population-dependent growth term in gi,t − pi = g⋆. Thereafter, GMP growth follows population-independent rate γ0, starting at a value g⋆+pi and thus scaling linearly with pi. After a very long time, we expect all cities’ GMP to show linear scaling with population. This apparently simple relationship is prone to speculation.Here, the population size-dependent term in the growth rate also depends on the city’s β = 1), as observed for WEu cities, may reflect the economically mature and politically stable conditions of the European Union. Linear scaling may be also a consequence of European redistribution of incomes to reduce disparities between small and large cities. Recent investments in internet connectivity and international high-speed trains have consequently increased the proximity of European capital cities, and for WEu economic activity, an average individual may be able to increasingly interact with out-of-town economic agents. As a result, the GMPs of different cities can become too inextricably interconnected to satisfy the assumption of independent LUZs (or MSAs). In the limit where an individual can interact with anyone in the larger economy of a state, his/her economic output will depend on the total population of the larger economic basin  and is thus independent of the population of the city where she/he resides. This leads to the traditional linear GDP vs population size relationship. The linearity of the GMP − pop relation as a signature of mature economies is also indicated by the exponents of the richer Eastern cities. The exponents for these cities are closer to 1 than are those for the Eastern Bloc as a whole  on the wealth of the city. The situation in Eastern cities tells a different story. After the collapse of the USSR, these economies developed rapidly, and more populous cities have been developing increasingly quickly over time. As expected, more populous cities have seen higher rates of growth, causing the exponent β to increase from 1.25 to 1.42. An increasing β indicates improving economic performance but may also indicate increasing inequality between cities. In a way, the observed hyper-linear scaling reflects an unbalanced situation of rapid growth of large cities and economic segregation of small ones, which makes the redistribution of income increasingly difficult. This finding is consistent with recent observations about the economic conditions and increasing economic inequality of Eastern European countries [In our opinion, linear scaling  the universality of scaling laws for cities and, consequently, on ii) on consistence and dependences between different types of urbanization economies.GMP/pop scaling is a powerful tool for observing economic fluctuations and convergences even over shorter periods. However, the universality of super-linear GMP/pop scaling remains difficult to prove. This in turn suggests that increasing connections among cities, rather than solely within cities, play crucial roles in urban economies and cannot be neglected in understandings urban economic and social dynamics. It might be possible also that super-linear scaling is an effect of aggregating many cities from different locations in the same analysis. In general terms, one might criticize the application quantitative tools inspired by physics and the natural sciences to cities, which reduces cities to natural or physical systems. Such systems (of cities) must then follow universal laws as any gas must expand with higher temperatures. The appeal of human/nature metaphors and of physical determinism in the study of cities, as for any simple and universal solution, is unquestionable. The simple results presented in this paper suggests, however, that very existence of universal laws that govern cities is questionable. Cities and urban systems can be indeed described using quantitative methodologies as with physical systems, but this does not justify the equation of cities to pure physical systems. In this sense is important to highlight that economic transition, specially in the case of Europe, is a top-down and planned phenomena and it played a crucial role in the global economy framework. On the other hand, cities are highly complex and self-organizing systems and we cannot neglect this complexity. In our opinion, a balance between quantitative and qualitative analysis, as in the proposed study, is key to extracting information from the data and to producing a more objective understanding of cities.The hypothesis of urban scaling certainly represents a large step in the scientific understanding of cities. We have shown that GMP/pop scaling exponent. Other factors, such as monetary changes or economic regulations, played a role in creating this separation and they should be taken in consideration for such analysis. Regardless of the causes of economic separation and their dynamics, it is interesting to observe that the fast economic convergence increases inequalities between cities in developing countries. It is possible that fast economic convergence due to forced liberalization may trigger further separation of poor and rich cities. Coexistence of poor and rich cities in a unique monetary and economic system might also suggest the existence of different urbanization economies which are rooted not only in economic growth but on different, and maybe hidden, urbanization drivers. Considering that most of the next urbanization is taking place in developing countries, such hidden and non GMP-based urbanization drivers must be further investigate.Other observations arise from observing the coexistence of distinct economic groups within the EU’ s cities. The causes of this separation are certainly rooted in the history of Europe. The causes of economic convergence are due to the transition from totalitarianism to liberal economies experienced by the Eastern countries. Certainly, as for the well-known Baltic Tigers, the advantageous conditions of Western states and extra-EU investors trigger a virtuous cycle of out-of-town economic input, consequently contributing to the observed increase in the


Drosophila pseudoobscura from North America. We used 27 isolines from three populations and exposed them to four temperature regimes  to examine environmental, genetic and genotype-by-environment sources of variance in wing size.Populations of a species often differ in key traits. However, it is rarely known whether these differences are associated with genetic variation and evolved differences between populations, or are instead simply a plastic response to environmental differences experienced by the populations. Here we examine the interplay of plasticity and direct genetic control by investigating temperature-size relationships in populations of By far the largest contribution to variation in wing size came from rearing temperature, with the largest flies emerging from the coolest temperatures. However, we also found a genetic signature that was counter to this pattern as flies originating from the northern, cooler population were consistently smaller than conspecifics from more southern, warmer populations when reared under the same laboratory conditions.We conclude that local selection on body size appears to be acting counter to the environmental effect of temperature. We find no evidence that local adaptation in phenotypic plasticity can explain this result, and suggest indirect selection on traits closely linked with body size, or patterns of chromosome inversion may instead be driving this relationship.The online version of this article (doi:10.1186/s12862-015-0323-3) contains supplementary material, which is available to authorized users. Recent work has highlighted the role of phenotypic plasticity in evolutionary divergence -4. The