Jean-Louis Pham

Association Studies Identify Natural Variation at PHYC Linked to Flowering Time and Morphological Variation in Pearl Millet

The identification of genes selected during and after plant domestication is an important research topic to enhance knowledge on adaptative evolution. Adaptation to different climates was a key factor in the spread of domesticated crops. We conducted a study to identify genes responsible for these adaptations in pearl millet and developed an association framework to identify genetic variations associated with the phenotype in this species. A set of 90 inbred lines genotyped using microsatellite loci and AFLP markers was used. The population structure was assessed using two different Bayesian approaches that allow inbreeding or not. Association studies were performed using a linear mixed model considering both the population structure and familial relationships between inbred lines. We assessed the ability of the method to limit the number of false positive associations on the basis of the two different Bayesian methods, the number of populations considered and different morphological traits while also assessing the power of the methodology to detect given additive effects. Finally, we applied this methodology to a set of eight pearl millet genes homologous to cereal flowering pathway genes. We found significant associations between several polymorphisms of the pearl millet PHYC gene and flowering time, spike length, and stem diameter in the inbred line panel. To validate this association, we performed a second association analysis in a different set of pearl millet individuals from Niger. We confirmed a significant association between genetic variation in this gene and these characters.

Farmers’ use of wild relative and sexual reproduction in a vegetatively propagated crop. The case of yam in Benin

The impact of traditional farmers’ management on genetic diversity of vegetatively propagated crops is poorly documented. In this study, we analysed the impact of ennoblement of spontaneous yams, an original traditional farmers’ practice, on the genetic diversity of yam (Dioscorea sp.) in Benin. We used 11 microsatellite markers on yam tubers from a small village in northern Benin and demonstrated that wild × cultivated hybrids are spontaneously formed. Many of the spontaneous yams collected by farmers from surrounding savannah areas for ennoblement were shown to be of wild and hybrid genotypes. Moreover, we demonstrated that some yam varieties have a wild or hybrid signature. Lastly, we performed a broader ranging genetic analysis on yam material from throughout Benin and showed that this practice is used in different ecological and ethno‐linguistic regions. Through this practice, farmers create new varieties with new genetic combinations via sexual reproduction of wild and cultivated yams. This system, whereby a sexual cycle and asexual propagation are mixed, ensures potential large‐scale cultivation of the best genotypes while preserving the potential for future adaptation.

Genetic Nature of Yams (Dioscorea sp.) Domesticated by Farmers in Benin (West Africa)

Abstract‘Domestication’ is a traditional farmers’ practice reported for yams (Dioscorea sp.) in Benin (West Africa). It involves introducing ‘spontaneous’ (naturally occurring) yams, supposedly wild (D. abyssinica and D. praehensilis), in varieties of the D. cayenensis–D. rotundata cultivated species complex. In this study, we established the genetic nature of ‘predomesticated’ yam plants using the amplified fragment length polymorphism (AFLP) technique. A total of 213 accessions, consisting of 32 predomesticated yams, 70 D. cayenensis–D. rotundata, 86 D. abyssinica and 25 D. praehensilis yams were analysed. Using 91 AFLP markers, three groups of accessions were distinguished, broadly corresponding to the above botanical species. Of the 32 predomesticated accessions, 16% were clustered with D. praehensilis, 37% with D. abyssinica and the remaining 47% with D. cayenensis–D. rotundata. These results demonstrated the use of wild plants by farmers in their domestication process, and suggested that plants derived from intervarietal and interspecific hybridisation may also be subject to this process. This study has shown that through domestication farmers influence and increase the genetic diversity in yam by using sexual reproduction of wild and possibly cultivated yams.

Genetic basis of pearl millet adaptation along an environmental gradient investigated by a combination of genome scan and association mapping

Identifying the molecular bases of adaptation is a key issue in evolutionary biology. Genome scan is an efficient approach for identifying important molecular variation involved in adaptation. Association mapping also offers an opportunity to gain insight into genotype–phenotype relationships. Using these two approaches coupled with environmental data should help to come up with a refined picture of the evolutionary process underlying adaptation. In this study, we first conducted a selection scan analysis on a transcription factor gene family. We focused on the MADS‐box gene family, a gene family which plays a crucial role in vegetative and flower development. Twenty‐one pearl millet populations were sampled along an environmental gradient in West Africa. We identified one gene, i.e. PgMADS11, using Bayesian analysis to detect selection signatures. Polymorphism at this gene was also associated with flowering time variation in an association mapping framework. Finally, we found that PgMADS11 allele frequencies were closely associated with annual rainfall. Overall, we determined an efficient way to detect functional polymorphisms associated with climate variation in non‐model plants by combining genome scan and association mapping. These results should help monitor the impact of recent climatic changes on plant adaptation.

Segregation patterns of isozyme loci and microsatellite markers show the diploidy of African yam Dioscorea rotundata (2n=40)

The cultivated yam species Dioscorea rotundata (2n=40) has been considered by most authors as a tetraploid species with a basic chromosome number of ten. In this paper, we analysed the segregation of two isozyme loci and six microsatellite markers in the progeny of a self-fertilised monoecious plant. For the eight markers, segregation patterns could be explained by only two genetic models: diploidy or tetraploidy with two null alleles. Given the nature of studied markers, the most parsimonious hypothesis was that the parental plant was diploid. These results, data from a diversity survey and results of other authors led to the conclusion that D. rotundata is a diploid species.

