Marie-Christine Flamand

Genetic structure and assignment tests demonstrate illegal translocation of red deer (Cervus elaphus) into a continuous population

Molecular forensic methods are being increasingly used to help enforce wildlife conservation laws. Using multilocus genotyping, illegal translocation of an animal can be demonstrated by excluding all potential source populations as an individuals population of origin. Here, we illustrate how this approach can be applied to a large continuous population by defining the population genetic structure and excluding suspect animals from each identified cluster. We aimed to test the hypothesis that recreational hunters had illegally introduced a group of red deer into a hunting area in Luxembourg. Reference samples were collected over a large area in order to test the possibility that the suspect individuals might be recent immigrants. Due to isolation‐by‐distance relationships in the data set, inferring the number of genetic clusters using Bayesian methods was not straightforward. Biologically meaningful clusters were only obtained by simultaneously analysing spatial and genetic information using the program baps 4.1. We inferred the presence of three genetic clusters in the study region. Using partial Mantel tests, we detected barriers to gene flow other than distance, probably created by a combination of urban areas, motorways and a river valley used for viticulture. The four focal animals could be excluded with a high certainty from the three genetic subpopulations and it was therefore likely that they had been released illegally.

Comparative landscape genetic analyses show a Belgian motorway to be a gene flow barrier for red deer (Cervus elaphus), but not wild boars (Sus scrofa)

While motorways are often assumed to influence the movement behaviour of large mammals, there are surprisingly few studies that show an influence of these linear structures on the genetic make‐up of wild ungulate populations. Here, we analyse the spatial genetic structure of red deer (Cervus elaphus) and wild boars (Sus scrofa) along a stretch of motorway in the Walloon part of Belgium. Altogether, 876 red deer were genotyped at 13 microsatellite loci, and 325 wild boars at 14 loci. In the case of the red deer, different genetic clustering tools identified two genetic subpopulations whose borders matched the motorway well. Conversely, no genetic structure was identified in the case of the wild boar. Analysis of isolation‐by‐distance patterns of pairs of individuals on the same side and on different sides of the motorway also suggested that the road was a barrier to red deer, but not to wild boar movement. While telemetry studies seem to confirm that red deer are more affected by motorways than wild boar, the red deer sample size was also much larger than that of the wild boars. We therefore repeated the analysis of genetic structure in the red deer with randomly sub‐sampled data sets of decreasing size. The power to detect the genetic structure using clustering methods decreased with decreasing sample size.

Migration barriers protect indigenous brown trout (Salmo trutta) populations from introgression with stocked hatchery fish

Brown trout populations in the Belgian rivers Scheldt and Meuse have been intensively stocked in the past decades, often with material of uncertain origin. Moreover, the species’ habitat has become increasingly fragmented, preventing gene flow between neighboring populations. We assessed how this impacted genetic diversity and population structure by analyzing 12 wild populations (total n=309) and seven hatchery stocks (n=200) at the mitochondrial control region with SSCP and at 27 RAPD loci. Historical records indicate that brown trout from distant locations have been used to supplement hatchery stocks; nevertheless we detected non-Atlantic mitochondrial genomes in only one population of the Scheldt basin and in one hatchery. In general, the hatchery samples displayed a higher genetic diversity and differentiated less among each other (global FST(mtDNA)=0.311/FST(RAPD)=0.029) compared to the wild populations (global FST(mtDNA)=0.477/FST(RAPD)=0.204). This is due to frequent exchanges between hatcheries and regular supplementation from several indigenous populations. Gene pools present in most downstream sections from tributaries of the Meuse were similar to each other and to the hatchery samples, despite the presence of migration barriers. Assignment analyses indicated that the contribution of hatchery material to the upstream parts was limited or even completely absent in populations separated by a physical barrier. Intensive stocking and exchange between hatcheries has homogenized the downstream sections of the Meuse River, whereas the migration barriers preserved the indigenous upstream populations. As such, uncontrolled removal of barriers might result in an irreversible loss of the remnant indigenous gene pools.

Multicollinearity in spatial genetics: separating the wheat from the chaff using commonality analyses.

Direct gradient analyses in spatial genetics provide unique opportunities to describe the inherent complexity of genetic variation in wildlife species and are the object of many methodological developments. However, multicollinearity among explanatory variables is a systemic issue in multivariate regression analyses and is likely to cause serious difficulties in properly interpreting results of direct gradient analyses, with the risk of erroneous conclusions, misdirected research and inefficient or counterproductive conservation measures. Using simulated data sets along with linear and logistic regressions on distance matrices, we illustrate how commonality analysis (CA), a detailed variance‐partitioning procedure that was recently introduced in the field of ecology, can be used to deal with nonindependence among spatial predictors. By decomposing model fit indices into unique and common (or shared) variance components, CA allows identifying the location and magnitude of multicollinearity, revealing spurious correlations and thus thoroughly improving the interpretation of multivariate regressions. Despite a few inherent limitations, especially in the case of resistance model optimization, this review highlights the great potential of CA to account for complex multicollinearity patterns in spatial genetics and identifies future applications and lines of research. We strongly urge spatial geneticists to systematically investigate commonalities when performing direct gradient analyses.

A Mitochondrial Plasmid Specifically Associated With Male-sterility and its Relation With Other Mitochondrial Plasmids in Vicia-faba L

SummarySmall supercoiled DNA molecules of Vicia faba were cloned and characterized by restriction mapping and molecular hybridization. The 1700 by S plasmid has been found to be specifically associated with the male sterile cytoplasms and exhibited large homologies with the other 1700 by F plasmid as shown by molecular cross hybridization. In one line bearing the male sterile 350 cytoplasm, a deleted mitochondrial plasmid of 1540 bp was observed and this plasmid derives from the 1700 bp S plasmid. Our results indicate therefore that three of the four plasmids analysed are closely related between each other.

