<p>In this thesis the genetic variation of two grouse species, the Chinese grouse (<i>Bonasa sewersowi</i>) and the Black grouse (<i>Tetrao tetrix</i>) was examined with neutral genetic markers: microsatellites. Habitat fragmentation and isolation leads to structuring among and loss of genetic variation within populations.</p><p>The Chinese grouse in a small population in Lianhuasan nature reserve was found to have undergone a population bottleneck and as a result of isolation and possible inbreeding showed genetic impoverishment hereof.</p><p>The Black grouse populations in Europe face various different conditions from widely distributed areas of suitable habitat in the northern and eastern parts of its range to highly naturally and anthropogenically fragmented habitat landscapes in the west.</p><p>Structure among populations was found in Great Britain where Wales, Scotland and England showed characteristics of three different genetic entities, indicating very little or no geneflow between these populations. </p><p>The Dutch population showed signs of loss of genetic variation as to be expected from a population that has historically decreased in population size from several thousands to tens of individuals in a matter of decades. However the possibility to spot signs of a bottleneck was impaired due to the short time-window in which this can be observed in a population with such a low effective population size (N_E).</p><p>The sampled populations in Europe clustered into five different groups of genetic identities. The different clusters were: Great Britain-, the Netherlands-, Fenno-Scandian-, Alpine- and lowland German-Austrian populations. The level of genetic variation when compared over all these different populations decreased as a sign of isolation and small N_E. However it was not feasible to separate the impact of these two factors.</p>

Flowering dogwood (Cornus florida L.) populations have experienced severe declines caused by dogwood anthracnose in the past three decades. Mortality has ranged from 48 to 98%, raising the concern that genetic diversity of this native tree has been reduced significantly. Microsatellite data were used to evaluate the level and distribution of genetic variation throughout much of the native range of the tree. In the first conducted study, we found that genetic variation in areas affected by anthracnose was as high as or higher than areas without die-offs. We found evidence of four widespread, spatially contiguous genetic clusters. However, there was little relationship between geographic distance and genetic difference. These observations suggest that high dispersal rates and large effective population sizes have so far prevented rapid loss of genetic diversity. The effects of anthracnose on demography and community structure are likely to be far more consequential than short-term genetic effects. The second study examined levels and distribution of genetic variation of C. florida throughout Great Smoky Mountains National Park (GSMNP). Significant genetic structure at both landscape and local levels were found. We infer that two genetic clusters exist within the park, mostly separated by the main dividing ridge of the Great Smoky Mountains. The differentiation is statistically significant, but subtle, with gene flow evident through low-elevation corridors. It seems unlikely that recent demographic dynamics have resulted in a depletion of genetic variation in flowering dogwoods.

The population genetic structure of Atlantic yellowfin tuna (Thunnus albacares) has received little attention despite the substantial fishing mortality of juveniles caused by purse seining around fish aggregating devices in the Gulf of Guinea targeting multi-species schools that also include similarly sized skipjack tuna (Katsuwonus pelamis) and bigeye tuna (T. obesus). We used sequence data from 355 bp of the mitochondrial control region I as well as six microsatellite loci to examine: (1) population structure, and (2) to look for evidence of reproductive variance. We analyzed two samples of adults from the Gulf of Mexico (GOM) and one sample of early juveniles (20-50 mm) from the Gulf of Guinea (GOG). We found no evidence of geographic or temporal differentiation among the samples. Accordingly, the null hypothesis of panmixia for yellowfin tuna in the Atlantic Ocean could not be rejected. A sudden expansion analysis based on mtDNA control region I sequence data of yellowfin tuna was highly significant. Time estimates for expansion were between 40,000 and 80,000 years before present. The associated high levels of homoplasy could be masking any existing population structure. Additional sampling from additional locations and across several years will be needed to test the hypothesis of panmixia. We also provide preliminary evidence of the Allendorf-Phelps effect, which may contribute to reproductive variance. This is the first evidence of this effect in any other tuna or pelagic species. Data indicates that early juveniles sharing the same mtDNA control region I haplotype were caught in the same tow and had a significant probability of halfsibship status as calculated from their haplotype and genotype at one microsatellite locus through kinship analysis. Sampling throughout the spawning season and across several years, as well as analysis with additional microsatellite loci that have a more even distribution of alleles, will be needed to more fully identify the sibling status of larvae and early juveniles caught in the same tow as well as the extent of this reproductive variance.

