Evolutionary Ecology Research最新文献

Background: When threespine stickleback colonized fresh water, they repeatedly evolved reduced armour plating via changes in Eda allele frequency. This evolution is typically attributed to differential predation pressure between marine and freshwater environments. However, the chromosomal region containing Eda is associated with many other phenotypes, including schooling, antipredator behaviour, and immunity. Consequently, the evolution of armour plating may be driven by multiple selective pressures acting on Eda or linked genes.

Question: Is parasite infection associated with armour phenotype?

Hypothesis: Parasite load differs between stickleback armour plate morphs.

Organisms: An armour-polymorphic population of threespine stickleback (Gasterosteus aculeatus), and their parasites.

Field site: In June 2009 and 2012, we sampled stickleback from a single human-made salt-marsh pool in the Campbell River Estuary on Vancouver Island.

Methods: We counted macroparasites on approximately 100 fish per year and counted lateral armour plates. We used generalized linear models to test for correlations between armour morph and parasite load.

Results: Most parasite species were not associated with armour. The gill parasite Thersitina was more abundant on more fully armoured fish in both years. The nematode Eustrongylides also exhibited a marginally significant positive trend. If parasitic infections reduce stickleback fitness, this positive covariance between armour and infection would accelerate the loss of armour plating in stickleback colonizing fresh water.

Background: Stickleback fish are widely used to study the genetic and ecological basis of phenotypic evolution. Although several major loci have now been identified that contribute to evolutionary differences between wild populations, further study of the phenotypes associated with particular genes and mutations has been limited by the difficulty of generating targeted mutations at precise locations in the stickleback genome.

Approach and aims: We compared different methods of expressing single-guide RNAs (sgRNAs) and Cas9 activity in fertilized stickleback eggs. We used an easily scored pigmentation gene (SLC24A5) to screen for molecular lesions, phenotypic effects, and possible germline transmission of newly induced alleles. We then used the optimized CRISPR methods to target two major evolutionary loci in sticklebacks, KITLG and EDA. We hypothesized that coding region mutations in the KITLG gene would alter body pigmentation and possibly sex determination, and that mutations in the EDA gene would disrupt the formation of most armor plates, fin rays, spines, teeth, and gill rakers.

Results: Targeted deletions were successfully induced at each target locus by co-injecting one-cell stage stickleback embryos with either Cas9 mRNA or Cas9 protein, together with sgRNAs designed to protein-coding exons. Founder animals were typically mosaic for multiple mutations, which they transmitted through the germline at overall rates of 21 to 100%. We found that the copy of KITLG on the X chromosome (KITLGX) has diverged from the KITLG on the Y chromosome (KITLGY). Predicted loss-of-function mutations in the KITLGX gene dramatically altered pigmentation in both external skin and internal organ, but the same was not true for KITLGY mutations. Predicted loss-of-function mutations in either the KITLGX or KITLGY genes did not lead to sex reversal or prevent fertility. Homozygous loss-of-function mutations in the EDA gene led to complete loss of armor plates, severe reduction or loss of most soft rays in the dorsal, anal, and caudal fins, and severe reductions in tooth and gill raker number. In contrast, long dorsal and pelvic spines remained intact in EDA mutant animals, suggesting that common co-segregation of plate loss and spine reduction in wild populations is unlikely to be due to pleiotropic effects of EDA mutations.

Conclusion: CRISPR-Cas9 approaches can be used to induce germline mutations in key evolutionary loci in sticklebacks. Targeted coding region mutations confirm an important role for KITLG and EDA in skin pigmentation and armor plate reduction, respectively. They also provide new information about the functions of these genes in other body structures.

Background: Individual growth rates both comprise and determine life-history phenotypes. Despite decades of interest in understanding the relationship between individual growth and life history, chelonian longevity has limited our ability to robustly estimate individual growth curves that span the life of both sexes.

Questions: (1) Do patterns of growth in size and shape differ between the sexes of the painted turtle, Chrysemys picta? (2) Does individual variation in size and shape affect female reproductive effort?

