Environmental and Life Sciences

To investigate cytokinins (CKs) in nematodes, CK profiles of a free-living Caenorhabditis elegans and a plant parasitic Heterodera glycines (soybean cyst nematode, SCN) were determined at the egg and larval stages. SCN had higher total CK level than C. elegans; however, CKs in SCN were mostly inactive precursors, whereas C. elegans had more bioactive forms. This is the first study to show that methylthiols are present in nematodes and may affect plant infection. In infectious SCN larvae, methylthiol levels were much higher than in eggs or C. elegans larvae. Furthermore, The CK profiles of SCN-susceptible and resistant Glycine max cultivars at three developmental stages revealed that, regardless of the resistance level, SCN infection caused an increase in root CKs. One resistant cultivar, Pion 93Y05, showed significantly high levels of bioactive N6-isopentenyladenine (iP) in the non-infected roots which indicated a potential role of CKs in soybean resistance to SCN.

Author Keywords: Cytokinins, HPLC-MS/MS, Nematode, SCN resistance, Soybean

Interspecific hybridization rates can vary depending on genomic compatibilities between progenitors, while subsequent hybrid spread can vary depending on hybrid performance and habitat availability for hybrid establishment and persistence. As a result, hybridization rates and hybrid frequencies can vary across regions of parental sympatry. In areas around the Laurentian Great Lakes, Typha × glauca is an invasive plant hybrid of native Typha latifolia and introduced Typha angustifolia. In areas of parental sympatry in Atlantic Canada and outside of North America, T. × glauca has been reported as either rare or non-existent. I investigated whether the low frequencies of hybrids documented in Nova Scotia, Atlantic Canada, are influenced by reproductive barriers that prevent hybrid formation or environmental factors (salinity) that reduce hybrid performance. I identified an abundance of hybrids in the Annapolis Valley (inland) and a scarcity of hybrids in coastal wetlands through preliminary site surveys throughout Nova Scotia. In Annapolis Valley populations, flowering times of progenitor species overlapped, indicating that asynchronous flowering times do not limit hybrid formation in this region. Viable progeny were created from interspecific crosses of T. latifolia and T. angustifolia from Nova Scotia, indicating that there are no genomic barriers to fertilization and germination of hybrid seeds. Typha × glauca germination in high salinity was significantly lower than that of T. latifolia, but there was no difference at lower salinities. Therefore, while germination of hybrid seeds may be impeded in the coastal wetlands where salinity is high, inland sites have lower salinity and thus an environment conducive to hybrid germination. However, I found that once established as seedlings, hybrids appear to have greater performance over T. latifolia across all salinities through higher ramet production. Moreover, I found that T. latifolia sourced from Ontario had reduced germination and lower survivorship in high salinities compared to T. latifolia sourced from Nova Scotia, which could indicate local adaptation by T. latifolia to increased salinity. These findings underline that interactions between environment and local progenitor lineages can influence the viability and the consequent distribution and abundance of hybrids. This, in turn, can help explain why hybrids demonstrate invasiveness in some areas of parental sympatry but remain largely absent from other areas.

Author Keywords: flowering phenology, Hybridization, invasive species, physiology, pollen compatibility, salinity tolerance

Glucocorticoids (e.g., corticosterone (CORT)) are hypothesized to mediate decisions regarding reproductive investment during breeding, but the directionality of the relationship is not clear. The CORT-fitness hypothesis posits that high levels of CORT arise from challenging environmental conditions in which an individual will conserve resources for future reproduction or self-maintenance, and thus result in lower reproductive success (a negative relationship). In contrast, the CORT-adaptation hypothesis suggests that, during energetically demanding periods, CORT will mediate physiological or behavioural changes that result in increased reproductive investment and success (a positive relationship). Inconsistencies arise due to the various species and life-history stages studied, and the complex interactions between fitness and glucocorticoids. Using an experimental approach, I investigated the relationship between CORT and reproductive success by manipulating baseline CORT levels in female tree swallows Tachycineta bicolor prior to laying using silastic implants. Implants failed to raise CORT levels of females during either incubation or the nestling stage, and maternal treatment had no effect on indices of fitness at either stage. Using a correlative approach, partial support for the CORT-adaptation hypothesis was found: There was a positive relationship between CORT and hatching success. This only occurred when CORT was measured during incubation, when baseline CORT levels may stimulate increased reproductive effort and success. In contrast, during the nestling stage baseline CORT levels were not related to reproductive investment or success. Maternal CORT levels during incubation also did not influence nestling phenotype, although nestling stress CORT levels were higher in individuals that survived to fledging. In conclusion, CORT mediates reproductive effort and success during some breeding stages, but it is still unclear why this is the case and whether this same pattern will prevail in other contexts.

