The Roles of Phenotypic Plasticity and Plant-Microbe Interactions in the Evolution of Complex Traits in Boechera stricta

All organisms live in complex habitats that shape the course of their evolution by altering the phenotype expressed by a given genotype (a phenomenon known as phenotypic plasticity) and simultaneously by determining the evolutionary fitness of that phenotype. In some cases, phenotypic evolution may alter the environment experienced by future generations. This dissertation describes how genetic and environmental variation act synergistically to affect the evolution of glucosinolate defensive chemistry and flowering time in Boechera stricta, a wild perennial herb. I focus particularly on plant-associated microbes as a part of the plant’s environment that may alter trait evolution and in turn be affected by the evolution of those traits. In the first chapter I measure glucosinolate production and reproductive fitness of over 1,500 plants grown in common gardens in four diverse natural habitats, to describe how patterns of plasticity and natural selection intersect and may influence glucosinolate evolution. I detected extensive genetic variation for glucosinolate plasticity and determined that plasticity may aid colonization of new habitats by moving phenotypes in the same direction as natural selection. In the second chapter I conduct a greenhouse experiment to test whether naturally-occurring soil microbial communities contributed to the differences in phenotype and selection that I observed in the field experiment. I found that soil microbes cause plasticity of flowering time but not glucosinolate production, and that they may contribute to natural selection on both traits; thus, non-pathogenic plant-associated microbes are an environmental feature that could shape plant evolution. In the third chapter, I combine a multi-year, multi-habitat field experiment with high-throughput amplicon sequencing to determine whether B. stricta-associated microbial communities are shaped by plant genetic variation. I found that plant genotype predicts the diversity and composition of leaf-dwelling bacterial communities, but not root-associated bacterial communities. Furthermore, patterns of host genetic control over associated bacteria were largely site-dependent, indicating an important role for genotype-by-environment interactions in microbiome assembly. Together, my results suggest that soil microbes influence the evolution of plant functional traits and, because they are sensitive to plant genetic variation, this trait evolution may alter the microbial neighborhood of future B. stricta generations. Complex patterns of plasticity, selection, and symbiosis in natural habitats may impact the evolution of glucosinolate profiles in Boechera stricta.

Interactions between nematodes, potato and soil-borne micro-organisms and their effect on the management of potato cyst nematodes

Potato cyst nematodes (PCN) cause significant damage to the potato crop worldwide and growers experience economic losses related to yield loss and the cost of control measures. Experiments were set up to further elucidate the complex tritrophic PCNpotato-soil bacteria relationship. Bacterial strains isolated from the sugar beet rhizosphere were shown to be hatch active towards Globodera pallida and to be capable of successfully colonising the sugar beet rhizosphere when applied exogenously. A trap-crop system, based on these isolates, was proposed. Ridge and bulk soil taken from a commercial potato field were incubated with sterile potato root leachate (sPRL) and subsequent in vitro hatching assays showed that PCN hatch was influenced by microorganisms present in the ridge, but not in the bulk soil. Community level physiological profiling (CLPP) of ridge and bulk soil, using BIOLOG EcoplatesTM, demonstrated differences in bacterial functional diversity between the two soil types. An investigation of the inter-species competition between G. pallida and G. rostochiensis showed that G. pallida performed significantly better, in terms of multiplication rate, in competition with G. rostochiensis compared to its multiplication rate in single-species populations. Effectively removing the early hatch of G. rostochiensis in pot trials led to the removal of this competitive advantage of G. pallida suggesting that this advantage was due, at least in part, to morphological changes to the root caused by the early hatching of G. rostochiensis.

Host genotype and age shape the leaf and root microbiomes of a wild perennial plant.

