Identification and characterization of microsatellite loci in ash (<i>Fraxinus excelsior</i> L.) and their conservation in the olive family (Oleaceae)

Lefort, François; Brachet, Stéphanie; Frascaria‐Lacoste, Nathalie; Edwards, K. J.; Douglas, G. C.

doi:10.1046/j.1365-294x.1999.00655_8.x

Cited by 68 publications

(69 citation statements)

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“…LOD thresholds (TF) for ML paternity assignment and type I error of false rejection and type II error of false assignment were determined by means of 50 000 simulations in FAMOZ. Both SE and ML methods were applied to seeds genotyped at microsatellites FEMSATL2 , M2.30B and 1.19 (Lefort et al, 1999) that displayed a multilocus genotype compatible with the multilocus genotype of their mother, with no further transformation (RAW), after allele binning at all three loci (BIN), after additional inclusion of a generalized null allele at locus 1.19 (BIN3NULL1). Results highlighted in bold were retained for detailed analysis of genetic connectivity among remnants.…”

Section: Discussionmentioning

confidence: 99%

“…The complete sample of 88 trees and 483 seeds was genotyped for five microsatellite markers previously developed for F. excelsior, namely, M2-30B, 1.19 and 3.1 , and FEMSATL2 and FEMSATL5 (Lefort et al, 1999). DNA isolation, amplification by polymerase chain reaction (PCR) and electrophoretic separation of PCR products were carried out as described elsewhere .…”

Section: Sampling and Data Collectionmentioning

confidence: 99%

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Paternity analysis of pollen-mediated gene flow for Fraxinus excelsior L. in a chronically fragmented landscape

Bacles

Ennos

2008

Heredity

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Paternity analysis based on microsatellite marker genotyping was used to infer contemporary genetic connectivity by pollen of three population remnants of the wind-pollinated, wind-dispersed tree Fraxinus excelsior, in a deforested Scottish landscape. By deterministically accounting for genotyping error and comparing a range of assignment methods, individual-based paternity assignments were used to derive population-level estimates of gene flow. Pollen immigration into a 300 ha landscape represents between 43 and 68% of effective pollination, mostly depending on assignment method. Individual male reproductive success is unequal, with 31 of 48 trees fertilizing one seed or more, but only three trees fertilizing more than ten seeds. Spatial analysis suggests a fat-tailed pollen dispersal curve with 85% of detected pollination occurring within 100 m, and 15% spreading between 300 and 1900 m from the source.Identification of immigrating pollen sourced from two neighbouring remnants indicates further effective dispersal at 2900 m. Pollen exchange among remnants is driven by population size rather than geographic distance, with larger remnants acting predominantly as pollen donors, and smaller remnants as pollen recipients. Enhanced wind dispersal of pollen in a barren landscape ensures that the seed produced within the catchment includes genetic material from a wide geographic area. However, gene flow estimates based on analysis of non-dispersed seeds were shown to underestimate realized gene immigration into the remnants by a factor of two suggesting that predictive landscape conservation requires integrated estimates of post-recruitment gene flow occurring via both pollen and seed.

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Section: Discussionmentioning

confidence: 99%

Section: Sampling and Data Collectionmentioning

confidence: 99%

Paternity analysis of pollen-mediated gene flow for Fraxinus excelsior L. in a chronically fragmented landscape

Bacles

Ennos

2008

Heredity

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“…We genotyped the above mentioned samples using four microsatellite loci: M230 from , and FEMSATL4, FEMSATL11 FEMSATL16 from (Lefort et al, 1999). The segregation of alleles for these markers has already been validated using control-pollinated full-sibs and no deviation from Mendelian segregation or null alleles were detected (Morand-Prieur, 2003).…”

Section: Molecular Techniquesmentioning

confidence: 99%

What sampling is needed for reliable estimations of genetic diversity in Fraxinus excelsior L. (Oleaceae)?

et al. 2008

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-• Sample size is a critical issue for genetic diversity studies and conservation programs. However, sample size evaluation requires previous knowledge of allele frequencies estimated with precision and this is not often the case.• Here, we evaluated sample size requirements for accurate genetic diversity in adult trees and family arrays in a 12 ha plot of Fraxinus excelsior L. (Oleaceae) in a community forest in central France. Data consisted of 579 adult trees and 480 offspring from 24 families genotyped at four nuclear microsatellites.• Mean square errors (MSE) estimates performed on Monte Carlo simulations of resampled data indicated that several adult individuals (> 300) are necessary for accurate measures of allele richness. However, expected heterozygosity requires smaller samples (< 30). Seeds captured about 90% of adult allelic diversity requiring a sampling effort roughly 50% larger than that of adult trees (480 seeds vs. 300 adults) suggesting that seed sampling is heavily penalized for allele counts. Nevertheless, gene diversity of seeds was essentially identical to that of the adult population.• Extrapolation of these results to other ash tree populations appears feasible because of similar levels of diversity reported in the literature but it is not granted for species with significant selfing or high genetic structure. Fraxinus excelsior / microsatellite/ genetic variation/ samplingRésumé -Quel échantillonnage pour des estimations fiables de la diversité génétique chez Fraxinus excelsior L. (Oleaceae) ?• La taille d'un échantillonnage est un paramètre difficile à estimer a priori pour les études de diversité génétique ainsi que pour les programmes de conservation. En général, l'évaluation de cette taille nécessite au préalable une connaissance fine des fréquences alléliques ce qui n'est pas toujours le cas.• Dans cette étude, nous avons évalué sur une parcelle de 12 ha de Fraxinus excelsior L. (Oleaceae), dans un forêt domaniale, la taille de l'échantillon-nage nécessaire pour estimer de façon fiable la diversité génétique des arbres adultes ainsi que de leurs descendants. Un échantillon de 579 individus adultes et 480 graines issues de 24 arbres-mères ont été génotypés grâce à quatre marqueurs microsatellites nucléaires.• Des analyses d'erreurs quadratiques moyennes obtenues dans le cadre de simulations de type Monte Carlo indiquent que plus de 300 individus adultes sont nécessaires pour obtenir des mesures alléliques fiables. Par contre, l'hétérozygotie espérée est obtenue pour des échantillons plus petits (< 30). Les graines capturent 90 % de la diversité allélique des adultes indiquant que l'échantillonnage des graines doit être deux fois celui des adultes pour obtenir la même information (480 graines vs. 300 adultes). Par ailleurs, la diversité génétique est identique pour les deux échantillonnages.• L'extrapolation de ces résultats à d'autres espèces de frêne est possible compte tenu des niveaux de diversité observés dans la littérature mais n'est pas garantie pour des espèces qui s...