aconverse Bergmann cline, where individuals in cooler climates  mature at smaller sizes has also been observed [The well-known relationship between temperature and body size, known as the temperature-size rule, provides a useful framework for examining phenotypic plasticity in nature -15. Thisobserved . Conversobserved . Opposinobserved . Direct observed . For exaobserved . Body siobserved . Body siobserved ,26. TempDrosophila pseudoobscura. The geographic range of this fruit fly extends from Canada at the northern end of North America, through the USA into Guatemala . Previous work has shown that phenotypic plasticity in body size occurs in this species [Here we examine temperature-size relationships in  species . Experim species ,28. Addi species . One potWL = μ + Year + T + Pop + T. Pop + T. Year + ISO + T. ISO + ε.We measured wing sizes in a total of 439 flies from 27 isolines across four temperatures. Summary statistics for wing sizes at different rearing temperatures and in different isolines are given in Table μ is the mean, Year is a 2-level fixed factor , T is a linear effect of temperature in °C , Pop is a 3-level fixed factor  and Year.Pop and T.Year.Pop are interaction terms. Our mixed model analysis showed that, after controlling for a small but significant Year effect and a marginally non-significant T.Year interaction effect, there was evidence for plasticity and among-population genetic variance in wing length (Table T.pop interaction term). Likelihood ratio tests provided evidence of significant within-population (among isoline) genetic variance  and GxE . Under the full model (i.e. including GxEs) the (co) variance parameters (SE) were estimated as: VR = 0.153 (0.011), VISO_int = 0.064 (0.022), VISO_slp = 0.026 (0.001) and COVISO_int.slp = −0.007 (0.003), the latter scaling to an intercept-slope correlation rISO_int.slp of −0.571 (0.198). Note that since T was mean centred and the raw data scaled to unit variance, VISO_int can be directly interpreted as the proportion of observed phenotypic variance in wing length attributable to genetic differences among isolines at an average temperature  in wing sizes. However, an entirely unexpected feature to emerge from our analysis was evidence consistent with a boratory ,33. Oppoboratory . Given td-stress . In conter sites .D. subobscura also has chromosome inversions and yet shows the more common pattern of larger body sizes at lower temperatures due to selection for efficient wing loading at different body sizes [D. pseudoobscura, which may prevent convergence between environmental and genetic determination of body sizes.One possibility is that the unique system of third chromosome inversions in this species has influenced the genetic diversity available within populations that enables them to respond effectively to environmental conditions . In our dy sizes ,18. It idy sizes , but seedy sizes ,38 for cdy sizes . For examean plasticity has diverged among populations due to local selection. Interestingly, we observed from the reaction norms that the isolines in our experiment deviated most from each other at the coldest temperature  between isolines within, but not between, populations. In other words, looking among populations there is no more GxE than expected given each population itself was comprised of a unique mixture of isolines. Thus there is no evidence that e Figure , suggestD. pseudoobscura reared at 14°C, 21°C and 26°C [D. pseudoobscura are liable to show plastic responses to temperature and with only limited influence of GxEs. GxEs have been posited as a key component to the maintenance of genetic variation [D. pseudoobscura have not significantly diverged into separate ‘specialist’ populations.The relative contributions of phenotypic plasticity  and genetic variance to body size variation closely resemble work by Gupta & Lewontin, who compared the number of thoracic bristles in natural populations of and 26°C . They alariation . So, whiD. pseudoobscura and can result in significant differences in adult flies inhabiting different climates with flies reared in cooler temperatures being significantly larger. However, there is also significant genetic variation for body size at both among- and within-population levels. Among populations, genetic effects on body size run counter to the environmental influence as genotypes from the southern populations grew larger at all temperatures than those from the northern population. This could be driven by unique systems of chromosome inversions, or due to indirect selection on body size via correlated traits. Genetic variance in plasticity is present within populations, but there is no evidence of local adaptation in plasticity itself.Temperature is the strongest influence on body sizes in D. pseudoobscura from three locations in North America. We used banana baits to collect flies from Lewistown, Montana , Show Low, Arizona  and Chiricahua, Arizona . We completed collections during the late spring season when populations are highly abundant and used individual wild-caught females to establish isofemale lines. Isofemale lines are created by inbreeding the offspring of a single wild-caught female, which rapidly become homozygous at most alleles, making individuals within an isoline virtually genetically identical, and minimizing adaptation to laboratory conditions [Drosophila porridge medium containing water, oats, sugar, brewer’s yeast, and agar, plus a dilute solution of nipagin  and propionic acid to inhibit mould and bacteria growth.In 2008 and 2012 we collected wild nditions  . Hennditions . We alsonditions . We initD. pseudoobscura harbour a meiotic driving X chromosome that can impact on temperature sensitivity [We allowed several generations of inbreeding  to account for carry over effects from the field and drift within the lab. We then selected 27 isolines for the experiment , 12 from Show Low , and 6 from Chiricahua ). Some southern USA populations of sitivity . We checsitivity .D. pseudoobscura in our experiment  was scaled to standard deviation units prior to analysis and then modelled as:Wing length data for each isoline conformed well to a normal distribution. We used linear mixed effect models fitted by restricted maximum likelihood (REML) using the program ASREML to test the effects of genotype, environment (temperature), GxE on wing size . Note thμ is the mean, Year is a 2-level fixed factor , T is a linear effect of temperature in °C , and Pop is a 3-level fixed factor . Interaction terms of Year.Pop and T.Year.Pop were initially tested but being non-significant and not directly relevant to hypotheses being tested were not included in models shown here. We then included random intercept (ISO) and slope (T.ISO) terms to model within-population (among-isoline) genetic variance and GxE. Random slope and intercept terms were assumed to be normally distributed with mean of zero and a covariance structure to be estimated. This covariance structure contains the estimates of variance among isoline intercepts (VISO_int) and slopes (VISO_slp), and the among-isoline intercept-slope covariance (COVISO_int.slp). Residual errors (ε) are assumed to be normally distributed (with mean zero and variance VR) and uncorrelated across observations. We assessed the significance of fixed effects using conditional F-tests implemented in ASREML, and employed likelihood ratio tests for statistical inference on random effects. Specifically, to determine the significance of within-population GxE we compared the full model to one in which no T.ISO term was fitted. We then dropped the ISO term as well to test the significance of within-population genetic variance. We generated norms of reaction for all isolines using expected values from this model .