Bacteria living on and in leaves and roots influence many aspects of plant health, so the extent of a plant's genetic control over its microbiota is of great interest to crop breeders and evolutionary biologists. Laboratory-based studies, because they poorly simulate true environmental heterogeneity, may misestimate or totally miss the influence of certain host genes on the microbiome. Here we report a large-scale field experiment to disentangle the effects of genotype, environment, age and year of harvest on bacterial communities associated with leaves and roots of Boechera stricta (Brassicaceae), a perennial wild mustard. Host genetic control of the microbiome is evident in leaves but not roots, and varies substantially among sites. Microbiome composition also shifts as plants age. Furthermore, a large proportion of leaf bacterial groups are shared with roots, suggesting inoculation from soil. Our results demonstrate how genotype-by-environment interactions contribute to the complexity of microbiome assembly in natural environments.

Bridging the Scale Gap from Leaf to Canopy in Biosphere-Atmosphere Gas and Particle Exchanges

Terrestrial ecosystems, occupying more than 25% of the Earth's surface, can serve as

`biological valves' in regulating the anthropogenic emissions of atmospheric aerosol

particles and greenhouse gases (GHGs) as responses to their surrounding environments.

While the signicance of quantifying the exchange rates of GHGs and atmospheric

aerosol particles between the terrestrial biosphere and the atmosphere is

hardly questioned in many scientic elds, the progress in improving model predictability,

data interpretation or the combination of the two remains impeded by

the lack of precise framework elucidating their dynamic transport processes over a

wide range of spatiotemporal scales. The diculty in developing prognostic modeling

tools to quantify the source or sink strength of these atmospheric substances

can be further magnied by the fact that the climate system is also sensitive to the

feedback from terrestrial ecosystems forming the so-called `feedback cycle'. Hence,

the emergent need is to reduce uncertainties when assessing this complex and dynamic

feedback cycle that is necessary to support the decisions of mitigation and

adaptation policies associated with human activities (e.g., anthropogenic emission

controls and land use managements) under current and future climate regimes.

With the goal to improve the predictions for the biosphere-atmosphere exchange

of biologically active gases and atmospheric aerosol particles, the main focus of this

dissertation is on revising and up-scaling the biotic and abiotic transport processes

from leaf to canopy scales. The validity of previous modeling studies in determining

iv

the exchange rate of gases and particles is evaluated with detailed descriptions of their

limitations. Mechanistic-based modeling approaches along with empirical studies

across dierent scales are employed to rene the mathematical descriptions of surface

conductance responsible for gas and particle exchanges as commonly adopted by all

operational models. Specically, how variation in horizontal leaf area density within

the vegetated medium, leaf size and leaf microroughness impact the aerodynamic attributes

and thereby the ultrane particle collection eciency at the leaf/branch scale

is explored using wind tunnel experiments with interpretations by a porous media

model and a scaling analysis. A multi-layered and size-resolved second-order closure

model combined with particle

uxes and concentration measurements within and

above a forest is used to explore the particle transport processes within the canopy

sub-layer and the partitioning of particle deposition onto canopy medium and forest

oor. For gases, a modeling framework accounting for the leaf-level boundary layer

eects on the stomatal pathway for gas exchange is proposed and combined with sap

ux measurements in a wind tunnel to assess how leaf-level transpiration varies with

increasing wind speed. How exogenous environmental conditions and endogenous

soil-root-stem-leaf hydraulic and eco-physiological properties impact the above- and

below-ground water dynamics in the soil-plant system and shape plant responses

to droughts is assessed by a porous media model that accommodates the transient

water

ow within the plant vascular system and is coupled with the aforementioned

leaf-level gas exchange model and soil-root interaction model. It should be noted

that tackling all aspects of potential issues causing uncertainties in forecasting the

feedback cycle between terrestrial ecosystem and the climate is unrealistic in a single

dissertation but further research questions and opportunities based on the foundation

derived from this dissertation are also brie

y discussed.

