Genetic diversity and climate adaption in "Arabidopsis lyrata"

Applied fields of research such as the one on global climate change has heightened the interest to understand the adaptive evolution process and limits to adaptive evolution. Progress in the field depends on knowing of the traits under selection and their genetic variation. The goal of my PhD thesis was to generally assess genome-wide single nucleotide polymorphism (SNP) diversity across an entire species geographic distribution and to detect SNPs and genes linked to adaptation to climatic variables and substrate type within the herbaceous plant Arabidopsis lyrata subsp. lyrata (A. lyrata). For this work, DNA of 52 populations covering the whole geographic range of A. lyrata were analyzed by pooling DNA of multiple individuals of each population, sequencing the pools (Pool-seq) and revealing population SNP frequencies. In the first chapter the wet-lab protocol of Pool-seq and the bioinformatics pipeline were tested. In the second chapter the genetic diversity of different genomic regions was analyzed to trace the history of the populations of A. lyrata. In the third chapter, the climatic variables that determine the ecological niche limits of the species distribution were defined. And, in the fourth chapter the SNP frequencies were associated with climatic variables and substrate type to detect the genomic regions involved in adaptation to climate and edaphic conditions, highlighting potentially relevant genes and pathways

Osmolyte-related recovery of the opaque-6 lethal phenotype in Zea mays L

Endosperm growth and development is a complex phenomenon, driven by the coordinate expression of several genes. A series of endosperm mutants with altered timing and zein synthesis rate have been studied, allowing the partial unravelling of a multifarious system, integrating carbohydrate, amino acid, and storage protein me¬tabolisms, and operating during endosperm growth and development. The exact biological function of one of these loci, the Opaque-6 (O6) gene, remains to be acknowledged. The o6 locus determines a general reduction of 19- and 22 kDa zeins as well as a number of non-zein polypeptides present in the wild type endosperm. The o6 mutants present a collapsed, dull endosperm, leaf striations and early seedling death; however, o6 seedlings can survive when grown in the presence of exogenous proline. It has been suggested that, in mutant seeds and in con¬trast with the development of the normal seeds, proline does not reach the sites of protein synthesis in adequate amounts. Yet, it has been demonstrated that amino acids other than proline are also able to restore o6 seedling le¬thality, contradicting this hypothesis. In this paper, we explored the possibility that the observed proline-mediated rescue of o6 mutant seedling lethality regarded an osmolyte-mediated mitigation of aberrant protein folding rather than the restoration of a reduced proline flux needed for protein synthesis. This hypothesis was tested by means of in vitro cultivation of o6 seedlings in the presence of putative osmolytes including a series of amino acids, me¬thylamines, and polyols. Several osmolytes were identified, which were able to restore normal growth in o6 mutant seedlings. Root reestablishment required higher osmolyte concentrations than those necessary for the recovery of the aboveground plant parts. The results presented in this paper provide sufficient preliminary evidence to assume that proline-induced recovery of the o6 mutant phenotype depends on the osmolytic properties of this amino acid

ngsJulia: population genetic analysis of next-generation DNA sequencing data with Julia language [version 3; peer review: 1 approved, 2 approved with reservations]

A sound analysis of DNA sequencing data is important to extract meaningful information and infer quantities of interest. Sequencing and mapping errors coupled with low and variable coverage hamper the identification of genotypes and variants and the estimation of population genetic parameters. Methods and implementations to estimate population genetic parameters from sequencing data available nowadays either are suitable for the analysis of genomes from model organisms only, require moderate sequencing coverage, or are not easily adaptable to specific applications. To address these issues, we introduce ngsJulia, a collection of templates and functions in Julia language to process short-read sequencing data for population genetic analysis. We further describe two implementations, ngsPool and ngsPloidy, for the analysis of pooled sequencing data and polyploid genomes, respectively. Through simulations, we illustrate the performance of estimating various population genetic parameters using these implementations, using both established and novel statistical methods. These results inform on optimal experimental design and demonstrate the applicability of methods in ngsJulia to estimate parameters of interest even from low coverage sequencing data. ngsJulia provide users with a flexible and efficient framework for ad hoc analysis of sequencing data.ngsJulia is available from: https://github.com/mfumagalli/ngsJulia

