Haplotype estimation in polyploids using DNA sequence data

Polyploid organisms possess more than two copies of their core genome and therefore contain k>2 haplotypes for each set of ordered genomic variants. Polyploidy occurs often within the plant kingdom, among others in important corps such as potato (k=4) and wheat (k=6). Current sequencing technologies enable us to read the DNA and detect genomic variants, but cannot distinguish between the copies of the genome, each inherited from one of the parents. To detect inheritance patterns in populations, it is necessary to know the haplotypes, as alleles that are in linkage over the same chromosome tend to be inherited together. In this work, we develop mathematical optimisation algorithms to indirectly estimate haplotypes by looking into overlaps between the sequence reads of an individual, as well as into the expected inheritance of the alleles in a population. These algorithm deal with sequencing errors and random variations in the counts of reads observed from each haplotype. These methods are therefore of high importance for studying the genetics of polyploid crops. </p

High-throughput computational methods and software for quantitative trait locus (QTL) mapping

De afgelopen jaren zijn vele nieuwe technologieen zoals Tiling arrays en High throughput DNA sequencing een belangrijke rol gaan spelen binnen het onderzoeksveld van de systeem genetica. Voor onderzoekers is het extreem belangrijk om te begrijpen dat deze methodes hun manier van werken zullen gaan beinvloeden. Deit proefschrift beschrijft mogelijke oplossingen voor deze 'Big Data' lawine die systemen genetica heeft getroffen.Dit proefschrift beschrijft de werkzaamheden uitgevoerd aan het Groningen Bioinformatics Centre om slimmere en geoptimaliseerde algoritmen zoals Pheno2Geno en MQM te ontwikkelen en een systeem om 'collaborative' research mogelijk te maken genaamd xQTL werkbank om door middel van high-throughput systemen genetica data te analyseren.In recent years many new technologies such as tiling arrays and high-throughput sequencinghave come to play an important role in systems genetics research. For researchers it is ofthe utmost importance to understand how this affects their research. This work describespossible solutions to this ‘Big Data’ avalanche which has hit systems genetics.This thesis describes the work carried out during the author’s 4 year PHD project at theGroningen Bioinformatics Centre to develop smarter and more optimized algorithms suchas Pheno2Geno and MQM, and to use a collaborative approach such as xQTL workbench tostore and analyse high-throughput systems genetics data

Parsimony-based genetic algorithm for haplotype resolution and block partitioning

This dissertation proposes a new algorithm for performing simultaneous haplotype resolution and block partitioning. The algorithm is based on genetic algorithm approach and the parsimonious principle. The multiloculs LD measure (Normalized Entropy Difference) is used as a block identification criterion. The proposed algorithm incorporates missing data is a part of the model and allows blocks of arbitrary length. In addition, the algorithm provides scores for the block boundaries which represent measures of strength of the boundaries at specific positions. The performance of the proposed algorithm was validated by running it on several publicly available data sets including the HapMap data and comparing results to those of the existing state-of-the-art algorithms. The results show that the proposed genetic algorithm provides the accuracy of haplotype decomposition within the range of the same indicators shown by the other algorithms. The block structure output by our algorithm in general agrees with the block structure for the same data provided by the other algorithms. Thus, the proposed algorithm can be successfully used for block partitioning and haplotype phasing while providing some new valuable features like scores for block boundaries and fully incorporated treatment of missing data. In addition, the proposed algorithm for haplotyping and block partitioning is used in development of the new clustering algorithm for two-population mixed genotype samples. The proposed clustering algorithm extracts from the given genotype sample two clusters with substantially different block structures and finds haplotype resolution and block partitioning for each cluster

