Sequence variability of the MspI satellite DNA family of the pinewood nematode Bursaphelenchus xylophilus at different geographic scales

Tandemly repeated sequences known as satellite DNA (satDNA) generally exhibit complex evolutionary patterns of concerted evolution in which mutations are homogenized and fixed in a stochastic process of molecular drive. Here, the nucleotidic variability of the MspI satDNA family of the pinewood nematode Bursaphelenchus xylophilus is analyzed in order to understand the evolutionary dynamics of satDNA at the intraspecific level. A total of 425 MspI monomer units, either PCR-amplified from isolates of local (Peninsula of Setúbal, Portugal) or worldwide origin, or retrieved from the B. xylophilus genome sequence, were characterized and compared. Whatever their origin, sliding window analysis of sequence variability patterns among monomers revealed low, moderate and highly variant domains, indicating that variable levels of evolutionary constraint may act upon the entire monomers. The phylogenetic inference based on the different sets of MspI satDNA family for this species shows a broad polymorphism of the individual monomers, which were distributed into four main clusters. However, such clustering appeared indepen- dent from the geographic origin of the nematodes, and could not discriminate isolates or groups of geo- graphically close isolates. Rather, the formation of different phylogenetic groups within this satDNA family suggests an a priori embodying of a set of diverging repeats from a common ancestor satDNA library, which have been differently amplified along the evolutionary pathway of this species. The present work improves knowledge on the evolutionary dynamics of satDNA at the intraspecific level, and pro- vides new information on satDNA sequence variability among natural populations sampled at a local geo- graphic scale

Invasion of the pinewood nematode, Bursaphelenchus xylophilus

Dans ce projet, nous nous sommes intéressés à l’invasion du nématode du pin, Bursaphelenchus xylophilus, un ver microscopique responsable de la maladie du dépérissement du pin et causant la perte de millions d’arbres annuellement à travers le monde. Notre objectif global était d’affiner nos connaissances sur ce cas d’invasion et particulièrement d’établir l’histoire d’invasion des populations européennes. Nous avons analysé la diversité génétique neutre d’échantillons de populations naturelles de nématode du pin de l’aire native et de différentes zones envahies à l’aide de méthodes de génétique des populations. Ces analyses ont essentiellement mis en évidence que (i) l’aire native du nématode du pin est fortement structurée à fines échelles spatiales et présente une diversité génétique faible à modérée, suggérant un rôle important de la dérive génétique, non compensée par la dispersion, (ii) les populations européennes présentent une diversité génétique extrêmement faible, suggérant un seul évènement d’introduction en Europe, et que (iii) l’Amérique du Nord est la source la plus probable des populations envahissantes européennes, non le Japon et la Chine. Ce projet souligne l’intérêt que les nématologistes et les généticiens des populations ont à collaborer. La génétique des populations a en effet permis de clarifier l’histoire de l’invasion des populations européennes et ce modèle nématode, du fait de son cycle de vie complexe résultant en un cas extrême de faible diversité génétique, a permis de s’interroger sur des méthodes et analyses couramment utilisées en génétique des populations.In this project, we were interested in the invasion of the pinewood nematode, Bursaphelenchus xylophilus, a microscopic worm responsible for the pine wilt disease and causing the annual loss of millions of trees worldwide. Our global objective was to improve our knowledge on this case of invasion and particularly to clarify the invasion history of European populations. After developing the microsatellite markers needed for our study, we analysed the genetic diversity of samples of natural populations from both the native area and invaded areas thanks to population genetics methods. These analyses revealed that (i) the native area is strongly genetically structured at fine scales and displays a low to moderate genetic diversity, suggesting an important role of the genetic drift, not compensated by dispersal, (ii) the European populations exhibit an extremely low genetic diversity, indicating a unique event of introduction in Europe and (iii) North America is the most probable source of European populations compared to China and Japan. This project is finally a good example of the benefits of collaboration between nematologists and population geneticists. Population genetics allowed to clarify the invasion history of European populations and this “nematode” model, because of its complex life cycle resulting in an extreme case of low genetic diversity, provided the opportunity to question methodologies in population genetics that are routinely used

