Sample size for diversity studies in tetraploid alfalfa (Medicago sativa) based on codominantly coded SSR markers

The number of genotypes investigated per population is important for the reliability of diversity studies. The objective of this study was to determine the sample size for the identification of differences among populations of an outcrossing autotetraploid species, alfalfa (Medicago sativa), using codominantly coded SSR markers. One hundred and twenty genotypes from each of two closely related populations were analysed with two markers. Twenty random subsamples for each of three sample sizes (10, 20 and 40 genotypes) were built. Compared to the populations with 120 genotypes, alleles that were no longer present in subsamples with 40 genotypes were mainly rare, whereas abundant alleles were also excluded in subsamples with 10 genotypes. F ST values for pairs of subsamples between the two populations were always significantly different based on 40 genotypes, whereas for 10 genotypes more than half of the pairs were not significantly different. We concluded that 40 genotypes are a reasonable sample size for diversity studies with closely related populations of tetraploid alfalfa investigated with SSR markers. Twenty genotypes may be an economical alternative for large scale studies, but 10 genotypes were a too low number for reliable results

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

Genetic Diversity Among Alfalfa Cultivars Using SSR Markers

Alfalfa (Medicago sativa) is an autotetraploid, allogamous and heterozygous species. Cultivated varieties are synthetic cultivars, usually obtained through 3 or 4 generations of panmictic reproduction of a set of various numbers of parents. The parents can be clones, half-sib or full-sib families. The breeders apply selection pressure for some agronomic traits, to induce changes in the genetic background. The objective of this study was to investigate the differentiation level among seven cultivars originating from one breeding program, and between these cultivars and the breeding pool, with eight SSR markers

Construction of two genetic linkage maps in cultivated tetraploid alfalfa (Medicago sativa) using microsatellite and AFLP markers

BACKGROUND: Alfalfa (Medicago sativa) is a major forage crop. The genetic progress is slow in this legume species because of its autotetraploidy and allogamy. The genetic structure of this species makes the construction of genetic maps difficult. To reach this objective, and to be able to detect QTLs in segregating populations, we used the available codominant microsatellite markers (SSRs), most of them identified in the model legume Medicago truncatula from EST database. A genetic map was constructed with AFLP and SSR markers using specific mapping procedures for autotetraploids. The tetrasomic inheritance was analysed in an alfalfa mapping population. RESULTS: We have demonstrated that 80% of primer pairs defined on each side of SSR motifs in M. truncatula EST database amplify with the alfalfa DNA. Using a F1 mapping population of 168 individuals produced from the cross of 2 heterozygous parental plants from Magali and Mercedes cultivars, we obtained 599 AFLP markers and 107 SSR loci. All but 3 SSR loci showed a clear tetrasomic inheritance. For most of the SSR loci, the double-reduction was not significant. For the other loci no specific genotypes were produced, so the significant double-reduction could arise from segregation distortion. For each parent, the genetic map contained 8 groups of four homologous chromosomes. The lengths of the maps were 2649 and 3045 cM, with an average distance of 7.6 and 9.0 cM between markers, for Magali and Mercedes parents, respectively. Using only the SSR markers, we built a composite map covering 709 cM. CONCLUSIONS: Compared to diploid alfalfa genetic maps, our maps cover about 88–100% of the genome and are close to saturation. The inheritance of the codominant markers (SSR) and the pattern of linkage repulsions between markers within each homology group are consistent with the hypothesis of a tetrasomic meiosis in alfalfa. Except for 2 out of 107 SSR markers, we found a similar order of markers on the chromosomes between the tetraploid alfalfa and M. truncatula genomes indicating a high level of colinearity between these two species. These maps will be a valuable tool for alfalfa breeding and are being used to locate QTLs

Both selection and plasticity drive niche differentiation in experimental grasslands

Abstract The way species avoid each other in a community by using resources differently across space and time is one of the main drivers of species coexistence in nature1,2. This mechanism, known as niche differentiation, has been widely examined theoretically but still lacks thorough experimental validation in plants. To shape niche differences over time, species within communities can reduce the overlap between their niches or find unexploited environmental space3. Selection and phenotypic plasticity have been advanced as two candidate processes driving niche differentiation4,5, but their respective role remains to be quantified6. Here, we tracked changes in plant height, as a candidate trait for light capture7, in 5-year multispecies sown grasslands. We found increasing among-species height differences over time. Phenotypic plasticity promotes this change, which explains the rapid setting of differentiation in our system. Through the inspection of changes in genetic structure, we also highlighted the contribution of selection. Altogether, we experimentally demonstrated the occurrence of species niche differentiation within artificial grassland communities over a short time scale through the joined action of both plasticity and selection