Many large species have declined worldwide due to habitat fragmentation and poaching. The defaunation of large frugivores and the consequent reductions of seed dispersal services may have immediate effects on plant demography. Yet, the lasting effects of frugivore defaunation on microevolutionary processes of the plants they disperse remain understudied. We tested if the loss of large seed dispersers can lead to microevolutionary changes of a tropical palm. We show that frugivore defaunation is the main driver of changes in allelic frequency among populations. Turnover of alleles accounted for 100% of dissimilarity in allelic frequencies of individuals between defaunated and non-defaunated forests; and individuals from defaunated sites are 1.5 times more similar genetically than those found in pristine sites. Given that sizeable fractions of the palm fruit crops remain undispersed in defaunated sites due to lack of large-bodied frugivores, this distinct pattern of gene pool composition of early recruits may reveal strong dispersal limitation for specific genotypes, or collapses of gene flow between fragmented areas, or both. Because most of tropical tree species rely on seed dispersal by vertebrates, our results show that defaunation has a lasting effect on microevolutionary processes, with potential consequences for persistence under scenarios of environmental change. Many species and populations of animals are facing a dramatic decline worldwide, a phenomenon named “Anthropocene defaunation”236610Given the negative effects of defaunation on plant populations, we tested the hypothesis that the functional loss of large seed dispersers may lead to microevolutionary changes of the plants that rely on their dispersal. In addition to anthropogenic impacts, the distribution of genetic variability among plant populations over large spatial scales may be also due to a combination of historic and ongoing landscape effects that ultimately influence the joint action of gene flow, selection and genetic drift1314Euterpe edulis in 19 populations in the Brazilian Atlantic forest. This palm is a dominant species distributed across the Atlantic forest of South AmericaEuterpe edulis produces round fleshy fruits ranging from 8.3 to 14.1 mm in diameterProcnias nudicollis, toucans Ramphastos spp. and guans Penelope spp. and Aburria jacutinga) and medium-sized avian frugivores (Turdus spp.)E. edulis populations and has been a driver for a rapid evolutionary reduction of seed size in defaunated palm populationsHere, we analyzed recruits (seedlings) of the palm We tested four different hypothetical scenarios to assess variation in local patterns of genetic variability, i.e., to determine which scenario best fits the observed pattern of genetic variation. We thus contrasted genetic variability patterns across replicated local palm populations in different situations according with these scenarios.E. edulis stands due to a selection against large seed size and to the collapse of the long-distance seed dispersal events, impairing effective gene flow  defined by the previous hypotheses. We also tested the effects of sampling design as a potential influence for the observed genetic variability distribution, i.e., a situation where differences would emerge by chance effects related to the specific populations sampled.We sampled the Euterpe edulis (a priori groupings defined by the hypotheses (represented by kappa index), followed by forest fragmentation and sampling design  among defaunated and non defaunated sites or among sites from distinct biogeographic regions is due to alleles replacement  or richness differences driven to passive allelic loss βdiversity = 0.64, 95% CI = 0.63 to 0.64; Mean βsim = 0.64, 95% CI = 0.63 to 0.64; Mean βsne = 0.00, 95% CI = 0.00 to 0.00, βdiversity = 0.61, 95% CI = 0.60 to 0.62; Mean βsim = 0.61, 95% CI = 0.60 to 0.62; Mean βsne = 0.00, 95% CI = 0.00 to 0.00, β-allelic diversity and its components are lower among palm recruits in defaunated sites  using the forward selection method and found that the model including the effects of defaunation and biogeographical regions, but not spatial effects, was the best to explain variation in allele frequencies among sites .Finally, we estimated the contribution of spatial effects on genetic differentiation of Our results indicate that the defaunation of large seed dispersers has a distinct signal on large-scale genetic variability, potentially arising from microevolutionary changes in the palm populations. This supports and extends previous findings8Euterpe edulis populations. This could be a synergistic effect from a higher frequency of mating among closely-related individuals, a reduction of effective population size, limited long distance dispersal among populations1115E. edulis populations in defaunated areasE. edulis populations were consistently associated with scenarios of strong seed dispersal limitation12We found that defaunation leads to genetic changes in E. edulis populations, with allele replacement being the main driver of allelic richness differences between rainforest and semideciduous forests. These two biogeographical regions show marked differences in annual rainfall and predominant soil