Understanding reproductive isolation and divergence is the focus of speciation research. Recent evidence suggested that some Dendroctonus ponderosae populations produced hybrids with reproductive incompatibilities, a reproductive boundary undetected by phylogeographic analyses using molecular markers. Additionally, the unique bifurcated distribution of D. ponderosae and the proposed isolation-by-distance gene flow pattern around the Great Basin Desert provided a unique opportunity to investigate the evolution of postmating (postyzygotic) isolation while also understanding phenotypic divergence along latitudinal (climatic) gradients. First, I characterized the strength, biological pattern, and geographic pattern of postzygotic isolation in D. ponderosae by crossing increasingly divergent populations in a common garden environment. There was little evidence of hybrid inviability in these crosses, yet geographically distant crosses produced sterile males, consistent with expectations under Haldane's rule. Hybrid male sterility appeared at a threshold among increasingly divergent populations, was bidirectional (reciprocal crosses were affected), and less geographically distant crosses did not show significant gender-specific decreases in fitness. Second, a separate investigation of two critical phenotypic traits (body size and development time) was conducted on intrapopulation F2 generation offspring from a common garden experiment. Genetic differences contributing to phenotypic variance were interpreted within the context of the previously described reproductive incompatibilities, gene flow patterns, and latitudinal gradients. Genetic differences in development time were striking between faster developing and more synchronized northern populations and slower developing, less synchronized southern populations. Differences in development time were not detected between populations at similar latitudes. Body size, although more variable than developmental time, generally conformed to expectations, with northern populations being smaller than southern populations. Average adult size was found to be quite different between many populations and did vary between populations at similar latitudes, yet relative sexual size dimorphism was rather consistent. There was no evidence of correspondence between phenotypic traits (body size and development time) and either reproductive boundaries or gene flow patterns. The results suggest that latitudinally imposed climatic differences are likely driving phenotypic divergence between populations.

The study of speciation is key to understanding the origins of biodiversity. Model organisms, with a host of genetic and molecular tools, are crucial to dissecting the genetics of speciation. Crosses between the recently-discovered Caenorhabditis sp. 9 and the well-known C. briggsae produce fertile F1 hybrid females, opening the door to genetic studies of speciation for the first time within the genus. I crossed eight different isogenic strains of C. briggsae reciprocally with six inbred lines of C. sp. 9 to investigate: Haldane’s rule, systematic asymmetries in hybrid viability, and whether genetic variation for hybrid viability segregates within each species. Results of these crosses confirm Haldane’s rule and demonstrate strong asymmetric parent-of-origin effects. Furthermore, I observed genotype-dependent differences in the number of F1 hybrid progeny. This provides evidence for genetic variation for hybrid viability within both species, allowing insights into the genetic forces driving the evolution of incompatibility loci.

The study of speciation is key to understanding the origins of biodiversity. Model organisms, with a host of genetic and molecular tools, are crucial to dissecting the genetics of speciation. Crosses between the recently-discovered Caenorhabditis sp. 9 and the well-known C. briggsae produce fertile F1 hybrid females, opening the door to genetic studies of speciation for the first time within the genus. I crossed eight different isogenic strains of C. briggsae reciprocally with six inbred lines of C. sp. 9 to investigate: Haldane’s rule, systematic asymmetries in hybrid viability, and whether genetic variation for hybrid viability segregates within each species. Results of these crosses confirm Haldane’s rule and demonstrate strong asymmetric parent-of-origin effects. Furthermore, I observed genotype-dependent differences in the number of F1 hybrid progeny. This provides evidence for genetic variation for hybrid viability within both species, allowing insights into the genetic forces driving the evolution of incompatibility loci.

Charles University in Prague Faculty of Science Ph.D. study program: Molecular and Cellular Biology, Genetics and Virology Abstract Genetics and genomics of hybrid sterility Mgr. Tanmoy Bhattacharyya Supervisor: Prof. MUDr. Jiří Forejt, DrSc. Praha 2013 Abstract Male-limited hybrid sterility restricts gene flow between the related species, an important pre- requisite of speciation. The F1 hybrid males of PWD/Ph female (Mus m. musculus subspecies) and C57BL/6J or B6 male (Mus m. domesticus) are azoospermic and sterile (PB6F1), while the hybrids from the reciprocal (B6PF1) cross are semi fertile. A disproportionately large effect of the X chromosome (Chr) on hybrid male sterility is a widespread phenomenon accompanying the origin of new species. In the present study, we mapped two phenotypically distinct hybrid sterility loci Hstx1 and Hstx2 to a common 4.7 Mb region on Chr. X. Analysis of meiotic prophase I of PB6F1 sterile males revealed meiotic block at mid-late pachynema and the TUNEL assay showed apoptosis of arrested spermatocytes. In sterile males over 95% of pachytene spermatocytes showed one or more unsynapsed autosomes visualized by anti SYCP1, HORMAD2 and SYCP3 antibodies. The phosphorylated form of H2AFX histone, normally restricted only to XY chromosome containing sex body decorated unsynapsed...