Effects of vermicomposted spent mushroom compost on growing medium characteristics, plant growth, yield and abiotic stress tolerance

Treatment of agricultural biodegradable wastes and by-products can be carried out using composting or vermicomposting, or a combination of both treatment methods, to create a growing medium amendment suitable for horticultural use. When compared to traditional compost-maturation, vermicompost-maturation resulted in a more mature growing medium amendment i.e. lower C/N and pH, with increased nutrient content and improved plant growth response, increasing lettuce shoot fresh and dry weight by an average of 15% and 14%, respectively. Vermicomposted horse manure compost was used as a growing medium amendment for lettuce and was found to significantly increase lettuce shoot and root growth, and chlorophyll content. When used as a growing medium amendment for tomato fruit production, vermicomposted spent mushroom compost increased shoot growth and marketable yield, and reduced blossom end rot in two independent studies. Vermicompost addition to peat-based growing media increased marketable yield by an average of 21%. Vermicompost also improved tomato fruit quality parameters such as acidity and sweetness. Fruit sweetness, as measured using Brix value, was significantly increased in fruits grown with 10% or 20% vermicompost addition by 0.2 in truss one and 0.3 in truss two. Fruit acidity (% citric acid) was significantly increased in plants grown with vermicompost by an average of 0.65% in truss one and 0.68% in truss two. These changes in fruit chemical parameters resulted in a higher tomato fruit overall acceptability rating as determined by a consumer acceptance panel. When incorporated into soil, vermicomposted spent mushroom compost increased plant growth and reduced plant stress under conditions of cold stress, but not salinity or heat stress. The addition of 20% vermicompost to cold-stressed plants increased plant growth by an average of 30% and increased chlorophyll fluorescence by an average of 21%. Compared to peat-based growing medium, vermicompost had consistently higher nutrient content, pH, electrical conductivity and bulk density, and when added to a peat-based growing medium, vermicomposted spent mushroom compost altered the microbial community. Vermicompost amendment increased the microbial activity of the growing medium when incorporated initially, and this increased microbial activity was observed for up to four months after incorporation when plants were grown in it. Vermicomposting was shown to be a suitable treatment method for agricultural biodegradable wastes and by-products, with the resulting vermicompost having suitable physical, chemical and biological properties, and resulting in increased plant growth, marketable yield and yield quality, when used as an amendment in peat-based growing medium.

Field data on methane fluxes, temperature, soil gas profiles and microbial activities in ponds of the northeast Siberian polygonal tundra

The data files give the basic field and laboratory data on five ponds in the northeast Siberian Arctic tundra on Samoylov. The files contain water and soil temperature data of the ponds, methane fluxes, measured with closed chambers in the centres without vascular plants and the margins with vascular plants, the contribution of plant mediated fluxes on total methane fluxes, the gas concentrations (methane and dissolved inorganic carbon, oxygen) in the soil and the water column of the ponds, microbial activities (methane production, methane oxidation, aerobic and anaerobic carbon dioxide production), total carbon pools in the different horizons of the bottom soils, soil bulk density, soil substance density, and soil porosity.

n-Alkane content and compound-specific d13C values of soil samples, river samples, and marine surface samples

Southwestern Africa's coastal marine mudbelt, a prominent Holocene sediment package, provides a valuable archive for reconstructing terrestrial palaeoclimates on the adjacent continent. While the origin of terrestrial inorganic material has been intensively studied, the sources of terrigenous organic material deposited in the mudbelt are yet unclear. In this study, plant wax derived n-alkanes and their compound-specific d13C in soils, flood deposits and suspension loads from regional fluvial systems and marine sediments are analysed to characterize the origin of terrestrial organic material in the southwest African mudbelt. Soils from different biomes in the catchments of the Orange River and small west coast rivers show on average distinct n-alkane distributions and compound-specific d13C values reflecting biome-specific vegetation types, most notably the winter rainfall associated Fynbos Biome of the southwestern Cape. In the fluvial sediment samples from the Orange River, changes in the n-alkane distributions and compound-specific d13C compositions reveal an overprint by local vegetation along the river's course. The smaller west coast rivers show distinct signals, reflecting their small catchment areas and particular vegetation communities. Marine surface sediments spanning a transect from the northern mudbelt (29°S) to St. Helena Bay (33°S) reveal subtle, but spatially coherent, changes in n-alkane distributions and compound-specific d13C, indicating the influence of Orange River sediments in the northern mudbelt, the increasing importance of terrigenous input from the adjacent western coastal biomes in the central mudbelt, and contributions from the Fynbos Biome to the southern mudbelt. These findings indicate the different sources of terrestrial organic material deposited in the mudbelt, and highlight the potential the mudbelt has to preserve evidence of environmental change from the adjacent continent.

