Jm O'Reilly-Wapstra

A geographic mosaic of genetic variation within a foundation tree species and its community-level consequences

Knowledge of the manner in which genetic variation within a tree species affects associated communities and ecosystem processes across its entire range is important for understanding how geographic mosaics of genetic interactions might develop and support different communities. While numerous studies have investigated the community and ecosystem consequences of genetic variation at the hybrid cross type or genotype level within a species, none has investigated the community-level effects of intraspecific genetic variation across the geographic range of a widespread species. This is the scale at which geographic mosaics of coevolution are hypothesized to exist. Studies at this level are particularly important for foundation tree species, which typically support numerous microbial, fungal, plant, and animal communities. We studied genetic variation across eight geographical races of the forest tree Eucalyptus globulus representing its natural distribution across southeastern Australia. The study was conducted in a 15-year-old common garden trial based on families derived from single-tree open-pollinated seed collections from the wild. Neutral molecular genetic variation within E. globulus was also assessed and compared with genetic divergence in the phenotypic and community traits. Three major findings emerged. First, we found significant genetically based, hierarchical variation in associated communities corresponding to geographical races of E. globulus and families within races. Second, divergence in foliar communities at the racial level was associated with genetically based divergence in specific leaf morphological and chemical traits that have known defensive functions. Third, significant positive correlations between canopy community dissimilarity and both neutral molecular genetic and leaf quantitative genetic dissimilarity at the race level supported a genetic similarity rule. Our results argue that genetic variation within foundation tree species has the potential to be a significant driver of the geographical mosaics of variation typical of forest communities, which could have important ecological and evolutionary implications.

Genetic variation in resistance of Eucalyptus globulus to marsupial browsers

The evolution of plant defensive traits in response to selection pressures imposed by herbivores is central to co-evolutionary theory. To demonstrate the role of herbivores as selective agents on plant resistance there must be variability in plant resistance to herbivores within a plant population. This variability must be under genetic control, and the variability in plant resistant traits and consequently herbivore damage to plants must reflect variability in plant fitness. We used a common eucalypt species, Eucalyptus globulus, and two major mammalian herbivores, the common brushtail possum (Trichosurus vulpecula) and the red-bellied pademelon (Thylogale billardierii), as a system to investigate intraspecific variation in plant resistance to mammalian herbivores and to investigate if this variation has a genetic basis. We measured mammalian browsing damage on 2,302 individual trees of E. globulus, from 563 families derived from range-wide native stand seed collections of known pedigree and grown in a common environment field trial. Using a selection of trees from the field trial we then conducted a feeding trial with captive herbivores to assess if the genetic variation in plant resistance in the field was reflected in feeding preferences of captive animals, as measured by relative intake. Results from the field trial showed significant genetic variation in plant resistance amongst races, localities and amongst different families. These results were consolidated in the captive trial with similar trends in genetic variation among E. globulus localities. Dry matter intake of foliage by Trichosurus vulpecula was consistently greater than that by Thylogale billardierii; however, the intraspecific preferences of the two herbivores were significantly correlated.

Effects of nutrient variability on the genetic-based resistance of Eucalyptus globulus to a mammalian herbivore and on plant defensive chemistry

Plant resistance to herbivores can be influenced not only by the independent effects of plant genotype and environmental variation, but by interactions between the two. The main aim of this study was to assess the effects of environmental variability (nutrient treatment) on the known genetic-based expression of resistance and defensive chemistry of Eucalyptus globulus to browsing by the generalist mammalian herbivore Trichosurus vulpecula. In a captive feeding trial, we measured intake of seedlings from one relatively resistant (Blue Gum Hill) and one relatively susceptible (St Helens) population of E. globulus grown under two nutrient treatments (no fertiliser, plus fertiliser). There was a significant genotype×fertiliser interaction effect on intake of E. globulus foliage by T. vulpecula, and the predicted genetic-based resistance of the two populations was expressed only for the non-fertilised treatment. Expression of resistance largely reflected the combined and inverse effects of nitrogen and condensed tannin concentrations. The expression of plant secondary metabolite concentration differed between compounds, but in all cases the effects of plant genotype and fertiliser treatment were independent. The formylated phloroglucinol compounds differed significantly between genotypes but not between fertiliser treatments. In contrast, the effect of plant genotype on the expression of condensed tannins was weak but they were significantly reduced by fertiliser. Essential oils were influenced by both plant genotype and fertiliser treatment and were significantly higher in the fertilised seedlings than in the non-fertilised seedlings. This study highlights interactive effects of plant genotype and environment in influencing the phenotypic expression of resistance in a eucalypt species to a mammalian browser. It also demonstrates that this interactive effect is the net result of independent effects of genotype and environment on plant chemistry and finally, that different groups of compounds within a plant can respond very differently to variation in environmental conditions.