Population structure and genetic diversity of red deer (Cervus elaphus) in forest fragments in north-western France

Red deer have been subjected to anthropogenic interference for many centuries. Most populations are managed according to hunting schedules, some have been kept long-term in enclosures and other populations have been restocked with foreign deer. The red deer in the Brittany region of north-western France only occupy the largest forests in the region, reaching quite high densities in restricted areas. Here, we aimed to assess the extent of the genetic variability of the populations in four forest fragments and investigate their population genetic structure. We show that, despite relatively large expected heterozygosity values, these geographically isolated populations are genetically impoverished relative to individuals from large continuous forests in other parts of Western Europe. We provide evidence for population genetic structure with large genetic differentiation between geographically close populations, suggesting the absence of effective exchange between the forests. Using samples from the most likely source population, we show that at least two populations were non-indigenous. In order to limit further loss of genetic diversity, it should be a management objective to reduce isolation of the different forests, rather than further increase it by fences and hunting practices that could limit free movement of red deer.

Nucleotide-sequence and Transcriptional Analysis of a Mitochondrial Plasmid From a Cytoplasmic Male-sterile Line of Sunflower

SummaryA mitochondrial plasmid of 1,939 bp (P2) from a cytoplasmic male-sterile line of sunflower has been cloned and sequenced. It presents 437 bp of near-perfect homology to the 1.4-kb mitochondrial plasmid P1 from sunflower. Sequences homologous to P2 were found in nuclear DNA. P2 was transcribed into a major 980-nucleotide (nt) RNA molecule and two minor transcripts of 570 and 520 nt. They were all transcribed from the same strand and within the region nonhomologous to P1. A single 5′ boundary and three 3′ termini were determined for P2 transcripts. The 5′ end is similar to a consensus sequence for plant mitochondrial genes. No evidence of translation products can be provided.

Genetic Structure and Effective Population Sizes in European Red Deer (Cervus elaphus) at a Continental Scale: Insights from Microsatellite DNA

We analyzed more than 600 red deer (Cervus elaphus) from large parts of its European distribution range at 13 microsatellite loci, presenting the first continent-wide study of this species using nuclear markers. Populations were clearly differentiated (overall F ST = 0.166, Josts D est = 0.385), and the BAPS clustering algorithm yielded mainly geographically limited and adjacent genetic units. When forced into only 3 genetic clusters our data set produced a very similar geographic pattern as previously found in mtDNA phylogeographic studies: a western group from Iberia to central and parts of Eastern Europe, an eastern group from the Balkans to Eastern Europe, and a third group including the threatened relict populations from Sardinia and Mesola in Italy. This result was also confirmed by a multivariate approach to analyzing our data set, a discriminant analysis of principal components. Calculations of genetic diversity and effective population sizes (linkage disequilibrium approach) yielded the lowest results for Italian (Sardinia, Mesola; N e between 2 and 8) and Scandinavian red deer, in line with known bottlenecks in these populations. Our study is the first to present comparative nuclear genetic data in red deer across Europe and may serve as a baseline for future analyses of genetic diversity and structuring in this widespread ungulate.

Farmers’ unconscious incorporation of sexually-produced genotypes into the germplasm of a vegetatively-propagated crop (Oxalis tuberosa Mol.)

Clonal propagation and sexual reproduction have diametrically opposite effects on the genetic diversity of crops, permitting, respectively, the conservation of existing genotypes or the creation of new diversity. Oxalis tuberosa Mol. (“oca”) exhibits a heteromorphic self-incompatibility system and is traditionally propagated clonally but is capable of sexual reproduction. In this study we investigated the influence of sexual reproduction on the in situ genetic diversity of a vegetatively propagated crop, taking oca as a model plant. The occurrence of sexual reproduction in natural conditions and its use by farmers were studied through surveys and interviews, while the effect of sexual reproduction on the in situ genetic diversity was evaluated using microsatellite analysis. Plantlets issuing from sexual reproduction were encountered in five of the nine visited communities. The interviews indicated that six out of the nine interviewed farmers were likely, albeit unconsciously, incorporating clonal lineages issuing from sexual reproduction into their varieties. The microsatellite analysis indicated a strong effect of such incorporations on the genetic diversity of the plant varieties. Assignment tests confirmed the allogamy of oca. “Complex varieties”, made up of individuals exhibiting genetically differentiated genotypes, were most likely the result of recent crossing. Unconscious incorporation of new genotypes into the cultivated oca germplasm is therefore highly likely, raising the question of its potential occurrence in other vegetatively propagated crops. These findings could have an important effect on the adaptive potential of oca, especially in light of the genetic erosion that threatens the crop.

Biological Effects of Pseudomonas fuscovaginae Toxins on Rice Cells

Pseudomonas fuscovaginae, the causai agent of sheath brown rot of rice, produces in vitro phytotoxic lipopeptides among which the lipodepsinonapeptide syringotoxin and two more hydrophobic lipodepsipeptides FP-A and FP-B. Biological effects of purified extracts or of single components of the bacterial Phytotoxins were investigated on rice cells. The toxins per se induced rapid and high electrolyte leakage from calli. Syringotoxin alone induced a concentration dependent stimulation of H+-ATPase activity on right-side out rice plasma membranes, corroborating and extending previous findings with other plant species. In contrast, with inside out vesicles obtained by Brij58 treatment, the enzyme activity was inhibited in a concentration depending manner. Similar results were obtained with syringomycin E. FP-A and FP-B induced an inhibition of the ATPase activity no matter the orientation of the vesicles. Inhibition was increased in presence of syringotoxin or syringomycin E, suggesting a synergistic action by the two types of toxins.