Browsing by Author "Plomion, Christophe"
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- Comparison of genetic diversity estimates within and among populations of maritime pine using chloroplast simple-sequence repeat and amplified fragmentPublication . Ribeiro, M.M.A.; Mariette, Stéphanie; Vendramin, Giovanni Giuseppe; Szmidt, A.E.; Plomion, Christophe; Kremer, AntoineWe compared the genetic variation of Pinus pinaster populations using amplified fragment length polymorphism (AFLP) and chloroplast simple-sequence repeat (cpSSR) loci. Populations’ levels of diversity within groups were found to be similar with AFLPs, but not with cpSSRs. The high interlocus variance associated with the AFLP loci could account for the lack of differences in the former. Although AFLPs revealed much lower genetic diversity than cpSSRs, the levels of among-population differentiation found with the two types of marker were similar, provided that loci showing fewer than four null-homozygotes, in any population, were pruned from the AFLP data. Moreover, the French and Portuguese populations were clearly differentiated from each other, with both markers. The Mantel test showed that the genetic distance matrix calculated using the AFLP data was correlated with the matrix derived from the cpSSRs. Because of the concordance found between markers we conclude that gene flow was indeed the predominant force shaping nuclear and chloroplastic genetic variation of the populations within regions, at the geographical scale studied.
- Genetic resources in maritime pine (Pinus pinaster Aiton):molecular and quantitative measures of genetic variation and differentiation among maternal lineagesPublication . González Martínez, Santiago; Mariette, Stéphanie; Ribeiro, M.M.A.; Burban, Christian; Raffin, Annie; Chambel, Regina; Ribeiro, Carla Alexandra; Aguiar, Alexandre; Plomion, Christophe; Alía, Ricardo; Gil, Luis; Vendramin, Giovanni Giuseppe; Kremer, AntoinePinus pinaster is a conifer native to western Europe and northern Africa. Following on-going breeding programmes, provenance and progeny trials were established in some of the countries of the species’ range (France, Portugal and Spain) and quantitative traits were measured: growth, stem form, survival and pest and disease resistance, amongst others. Populations from the wide range of P. pinaster were recently screened with molecular markers in order to assess their genetic diversity. Data were obtained using allozymes, chloroplast (cpSSRs) and nuclear (nuSSRs) microsatellites and amplified fragment length polymorphisms (AFLPs). Based on mtDNA-RFLP analysis, three maternal lineages (named ‘‘western’’, ‘‘eastern’’ and ‘‘Moroccan’’) were identified and no population showed a mixed composition. In this study, the imprint that differentiation in putatively isolated glacial refugia (identified by the different maternal lineages) might have left on the nuclear genome was analysed using a wide range of molecular markers and adaptive traits. Multivariate ordination of populations based on nuclear molecular markers (allozymes and nuSSRs) showed a clear clustering of provenances sharing a given mtDNA lineage. However, that clustering was found to be less tight when only quantitative traits were investigated. In P. pinaster, the within-population estimates of gene diversity using different traits were not correlated. Therefore, caution is advisable when designing conservation strategies based on molecular marker studies or a limited number of populations. After these results, we recommend a conservation strategy characterised by gene flow consistent with the current population structure, careful seed transfer among maternal lineages (if any), selection of populations for conservation based on the originality of their allelic composition and definition of Management Units (MUs) based on adaptive traits..