types3139E. edulis to these different domains of the Atlantic forestE. edulis populations, remarkably differentiating the two scenarios despite the variation due to forest physiognomy. Remarkably, the magnitude of this effect appears as large as the magnitude of differentiation when comparing populations from the two biogeographic regions. The large-scale fragmentation and defaunation in Atlantic forest is a recent anthropogenic impact. Despite that, our results reveal that the strong selective pressure imposed by defaunation on the genetic pool of E. edulis can be comparable in terms of its effects size with signals left by biogeographical process. If we assume that smaller values of β-allelic diversity represent a higher homogenization of genetic pools among populations, ongoing defaunation is driving a stronger genetic homogenization than we would expect just from historical, phylogeographic dynamics.Our results also showed that biogeographical origin has a signature on the genetic structure of E. edulisOur results provide a rigorous test, within an adequately replicated setup of distinct landscape types, of the hypothesis that contemporary defaunation is affecting the diversity of genetic pools of animal-dispersed tropical plants. This supports previous findings10193341The Brazilian Atlantic forest is an ideal system to test the effects of antrophocene defaunation on ecological and evolutionary processes. The Atlantic forest originally covered from northeast to south of the Brazilian territory and parts of Argentina and ParaguayEuterpe edulis, Arecaceae), a threatened Atlantic forest species, was once one of the dominant palms in this ecosystemE. edulis is a self-compatible monoecious species, but with predominant outcrossed reproductionTrigona spinipes). Their fruits are eaten by more than 58 birds and 20 mammalian species but are dispersed mostly by a reduced subset of large frugivorous birds and thrushes , following PCR protocol described by Gaiotto a priori groupings of the local palm populations according to defaunation status, biogegraphical region, forest fragmentation status and sampling) maximized the differences among groups (clusters). The sampled palm populations were defined according to a replicated design across the different strata defined by each hypothesis . Then, we conducted a DAPC and calculated the proportion of correct assignment of individuals into each hypothesis . To avoid over-fitting, we retained 50 principal components, corresponding to 82% of the total genetic information. We carried out the DAPC using the R package adegenetWe used Discriminant Analysis of Principal Component DAPC to find esis see . DAPC reHe - expected heterozygosity under Hardy-Weinberg equilibrium, following NeiARf, obtained from analysis of variance of allelic frequencyNe), using the software NeEstimatorWe assessed the influence of defaunation and biogegraphical regions on genetic variability and contemporary effective population size across the study sites. We estimated genetic diversity  and estimate the β-allelic diversity and its components among these groups . This procedure were carried out 499 times (bootstrapping) and we obtained the mean, standard deviation and 95% confidence interval for each β diversity component. We constructed a null model with 1000 permutation and a 95% confidence interval to verify whether the results of the β diversity analysis for each hypothesis differed from random. With this analysis we could estimate the importance of each β-allelic diversity component for the overall allelic richness dissimilarity between defaunated and non-defaunated sites or between rain forest and semideciduous forest. Second, we randomly sampled all palm recruits from two sites of each group (i.e. two sites from defaunated sites) and estimate the β-allelic diversity and its components among the sites from each group. We carried out this procedure 499 times (bootstrapping) and obtained the mean, standard deviation and 95% confidence interval for each β diversity component. The same was done for all groups of all hypotheses . With this analysis we could compare and contrast the β-allelic diversity results between defaunated and non-defaunated sites or between rain forest and semideciduous forest. For example, we expected that β diversity might be lower among individuals in defaunated sites compared to non-defaunated sites.The difference in allelic richness among sites may be mainly due to allele replacement (turnover) or richness differences  is exceeded. We used the adegenet R packagepackforveganThe spatial distribution of individuals and populations are also important factors that affect genetic differentiation patterns. For example, some populations can follow an isolation by distance model while others may follow an island model. To deal with spatial effects on genetic differentiation, we conducted a Redundancy Analysis (RDA). RDA is a multivariate method that assesses the influence of a matrix of independent variables (e.g. geographic coordinates) on a matrix of dependent variables How to cite this article: Carvalho, C. S. et al. Defaunation leads to microevolutionary changes in a tropical palm. Sci. Rep.6, 31957; doi: 10.1038/srep31957 (2016).