On the origin of the widespread self-compatible allotetraploid Capsella bursa-pastoris (Brassicaceae)

Polyploidy, or whole-genome duplication, is a common speciation mechanism in plants. An important barrier to polyploid establishment is a lack of compatible mates. Because self-compatibility alleviates this problem, it has long been hypothesized that there should be an association between polyploidy and self-compatibility (SC), but empirical support for this prediction is mixed. Here, we investigate whether the molecular makeup of the Brassicaceae self-incompatibility (SI) system, and specifically dominance relationships among S-haplotypes mediated by small RNAs, could facilitate loss of SI in allopolyploid crucifers. We focus on the allotetraploid species Capsella bursa-pastoris, which formed similar to 300 kya by hybridization and whole-genome duplication involving progenitors from the lineages of Capsella orientalis and Capsella grandiflora. We conduct targeted long-read sequencing to assemble and analyze eight full-length S-locus haplotypes, representing both homeologous subgenomes of C. bursa-pastoris. We further analyze small RNA (sRNA) sequencing data from flower buds to identify candidate dominance modifiers. We find that C. orientalis-derived S-haplotypes of C. bursa-pastoris harbor truncated versions of the male SI specificity gene SCR and express a conserved sRNA-based candidate dominance modifier with a target in the C. grandiflora-derived S-haplotype. These results suggest that pollen-level dominance may have facilitated loss of SI in C. bursa-pastoris. Finally, we demonstrate that spontaneous somatic tetraploidization after a wide cross between C. orientalis and C. grandiflora can result in production of self-compatible tetraploid offspring. We discuss the implications of this finding on the mode of formation of this widespread weed

Genomic analyses of the Linum distyly supergene reveal convergent evolution at the molecular level

Supergenes govern multi-trait-balanced polymorphisms in a wide range of systems; however, our understanding of their origins and evolution remains incomplete. The reciprocal placement of stigmas and anthers in pin and thrum floral morphs of distylous species constitutes an iconic example of a balanced polymorphism governed by a supergene, the distyly S-locus. Recent studies have shown that the Primula and Turnera distyly supergenes are both hemizygous in thrums, but it remains unknown whether hemizygosity is pervasive among distyly S-loci. As hemizygosity has major consequences for supergene evolution and loss, clarifying whether this genetic architecture is shared among distylous species is critical. Here, we have characterized the genetic architecture and evolution of the distyly supergene in Linum by generating a chromosome-level genome assembly of Linum tenue, followed by the identification of the S-locus using population genomic data. We show that hemizygosity and thrum-specific expression of S-linked genes, including a pistil-expressed candidate gene for style length, are major features of the Linum S-locus. Structural variation is likely instrumental for recombination suppression, and although the non-recombining dominant haplotype has accumulated transposable elements, S-linked genes are not under relaxed purifying selection. Our findings reveal remarkable convergence in the genetic architecture and evolution of independently derived distyly supergenes, provide a counterexample to classic inversion-based supergenes, and shed new light on the origin and maintenance of an iconic floral polymorphism.European Research Council (ERC) 757451Swedish Research Council 2019-04452, 2018-0597

ngsJulia : population genetic analysis of next-generation DNA sequencing data with Julia language

