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Single primer enrichment technology (SPET) is a novel high-throughput genotyping method based on short-read sequencing of specific genomic regions harboring polymorphisms. SPET provides an efficient and reproducible method for genotyping target loci, overcoming the limits associated with other reduced representation library sequencing methods that are based on a random sampling of genomic loci. The possibility to sequence regions surrounding a target SNP allows the discovery of thousands of closely linked, novel SNPs. In this work, we report the design and application of the first SPET panel in lettuce, consisting of 41,547 probes spanning the whole genome and designed to target both coding (~96%) and intergenic (~4%) regions. A total of 81,531 SNPs were surveyed in 160 lettuce accessions originating from a total of 10 countries in Europe, America, and Asia and representing 10 horticultural types. Model ancestry population structure clearly separated the cultivated accessions (Lactuca sativa) from accessions of its presumed wild progenitor (L. serriola), revealing a total of six genetic subgroups that reflected a differentiation based on cultivar typology. Phylogenetic relationships and principal component analysis revealed a clustering of butterhead types and a general differentiation between germplasm originating from Western and Eastern Europe. To determine the potentiality of SPET for gene discovery, we performed genome-wide association analysis for main agricultural traits in L. sativa using six models (GLM naive, MLM, MLMM, CMLM, FarmCPU, and BLINK) to compare their strength and power for association detection. Robust associations were detected for seed color on chromosome 7 at 50 Mbp. Colocalization of association signals was found for outer leaf color and leaf anthocyanin content on chromosome 9 at 152 Mbp and on chromosome 5 at 86 Mbp. The association for bolting time was detected with the GLM, BLINK, and FarmCPU models on chromosome 7 at 164 Mbp. Associati

Rapeseed is one of the most important agricultural crops and is used in many ways. Due to the advancing climate crisis, the yield potential of rapeseed is increasingly impaired. In addition to changing environmental conditions, the expansion of cultivated areas also favours the infestation of rapeseed with various pests and pathogens. This results in the need for continuous further development of rapeseed varieties. To this end, the potential of the rapeseed gene pool should be exploited, as the various species included in it contain promising resistance alleles against pests and pathogens. In general, the biodiversity of crops and their wild relatives is increasingly endangered. In order to conserve them and to provide impulses for breeding activities as well, strategies for the conservation of plant genetic resources are necessary. In this study, we investigated to what extent the different species of the rapeseed gene pool are conserved in European genebanks and what gaps exist. In addition, a niche modelling approach was used to investigate how the natural distribution ranges of these species are expected to change by the end of the century, assuming different climate change scenarios. It was found that most species of the rapeseed gene pool are significantly underrepresented in European genebanks, especially regarding representation of the natural distribution areas. The situation is exacerbated by the fact that the natural distributions are expected to change, in some cases significantly, as a result of ongoing climate change. It is therefore necessary to further develop strategies to prevent the loss of wild relatives of rapeseed. Based on the results of the study, as a first step we have proposed a priority list of species that should be targeted for collecting in order to conserve the biodiversity of the rapeseed gene pool in the long term.

Lactuca saligna L. is a wild relative of cultivated lettuce (Lactuca sativa L.), with which it is partially interfertile. Hybrid progeny suffer from hybrid incompatibilities (HI), resulting in reduced fertility and distorted transmission ratios. Lactuca saligna displays broad spectrum resistance against lettuce downy mildew caused by Bremia lactucae Regel and is considered a non-host species. This phenomenon of resistance in L. saligna is called non-host resistance (NHR). One possible mechanism behind this NHR is through the plant–pathogen interaction triggered by pathogen-recognition receptors, including nucleotide-binding leucin-rich repeats (NLRs) and receptor-like kinases (RLKs). We report a chromosome-level genome assembly of L. saligna (accession CGN05327), leading to the identification of two large paracentric inversions (>50 Mb) between L. saligna and L. sativa. Genome-wide searches delineated the major resistance clusters as regions enriched in NLRs and RLKs. Three of the enriched regions co-locate with previously identified NHR intervals. RNA-seq analysis of Bremia infected lettuce identified several differentially expressed RLKs in NHR regions. Three tandem wall-associated kinase-encoding genes (WAKs) in the NHR8 interval display particularly high expression changes at an early stage of infection. We propose RLKs as strong candidate(s) for determinants for the NHR phenotype of L. saligna.