Methods: Using 30 years of field data on shell morphology of a single population of painted turtles, we used principal components analysis to summarize multivariate size and shape. We assessed the ability of three non-linear growth models - the logistic, Gompertz, and von Bertalanffy - to predict size-at-age and used model comparison to justify sex-specific model fits. We correlated age-specific size and shape of females with their reproductive efforts.

Results: Model comparison supported separate fits of the von Bertalanffy growth function for each sex; non-overlapping confidence intervals imply differences in sex-specific asymptotic size, but not growth rate. Higher-order axes of variation in shell morphology described significant sexual dimorphism in shell shape related to the sphericity and curviness of the shell. Shell sphericity of females covaried with clutch size, mean egg mass, and total clutch mass. Irrespective of shell morphology, we found evidence of an egg number versus egg mass trade-off. Yet, females who matured at a larger size produced greater reproductive efforts.

Background: Threespine stickleback are an important model for behaviour and evolutionary studies. A growing number of quantitative trait loci (QTL) and gene expression studies are identifying genes related to ecologically important traits in sticklebacks. In order to visualize the expression of candidate genes, we developed a fluorescence in situ hybridization (FISH) protocol.

Methods: We present a protocol for FISH on fresh or flash-frozen dissected tissue, using either cryo- or paraffin embedding. The protocol covers probe design guidelines and synthesis, sample embedding, sectioning, and the hybridization process. The protocol is optimized for brain tissue. Key steps for modifying the protocol for other tissues are noted.

Results: The FISH protocol resulted in specific labelling under all combinations of dissection and embedding conditions. Paraffin embedding preserved morphology better than cryo-embedding. We provide representative results showing the expression of glial fibrillary acidic protein (GFAP), oxytocin receptor (OXTR), and tyrosine hydroxylase (TH) in the brain.

Background: Sexual size dimorphism (SSD) in insects often accompanies a sexual difference in development time, sexual bimaturism (SBM).

Goal: To determine whether three Aedes mosquito species have similar plasticity in SSD, attain sexual dimorphism through similar strategies, and whether SSD and SBM are associated.

Organisms: Aedes albopictus, Aedes aegypti, and Aedes triseriatus (Diptera: Culicidae).

Methods: In four different food availability environments, we quantified plastic responses of relative growth rate (RGR), development time, and adult body size in individually reared males and females.

Results: Food availability affected RGR differently for the sexes for all three species. The RGR of males and females differed significantly in the 0.1 g/L food treatment. This difference did not account for observed SSD. Food levels over which the largest changes in RGR were observed differed among the species. Male and female adult mass and development time were jointly affected by food availability in a pattern that differed among the three species, so that degree of SSD and SBM changed differentially with food availability for all three species. Development time was generally less sexually dimorphic than mass, particularly in A. albopictus. At lower food levels, A. aegypti and A. triseriatus had accentuated dimorphism in development time. These results, combined with our knowledge of mosquito life history, suggest that a direct benefit of SBM is improbable for mosquitoes and that the observed intersexual differences in development time are more likely byproducts of selection for SSD.

Background: A host obtains symbionts by horizontal transmission when infected from the environment or contagiously from other hosts in the same generation. In contrast, vertical transmission occurs when a host obtains its symbionts directly from its parents. Either vertical or horizontal transmission can sustain an association between a host and its symbiont.

Questions: What evolutionary forces are necessary to evolve from an ancestral state of horizontal transmission to a derived state of vertical transmission?

Mathematical methods: We explore a general model of fitness interaction, including both additive and epistatic effects, between host and symbiont genes. Recursion equations allow us to analyse the short-term behaviour of the model and to study long-term deterministic effects with numerical iterations.

Key assumptions: Obligate interaction between a symbiont and a single host species with genetically determined horizontal and vertical transmission. No free-living symbionts or uninfected hosts and each host is infected by only a single symbiont genetic lineage (no multiple infections). No population structure.

Conclusions: Epistasis for fitness between host and symbiont genes, like that in a matching alleles model, is a necessary condition for the evolution of vertical from horizontal transmission. Stochastic individual-based simulations show that (1) mutation facilitates the switch to vertical transmission and (2) vertical transmission is a stable evolutionary endpoint for a matching alleles model.