Small populations are facing a particular risk of extinction due to a lack of appropriate genetic diversity and associated negative effects, factors dealt with in the discipline of conservation genetics. Many orchid species exhibit characteristics that make them a perfect study object in the scope of conservation genetics. The aim with this thesis was to investigate genetic structure at different levels in two orchid species Gymnadenia conopsea, geographically widespread, although diminishing and G. odoratissima with a long history of being rare. Microsatellite markers, developed in and used in studies of G. conopsea were also used in the study of G. odoratissima. Populations of G. conopsea expressed high levels of genetic variation and a certain amount of gene flow, although investigated mating pattern in a small population indicated non-random mating among individuals, with the majority of pollen exchange between near neighbours, and noticeable levels of geitonogamous pollinations. Further a pronounced year to year variation in flowering frequency among individuals was found. It was also discovered that flowering time variants (early and late) within the species G. conopsea were highly differentiated and seem to have had a more ancient historical separation than the separation between the two different species, G. conopsea and G. odoratissima. Levels of genetic variation in the rare congener, G. odoratissima differed between island and mainland populations where the more numerous island populations expressed larger levels of genetic variation and were less differentiated compared to the few remaining and genetically depauperated mainland populations. Uppsala University Library, Box 510, 75120, Uppsala, Sweden

In this thesis the genetic variation of two grouse species, the Chinese grouse (Bonasa sewersowi) and the Black grouse (Tetrao tetrix) was examined with neutral genetic markers: microsatellites. Habitat fragmentation and isolation leads to structuring among and loss of genetic variation within populations. The Chinese grouse in a small population in Lianhuasan nature reserve was found to have undergone a population bottleneck and as a result of isolation and possible inbreeding showed genetic impoverishment hereof. The Black grouse populations in Europe face various different conditions from widely distributed areas of suitable habitat in the northern and eastern parts of its range to highly naturally and anthropogenically fragmented habitat landscapes in the west. Structure among populations was found in Great Britain where Wales, Scotland and England showed characteristics of three different genetic entities, indicating very little or no geneflow between these populations. The Dutch population showed signs of loss of genetic variation as to be expected from a population that has historically decreased in population size from several thousands to tens of individuals in a matter of decades. However the possibility to spot signs of a bottleneck was impaired due to the short time-window in which this can be observed in a population with such a low effective population size (NE). The sampled populations in Europe clustered into five different groups of genetic identities. The different clusters were: Great Britain-, the Netherlands-, Fenno-Scandian-, Alpine- and lowland German-Austrian populations. The level of genetic variation when compared over all these different populations decreased as a sign of isolation and small NE. However it was not feasible to separate the impact of these two factors.

The aims of this thesis were to evaluate the effects of agriculture on amphibians in terms of (i) population genetic consequences of agriculture-induced spatial changes of the landscape and (ii) local adaptation and tolerance to frequently used agrochemicals. The study was performed using the common frog Rana temporaria as a model. Abundance, occurrence, genetic diversity and gene flow were negatively affected by agriculture in southern Sweden, but unaffected or even positively affected by agriculture in the central and northern regions, respectively. These test parameters correlated positively with landscape diversity both in the south and in the north. Moreover, the size and occurrence of R. temporaria populations decreased towards the north i.e. the margin of the species’ distribution range. In accordance with theoretical expectations, genetic variability decreased and population substructuring increased as a negative function of (effective) population size. Southern Swedish common frogs are naturally exposed to higher levels of nitrates, and thus have a higher tolerance to high nitrate levels than their northern conspecifics. This suggests local adaptation to naturally varying nitrate levels. Consequently, increased anthropogenic supplementation of nitrate could impact more the northern than the southern Swedish common frog populations. Exposure to the pesticides azoxystrobin, cyanazine and permethrin at ecologically relevant concentrations had small or no effects on R. temporaria tadpoles. The populations with lowest microsatellite variation (fragmented populations) in southern Sweden had considerably lower fitness in terms of survival and growth as compared to those with the highest genetic variability (non-fragmented populations). The results indicate that populations with low levels of neutral genetic variability were phenotypically less differentiated than populations with higher levels of variability. One possible explanation for this is that the degree of population differentiation in low variability populations has been constrained due to lack of suitable genetic variation or inefficiency of selection relative to genetic drift.

In this thesis, molecular genetic methods were used to study a threatened species, the Eurasian otter. Estimates of population size and population dynamics parameters were obtained, the genetic effects of a restocking program was evaluated, and a population viability analysis was conducted to assess which demographic parameters are most important for the future viability of an otter population. Many of the studies were based on noninvasive genetic sampling of faeces. In the genetic evaluation of the restocking program, it was found that the released otters had contributed to subsequent generations. However, the effects were to a large degree limited to the near surroundings of the release areas. Comparison of two census methods, snow-tracking and noninvasive genetic census based on faeces, showed that approximately only half of the otters detected with the genetic census were found with the snow-tracking census. It is recommended to combine these two methods to obtain the most reliable estimates of population size. A short-term study on population dynamics in otters showed that apparent survival was higher in females than in males and that the rate of addition was also high and likely influenced by migration. The population viability analysis incorporated both genetics and demography and revealed that survival to first reproduction was the most crucial demographic parameter affecting the viability of the study population. This result suggests that conservation efforts should be focused on protocols that enhance the survival prospects of young females. Environmental stochasticity was also found to have large effects on the probability of extinction of this population.