The collection was developed within the framework of the TKI project, entitled ‘International Lactuca Genomics Consortium (ILGC)’ (Project number:1406-039) . For this project a cross section of the lettuce gene pool, comprising nearly 500 accessions from theCGN (Centre for Genetic Resources, the Netherlands) collection, was selected for DNA resequencing purposes. Resequencing of 100 lines was initiated in 2017 within the framework of the ILGC., The collection was developed within the framework of the TKI project, entitled ‘International Lactuca Genomics Consortium (ILGC)’ (Project number:1406-039) . For this project a cross section of the lettuce gene pool, comprising nearly 500 accessions from theCGN (Centre for Genetic Resources, the Netherlands) collection, was selected for DNA resequencing purposes. Resequencing of 100 lines was initiated in 2017 within the framework of the ILGC.

Introduction: Lettuce (Lactuca sativa L.) is generally not specifically acknowledged for its taste and nutritional value, while its cultivation suffers from limited resistance against several pests and diseases. Such key traits are known to be largely dependent on the ability of varieties to produce specific phytochemicals. Objectives: We aimed to identify promising genetic resources for the improvement of phytochemical composition of lettuce varieties. Methods: Phytochemical variation was investigated using 150 Lactuca genebank accessions, comprising a core set of the lettuce gene pool, and resulting data were related to available phenotypic information. Results: A hierarchical cluster analysis of the variation in relative abundance of 2026 phytochemicals, revealed by untargeted metabolic profiling, strongly resembled the known lettuce gene pool structure, indicating that the observed variation was to a large extent genetically determined. Many phytochemicals appeared species-specific, of which several are generally related to traits that are associated with plant health or nutritional value. For a large number of phytochemicals the relative abundance was either positively or negatively correlated with available phenotypic data on resistances against pests and diseases, indicating their potential role in plant resistance. Particularly the more primitive lettuces and the closely related wild relatives showed high levels of (poly)phenols and vitamin C, thus representing potential genetic resources for improving nutritional traits in modern crop types. Conclusion: Our large-scale analysis of phytochemical variation is unprecedented in lettuce and demonstrated the ample availability of suitable genetic resources for the development of improved lettuce varieties with higher nutritional quality and more sustainable production.

thesis Ximena Cadima Fuentes (to be defended on 8 Dec 2014): Conserving the genetic diversity of Bolivian wild potatoes The wild relatives of potatoes (Solanum sect. Petota) form the genetic reservoir for the improvement of the cultivated potato. Bolivia harbours 39 wild taxa of these wild potatoes, 21 of which are endemic species. This study aimed to evaluate to what level the current ex situ and in situ management efforts have conserved the genetic diversity of Bolivian wild potato species, and what recommendations can be formulated for improvement. The current conservation status of Bolivian endemic wild potato species was assessed using both the globally accepted IUCN criteria and a methodology developed within the framework of the UNEP/GEF-Crop Wild Relative Project (CWR Project). These two methods led to different estimates of threat status for some of the species. Spatial analysis allowed to distinguish eight priority areas for in situ conservation of the 21 Bolivian endemic wild potato species. These areas represent a high concentration of endemic species and have a relatively low level of threat, but only one of them has a conservation status. This is a first step to direct the conservation efforts for wild potato species. The genetic stability and diversity of material from different species under ex situ management was evaluated using microsatellite markers. The analysis was performed on accessions that went through a process of seed regeneration and multiplication during ex situ conservation. Genetic changes between different generations of ex situ germplasm were observed for the majority, but not all, of the investigated species. Potential causes of these changes include genetic drift and contamination resulting from human error during regeneration. The populations generated under ex situ conditions were also compared with re-collected in situ populations from the same location or area as the original collection. The results showed highly signi