Collection of aboveground community and species-specific plant biomass from the Jena Experiment (time series since 2002).

This data set comprises time series of aboveground community plant biomass (Sown plant community, Weed plant community, Dead plant material, and Unidentified plant material; all measured in biomass as dry weight) and species-specific biomass from the sown species of several experiments at the field site of a large grassland biodiversity experiment (the Jena Experiment; see further details below). Aboveground community biomass was normally harvested twice a year just prior to mowing (during peak standing biomass twice a year, generally in May and August; in 2002 only once in September) on all experimental plots in the Jena Experiment. This was done by clipping the vegetation at 3 cm above ground in up to four rectangles of 0.2 x 0.5 m per large plot. The location of these rectangles was assigned by random selection of new coordinates every year within the core area of the plots. The positions of the rectangles within plots were identical for all plots. The harvested biomass was sorted into categories: individual species for the sown plant species, weed plant species (species not sown at the particular plot), detached dead plant material (i.e., dead plant material in the data file), and remaining plant material that could not be assigned to any category (i.e., unidentified plant material in the data file). All biomass was dried to constant weight (70°C, >= 48 h) and weighed. Sown plant community biomass was calculated as the sum of the biomass of the individual sown species. The data for individual samples and the mean over samples for the biomass measures on the community level are given. Overall, analyses of the community biomass data have identified species richness as well as functional group composition as important drivers of a positive biodiversity-productivity relationship. The following series of datasets are contained in this collection: 1. Plant biomass form the Main Experiment: In the Main Experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). 2. Plant biomass from the Dominance Experiment: In the Dominance Experiment, 206 grassland plots of 3.5 x 3.5 m were established from a pool of 9 species that can be dominant in semi-natural grassland communities of the study region. In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 3, 4, 6, and 9 species). 3. Plant biomass from the monoculture plots: In the monoculture plots the sown plant community contains only a single species per plot and this species is a different one for each plot. Which species has been sown in which plot is stated in the plot information table for monocultures (see further details below). The monoculture plots of 3.5 x 3.5 m were established for all of the 60 plant species of the Jena Experiment species pool with two replicates per species like the other experiments in May 2002. All plots were maintained by bi-annual weeding and mowing.

Collection of vegetation and soil surface cover in the Jena Experiment (time series since 2002)

This collection contains measurements of vegetation and soil surface cover measured on the plots of the different sub-experiments at the field site of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. The following series of datasets are contained in this collection: 1. Measurements of vegetation cover, i.e. the proportion of soil surface area that is covered by different categories of plants per estimated plot area. Data was collected on the plant community level (sown plant community, weed plant community, dead plant material, and bare ground) and on the level of individual plant species in case of the species that have been sown into the plots to create the gradient of plant diversity.

Collection of measurements on physical soil properties in plots of the Jena Experiment (time series since 2002)

This collection contains measurements on physical soil properties of the plots of the different sub-experiments at the field site of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing

Aboveground plant community and species-specific vegetation cover from the Jena Experiment (Main Experiment, year 2009)

This data set contains information on vegetation cover, i.e. the proportion of soil surface area that is covered by different categories of plants per estimated plot area. Data was collected on the plant community level (sown plant community, weed plant community, dead plant material, and bare ground) and on the level of individual plant species in case of the sown species. Data presented here is from the Main Experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. In 2009, vegetation cover was estimated twice in May and August just prior to mowing (during peak standing biomass) on all experimental plots of the Main Experiment. Cover was visually estimated in a central area of each plot 3 by 3 m in size (approximately 9 m²) using a decimal scale (Londo). Cover estimates for the individual species (and for target species + weeds + bare ground) can add up to more than 100% because the estimated categories represented a structure with potentially overlapping multiple layers. In 2009, in addition to the four community level cover estimates, cover of the moss layer was estimated.