Genetic diversity analysis of yam cultivars (Dioscorea rotundata Poir.) in Benin using simple sequence repeat (SSR) markers

Abstract Guinea yam (Dioscorea rotundata Poir.) is a dioecious vegetatively propagated tuber crop. It iswidely cultivated by traditional techniques in West Africa, its area of origin. The genetic diver-sity of 146 accessions from Benin was analysed using 10 polymorphic simple sequence repeat(SSR) nuclear markers and agromorphological traits. An average of 8.4 alleles per locus wasdetected. The mean heterozygosity was 0.57 and the mean polymorphism information content(PIC) for polymorphic markers was 0.51. Some cultivars (23%) were found to have an identicalgenotype for the 10 markers. The structure of the genetic diversity observed in Benin is theresult of farmers’ crop management practices and their know-how. The cultivar diversityhad a geographical component. We also noted major differentiation between early and latecultivars, with higher diversity in the early ones. Cultivars from northern Benin and early cul-tivars had the greatest allelic richness. SSR markers proved to be powerful tools for fingerprint-ing each cultivar and analysing their genetic relationships. The results of this study could beuseful for defining a strategy for the conservation of genetic diversity in yams.

Natural hazards and genetic diversity in rice

Rice crop diversity hasdecreased dramatically in the recent past.Understanding the causes that underlie theevident genetic erosion is critical for thefood security of subsistence rice farmers andbiodiversity. Our study shows that farmers inthe northeastern Philippines had a markedreduction in rice diversity from 1996 to 1998.The ultimate causes were a drought resultingfrom the El Niño phenomenon in 1997 andflooding due to two successive typhoons in1998. The proximate causes, however, includedlocal water control factors, limitations in thehousehold and village-level seedinfrastructure, farm location in relation tothe goods and services necessary to obtainseeds, policies and programs of the Departmentof Agriculture, and the characteristics of therice varieties themselves. The implications ofour study are that genetic erosion is notalways the result of purposeful acts by farmersnor is it necessarily gradual. Improvingon-farm seed technology will stabilize the seedproduction, distribution, and use system andthereby enhance household food security.Ultimately, rice diversity will be improvedonly if diversity is a safe and viable optionfor farmers. Therefore, public policy thatsupports farmers who maintain a diverse set ofcultivars is critical for any on-farmconservation strategy.

Genetic structure of farmer-managed varieties in clonally-propagated crops

The relative role of sexual reproduction and mutation in shaping the diversity of clonally propagated crops is largely unknown. We analyzed the genetic diversity of yam—a vegetatively-propagated crop—to gain insight into how these two factors shape its diversity in relation with farmers’ classifications. Using 15 microsatellite loci, we analyzed 485 samples of 10 different yam varieties. We identified 33 different genotypes organized in lineages supported by high bootstrap values. We computed the probability that these genotypes appeared by sexual reproduction or mutation within and between each lineage. This allowed us to interpret each lineage as a product of sexual reproduction that has evolved by mutation. Moreover, we clearly noted a similarity between the genetic structure and farmers’ classifications. Each variety could thus be interpreted as being the product of sexual reproduction having evolved by mutation. This highly structured diversity of farmer-managed varieties has consequences for the preservation of yam diversity.

Wild crop relative populations hot-spots of diversity are hot-spots of introgression in the case of pearl millet

Wild crop relatives are unique genetic resources for crop adaptation. Increasing pressure from agriculture threatens these populations both by reducing their habitats and by creating opportunities for wild-cultivated hybridization. In this study, we assessed the diversity of 38 wild pearl millet populations covering the whole known distribution of the species in Africa, which extends from Senegal to Sudan. Using genetic analyses of 10 cultivated varieties as control, we demonstrate the high diversity harbored by these wild populations. Diversity patterns suggest a diversity hot-spot in the southern part of the wild population’s range. However, this high wild genetic diversity could partly be explained by introgression from cultivated varieties. Such introgression is widespread in the Sahel. We validate the impact of cultivated introgression on the diversity of the wild population using a genetic introgression model. The introgression distorts the real assessment of the diversity of the wild population, and the burden of this gene flow compromises the long term survival of the wild populations’ original genome. Our study also questions the long term survival of the crop’s wild relatives.

Spatial and Temporal Variation in Selection of Genes Associated with Pearl Millet Varietal Quantitative Traits In situ

Ongoing global climate changes imply new challenges for agriculture. Whether plants and crops can adapt to such rapid changes is still a widely debated question. We previously showed adaptation in the form of earlier flowering in pearl millet at the scale of a whole country over three decades. However, this analysis did not deal with variability of year to year selection. To understand and possibly manage plant and crop adaptation, we need more knowledge of how selection acts in situ. Is selection gradual, abrupt, and does it vary in space and over time? In the present study, we tracked the evolution of allele frequency in two genes associated with pearl millet phenotypic variation in situ. We sampled 17 populations of cultivated pearl millet over a period of 2 years. We tracked changes in allele frequencies in these populations by genotyping more than seven thousand individuals. We demonstrate that several allele frequencies changes are compatible with selection, by correcting allele frequency changes associated with genetic drift. We found marked variation in allele frequencies from year to year, suggesting a variable selection effect in space and over time. We estimated the strength of selection associated with variations in allele frequency. Our results suggest that the polymorphism maintained at the genes we studied is partially explained by the spatial and temporal variability of selection. In response to environmental changes, traditional pearl millet varieties could rapidly adapt thanks to this available functional variability.