Author Keywords: corticosterone, HPA axis, maternal effects, reproductive success, tree swallow

Ustilago maydis (D.C.) Corda is a biotrophic pathogen that secretes effectors to establish and maintain a relationship with its host, Zea mays. In this pathosystem, the molecular function of effectors is well-studied, but the regulation of effector gene expression remains largely unknown. This study characterized Zfp1, a putative U. maydis Zn(II)2Cys6 transcription factor, as a modulator of effector gene expression. The amino acid sequence of Zfp1 indicated the presence of a GAL4-like zinc binuclear cluster as well as a fungal specific transcription factor domain. Nuclear localization was confirmed by tagging Zfp1 with enhanced green fluorescent protein. Deletion of zfp1 resulted in attenuated hyphal growth, reduced infection frequency, an arrest in pathogenic development, and decreased anthocyanin production. This phenotype can be attributed to the altered transcript levels of genes encoding predicted and confirmed U. maydis effectors in the zfp1 deletion strain during pathogenic growth. Complementation of zfp1 deletion strain with tin2, an effector involved in anthocyanin induction, suggested this effector is downstream of Zfp1 and its expression is influenced by this transcription factor during in planta growth. When wild-type zfp1 was ectopically inserted in the zfp1 deletion strain, pathogenesis and virulence were partially restored. This, coupled with zfp1 over-expression strains having a similar phenotype as the deletion strains, suggested Zfp1 may interact with other proteins for full function. These findings show that Zfp1, in conjunction with one or more binding partners, contributes to U. maydis pathogenesis, virulence, and anthocyanin production through the regulation of effector gene expression.

Author Keywords: effector, pathogenesis, transcription factor, Ustilago maydis, Zea mays, zinc finger

Coldwater streams are becoming increasingly impacted due to urbanization. Using environmental surveys, mark-recapture and telemetry, I assessed factors influencing seasonal brook trout (Salvelinus fontinalis) habitat use and movement in urban headwater streams in central Ontario between 2017-18. Generalized additive models were used to assess which habitat variables best explained seasonal yearling and older brook trout abundance, while generalized least squares models were used to assess overall trends in radio-tagged brook trout movement. My research demonstrated dynamic patterns in habitat use and movement by urban stream-dwelling brook trout. Yearlings were primarily influenced by water quality (stream temperature, conductivity), while older brook trout were most strongly influenced by stream morphology (depth, undercut bank). Movement occurred disproportionately around the spawning season and was more limited in the smaller, more altered stream. These findings may be used to inform fisheries managers on crucial timing and location of brook trout habitat refugia within urbanized environments.

Author Keywords: Brook trout, coldwater stream, groundwater, habitat use, radiotelemetry, urbanization

Invasive Phragmites australis ssp. australis (herein "Phragmites") has established and rapidly spread throughout many coastal areas of the Great Lakes. Known to displace native vegetation communities as it forms large, monotypic stands, Phragmites has a bad reputation when it comes to losses of biodiversity and habitat provision for wildlife. However, the extent to which Phragmites provides habitat for fish and invertebrates in coastal freshwater wetlands remains relatively unquantified. Thus, this study assessed whether fish assemblages and invertebrate communities in stands of Phragmites differ from those in stands of two native emergent vegetation communities, Typha spp. and Schoenoplectus spp. The findings showed significant differences in habitat variables among the vegetation communities in terms of water depth, macrophyte species richness, stem density and water quality. While abundance of the functional feeding group filterer-collectors was found to be significantly less in stands of Phragmites when compared to Schoenoplectus, no difference was observed in invertebrate taxa richness among vegetation communities. Lastly, no difference in fish assemblage or invertebrate community was detected when using multivariate analyses, implying that invasive Phragmites provides habitat that appears to be as valuable for fish and invertebrates as other emergent vegetation types in the St. Clair River Delta. The findings of this study will ultimately benefit the literature on invasive Phragmites and its role as fish habitat in freshwater wetlands, and aid management agencies in decisions regarding control of the invasive species.