- O impacto do homem na estrutura de uma espécie: Portugal, um paradigmaPublication . Ribeiro, M.M.A.; González-Martínez, Santiago; Burban, Christian; Plomion, Christophe; Vendramin, Giovanni GiuseppeA bacia do Mediterrâneo é caracterizada pelo tradicional desaparecimento da floresta devido à acção do Homem e o impacto humano tem tido influência na alteração da diversidade genética. O pinheiro bravo ocorre em populações fragmentadas na região oeste da bacia do Mediterrâneo e a sua área de distribuição tem sido alterada nos últimos séculos devido a intensa florestação, em particular no SW de França e no Noroeste da Península Ibérica e, também, devido ao comportamento invasivo desta espécie em áreas perturbadas do ponto de vista ecológico. Nesta apresentação pretendemos sintetizar e discutir o resultado de estudos que utilizaram técnicas moleculares para revelar a diversidade do pinheiro bravo e, também, o impacto humano na sua estrutura genética; que é demonstrado paradigmaticamente em Portugal. Os parâmetros genéticos estimados com base em microsatélites do cloroplasto (cpSSR) revelaram que a diversidade genética é muito elevada ao nível da distribuição da espécie. Observou-se uma clara e significativa diferenciação entre grupos de populações de diferente origem geográfica (Portugal, França, Itália, Espanha e Marrocos). Pelo contrário, o grau de divergência dentro dos países é geralmente muito baixo ou próximo de zero, o que indica uma variação homogénea dentro dos grupos. A presença de diferenciação entre grupos de populações de diferentes áreas reflecte a existência de diferentes refúgios durante o Quaternário. Em França, o padrão haplotípico sugere uma mistura de material proveniente de diferentes origens. Só foram encontrados 3 mitótipos, usando marcadores mitocondriais, marcador de herança maternal, o que proporciona uma imagem clara de áreas colonizadas a partir dos diferentes refúgios; nem uma só população possui uma composição mista. Uma análise filogenética feita com base em isoenzimas mostrou que a estrutura geográfica do pinheiro bravo na Península Ibérica (PI) é muito elevada. As populações de Noroeste formam um grupo e as de Sudeste outro. Observou-se níveis elevados de diversidade nas populações de Este e Sul e uma redução importante da variabilidade em populações da região Noroeste da PI. No entanto, pode ter existido em Portugal um refúgio, pois o pinheiro bravo pode ter sobrevivido durante a última glaciação em zonas abrigadas e de baixa altitude junto ao Oceano Atlântico, o que parece também ser evidente devido a descobertas de pólen e carvão fóssil. A distribuição da variação genética do pinheiro bravo em Portugal, observada através de cpSSR indica que a diferenciação entre populações é baixa e que a diversidade existe principalmente dentro das populações. Não se observa nenhum padrão geográfico, mas as evidências existentes de uma forte influência antrópica antropogénica associada a um fluxo genético extensivo poderiam explicar esse resultado.
- Origin identification of maritime pine stands in France using chloroplast simple-sequence repeatsPublication . Ribeiro, M.M.A.; Le Provost, Grégoire; Gerber, Sophie; Vendramin, Giovanni Giuseppe; Anzidei, Maria; Decroocq, Stéphane; Marpeau, Anne; Mariette, Stéphanie; Plomion, ChristopheMaritime pine seed-lots from north-western Iberian regions (Portugal and Galicia) were introduced in the 1950s to the southwest of France (Aquitaine region), and the stands they formed suffered considerable frost damage. In the mid 1980s, a biochemical test was developed to test the putative origin of adult stands in Aquitaine, before seeds could be distributed for commercial purposes in France. In this paper, we describe a new test employing chloroplast simple-sequence repeats (cpSSRs) to facilitate identification of stand origin based on randomisation tests. The origin of five stands of unknown origin was determined with both the cpSSR and biochemical (terpene profile analysis) tests. The results from the two tests were concordant, but the DNA-based test gave faster and more accurate results. Use of this test should help when determining the origin of maritime pine stands in the Aquitaine region of France.