Bactrocera dorsalis s.s. (Hendel) and B. papayae Drew & Hancock, are invasive pests belonging to the B. dorsalis complex. Their species status, based on morphology, is sometimes arguable. Consequently, the existence of cryptic species and/or population isolation may decrease the effectiveness of the sterile insect technique (SIT) due to an unknown degree of sexual isolation between released sterile flies and wild counterparts. To evaluate the genetic relationship and current demography in wild populations for guiding the application of area-wide integrated pest management using SIT, seven microsatellite-derived markers from B. dorsalis s.s. and another five from B. papayae were used for surveying intra- and inter-specific variation, population structure, and recent migration among sympatric and allopatric populations of the two morphological forms across Southern Thailand and West Malaysia.P > 0.05). Nonetheless, two sets of microsatellite markers showed significantly different levels of polymorphisms. Genetic differentiation between intra- and inter-specific differences was significant, but low. Seventeen populations revealed three hypothetical genetic clusters (K = 3) regardless of forms and geographical areas. The genetic structure of sympatric populations slightly changed during the different years of collection. Recent gene flow (m ≥ 0.10) was frequently detected whether samples were sympatric or allopatric. Ninety-five of 379 individuals distributed across the given area were designated as recent migrants or of admixed ancestry. As a consequence of substantial migration, no significant correlation between genetic and geographic distances was detected .Basic genetic variations were not significantly different among forms, populations, and geographical areas (B. dorsalis s.s. and B. papayae forms in Southern Thailand and West Malaysia. Both forms can be treated as a single target pest for the SIT program in an area-wide sense. Additionally, the result of species identification based on molecular data and morphological character are not congruent. The use of independent, multiple approaches in the characterization of the target population may ensure the effectiveness and feasibility of SIT-based control in the target area.According to the 12 microsatellite variations, weak population structure and recent gene flow suggest that there is no status for cryptic species between  The sterile insect technique (SIT) is a powerful biological control method. It relies on mating between sterile insects  and their wild counterparts in order to reduce reproductive potential. SIT is effective because it is a species-specific approach and one of the most environmentally friendly solutions to insect pest management. However, there are several steps: mass-rearing, sex-separation for male-only release, sterilization, marking, and mass-releasing . SIT is Problems such as potential sexual isolation due to the existence of cryptic species and/or population isolation must be investigated before SIT application . CrypticBactrocera dorsalis species complex, a large group of nominal tephritid fruit flies, is of interest to scientists in several fields of study. Regarding morphological characters and host preferences, the B. dorsalis complex consists of almost 100 similar species . When partitioning either morphological form B. dorsalis s.s.  or morphological form B. papayae  samples, no significant IBD is confirmed. These results are similar to those from the migration analysis, indicating no limitation of gene flow across populations of morphological forms.Isolation by distance (IBD): All 17 B. dorsalis s.s. and B. papayae - was carried out in areas spanning from the top of Southern Thailand to the tip of the Malay Peninsula using high-resolution microsatellite DNA markers derived from each form. It can be inferred that these two morphological forms comprise a panmictic unit. Genetic variation, population structure, and recent population demography suggest a high feasibility for area-wide integrated pest management using sterile insect technique (AW-IPM-SIT) control programs. A combination of population genetic tools and morphological characterization may be necessary to better understand target pest populations.A fine-scale population genetics study of two morphological forms - B. papayae-derived loci were less variable than B. dorsalis s.s. for all parameters. The explanation involves several factors that influenced microsatellite mutation rate, such as repeat number, repeat type, flanking sequence (GC content), chromosome location, and base substitutions in the microsatellite arrays [B. dorsalis s.s.-derived loci  are relatively lower than B. papayae-derived loci . It is possible that B. dorsalis s.s.-derived loci tend to provide a higher variability of microsatellite. Likewise, motifs of B. dorsalis s.s.-derived loci showed relatively higher base substitution than the other. This also supports the high variability in B. dorsalis s.s.-derived loci. In addition, no significant genetic differences were observed among all populations, the two forms, the two countries, and also the interaction between marker derivation and species factors. Consequently, it can be deduced that the genetic polymorphisms of all tested loci may be shaped by the issue of population histories, not by ascertainment bias. Therefore, using two different species-derived sets of microsatellite loci, if available, could help us to clarify the source of genetic variability as well as to avoid biasing the data. This may help us to not undermine the determination of sympatric species that have no interbreeding between groups [The factors affecting microsatellite DNA variability include locus- and genome-specific mutation rates . Microsae arrays . In our e arrays . In thisB. dorsalis s.s. and B. papayae forms are cryptic species in the investigated areas. Independent studies using different samples and sets of microsatellite markers ) present a comparable genetic variation between two morphological forms. These facts imply that FST of the given species cannot be resolved. A plausible explanation is that both forms comprise a panmictic unit or recently diverged, and/or they are connected through a high level of hybridization [cox1, nad4-3′, CAD, period, ITS1, ITS2 [Studies using high-resolution microsatellite markers had been successfully used to delineate the limits of other species complexes e.g., ,19,59); ,59; 19,5lis s.s.  and a codization . If bothdization , cox1, nS1, ITS2 , EF-1α, S1, ITS2 , and othS1, ITS2 , and matS1, ITS2 , along wS1, ITS2 ,14, provFST between two morphological forms and among populations was significant, but quite low. Approximately 2% to 18% of genetic diversity is the result of genetic differentiation among B. dorsalis s.s. and B. papayae populations. A high