Inheritance Of Resistance to Mammalian Herbivores and of Plant Defensive Chemistry in an Eucalyptus Species

Hybridization in plants provides an opportunity to investigate the patterns of inheritance of hybrid resistance to herbivores, and of the plant mechanisms conferring this resistance such as plant secondary metabolites. We investigated how inter-race differences in resistance of Eucalyptus globulus to a generalist mammalian herbivore, Trichosurus vulpecula, are inherited in their F1 hybrids. We assessed browsing damage of 3-year-old trees in a common environment field trial on four hybrid types of known progeny. The progeny were artificial intra-race crosses and reciprocal inter-race F1 hybrids of two geographically distinct populations (races) of E. globulus north-eastern Tasmania and south-eastern Tasmania. Populations of trees from north-eastern Tasmania are relatively susceptible to browsing by T. vulpecula, while populations from south-eastern Tasmania are more resistant. We assessed the preferences of these trees in a series of paired feeding trials with captive animals to test the field trial results and also investigated the patterns of inheritance of plant secondary metabolites. Our results demonstrated that the phenotypic expression of resistance of the inter-race F1 hybrids supported the additive pattern of inheritance, as these hybrids were intermediate in resistance compared to the pure parental hybrids. The expression of plant secondary metabolites in the F1 hybrids varied among major groups of individual compounds. The most common pattern supported was dominance towards one of the parental types. Together, condensed tannins and essential oils appeared to explain the observed patterns of resistance among the four hybrid types. While both chemical groups were inherited in a dominant manner in the inter-race F1 hybrids, the direction of dominance was opposite. Their combined concentration, however, was inherited in an additive manner, consistent with the phenotypic differences in browsing.

Assessment and Implications of Intraspecific and Phenological Variability in Monoterpenes of Scots Pine (Pinus sylvestris) Foliage

Scots pine populations contain individuals with widely differing amounts and composition of monoterpenes and exist as one of two chemotypes: with or without Δ3-carene. We investigated the significance for ecological studies of two types of variation in monoterpenes: (1) the inherent variability in the concentration of monoterpenes or their relative amounts in needles of seedlings, saplings, and mature trees; and (2) phenological variation in developing needles. The relative composition of needle monoterpenes in 5-year-old saplings changed during the needle development period until the final composition was reached upon needle maturity. Changes in composition depended on chemotype. Needles of the “no-Δ3-carene” chemotype had higher absolute concentrations of α-pinene, β-pinene, camphene, and total monoterpenes than “Δ3-carene” chemotype. For the “Δ3-carene” chemotype, the relative concentration of Δ3-carene during the needle growing season and immediately after emergence of seedlings was higher compared to that reached at needle maturity. Repeated removal of single needles (at weekly intervals during growth) from 5-year-old saplings did not influence the composition of monoterpenes. Within a natural Scots pine dominated woodland, 18% of mature Scots pines (N = 574) belonged to the “no-Δ3-carene” chemotype. Chemotypic variation within populations means that the statistical power with which differences in monoterpene concentrations can be detected is lower when sampling from the whole population compared to sampling within chemotypes. Reduction of this background variation and accounting for chiral variation if present, would significantly aid efficiency, interpretation, and understanding of processes in chemical and ecological research. One method for achieving this is the screening of plants for chemotypes before the establishment of experiments or field sampling regimes. We present a summary of suitable analytical methods for needle tissue that facilitates this prior screening.

Do multiple herbivores maintain chemical diversity of Scots pine monoterpenes

A central issue in our understanding of the evolution of the diversity of plant secondary metabolites (PSMs) is whether or not compounds are functional, conferring an advantage to the plant, or non-functional. We examine the hypothesis that the diversity of monoterpene PSMs within a plant species (Scots pine Pinus sylvestris) may be explained by different compounds acting as defences against high-impact herbivores operating at different life stages. We also hypothesize that pairwise coevolution, with uncorrelated interactions, is more likely to result in greater PSM diversity, than diffuse coevolution. We tested whether up to 13 different monoterpenes in Scots pine were inhibitory to herbivory by slugs (Arion ater), bank voles (Clethrionomys glareolus), red deer (Cervus elaphus) and capercaillie (Tetrao urogallus), each of which attack trees at a different life stage. Plants containing more α-pinene were avoided by both slugs and capercaillie, which may act as reinforcing selective agents for this dominant defensive compound. Herbivory by red deer and capercaillie were, respectively, weakly negatively associated with δ3-carene, and strongly negatively correlated with the minor compound β-ocimene. Three of the four herbivores are probably contributory selective agents on some of the terpenes, and thus maintain some, but by no means all, of the phytochemical diversity in the species. The correlated defensive function of α-pinene against slugs and capercaillie is consistent with diffuse coevolutionary processes.