Estimation of the Genetic Diversity in Tetraploid Alfalfa Populations Based on RAPD Markers for Breeding Purposes

Alfalfa is an autotetraploid, allogamous and heterozygous forage legume, whose varieties are synthetic populations. Due to the complex nature of the species, information about genetic diversity of germplasm used in any alfalfa breeding program is most beneficial. The genetic diversity of five alfalfa varieties, involved in progeny tests at Institute of Field and Vegetable Crops, was characterized based on RAPD markers. A total of 60 primers were screened, out of which 17 were selected for the analysis of genetic diversity. A total of 156 polymorphic bands were generated, with 10.6 bands per primer. Number and percentage of polymorphic loci, effective number of alleles, expected heterozygosity and Shannon's information index were used to estimate genetic variation. Variety Zuzana had the highest values for all tested parameters, exhibiting the highest level of variation, whereas variety RSI 20 exhibited the lowest. Analysis of molecular variance (AMOVA) showed that 88.39% of the total genetic variation was attributed to intra-varietal variance. The cluster analysis for individual samples and varieties revealed differences in their population structures: variety Zuzana showed a very high level of genetic variation, Banat and Ghareh were divided in subpopulations, while Pecy and RSI 20 were relatively uniform. Ways of exploiting the investigated germplasm in the breeding programs are suggested in this paper, depending on their population structure and diversity. The RAPD analysis shows potential to be applied in analysis of parental populations in semi-hybrid alfalfa breeding program in both, development of new homogenous germplasm, and identification of promising, complementary germplasm

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

Utilisation des marqueurs moléculaires pour la création de variétés synthétiques (Exemple du ray-grass anglais (Lolium perenne L.))

La Sélection Assistée par Marqueurs (SAM) est un outil d'aide à la sélection largement étudié pour l'obtention des variétés lignées ou hybrides mais peu d'auteurs ont traité de l'obtention des variétés synthétiques. Ces dernières sont sélectionnées à partir de descendances de polycross qui sont en fait des populations biparentales connectées. L'objectif de cette thèse est de développer des méthodologies pour utiliser la SAM dans le schéma de sélection du ray-grass anglais en prenant comme exemples de caractères la hauteur de plante et la résistance à la rouille couronnée. Dans une première partie, à partir de trois populations biparentales connectées par un parent commun, l'analyse multipopulations connectées du logiciel MCQTL s'est révélée plus performante pour identifier des allèles favorables que la détection réalisée dans chacune des populations. Cette méthode a permis de détecter un grand nombre de QTL (Quantitative Trait Loci) malgré une base génétique étroite du matériel élite utilisé. Dans une deuxième partie, ce type d'analyse a permis de détecter des QTL directement à partir du matériel de sélection tout en mettant en évidence les contraintes liées à ce type de matériel. L'utilisation des QTL au sein des schémas de sélection pose la question de leur stabilité dans différents fonds génétiques. Ainsi dans une dernière partie, il a été montré que seuls les QTL ayant des effets forts sont stables. Ces résultats ont permis d'apporter des propositions d'implémentation de la SAM chez les espèces commercialisées sous forme de variétés synthétiques, en plaçant les phases de détection de QTL directement dans le schéma de sélection.Marker Assisted Selection (MAS) is a widely studied tool for the creation of line and hybrid varieties but few authors analyzed its implementation for the creation of synthetic varieties. Synthetic varieties are selected from polycross progenies which are biparental connected populations. The objective of this thesis consisted in developping methodologies to implement MAS in the breeding scheme of perennial ryegrass, taking plant height and crown rust resistance as example traits. In the first part, based on three biparental populations connected by a common parent, the multipopulation connected analysis from MCQTL software was more powerful for the identification of favorable alleles than the detection within each population. Numerous QTL (Quantitative Trait Loci) were detected by using this method, in spite of a narrow genetic basis of the élite studied material. In the second part, the same multipopulation connected analysis enabled us to detect QTL directly in breeding material, while revealing the constraints by using this type of material. The use of QTL in breeding schemes raises the question of their stability across different genetic backgrounds. Thus, in the last part, it was shown that only QTL with strong effects were stable over different genetic backgrounds. As a result of this thesis, a strategy, based on QTL detection directly in the breeding material is suggested in order to implement MAS in species commercialized as synthetic varieties.POITIERS-BU Sciences (861942102) / SudocSudocFranceF