level of gene flow between populations or recent species divergence is the most likely explanation. Considering the genetic diversity of the single species B. dorsalis s.s., using at least the same seven microsatellite loci as the current study, we found that approximately one to six percent (1-6%) and up to 30% of genetic diversity are the consequence of genetic differentiation among the populations at the level of micro-geography (populations collected from Thailand) and macro-geography , respectively [B. dorsalis s.s., the FST of former falls between micro- and macro-geographical values. However, this interpretation must be viewed with caution because the FST value is independent of the particular characteristics of individual loci or alleles and is influenced by the geographical differences of sampling locations [Other evidence of ectively . Compariocations . Finallyocations -16,60 orB. dorsalis s.s. and B. papayae[B. dorsalis s.s. and B. papayae in our studied areas. The data shows a high level of genetic variation, a low level of genetic differentiation, and recent asymmetric migration to almost all populations. Similarly, Ratchaburi was previously proposed as a genetic source of B. dorsalis s.s. in Thailand when samples from Southern Thailand and West Malaysia were not taken [vice versa. The genetic ancestry of a few fruit fly samples (1.32%) cannot be traced back to any investigated populations, although nearby populations were no more than 200 km away. Adequate representative populations could be sampled in order to expand on this type of population dynamics detail, which may help the implementation of SIT programs.We inferred population dynamics in allopatric and sympatric areas with regard to recent migration and population expansion/contraction. The narrowest part of the Thai-Malay Peninsula - Kra Isthmus - is located between Ranong and Chumphon Provinces, Thailand. Despite the fact that this area was proposed to be a biogeographic transition zone for . papayae,11, our . papayae). In addot taken . Howeverot taken  may be tB. dorsalis s.s., seem to maintain their stable populations, which is inferred from the comparable population size and genetic variability. On the other hand, populations of the other morphological form appear to be expanding. The first collection, PK1b, has a small population size (N < 10); however, two years later, PK2b was observed to have increased in population size as well as in genetic variability. According to inferences made from allopatric and sympatric populations, the fruit fly population dynamics may not be at equilibrium in Southern Thailand and West Malaysia. This demonstrates that population genetic study using multiple-time-point sample collection is encouraged to infer delicate population dynamics processes.Population dynamics is also illustrated using the data from two different collections at two different time points, but in the same area of Prachub Kirikan Province. Populations PK1a and PK2a, the same morphological form B. dorsalis s.s. and B. papayae forms, it is consistent with other independent identification approaches using non-morphological characteristics and morphometric analyzes (as mentioned before). This implies that morphological variation is not a standalone indicator of species boundaries. Extreme environments can impose stabilizing selection on morphological characters which have nothing to do with the species differentiation process. In contrast, selection of non-morphological traits such as behavior, molecular markers, or reproductive character, can accompany speciation [Fruit fly surveillance using a combination of tools  is a key concept for planning and implementation of an AW-IPM-SIT program. Utilization of the traditional approach based on morphology and type of fruit fly hosts for characterization of pest species may result in misidentification. Sample collection from significantly isolated geographical areas without intermediate sites could provide samples with more discrete morphological characters than geographically closer sites . Althougeciation . IncorpoCeraitis capitata (Vienna 8) [Bactrocera dorsalis  [Anastrepha ludens (Tapachula-7) [B. dorsalis s.s. and B. papayae is possible, in an area-wide sense, in Southern Thailand and West Malaysia. Several reliable lines of evidence  against B. carambolae in Suriname had been reported [The release of sterile male only is a very crucial step to improve SIT, in terms of the cost and biological efficiency in the field . Developienna 8) , BactrocSalaya1) , and Anachula-7) . Therefoge tests  and estireported .However, we detect ongoing delicate population dynamics processes such as population reduction/expansion and migration within studied areas. The phenomena could produce subpopulations by various scenarios and subsequently impact the effectiveness of AW-IPM-SIT. For example, new immigrants may be introduced into a new microhabitat which may form a pocket population. When the SIT activity is less intense or no longer practiced, they may be founders for an incursion scenario . This reB. dorsalis s.s. and B. papayae - in Southern Thailand and West Malaysia based on the variations of microsatellite DNA markers derived from both species. Hence, both forms may be treated as a single target pest for an SIT control program. However, resolution of genetic isolation and morphology are not congruent in species identification. The characterization of a pest population using multiple approaches may ensure the effectiveness and feasibility of the SIT-based method.In summary, there is no status for cryptic species between two morphological forms - The authors declare that they have no competing interests.NA and ST conceived and designed the research project; NA and SI performed the genotyping; NA conducted the population genetic analyzes; NA wrote the manuscript. All authors reviewed the results from the data analysis and approved the final manuscript.Description of each primer used in the current study.Click here for fileResults of migration analyses using STRUCTUREClick here for file