A sound analysis of DNA sequencing data is important to extract meaningful information and infer quantities of interest. Sequencing and mapping errors coupled with low and variable coverage hamper the identification of genotypes and variants and the estimation of population genetic parameters. Methods and implementations to estimate population genetic parameters from sequencing data available nowadays either are suitable for the analysis of genomes from model organisms only, require moderate sequencing coverage, or are not easily adaptable to specific applications. To address these issues, we introduce ngsJulia, a collection of templates and functions in Julia language to process short-read sequencing data for population genetic analysis. We further describe two implementations, ngsPool and ngsPloidy, for the analysis of pooled sequencing data and polyploid genomes, respectively. Through simulations, we illustrate the performance of estimating various population genetic parameters using these implementations, using both established and novel statistical methods. These results inform on optimal experimental design and demonstrate the applicability of methods in ngsJulia to estimate parameters of interest even from low coverage sequencing data. ngsJulia provide users with a flexible and efficient framework for ad hoc analysis of sequencing data.ngsJulia is available from: https://github.com/mfumagalli/ngsJuli

Demographic processes linked to genetic diversity and positive selection across a species' range

Demography determines the strength of genetic drift, which generally reduces genetic variation and the efficacy of selection. Here, we disentangled the importance of demographic processes at a local scale (census size and mating system) and at a species-range scale (old split between population clusters, recolonization after the last glaciation cycle, and admixture) in determining within-population genomic diversity and genomic signatures of positive selection. Analyses were based on re-sequence data from 52 populations of North American Arabidopsis lyrata collected across its entire distribution. The mating system and range dynamics since the last glaciation cycle explained around 60% of the variation in genomic diversity among populations and 52% of the variation in the signature of positive selection. Diversity was lowest in selfing compared with outcrossing populations and in areas further away from glacial refugia. In parallel, reduced positive selection was found in selfing populations and in populations with a longer route of postglacial range expansion. The signature of positive selection was also reduced in populations without admixture. We conclude that recent range expansion can have a profound influence on diversity in coding and non-coding DNA, similar in magnitude to the shift toward selfing. Distribution limits may in fact be caused by reduced effective population size and compromised positive selection in recently colonized parts of the range

Accumulation of Mutational Load at the Edges of a Species Range

Why species have geographically restricted distributions is an unresolved question in ecology and evolutionary biology. Here, we test a new explanation that mutation accumulation due to small population size or a history of range expansion can contribute to restricting distributions by reducing population growth rate at the edge. We examined genomic diversity and mutational load across the entire geographic range of the North American plant Arabidopsis lyrata, including old, isolated populations predominantly at the southern edge and regions of postglacial range expansion at the northern and southern edges. Genomic diversity in intergenic regions declined toward distribution edges and signatures of mutational load in exon regions increased. Genomic signatures of mutational load were highly linked to phenotypically expressed load, measured as reduced performance of individual plants and lower estimated rate of population growth. The geographic pattern of load and the connection between load and population growth demonstrate that mutation accumulation reduces fitness at the edge and helps restrict species’ distributions. distribution limits, genetic drift, genetic load, geographic expansion, leading edge, mutation accumulation, range limits, trailing edg

Box plot illustrating the distribution of the absolute difference in SNP frequency estimates between Pool-seq and GBS.

<p>The upper panel (dark grey) shows distributions when SNPs were called with VarScan for Pool-seq, the lower panel (light grey) shows distributions with Snape. Library names contain information on: the population (A or B), and the sequencing depth by Pool-seq. The band inside each box shows the median, while the lower and upper ends indicate the first and third quartile, respectively. The lower whisker is -1.5x the interquartile range from the first quartile, while the upper whisker is +1.5x the interquartile range from the third quartile. The diamonds represent outliers.</p

Histogram of minor allele frequency of GBS.

<p>The grey bars represent the SNPs present only in GBS. The striped bars represent the SNPs sequenced in the GBS and Pool-seq samples. The 10 panels show the results for the various Pool-seq library/lane combinations and the two SNP callers. The name of a library/lane combination contains information on: the population (A or B), sequencing depth per individual by Pool-seq, and the software used to detect SNPs for Pool-seq (either VarScan or Snape; for GBS, only VarScan was used).</p