Our ability to map and sequence whole genomes is one of the most important developments in biological science. It will provide us with an unprecedented insight into the genetic background of phenotypic traits, such as disease resistance, reproduction and growth and also makes it feasible to study the processes of genome evolution. The main focus of this thesis has been to develop a linkage map of the horse (Equus caballus) genome. A secondary aim was to expand the number of physically mapped genes in the horse, allowing comparative analyses with data from the human genome map. Finally, attempts were made to identify single nucleotide polymorphisms (SNPs) on the horse Y chromosome. The development of a genome map relies on the information generated by both linkage and cytogenetical studies. To integrate genetical and physical assignments in the very early phase of equine genome map construction, 19 polymorphic microsatellite markers were isolated from lambda phage clones which, in parallel, were physically assigned to chromosomes by fluorescent in situ hybridization (FISH). The microsatellites were simultaneously mapped by linkage analysis in a Swedish reference pedigree. A first primary male autosomal linkage map of the domestic horse was constructed by segregation analysis of 140 genetic markers within eight half-sib families with, in total, 263 offspring. One hundred markers were arranged into 25 linkage groups, 22 of which could be assigned physically to 18 different chromosomes. The total map distance contained within linkage groups was 679 cM. The presented map provides an important framework for future genome mapping in the horse. Our contribution to the comparative horse genome map, was the presentation of map data for 12 novel genes using FISH and somatic cell hybrid mapping. AD chromosomal assignments except one were in agreement with human-horse Zoo-FISH data. The exception concerned the CLU gene which was mapped by synteny to ECA2 while human-horse Zoo-FISH data predicted that it would be located on ECA9. The level of SNPs on the horse Y chromosome was also investigated by DNA sequencing and denaturing high performance liquid chromatography (DHPLC) of Y chromosome-specific fragments derived mainly from BAC clone subcloning. The amount of genetic variability was found to be very low, consistent with low male effective population size.

The population structure of a lichen, Thamnolia subuliformis and a moss, Dicranum elongatum, was examined in northeastern regions of Wapusk National Park. With the use of microsatellite molecular markers, it was reported that the sterile lichen had a level of variation similar to that of a sexually reproducing species. The variation reported for the moss, where sporophytes were not encountered, supported subdivided populations. It was concluded that these species have the genetic resources necessary to adapt to changing environmental conditions. / February 2007

Reintroduction of Anatolian mouflon population at Bozdag Protection &amp / Breeding Station to its former habitats(Emremsultan Wildlife Development Area in Ankara-Nallihan, and Karadag in Karaman) started in 2004. The magnitude of genetic change among Bozdag and reintroduced populations was evaluated by 11 microsatellite loci. Study populations revealed close results (&plusmn / st.dev.) &ndash / Bozdag population: nk = 2.9091 (&plusmn / 1.1362), AE = 2.0250 (&plusmn / 0.9537), Ho = 0.3830 (&plusmn / 0.2717), He = 0.3956 (&plusmn / 0.2746) / Nallihan population: nk = 2.9091 (&plusmn / 1.1362), AE = 2.0592 (&plusmn / 0.9451), Ho = 0.4086 (&plusmn / 0.2977), He = 0.4052 (&plusmn / 0.2767) / and Karadag population: nk = 2.5455 (&plusmn / 1.1282), AE = 1.8809 (&plusmn / 0.8758), Ho = 0.3388 (&plusmn / 0.2775), He = 0.3607 (&plusmn / 0.2716). Population differences for major genetic parameters were not significant (p &gt / 0.05) by comparisons with paired t-test. Also, temporal change in genetic diversity for Bozdag population was investigated by comparison with temporal data. Temporal changes in genetic parameters were found to be not significant and possible causes for differences were argued. Additionally, genetic diversity and PI computations for different traps were verified and compared to uncover any potential bias due to the catching method. Comparisons did not reveal significant differences illustrating the homogeneity among traps. On the other hand, simulations detected the higher sensitivity of allelic diversity (A) to founder events than P and heterozygosity (Ho &amp / He) levels which supports heterozygosity excess method for bottleneck analysis. With the same simulation analysis, observed genetic diversity within reintroduced samples were found to be in the ranges of expectation (99% CI) indicating that translocated individuals were chosen randomly. Bottleneck analysis based on heterozygosity excess method (one-tailed test for heterozygosity excess: pSMM = 0.28515, pTPM = 0.06445, pIAM = 0.02441) and allele frequency distributions method (normal L-shaped) could not detect a recent genetic bottleneck for Bozdag population. However, simulations determined that these two methods are prone to type II error. Bottleneck detection failure for the study population is probably due to type II error instead of other sources of error like violations of model assumptions.