Since the neoplastic phenotype of a cell is largely driven by aberrant gene expression patterns, increasing attention has been focused on transcription factors that regulate critical mediators of tumorigenesis such as signal transducer and activator of transcription 3 (STAT3). As proteins that interact with STAT3 may be key in addressing how STAT3 contributes to cancer pathogenesis, we took a proteomics approach to identify novel STAT3-interacting proteins. We performed mass spectrometry-based profiling of STAT3-containing complexes from breast cancer cells that have constitutively active STAT3 and are dependent on STAT3 function for survival. We identified granulin (GRN) as a novel STAT3-interacting protein that was necessary for both constitutive and maximal leukemia inhibitory factor (LIF)induced STAT3 transcriptional activity. GRN enhanced STAT3 DNA binding and also increased the time-integrated amount of LIF-induced STAT3 activation in breast cancer cells. Furthermore, silencing GRN neutralized STAT3-mediated tumorigenic phenotypes including viability, clonogenesis, and migratory capacity. In primary breast cancer samples, GRN mRNA levels were positively correlated with STAT3 gene expression signatures and with reduced patient survival. These studies identify GRN as a functionally important STAT3-interacting protein that may serve as an important prognostic biomarker and potential therapeutic target in breast cancer. Breast cancer is the most frequently diagnosed cancer and second leading cause of cancer death in American women, accounting for about 230,000 new cases and 40,000 deaths per year . Triple-Since cancer is associated with aberrant gene expression patterns, increasing attention has been focused on transcription factors like signal transducer and activator of transcription 3 (STAT3) which lies at the convergence points of many oncogenic signaling pathways. STAT3 is one of a family of transcription factors that mainly reside in the cytoplasm until activated by phosphorylation on a conserved tyrosine residue 705 (PY-STAT3) by growthfactor receptor tyrosine kinases, cytokine-receptorassociated Janus kinases (JAKs), or non-receptor tyrosine kinases . AlthougSTAT3 constitutes an important therapeutic target because cancer cells depend on the inappropriate activity of this oncogenic transcription factor while normal cells can often tolerate disruption of these proteins with little toxicity due to redundancies in normal signaling pathways . HoweverSeveral proteins have been reported to interact with STAT3 and modify its function, including STAT3- interacting protein 1 (StIP1) which preferentially associates with unphosphorylated STAT3 to promote STAT3 interaction with upstream kinases , transcrTo identify novel STAT3-interacting proteins that modulate STAT3 activity, we utilized the TNBC cell lines MDA-MB-468 and SUM159PT, which display constitutively active STAT3 signaling including tyrosine phosphorylation of STAT3 at the 705 residue (PY-STAT3), and are dependent on STAT3 function for growth and survival  with phosphatase and complete protease inhibitors . Five percent of the total protein lysate (approximately 12.5 μg of 250 μg) was saved as an immunoprecipitation input loading control, then the remaining lysate was divided into equal volumes for overnight incubation at 4 °C with 1 μg of an antibody to STAT3 , GRN , or isotype control IgG . The lysate-antibody mixture was incubated for 4 hours at 4 °C with protein A/G PLUS agarose beads (Santa Cruz) blocked with 1% bovine serum albumin (Sigma-Aldrich) for 1 hour at 4 °C. The antibody-bead mixture was washed 3 times in lysis buffer, and eluted in sample buffer with10% β-mercaptoethanol.Immunoprecipitations were performed as previously described . Brieflyg for 5 minutes, then re-suspended in 200 μL of 10% acetic acid and 0.1% trifluoroacetic acid (TFA) (pH ~2) and incubated at 65°C for 10 minutes. Elution buffer was collected and dried using a SpeedVac. Eluted complexes were reconstituted in 5 mM TCEP  and 25 mM ammonium bicarbonate (Sigma-Aldrich) pH 8.0, then incubated at 37 °C for 30 minutes. Iodoacetoamide (Sigma-Aldrich) was added to 25 mM and the samples incubated at 37 °C for 1 hour. The same amount of TCEP was added and the samples incubated at room temperature for 1 hour. Digestion was carried out by sequencing grade trypsin  at 37 °C for 24 hours as described  with phosphatase and complete protease inhibitors (Roche). Blots were probed with antibodies to STAT3 (sc-482), hsp90 (sc-13119) from Santa Cruz; phospho-STAT3 (Tyr705) (9131), PARP (9542S), and His-Tag (2365) from Cell Signaling Technology, Inc. ; GRN  from Invitrogen; FLAG M2 (F1804) and tubulin (T5168) from Sigma-Aldrich; GRN  from Thermo Fisher Scientific; turboGFP (TA150041) from Origene ; and phospho-STAT3 (Ser727) previously generated in rabbits [Cells (5 × 10 rabbits . Immunob4) were transfected with 1 μg of a STAT3-dependent reporter  or B-luc [Renilla luciferase transfection control phRL TK-luc (Promega) using Lipofectamine 2000 (Invitrogen). Twenty-four hours after transfection, cells were stimulated with cytokine or lysed directly and quantitated using dual-luciferase reagents (Promega) on a Luminoskan Ascent luminometer . STAT3-dependent luciferase production was normalized to Renilla luciferase.Cells (5 × 10or B-luc ) and 0.1STAT3 luciferase reporter cells stimulated with IL-6 and PGRN for 6 h were analyzed by Bright-Glo luciferase assay (Promega) on a Luminoskan Ascent luminometer (ThermoLab Systems).5) using RNeasy Mini kits  and reverse transcribed with TaqMan kits . Quantitative PCR was performed in triplicate using SYBR select master mix (Applied Biosystems) on a 7300 or 7500 real-time PCR system (Applied Biosystems). Specificity of amplification was confirmed by melt curve analysis. Cycle threshold (C ) values for target isoforms were normalized to the indicated endogenous reference gene. Primer sequences  were fixed in 1% formaldehyde for 10 minutes, sonicated in 15 second pulses using a Fisher Scientific Sonic Dismembranator Model 500 PDQ on setting 15, and lysates immunoprecipitated overnight at 4°C with an antibody for STAT3 (sc-482) from Santa Cruz. Quantitative PCR was performed using the indicated primers  were transfected with siRNA targeting GRN, STAT3, or a non-targeting control in triplicate. Seventy-two hours later, viable cell number was quantitated by measuring intracellular ATP using Cell Titer-Glo (Promega) on a Luminoskan Ascent luminometer.Cells (1 × 104) were transfected with siRNA targeting GRN, STAT3, or a non-targeting control for 24 hours, then trypsinized and seeded at 1000 cells/well. After 7 days, colonies were washed in PBS, then fixed and stained in 0.5% crystal violet and 6% glutaraldehyde. Stains were solubilized in 10% acetic acid and measured at OD 590nm on a Spectramax M3 .SUM159PT cells (5 × 106) were transfected with siRNA targeting GRN, STAT3, or a non-targeting control for 48 hours, then the confluent monolayer was disrupted with a linear scratch made with a sterile P200 pipette tip. Cells were washed in PBS, incubated in serum-free media to prevent proliferation, and imaged at 24 hour intervals.SUM159PT cells  and a rabbit anti-GRN antibody  overnight at 4C, followed by Alexa Fluor 568-conjugated goat anti-mouse secondary antibody  and Alexa Fluor 488-conjugated donkey anti-rabbit secondary antibody  for 1 hour. Cover slips were mounted onto glass slides using DAPI-containing Vectashield mounting medium . Images were acquired using an Olympus BX51 fluorescence microscope with attached DP71 camera and DP Manager Software. Three microscopic fields/slide were photographed (40x).Cells grown on glass cover slips were fixed with 4% paraformaldehyde/PBS for 20 min, permeabilized with 0.5% Triton X-100/HLevel 3 mRNA expression Agilent 244k array) and clinical parameters were downloaded from The Cancer Genome Atlas (TCGA) breast carcinoma dataset on October 18, 2013. A second set of previously published gene expression data for 129 breast tumors profiled using Affymetrix U133p2.0 microarrays were downloaded from the Gene Expression Omnibus (GSE5460). PhosphoSTAT3 immunohistochemistry for the corresponding tissue microarrays were previously reported [k array ap < 0.05; **, p < 0.01; and ***, p < 0.001). Data are presented as mean ± SD for the indicated number of independent experiments . Values of p < 0.05 were considered significant (*, nts Fig. , otherwi


