Understanding complex traits is a fundamental challenge in plant genetics and a prerequisite for molecular breeding. Tools for trait dissection are markers, maps, and quantitative trait locus (QTL) analysis. Marker-assisted selection (MAS) is an application that integrates these tools. In this thesis research, a new sequence-based marker was evaluated, maps were constructed and used, and QTLs were detected using two types of populations. Marker-assisted selection was used to develop a novel class of barley. Restriction-site Associated DNA (RAD), a sequence based-marker technology, allows for simultaneous high-density single nucleotide polymorphism (SNP) discovery and genotyping. We assessed the value of RAD markers for linkage map construction using the Oregon Wolfe Barley (OWB) mapping population. We compared a RAD-based map to a map generated using Illumina GoldenGate Assay (EST-based SNPs). The RAD markers generated a high quality map with complete genome coverage. We then used the RAD map to locate QTL for agronomic fitness traits. A paper describing this research was published (Chutimanitsakun et al., 2011). Marker-assisted selection was used to rapidly develop fall-sown barley germplasm for human food uses. The target traits were high grain β-glucan, vernalization sensitivity (VS) and low temperature tolerance (LTT). The target loci were WX and VRN-H2. Marker-assisted selection was effective in fixing target alleles at both loci and waxy starch led to increase in grain β-glucan. Unexpected segregation at VRN-H1 and VRN-H3, revealed by genome-wide association mapping (GW-AM), led to unanticipated phenotypic variation in VS and LTT. We found that GW-AM is an efficient and powerful method for identifying the genome coordinates of genes determining target traits. Precise information is obtained with perfect markers; additional research may be needed when multiple alleles are segregating at target loci and significant associations are with markers in linkage disequilibrium (LD) with the target loci. A paper describing this research will be submitted for publication. / Graduation date: 2012

An important goal of biology is to understand the key mechanisms of evolution underlying the diversity of living organisms on Earth. In that respect, the recent innovations in the field of new generation sequencing technologies (NGS) are bringing new and exciting opportunities. This thesis presents results obtained with these tools in the specific context of the study of two sister species of cold-adapted leaf beetles, Gonioctena intermedia and G. quinquepunctata. More specifically, this work is focused around four research directions: the two first explore methods of statistical inference using a spatially explicit model of coalescence, by (1) evaluating the potential of various summary statistics to discriminate phylogeographic hypotheses, and (2) investigating the dispersal abilities of a montane leaf beetle, G. quinquepunctata, using an original method that avoids using summary statistics. The third research direction focuses on the adaptation to cold conditions in this montane leaf beetle, by testing the association between genetic polymorphism across tens of thousands of genetic markers and altitude in samples collected at various elevation levels in the Vosges (France). Finally, the fourth, and last, research axis presents the discovery of mitochondrial heteroplasmy, i.e. the presence in an individual of multiple copies of the mitochondrial genome, in natural populations of G. intermedia. We illustrate, here, how NGS technologies could help identify this phenomenon, probably underestimated in animals, on a large scale. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Chickpea (Cicer arietinum L.) was recently introduced to the Canadian prairies, a region which has a short growing season in which crop maturation often occurs under cool and wet conditions. To improve the yield of chickpea, crop duration must closely match the available growing season. The objectives of this study were to: i) examine the days to flowering of diverse chickpea accessions grown in either long or short-days; ii) examine the days to flowering of selected chickpea accessions grown in a range of thermal regimes combined with either long or short days and to examine the interaction between photoperiod and day and night temperatures on crop duration; iii) determine the timing and duration of the photoperiod-sensitive phase in selected chickpea accessions, and vi) determine the genetic basis of the association between flowering time and reaction to ascochyta blight in chickpea. A wide variation was observed in chickpea accessions for their response to flowering under long (16/8 hours day /night) and short days (10/14 hours day/night). Earlier flowering was observed under long photoperiod regimes compared with the short photoperiod regimes. Variability was detected among chickpea accessions for their flowering responses when different temperatures were combined with different photoperiods. Earlier flowering was observed under long days (16/8 hours day/night) coupled with high to moderate temperature regimes (24/16 ºC and 20/12 ºC, day and night respectively) compared to short-days (10/14 hours day and night) and moderate to low temperature regimes (20/12 ºC and 16/8 ºC day and night, respectively). Those chickpea accessions such as ICC 6821 and ICCV 96029 which originated from the lower latitudes of Ethiopia and India, respectively, flowered earlier compared to accessions such as CDC Corinne and CDC Frontier which originated from the higher latitudes and cooler temperate environments of western Canada. Photoperiod sensitivity phases were detected in chickpea accessions adapted to the cold environments of western Canada, whereas no photoperiod sensitivity phase was identified in the extra-early flowering cultivar ICCV 96029. The duration of the photoperiod sensitive phase in the chickpea accessions was longer under short days compared to long days. Field and growth chamber evaluation of a chickpea RIL population (CP-RIL-1) revealed the presence of variability among the lines and the two parents for their days to flowering and level of resistance to ascochyta blight. Broad sense heritability across different site-years for days to flower 0.45 to 0.78, plant height 0.48 to 0.78, ascochyta blight resistance 0.14 to 0.68, days to maturity 0.26, photoperiod sensitivity 0.83 and nodes number of first flowering 0.37 to 0.75 were estimated. Days to flower and photoperiod sensitivity were significantly r = -0.21 to -0.58 (P ≤ 0.05 to 0.001) and -0.28 to -0.41 (P ≤ 0.01 to 0.001), respectively and negatively correlated with ascochyta blight resistance in the CP-RIL-1 population. A genetic linkage map consisting of eight linkage groups was developed using 349 SNP markers. Seven QTLs were identified for days to flowering under growth chamber and field conditions on chromosomes 3, 5, 6 and 8 each and 3 QTLs on chromosome 4. The total phenotypic variation explained by QTLs for days to flowering ranged from 7 to 44%. Two QTLs for days to maturity were identified on chromosomes 3 and 8. Three QTLs, one each on chromosomes 3, 4 and 5 were identified for photoperiod sensitivity. The total phenotypic variation explained by each QTL for photoperiod sensitivity ranged from 7 to 41%. A total of three QTL for node of first flowering, one on chromosomes 3 and 8 each, and two on chromosome 4 were identified. The two QTL on chromosome 4 explained total phenotypic variations of 11 and 32%, respectively. Ten QTLs distributed across all chromosomes, except chromosomes 2 and 5, were identified for ascochyta blight resistance. The phenotypic variability explained by each QTL for ascochyta blight resistance ranged from 7 to 17%. The molecular markers associated with these QTLs have potential for use in chickpea breeding.