Structuration de la diversité génétique chez la luzerne cultivée, conséquence pour l'identification de gènes liés à des caractères agronomiques

La luzerne, espèce allogame et autotétraploïde, est une légumineuse fourragère dont l’intérêt connaît un renouveau certain grâce à ses atouts pour le développement durable de l’agriculture. L’amélioration des variétés nécessite de mieux connaître les ressources génétiques, pour les exploiter, que ce soit en sélection classique ou en utilisant les outils moléculaires pour implémenter la sélection assistée par marqueurs. En étudiant 10 variétés européennes, nous avons montré que la diversité à l’intérieur des variétés est grande (hétérozygotie attendue de 0.75, déviation standard de 0.80, pour des caractères phénotypiques et des marqueurs moléculaires neutres, respectivement). La différenciation entre variétés est notable pour les caractères phénotypiques. Cette structuration de la diversité est favorable à l’utilisation de la génétique d’association basée sur des gènes candidats pour identifier des gènes liés à des caractères agronomiques. Une telle étude a permis de montrer que le gène Constans-like contribue à expliquer les différences de longueur de tiges chez la luzerne, une composante du rendement fourrager. Alfalfa, an allogamous and autotetraploid species, is a forage legume for which a renewed interest has raised because of its advantages for a sustainable agriculture. Variety improvement requires a better knowledge of the genetic resources in order to exploit them, with classical breeding methods or with molecular tools to implement marker assisted selection. We have described 10 European varieties and found a large within-variety diversity (expected heterozygozity of 0.75, standard deviation of 0.80, with phenotypic traits and neutral molecular markers, respectively). Differentiation between varieties was noticeable for phenotypic traits. This genetic structure of diversity is favourable to the use of association genetics based on candidate genes to identify genes related to agronomic traits. Such a study showed that Constans-like gene contributes to explain differences in alfalfa stem length, a component of forage yield

Population structure and linkage disequilibrium unravelled in tetraploid potato

Association mapping is considered to be an important alternative strategy for the identification of quantitative trait loci (QTL) as compared to traditional QTL mapping. A necessary prerequisite for association analysis to succeed is detailed information regarding hidden population structure and the extent of linkage disequilibrium. A collection of 430 tetraploid potato cultivars, comprising two association panels, has been analysed with 41 AFLP® and 53 SSR primer combinations yielding 3364 AFLP fragments and 653 microsatellite alleles, respectively. Polymorphism information content values and detected number of alleles for the SSRs studied illustrate that commercial potato germplasm seems to be equally diverse as Latin American landrace material. Genome-wide linkage disequilibrium (LD)—reported for the first time for tetraploid potato—was observed up to approximately 5 cM using r2 higher than 0.1 as a criterion for significant LD. Within-group LD, however, stretched on average twice as far when compared to overall LD. A Bayesian approach, a distance-based hierarchical clustering approach as well as principal coordinate analysis were adopted to enquire into population structure. Groups differing in year of market release and market segment (starch, processing industry and fresh consumption) were repeatedly detected. The observation of LD up to 5 cM is promising because the required marker density is not likely to disable the possibilities for association mapping research in tetraploid potato. Population structure appeared to be weak, but strong enough to demand careful modelling of genetic relationships in subsequent marker-trait association analyses. There seems to be a good chance that linkage-based marker-trait associations can be identified at moderate marker densities

Comparison of morphological traits and molecular markers to analyse diversity and structure of alfalfa (Medicago sativa L.) cultivars

Diversity and structure of populations may differ substantially between morphological traits and molecular markers. Explanations of such discrepancies are crucial for further progress in breeding as well as for the maintenance of genetic resources. Our objective was to compare indices of among-cultivars differentiation for morphological traits (QST) and molecular markers (FST) in alfalfa (Medicago sativa), a legume forage species. Ten cultivars representing the Northern and Southern types commonly grown in Europe were investigated. For each cultivar, 40 genotypes were analysed with 16 SSR markers and four morphological traits measured in two locations and several cuts. QST values were in general high (0.02–0.39) compared to the differentiation observed with molecular markers (FST = 0.01), especially for growth habit, indicating that morphological traits were more efficient to structure the diversity than molecular markers. For morphological traits, a clear separation of Northern and Southern cultivars was observed, whereas for molecular markers, no clear structure was detected. Nevertheless, the grazing type cultivar Luzelle was separated from the rest of the cultivars for both morphological traits and molecular markers. Although pairwise differences between cultivars were significant for both morphological traits and molecular markers, the main part of the variation was found within cultivars. This large within-cultivar variation may be explained, besides the outcrossing reproductive mode and autotetraploid genetics, by the recent history of M. sativa domestication in Europe and the frequent seed exchanges. Selection for morphological traits (QST > FST) was achieved without modification of within-cultivar neutral diversity

Genome-wide SNP discovery in tetraploid alfalfa using 454 sequencing and high resolution melting analysis

<p>Abstract</p> <p>Background</p> <p>Single nucleotide polymorphisms (SNPs) are the most common type of sequence variation among plants and are often functionally important. We describe the use of 454 technology and high resolution melting analysis (HRM) for high throughput SNP discovery in tetraploid alfalfa (<it>Medicago sativa </it>L.), a species with high economic value but limited genomic resources.</p> <p>Results</p> <p>The alfalfa genotypes selected from <it>M. sativa </it>subsp. <it>sativa </it>var. 'Chilean' and <it>M. sativa </it>subsp. <it>falcata </it>var. 'Wisfal', which differ in water stress sensitivity, were used to prepare cDNA from tissue of clonally-propagated plants grown under either well-watered or water-stressed conditions, and then pooled for 454 sequencing. Based on 125.2 Mb of raw sequence, a total of 54,216 unique sequences were obtained including 24,144 tentative consensus (TCs) sequences and 30,072 singletons, ranging from 100 bp to 6,662 bp in length, with an average length of 541 bp. We identified 40,661 candidate SNPs distributed throughout the genome. A sample of candidate SNPs were evaluated and validated using high resolution melting (HRM) analysis. A total of 3,491 TCs harboring 20,270 candidate SNPs were located on the <it>M. truncatula </it>(MT 3.5.1) chromosomes. Gene Ontology assignments indicate that sequences obtained cover a broad range of GO categories.</p> <p>Conclusions</p> <p>We describe an efficient method to identify thousands of SNPs distributed throughout the alfalfa genome covering a broad range of GO categories. Validated SNPs represent valuable molecular marker resources that can be used to enhance marker density in linkage maps, identify potential factors involved in heterosis and genetic variation, and as tools for association mapping and genomic selection in alfalfa.</p