Lens culinaris Medik.) collection consisting of 352 accessions originating from 54 diverse countries to estimate genetic diversity and genetic structure using 1194 polymorphic single nucleotide polymorphism (SNP) markers which span the lentil genome. Using principal coordinate analysis, population structure analysis and UPGMA cluster analysis, the accessions were categorized into three major groups that prominently reflected geographical origin . The three clusters complemented the origins, pedigrees, and breeding histories of the germplasm. The three groups were (a) South Asia , (b) Mediterranean, and (c) northern temperate. Based on the results from this study, it is also clear that breeding programs still have considerable genetic diversity to mine within the cultivated lentil, as surveyed South Asian and Canadian germplasm revealed narrow genetic diversity.Assessment of genetic diversity and population structure of germplasm collections plays a critical role in supporting conservation and crop genetic enhancement strategies. We used a cultivated lentil ( Lens culinaris Medikus ssp. culinaris) is the third most important cool-season grain legume in the world after chickpea (Cicer arietinum L.) and pea (Pisum sativum L.)  from nearly 4.3 million ha (Mha) in 2013. Canada was the largest producer, contributing 38% of the world's production, followed by India, Turkey, and Australia  FAO, . Legumeslia FAO, . Lentil lia FAO, . Lentil lia FAO, . It is alia FAO, . Globalllia FAO, . These zLens accessions held in various genebanks worldwide .Success in crop breeding is a function of heritability, genetic diversity, and selection. Natural agro-biodiversity stored in genebanks can be used to expand the diversity in crops. These collections are a vital source for discovering useful genes/alleles, which serve as a cornerstone for any pre-breeding program. There are currently 58,405 ide FAO, . InternaLens genetic resources for agro-morphological and phenological characteristics  collection in Saskatoon, Canada, from ICARDA, and the USDA-ARS. The accession numbers and origins of the selected accessions are given in Supplementary Table Canadian lines, along with ICARDA germplasm, were grown in Saskatoon, Canada and leaf tissues from at least five different plants were collected for genomic DNA extraction using a modified CTAB extraction method  described by Sharpe et al.  was usedThe SNP marker data were analyzed using PowerMarker v. 3.25  in the panel. Five independent runs were conducted for each K ranging from 1 to 10 with both a burn-in time and Markov Chain Monte Carlo (MCMC) replication number of 500,000 using an admixture model. Selection of the best K-value was based on the procedure presented in Evanno et al.  and ΔK-value  using GenAlEx v. 6.5  and Supplementary Table The sequences of all the markers used in this study are described in Sharpe et al. . All datOf 1400 SNP markers, 5.46% were monomorphic among the accessions, 2.14% generated ambiguous products, and 7.21% were rejected on the grounds of excess missing data points (>30% missing data per marker). This filtering resulted in 1194 high quality polymorphic SNP markers for use in the clustering analyses. Around 40% of these SNP markers were previously mapped and shown to be evenly distributed throughout the genome  from 1 to 10 on the entire set of accessions. The estimated log probability of the data [LnP(K)] for each K plateaued at K = 3. The maximum ΔK-value was also reached at K = 3 . Generally, South Asian and Middle Eastern  germplasm grouped together, however, some of the Iranian and Turkish germplasm skewed to those from Mediterranean and northern climates. Within the second group, germplasm from Chile exhibited the closest relationship to germplasm from Morocco (0.0828). Canadian cultivars and breeding lines, representative of group 3, were closely related to much of the “W6” numbered lines which used in USDA lentil breeding program as parents in recombinant inbred lines development  and Russian (0.0961) germplasm  and relatively high marker density, this study identified three major clusters of germplasm reflecting the geographical origins, pedigrees, and breeding history of accessions. We categorized these clusters as (a) subtropical, South Asian, (b) Mediterranean, including Southern Europe and North Africa, and (c) northern temperate climates. These three groups reflect the main climatic regions in which lentil is widely grown as described by Tullu et al. .Lentil accessions from South Asia  have a narrow genetic base and are genetically more isolated relative to other origins included in our study. This is likely a result of their specific phenological adaptation to the sub-tropical savannah environment and a potential genetic bottleneck during a time when lentil was introduced to South Asia around 2000 BC  and Australia (Mediterranean climate) FAO, , regionsLentil domestication already has led to ~40% loss in genetic diversity  sequences from nine e et al.  can provGlobal cultivated lentil germplasm selected for this study clustered primarily based on eco-geographical origin into three basic groups: subtropical savannah, Mediterranean, and northern temperate. The narrow genetic base of some groups of germplasm  raises concern over the loss/penalty in yield due to biotic and abiotic stresses, particularly with the threat to global food security from climate change. This highlights the importance of harnessing the potential of lentil wild species in breeding programs by introgression of favorable genes from other regions. Based on the results from this study, it is also clear that breeding programs still have a lot of genetic diversity to mine within the cultivated species.KB and AV designed the research; HK, and CTC analyzed data; MD and MF contributed to data generation; CJC and RM contributed germplasm and reagents; HK, CTC, KB, and AV contributed to writing of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.