Aboveground plant community and species-specific vegetation cover from the Jena Experiment (Main Experiment, year 2010)

This data set contains information on vegetation cover, i.e. the proportion of soil surface area that is covered by different categories of plants per estimated plot area. Data was collected on the plant community level (sown plant community, weed plant community, dead plant material, and bare ground) and on the level of individual plant species in case of the sown species. Data presented here is from the Main Experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. In 2010, vegetation cover was estimated twice in May and August just prior to mowing (during peak standing biomass) on all experimental plots of the Main Experiment. Cover was visually estimated in a central area of each plot 3 by 3 m in size (approximately 9 m²) using a decimal scale (Londo). Cover estimates for the individual species (and for target species + weeds + bare ground) can add up to more than 100% because the estimated categories represented a structure with potentially overlapping multiple layers.

Aboveground plant community and species-specific vegetation cover from the Jena Experiment (Main Experiment, year 2013)

This data set contains information on vegetation cover, i.e. the proportion of soil surface area that is covered by different categories of plants per estimated plot area. Data was collected on the plant community level (sown plant community, weed plant community, dead plant material, and bare ground) and on the level of individual plant species in case of the sown species. Data presented here is from the Main Experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. In 2013, vegetation cover was estimated twice in May and August just prior to mowing (during peak standing biomass) on all experimental plots of the Main Experiment. Cover was visually estimated in a central area of each plot 3 by 3 m in size (approximately 9 m²) using a decimal scale (Londo). Cover estimates for the individual species (and for target species + weeds + bare ground) can add up to more than 100% because the estimated categories represented a structure with potentially overlapping multiple layers.

Aboveground plant community and species-specific vegetation cover from the Jena Experiment (Main Experiment, year 2008)

This data set contains information on vegetation cover, i.e. the proportion of soil surface area that is covered by different categories of plants per estimated plot area. Data was collected on the plant community level (sown plant community, weed plant community, dead plant material, and bare ground) and on the level of individual plant species in case of the sown species. Data presented here is from the Main Experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. In 2008, vegetation cover was estimated twice in May and August just prior to mowing (during peak standing biomass) on all experimental plots of the Main Experiment. Cover was visually estimated in a central area of each plot 3 by 3 m in size (approximately 9 m²) using a decimal scale (Londo). Cover estimates for the individual species (and for target species + weeds + bare ground) can add up to more than 100% because the estimated categories represented a structure with potentially overlapping multiple layers.

Aboveground plant community and species-specific vegetation cover from the Jena Experiment (Main Experiment, year 2002)

This data set contains information on vegetation cover, i.e. the proportion of soil surface area that is covered by different categories of plants per estimated plot area. Data was collected on the plant community level (sown plant community, weed plant community, dead plant material, and bare ground) and on the level of individual plant species in case of the sown species. Data presented here is from the Main Experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. In 2002, vegetation cover was estimated only once in Septemper just prior to mowing (during peak standing biomass) on all experimental plots of the Main Experiment. Cover was visually estimated in a central area of each plot 3 by 3 m in size (approximately 9 m²) using a decimal scale (Londo). Cover estimates for the individual species (and for target species + weeds + bare ground) can add up to more than 100% because the estimated categories represented a structure with potentially overlapping multiple layers. In 2002, cover on the community level was only estimated for the sown plant community, weed plant community and bare soil. In contrast to later years, cover of dead plant material was not estimated.