Breeding Innovations in Underutilized Temperate Fruit Trees

The recent growing interest in minor species (i.e., fig, pomegranate, feijoa, etc.) has recently driven new research on breeding and genetics to address producer and consumer traits. Since these species have received little attention from the scientific community, they were less improved via conventional breeding, and lacked detailed genomic information on important traits. This lack of data, together with a general poor genetic knowledge of these species, has limited a wider cultivation of varieties with improved characteristics. For these reasons, and with the objective to increase the interest of scientists, farmers, and consumers for these fruits, this Research Topic “Breeding Innovations in Underutilized Temperate Fruit Trees” comprises biochemical, morphological, and genetic studies on some minor species regarding fruit trait variation, resistance, evolution, or sex determination. In this context, Marcotuli et al. examined the mechanisms behind the bud evolution toward breba or main crop in fig (Ficus carica L.), since this aspect remains unclear. The X-ray images of buds showed a great structural similarity between breba and the main crop during the initial stages of development, but breba inflorescence differentiation was completed in two seasons whereas that of main crop started at the end of winter and was completed within 2–3 months in the same season. The higher expression of floral homeotic protein AGAMOUS in breba compared to the main crop may indicate a role of these fruits on staminate flowers’ production for pollination of the main crop, as profichi act in the caprifig. Within the same species (Ficus carica L.) and for sexual determination, Ikegami et al. analyzed the FcRAN1 gene (during a breeding program for the selection of female plants) strongly associated with the sex phenotype. A male-biased segregation ratio distortion was obtained in 12 F1 populations, suggesting some genetic factor(s) affecting it. A comparison between the annotated genes and the genes required for normal embryo or gametophyte development and function identified several candidate genes responsible for the segregation distortion in fig. Following the same topic, Wang et al. hypothesized an early sex-identification method to improve breeding efficiency. The use of a deletion as a forward primer, a newly established AG-Marker, was as accurate as the RAN1-Marker, and provided the identification of male plants, giving new clues to understanding Ficus sex determination. Moving toward another attracting species, i.e., pomegranate (Punica granatum L.), Trainin et al. investigated the black peel color of some pomegranate varieties. Biochemical analysis revealed that delphinidin is highly abundant in the peel of black varieties and the pattern of anthocyanin accumulation is different from that of other pomegranates with red or pink colors of the peel. Genetic analysis of an F2 population segregating for the black phenotype revealed that it is determined by a single recessive gene. Pomegranate was also studied by Goudappa Patil et al. with regards to the SSR of “Tunisia” pomegranate variety. There was a positive trend in chromosome length and the SSR abundance, as marker density, enhanced with a shorter chromosome length. Examination of the distribution of SSR motif types within a chromosome suggested the abundance of hexanucleotide repeats in each chromosome followed by dinucleotides. A comprehensive set of highly polymorphic genome-wide SSRs was successively developed and tested. These chromosome-specific SSRs could serve as a powerful genomic tool to leverage future genetic studies, germplasm management, and genomics-assisted breeding in pomegranate varieties. Some evolutionary aspects of pear were investigated by Kumar et al. who put a light on runs of homozygosity (ROH) in self-incompatible plants, in particular Asian pears, European pears, and interspecific hybrids using genotyping-by-sequencing. The observed ROH patterns suggested that systematic breeding of European pears would have started earlier than Asian pears. Fruit trait variation in Persian walnut (Juglans regia L.) was addressed by Bernard et al. who conducted a genome-wide association study (GWAS) using multi-locus models in a panel of 170 accessions of J. regia to elucidate the genetic determinants of fruit quality traits in walnut toward the breeding of new varieties. The authors proposed several candidate genes involved in nut characteristics, such as a gene coding for a beta-galactosidase linked to several size-related traits and known to also be involved in fruit development in other species. With regards of fruit traits, Kyratzis et al. investigated the germplasm of an ancient species, the carob (Ceratonia siliqua L.), on the island of Cyprus. The domestic germplasm varies both in terms of pod morphology and composition, reflecting the genetic and physiological characteristics of both grafted and non-grafted accessions, and possibly the impact of agro-environmental conditions. Morphological traits, such as seeds-to-pod weight ratio, pod width, and thickness, were principally under genetic control. Contrarily, chemical compounds, particularly total phenolic content, including condensed tannins, in vitro antioxidant capacity, and to a lesser extent gallic acid, organic acids, sugars (glucose and fructose), and minerals were more under agro-environmental control. In the Southern Hemisphere, Quezada et al. worked on feijoa (Acca sellowiana Berg.), a fruit tree species native to Uruguay and Brazil. A high-density composite genetic linkage map of feijoa was constructed using two genetically populations. Genotyping by sequencing (GBS) approach was successfully applied for developing single nucleotide polymorphism (SNP) markers. They used both the reference genome of the closely related species Eucalyptus grandis and a de novo pipeline to construct a composite map. A novel approach for the construction of composite maps where the meiosis information of individuals of two connected populations is captured in a single estimator is described. The topic of resistance was carried out by Alves et al. in order to find sources of genetic resistance to Huanglongbing (HLB)-associated “Candidatus Liberibacter asiaticus” (Las), one of the most destructive diseases of citrus. Some genotypes from subtribe Citrinae, sexually incompatible but graft-compatible with Citrus, may provide new rootstocks able to restrict bacterial titer in the canopy. Authors tested for Las resistance a wide collection of graft-compatible Citrinae species using an aggressive and consistent challenge-inoculation and evaluation procedure. Eremocitrus glauca and Papua/New Guinea Microcitrus species as well as their hybrids resulted in full resistance, opening the way for using these underutilized genotypes as Las resistance sources in breeding programs

Data analysis methods for copy number discovery and interpretation

Copy number variation (CNV) is an important type of genetic variation that can give rise to a wide variety of phenotypic traits. Differences in copy number are thought to play major roles in processes that involve dosage sensitive genes, providing beneficial, deleterious or neutral modifications to individual phenotypes. Copy number analysis has long been a standard in clinical cytogenetic laboratories. Gene deletions and duplications can often be linked with genetic Syndromes such as: the 7q11.23 deletion of Williams-­‐Bueren Syndrome, the 22q11 deletion of DiGeorge syndrome and the 17q11.2 duplication of Potocki-­‐Lupski syndrome. Interestingly, copy number based genomic disorders often display reciprocal deletion / duplication syndromes, with the latter frequently exhibiting milder symptoms. Moreover, the study of chromosomal imbalances plays a key role in cancer research. The datasets used for the development of analysis methods during this project are generated as part of the cutting-­‐edge translational project, Deciphering Developmental Disorders (DDD). This project, the DDD, is the first of its kind and will directly apply state of the art technologies, in the form of ultra-­‐high resolution microarray and next generation sequencing (NGS), to real-­‐time genetic clinical practice. It is collaboration between the Wellcome Trust Sanger Institute (WTSI) and the National Health Service (NHS) involving the 24 regional genetic services across the UK and Ireland. Although the application of DNA microarrays for the detection of CNVs is well established, individual change point detection algorithms often display variable performances. The definition of an optimal set of parameters for achieving a certain level of performance is rarely straightforward, especially where data qualities vary ... [cont.]