Invasion du nématode du pin Bursaphelenchus xylophilus

In this project, we were interested in the invasion of the pinewood nematode, Bursaphelenchus xylophilus, a microscopic worm responsible for the pine wilt disease and causing the annual loss of millions of trees worldwide. Our global objective was to improve our knowledge on this case of invasion and particularly to clarify the invasion history of European populations. After developing the microsatellite markers needed for our study, we analysed the genetic diversity of samples of natural populations from both the native area and invaded areas thanks to population genetics methods. These analyses revealed that (i) the native area is strongly genetically structured at fine scales and displays a low to moderate genetic diversity, suggesting an important role of the genetic drift, not compensated by dispersal, (ii) the European populations exhibit an extremely low genetic diversity, indicating a unique event of introduction in Europe and (iii) North America is the most probable source of European populations compared to China and Japan. This project is finally a good example of the benefits of collaboration between nematologists and population geneticists. Population genetics allowed to clarify the invasion history of European populations and this “nematode” model, because of its complex life cycle resulting in an extreme case of low genetic diversity, provided the opportunity to question methodologies in population genetics that are routinely used.Dans ce projet, nous nous sommes intéressés à l’invasion du nématode du pin, Bursaphelenchus xylophilus, un ver microscopique responsable de la maladie du dépérissement du pin et causant la perte de millions d’arbres annuellement à travers le monde. Notre objectif global était d’affiner nos connaissances sur ce cas d’invasion et particulièrement d’établir l’histoire d’invasion des populations européennes. Nous avons analysé la diversité génétique neutre d’échantillons de populations naturelles de nématode du pin de l’aire native et de différentes zones envahies à l’aide de méthodes de génétique des populations. Ces analyses ont essentiellement mis en évidence que (i) l’aire native du nématode du pin est fortement structurée à fines échelles spatiales et présente une diversité génétique faible à modérée, suggérant un rôle important de la dérive génétique, non compensée par la dispersion, (ii) les populations européennes présentent une diversité génétique extrêmement faible, suggérant un seul évènement d’introduction en Europe, et que (iii) l’Amérique du Nord est la source la plus probable des populations envahissantes européennes, non le Japon et la Chine. Ce projet souligne l’intérêt que les nématologistes et les généticiens des populations ont à collaborer. La génétique des populations a en effet permis de clarifier l’histoire de l’invasion des populations européennes et ce modèle nématode, du fait de son cycle de vie complexe résultant en un cas extrême de faible diversité génétique, a permis de s’interroger sur des méthodes et analyses couramment utilisées en génétique des populations

Data from: No clear effect of admixture between two European invading outbreaks of Diabrotica virgifera virgifera in natura

In this study, we challenged the hypothesis that admixture may have had a positive impact in the context of the European invasion of the western corn rootworm (WCR), Diabrotica virgifera virgifera, LeConte. This beetle was introduced in Europe from the USA several times since the 1980’s. The multiple introductions of this major pest of cultivated corn led to the formation of two major outbreaks in North Western (NW) Italy and in Central and South Eastern (CSE) Europe that eventually merged into a secondary contact zone where insects from both outbreaks interbreed. We collected about 600 insects from this contact zone and genotyped them using 13 microsatellite markers. Three types of information were obtained from the collected individuals: (i) their survival under starvation; (ii) their admixed status, determined through a Bayesian method of genetic clustering and (iii) their mating probability, studied via the detection, isolation and genotyping of sperm in female spermathecae. Twenty six % and 12% of the individuals were assigned to the NW Italy or the CSE Europe parental types, respectively, and 23% and 39% to the F1 and backcross hybrid types, respectively. Globally, our results do not reveal any significant impact of the admixed status on the mating probability and on the choice of mating partners. However the admixed status had a sex- and sampling site-dependent effect on survival in adults under starvation. In addition sex had an effect on survival, with mortality hazard about 3 times larger in males than in females. The consequences of these findings for the evolution of the admixture zone of northern Italy are discussed

Data from: No clear effect of admixture between two European invading outbreaks of Diabrotica virgifera virgifera in natura

In this study, we challenged the hypothesis that admixture may have had a positive impact in the context of the European invasion of the western corn rootworm (WCR), Diabrotica virgifera virgifera, LeConte. This beetle was introduced in Europe from the USA several times since the 1980’s. The multiple introductions of this major pest of cultivated corn led to the formation of two major outbreaks in North Western (NW) Italy and in Central and South Eastern (CSE) Europe that eventually merged into a secondary contact zone where insects from both outbreaks interbreed. We collected about 600 insects from this contact zone and genotyped them using 13 microsatellite markers. Three types of information were obtained from the collected individuals: (i) their survival under starvation; (ii) their admixed status, determined through a Bayesian method of genetic clustering and (iii) their mating probability, studied via the detection, isolation and genotyping of sperm in female spermathecae. Twenty six % and 12% of the individuals were assigned to the NW Italy or the CSE Europe parental types, respectively, and 23% and 39% to the F1 and backcross hybrid types, respectively. Globally, our results do not reveal any significant impact of the admixed status on the mating probability and on the choice of mating partners. However the admixed status had a sex- and sampling site-dependent effect on survival in adults under starvation. In addition sex had an effect on survival, with mortality hazard about 3 times larger in males than in females. The consequences of these findings for the evolution of the admixture zone of northern Italy are discussed

Inference of the worldwide invasion routes of the pinewood nematode, Bursaphelenchus xylophilus, using ABC analysis on microsatellite data

Population genetics have been greatly beneficial to improve knowledge about biological invasions. Quantitative genetic methods, such as Approximate Bayesian Computation (ABC), have brought this improvement up a notch and are now essential tools to decipher the invasion routes of any invasive species. In this paper, we performed ABC analyses to shed light on the pinewood nematode (PWN) worldwide invasion routes and identify the source of the European populations. Originating from North America, this microscopic worm has been invading Asia since 1905 and Europe since 1999, causing tremendous damage on pine forests. Using microsatellite data, we demonstrated the existence of multiple independent events of introduction in Japan (one from the USA and one with an unknown origin) and China (one from the USA and one from Japan). We also found an American origin of the Portuguese samples. Although, we observed some discrepancies between traditional genetic methods and the ABC method, which are worth investigating and are discussed here, the ABC approach definitely helped clarify the worldwide history of the PWN invasion

Phenotypic_data_Dryad

Phenotypic_data_Drya