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“…Preliminary PCR tests were performed with 10 individuals of each species in 20 µl solution containing 0.5 units of Q-BIOgene Taq DNA Pol, 100 µM of each dNTP, 1. Second, primers were defined on the flanking regions of the nuclear microsatellite locus Femsatl 19 (GenBank AN: AF020400 [7]) with Primer3 software, and the same PCR tests were carried out with an annealing temperature of 55 • C.…”

Section: Marker Developmentmentioning

confidence: 99%

New insights in the recognition of the European ash speciesFraxinus excelsiorL. andFraxinus angustifoliaVahl as useful tools for forest management

et al. 2006

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-Common ash (Fraxinus excelsior L.) and narrow-leaved ash (F. angustifolia Vahl) are the most common ash species in Western Europe. The former is considered to be a highly valuable timber tree and contamination of its seed lots by the latter is strongly undesirable. We studied molecular, physiological and morphological characteristics that can help to detect the presence of F. angustifolia at the population level, either in seed and/or seedling samples, or in adult ash leave samples from natural or managed populations. First we developed two molecular markers, which correspond to a RAPD-SCAR marker and a nuclear microsatellite-derived marker. Results indicate that these markers are almost specific to one or the other species for a set of populations sampled across Europe. Second, first year dormancy was studied using germination tests without stratification. F. excelsior seeds did not germinate at all, while germinations were observed in samples of F. angustifolia and introgressed populations after 16 weeks. In addition, F. angustifolia embryo/seed length ratios at the dispersal stage were significantly higher than those from F. excelsior populations. This study provides straightforward and robust tools for avoiding commercial problems of impurity of seed lots and can help forest managers to certify common ash stands.ashes / RAPD-SCAR / dormancy / discriminant markers / hybridization Résumé -De nouveaux outils pour la reconnaissance des frênes européens Fraxinus excelsior L. et F. angustifolia Vahl : une aide au diagnostic. Le frêne commun (Fraxinus excelsior L.) et le frêne oxyphylle (F. angustifolia Vahl) sont les deux espèces de frêne les plus répandues en Europe occidentale. Le frêne commun étant le seul à posséder une importante valeur commerciale, la contamination de lots de graines par du frêne oxyphylle est fortement indésirable. Nous avons étudié des caractéristiques moléculaires, morphologiques et physiologiques qui pourront se révéler très utiles à la détection de la présence de F. angustifolia à l'échelle populationnelle, dans des lots de graines ou de semis, ainsi que dans des échantillons de feuilles d'arbres adultes issus de populations naturelles ou exploitées. Nous avons développé deux marqueurs moléculaires, un SCAR-RAPD et un marqueur issu d'un locus microsatellite nucléaire. Testés sur un ensemble de populations européennes, ces marqueurs se sont révélés quasi-spécifiques de l'une ou l'autre des deux espèces. D'autre part, nous avons étudié la dormance des graines en première année par des tests de germination sans stratification. Aucune germination n'a été observée sur un ensemble de descendances de frêne commun, alors que des germinations ont été observées sur toutes les descendances de frêne oxyphylle après seize semaines, ainsi que sur certaines descendances issues de populations introgressées. Enfin, nous avons montré que les embryons de F. excelsior à maturité dispersive occupent significativement moins de place dans la graine que ceux de F. angustifolia. Cette étude permet de ...

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Identification and characterization of microsatellite loci in ash (Fraxinus excelsior L.) and their conservation in the olive family (Oleaceae)

Cited by 68 publications

References 2 publications

Paternity analysis of pollen-mediated gene flow for Fraxinus excelsior L. in a chronically fragmented landscape

Paternity analysis of pollen-mediated gene flow for Fraxinus excelsior L. in a chronically fragmented landscape

What sampling is needed for reliable estimations of genetic diversity in Fraxinus excelsior L. (Oleaceae)?

New insights in the recognition of the European ash speciesFraxinus excelsiorL. andFraxinus angustifoliaVahl as useful tools for forest management

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