Discrimination between seedlings of Eucalyptus globulus, E. nitens and their F1 hybrid using near-infrared reflectance spectroscopy and foliar oil content

Summary Identification of plant hybrids produced from closely related species can be difficult using morphological characteristics alone, particularly when identifying young seedlings. In this study, we compared the performance of three calibration models developed to discriminate between seedlings of Eucalyptus globulus, E. nitens and their first-generation hybrid using either foliar oil chemistry or near-infrared reflectance spectral data from fresh, whole leaves. Both oil and near-infrared reflectance spectroscopy (NIRS) models were developed using partial least-squares discriminant analysis and showed high classification accuracy, all correctly classifying more than 91% of samples in cross-validation. Additionally, we developed a larger, “global” and independently validated NIRS model specifically to discriminate between E. globulus and F1 hybrid seedlings of different ages. This model correctly classified 98.1% of samples in cross-validation and 95.1% of samples from an independent test set. These results show that NIRS analysis of fresh, whole leaves can be used as a rapid and accurate alternative to chemical analysis for the purpose of hybrid identification.

Responses to mild water deficit and rewatering differ among secondary metabolites but are similar among provenances within Eucalyptus species

Water deficit associated with drought can severely affect plants and influence ecological interactions involving plant secondary metabolites. We tested the effect of mild water deficit and rewatering on physiological, morphological and chemical traits of juvenile Eucalyptus globulus Labill. and Eucalyptus viminalis Labill. We also tested if responses of juvenile eucalypts to water deficit and rewatering varied within species using provenances across a rainfall gradient. Both species and all provenances were similarly affected by mild water deficit and rewatering, as only foliar abscisic acid levels differed among provenances during water deficit. Across species and provenances, water deficit decreased leaf water potential, above-ground biomass and formylated phloroglucinol compound concentrations, and increased condensed tannin concentrations. Rewatering reduced leaf carbon : nitrogen, and total phenolic and chlorogenic acid concentrations. Water deficit and rewatering had no effect on total oil or individual terpene concentrations. Levels of trait plasticity due to water deficit and rewatering were less than levels of constitutive trait variation among provenances. The overall uniformity of responses to the treatments regardless of native provenance indicates limited diversification of plastic responses when compared with the larger quantitative variation of constitutive traits within these species. These responses to mild water deficit may differ from responses to more extreme water deficit or to responses of juvenile/mature eucalypts growing at each locality.

Effectiveness of repellents for reducing damage to eucalypt seedlings by browsing mammals

Summary Marsupial herbivores cause significant damage to seedlings in forestry operations. Damage can lead to changes in tree form, reduced growth rates and seedling death. Non-lethal tactics, as a component of an integrated browsing management strategy, are currently sought to assist in reducing this damage. One such tactic involves making seedlings deterrent or unpalatable through the application of chemical repellents. We investigated the effect of three chemical repellents, Plant Plus, Sentree and Hot Shot, upon browsing of Eucalyptus nitens seedlings by two marsupial herbivores known to browse newly established plantations in Tasmania: the common brushtail possum and the red-bellied pademelon. One repellent was designed to deter herbivores through an unpleasant odour (Plant Plus), one to be unpalatable (Hot Shot), and the third to be both deterrent and unpalatable (Sentree). We ran a combination of paired feeding trials (one repellent + control) and cafeteria trials (all three repellents at once with no control) with captive possums and pademelons, for three nights per trial. Both species consumed significantly more foliage from control seedlings than those treated with Plant Plus or Sentree repellents. Pademelons also preferred controls to Hot Shot treated seedlings but, interestingly, possums consumed significantly more foliage from seedlings treated with Hot Shot than from control seedlings. The most effective repellent against both herbivores was Sentree, and the marked reduction in browsing indicates that further testing in the field is warranted.

Evolutionary history and novel biotic interactions determine plant responses to elevated CO2 and nitrogen fertilization.

A major frontier in global change research is predicting how multiple agents of global change will alter plant productivity, a critical component of the carbon cycle. Recent research has shown that plant responses to climate change are phylogenetically conserved such that species within some lineages are more productive than those within other lineages in changing environments. However, it remains unclear how phylogenetic patterns in plant responses to changing abiotic conditions may be altered by another agent of global change, the introduction of non-native species. Using a system of 28 native Tasmanian Eucalyptus species belonging to two subgenera, Symphyomyrtus and Eucalyptus, we hypothesized that productivity responses to abiotic agents of global change (elevated CO2 and increased soil N) are unique to lineages, but that novel interactions with a non-native species mediate these responses. We tested this hypothesis by examining productivity of 1) native species monocultures and 2) mixtures of native species with an introduced hardwood plantation species, Eucalyptus nitens, to experimentally manipulated soil N and atmospheric CO2. Consistent with past research, we found that N limits productivity overall, especially in elevated CO2 conditions. However, monocultures of species within the Symphyomyrtus subgenus showed the strongest response to N (gained 127% more total biomass) in elevated CO2 conditions, whereas those within the Eucalyptus subgenus did not respond to N. Root:shoot ratio (an indicator of resource use) was on average greater in species pairs containing Symphyomyrtus species, suggesting that functional traits important for resource uptake are phylogenetically conserved and explaining the phylogenetic pattern in plant response to changing environmental conditions. Yet, native species mixtures with E. nitens exhibited responses to CO2 and N that differed from those of monocultures, supporting our hypothesis and highlighting that both plant evolutionary history and introduced species will shape community productivity in a changing world.