Author Keywords: aquatic invasive species, aquatic macroinvertebrates, freshwater fish, freshwater wetlands, nMDS, Phragmites

Syrphids, also known as hover flies (Diptera: Syrphidae) are a diverse and widespread family of flies. Here, I report on their distributions from a previously understudied region, the far north of Ontario, as well as Akimiski Island, Nunavut. I used samples collected through a variety of projects to update known range and provincial records for over a hundred species, bringing into clearer focus the distribution of syrphids throughout this region. I also analysed a previously un-tested trap type for collecting syrphids (Nzi trap), and report on results of DNA analysis for a handful of individuals, which yielded a potential new species.

Author Keywords: Diptera, Ontario, range extension, Syrphidae

Competition is known to impact population dynamics through both indirect and direct interactions, and direct interactions can often lead to injury in one or both parties. As such, response to injury through tissue regeneration can be important for surviving post-competitive interaction. However, the impacts of outside factors like temperature and genome size (e.g. polyploidy) are not well studied, especially in syntopic systems. We addressed this knowledge gap by comparing regeneration rates of diploid Ambystoma laterale and triploid unisexual Ambystoma at two ecologically-relevant temperatures. Environmental factors appeared to have stronger effects on regeneration than ploidy level, but overall mass was impacted more strongly by ploidy level. Interestingly, there was an interaction between temperature and time within unisexuals that was absent when comparing different ploidy levels, implying temperature has a more complex effect on polyploids. This study supports the hypothesis that polyploid organisms are better equipped to respond to shifts in their environments, which can give them a competitive advantage at the northern range limit of this species complex.

Author Keywords: Ambystoma, Genome dosage, Hybrid vigor, Polyploidy, Thermal optimum, Tissue regeneration

Predation risk can elicit a range of responses in prey, but to date little is known about breadth of potential responses that may arise under realistic field conditions and how such responses are linked, leaving a fragmented picture of risk-related consequences on individuals. We increased predation risk in free-ranging snowshoe hares (Lepus americanus) during two consecutive summers by simulating natural chases using a model predator (i.e., domestic dog), and monitored hare stress physiology, energy expenditure, behaviour, condition, and habitat use. We show that higher levels of risk elicited marked changes in physiological stress metrics including sustained high levels of free plasma cortisol which had cascading effects on glucose, and immunology, but not condition. Risk-augmented hares also had lowered daily energy expenditure, spent more time foraging, and decreased rest, vigilance, and travel. It is possible that these alterations allowed risk-exposed hares to increase their condition at the same rate as controls. Additionally, risk-augmented hares selected, had high fidelity to, and were more mobile in structurally dense habitat (i.e., shrubs) which provided them additional cover from predators. They also used more open habitat (i.e., conifer) differently based on locale within the home range, using familiar conifer areas within cores for rest while moving through unfamiliar conifer areas in the periphery. Overall, these findings show that prey can have a multi-faceted, highly plastic response in the face of risk and can mitigate the effects of their stress physiology given the right environmental conditions.

Author Keywords: behaviour, condition, daily energy expenditure, predator-prey interactions, snowshoe hare, stress physiology

Understanding the complex genomic architecture underlying quantitative traits can provide valuable insight for the conservation and management of wildlife. Despite improvements in sequencing technologies, few empirical studies have identified quantitative trait loci (QTL) via whole genome sequencing in free-ranging mammal populations outside a few well-studied systems. This thesis uses high-depth whole genome pooled re-sequencing to characterize the molecular basis of the natural variation observed in two sexually selected, heritable traits in white-tailed deer (Odocoileus virginianus, WTD). Specifically, sampled individuals representing the phenotypic extremes from an island population of WTD for antler and body size traits. Our results showed a largely homogenous genome between extreme phenotypes for each trait, with many highly differentiated regions throughout the genome, indicative of a quantitative model for polygenic traits. We identified and validated several potential QTL of putatively small-to-moderate effect for each trait, and discuss the potential for real-world application to conservation and management.

Author Keywords: evolution, extreme phenotypes, genetics, genomics, quantitative traits, sexual selection