- Range-wide phylogeography and gene zones in Pinus pinaster Ait. revealed by chloroplast microsatellite markersPublication . Bucci, Gabriele; González-Martínez, Santiago; Le Provost, Grégoire; Plomion, Christophe; Ribeiro, M.M.A.; Sebastiani, Frederico; Alía, Ricardo; Vendramin, Giovanni GiuseppeSome 1339 trees from 48 Pinus pinaster stands were characterized by five chloroplast microsatellites, detecting a total of 103 distinct haplotypes. Frequencies for the 16 most abundant haplotypes (pk0.01) were spatially interpolated over a lattice made by 430 grid points. Fitting of spatially interpolated values on raw haplotype frequencies at the same geographical location was tested by regression analysis. A range-wide ‘diversity map’ based on interpolated haplotype frequencies allowed the identification of one ‘hotspot’ of diversity in central and southeastern Spain, and two areas of low haplotypic diversity located in the western Iberian peninsula and Morocco. Principal component analysis (PCA) carried out on haplotypes frequency surfaces allowed the construction of a colour-based ‘synthetic’ map of the first three PC components, enabling the detection of the main range-scale genetic trends and the identification of three main ‘gene pools’ for the species: (i) a ‘southeastern’ gene pool, including southeastern France, Italy, Corsica, Sardinia, Pantelleria and northern Africa; (ii) an ‘Atlantic’ gene pool, including all the western areas of the Iberian peninsula; and (iii) a ‘central’ gene pool, located in southeastern Spain. Multivariate andamova Analyses carried out on interpolated grid point frequency values revealed the existence of eight major clusters (‘gene zones’), whose genetic relationships were related with the history of the species. In addition, demographic models showed more ancient expansions in the eastern and southern ranges of maritime pine probably associated to early postglacial recolonization. The delineation of the gene zones provides a baseline for designing conservation areas in this key Mediterranean pine.
- Variation in chloroplast single-sequence repeats in Portuguese maritime pine (Pinus pinaster Ait.)Publication . Ribeiro, M.M.A.; Plomion, Christophe; Petit, R.; Vendramin, Giovanni GiuseppeGenetic variation in 12 Pinus pinaster (maritime pine) populations spanning most of the distribution range of the species in Portugal was evaluated using six polymorphic chloroplast microsatellite (cpSSR) loci. Thirty-two haplotypes were found. There were indications of very weak differentiation among populations (Weir’s coefficient, 0.023), and the RST value, derived from the stepwise mutation model (SMM), was not significantly diferente from zero. The pattern, in which similarities in allele size, in base pairs, do not contribute to the genetic structure, may be due to the recent mixing of genetic material from different stands through plantations. Overall, a high level of haplotypic variation within populations was detected. Using the SMM estimator (mean genetic distance of individuals within populations, D2sh) we divided the populations into two groups, with above and below average values. The first group contained 5 populations, mainly from the central part of the country, which possess, in general, high levels of haplotypic diversity. Among them, 2 populations were divergent from the others based on the pair-wise Nei’s distance. The results indicate that there is no discernible geographic genetic pattern for the Portuguese populations of P. pinaster investigated. The history of expansion of the species range in Portugal during the twentieth century (mainly due to human activity) and extensive gene flow among populations associated with the expansion could explain this finding.
- What can nuclear microsatellites tell us about maritime pine genetic resources conservation and provenance certification strategies?Publication . Dérory, Jérémy; Mariette, Stéphanie; González-Martínez, Santiago; Chagné, David; Madur, Delphine; Gerber, Sophie; Brach, Jean; Persyn, François; Ribeiro, M.M.A.; Plomion, ChristopheMaritime pine (Pinus pinaster Ait.) is the first conifer used for reforestation in France and now covers 2.4 million ha of the Iberian Peninsula. In order to preserve the genetic resources of this economically and ecologically important species prior knowledge of the distribution of genetic diversity is needed. In this paper, a genetic diversity study was performed using nuclear simple sequence repeats (SSRs or microsatellites). Classical parameters of diversity (allelic richness and heterozygosity) and differentiation were estimated for 47 populations of P. pinaster. Most of the populations (40) were collected in France, six populations were also collected in the Iberian Peninsula and one Moroccan population was also included in the study. The population genetic parameters indicated that some populations should be a focus of conservation efforts (higher level of diversity, higher allelic richness and presence of rare alleles). A diagnostic test for sample origin was developed to distinguish Corsican from Landes populations.