A mathematical model of glioblastoma tumor spheroid invasion in a three-dimensional in vitro experiment

Glioblastoma, the most malignant form of brain cancer, is responsible for 23% of primary brain tumors and has extremely poor outcome. Confounding the clinical management of glioblastomas is the extreme local invasiveness of these cancer cells. The mechanisms that govern invasion are poorly understood. To gain insight into glioblastoma invasion, we conducted experiments on the patterns of growth and dispersion of U87 glioblastoma tumor spheroids in a three-dimensional collagen gel. We studied two different cell lines, one with a mutation to the EGFR (U87DeltaEGFR) that is associated with increased malignancy, and one with an endogenous (wild-type) receptor (U87WT). We developed a continuum mathematical model of the dispersion behaviors with the aim of identifying and characterizing discrete cellular mechanisms underlying invasive cell motility. The mathematical model quantitatively reproduces the experimental data, and indicates that the U87WT invasive cells have a stronger directional motility bias away from the spheroid center as well as a faster rate of cell shedding compared to the U87DeltaEGFR cells. The model suggests that differences in tumor cell dispersion may be due to differences in the chemical factors produced by cells, differences in how the two cell lines remodel the gel, or different cell-cell adhesion characteristics.     

The impact of phenotypic switching on glioblastoma growth and invasion

The brain tumour glioblastoma is characterised by diffuse and infiltrative growth into surrounding brain tissue. At the macroscopic level, the progression speed of a glioblastoma tumour is determined by two key factors: the cell proliferation rate and the cell migration speed. At the microscopic level, however, proliferation and migration appear to be mutually exclusive phenotypes, as indicated by recent in vivo imaging data. Here, we develop a mathematical model to analyse how the phenotypic switching between proliferative and migratory states of individual cells affects the macroscopic growth of the tumour. For this, we propose an individual-based stochastic model in which glioblastoma cells are either in a proliferative state, where they are stationary and divide, or in motile state in which they are subject to random motion. From the model we derive a continuum approximation in the form of two coupled reaction-diffusion equations, which exhibit travelling wave solutions whose speed of invasion depends on the model parameters. We propose a simple analytical method to predict progression rate from the cell-specific parameters and demonstrate that optimal glioblastoma growth depends on a non-trivial trade-off between the phenotypic switching rates. By linking cellular properties to an in vivo outcome, the model should be applicable to designing relevant cell screens for glioblastoma and cytometry-based patient prognostics.     

Increased Phenotypic Plasticity to Climate May Have Boosted the Invasion Success of Polyploid Centaurea stoebe

Phenotypic plasticity may allow organisms to cope with altered environmental conditions as e.g. after the introduction into a new range. In particular polyploid organisms, containing more than two sets of chromosomes, may show high levels of plasticity, which could in turn increase their environmental tolerance and invasiveness. Here, we studied the role of phenotypic plasticity in the invasion of Centaurea stoebe (Asteraceae), which in the native range in Europe occurs as diploids and tetraploids, whereas in the introduced range in North America so far only tetraploids have been found. In a common garden experiment at two sites in the native range, we grew half-sibs of the three geo-cytotypes (native European diploids, European tetraploids and invasive North American tetraploids) from a representative sample of 27 populations. We measured the level and the adaptive significance of phenotypic plasticity in eco-physiological and life-history traits in response to the contrasting climatic conditions at the two study sites as well as three different soil conditions in pots, simulating the most crucial abiotic differences between the native and introduced range. European tetraploids showed increased levels of phenotypic plasticity as compared to diploids in response to the different climatic conditions in traits associated with rapid growth and fast phenological development. Moreover, we found evidence for adaptive plasticity in these traits, which suggests that increased plasticity may have contributed to the invasion success of tetraploid C. stoebe by providing an advantage under the novel climatic conditions. However, in invasive tetraploids phenotypic plasticity was similar to that of native tetraploids, indicating no evolution of increased plasticity during invasions. Our findings provide the first empirical support for increased phenotypic plasticity associated with polyploids, which may contribute to their success as invasive species in novel environments.     

In silico identification of functional divergence between the multiple groEL gene paralogs in Chlamydiae

Background

RNA molecules, through their dual appearance as sequence and structure, represent a suitable model to study evolutionary properties of quasispecies. The essential ingredient in this model is the differentiation between genotype (molecular sequences which are affected by mutation) and phenotype (molecular structure, affected by selection). This framework allows a quantitative analysis of organizational properties of quasispecies as they adapt to different environments, such as their robustness, the effect of the degeneration of the sequence space, or the adaptation under different mutation rates and the error threshold associated.

Results

We describe and analyze the structural properties of molecular quasispecies adapting to different environments both during the transient time before adaptation takes place and in the asymptotic state, once optimization has occurred. We observe a minimum in the adaptation time at values of the mutation rate relatively far from the phenotypic error threshold. Through the definition of a consensus structure, it is shown that the quasispecies retains relevant structural information in a distributed fashion even above the error threshold. This structural robustness depends on the precise shape of the secondary structure used as target of selection. Experimental results available for natural RNA populations are in qualitative agreement with our observations.

Conclusion

Adaptation time of molecular quasispecies to a given environment is optimized at values of the mutation rate well below the phenotypic error threshold. The optimal value results from a trade-off between diversity generation and fixation of advantageous mutants. The critical value of the mutation rate is a function not only of the sequence length, but also of the specific properties of the environment, in this case the selection pressure and the shape of the secondary structure used as target phenotype. Certain functional motifs of RNA secondary structure that withstand high mutation rates (as the ubiquitous hairpin motif) might appear early in evolution and be actually frozen evolutionary accidents.     

Phenotypic variation in invasive and biocontrol populations of the harlequin ladybird, Harmonia axyridis

Despite numerous releases for biological control purposes during more than 20 years in Europe, Harmonia axyridis failed to become established until the beginning of the 21st century. Its status as invasive alien species is now widely recognised. Theory suggests that invasive populations should evolve toward greater phenotypic plasticity because they encounter differing environments during the invasion process. On the contrary, populations used for biological control have been maintained under artificial rearing conditions for many generations; they are hence expected to become specialised on a narrow range of environments and show lower phenotypic plasticity. Here we compared phenotypic traits and the extent of adaptive phenotypic plasticity in two invasive populations and two populations commercialized for biological control by (i) measuring six phenotypic traits related to fitness (eggs hatching rate, larval survival rate, development time, sex ratio, fecundity over 6 weeks and survival time of starving adults) at three temperatures (18, 24 and 30°C), (ii) recording the survival rate and quiescence aggregation behaviour when exposed to low temperatures (5, 10 and 15°C), and (iii) studying the cannibalistic behaviour of populations in the absence of food. Invasive and biocontrol populations displayed significantly different responses to temperature variation for a composite fitness index computed from the traits measured at 18, 24 and 30°C, but not for any of those traits considered independently. The plasticity measured on the same fitness index was higher in the two invasive populations, but this difference was not statistically significant. On the other hand, invasive populations displayed significantly higher survival and higher phenotypic plasticity when entering into quiescence at low temperatures. In addition, one invasive population displayed a singular cannibalistic behaviour. Our results hence only partly support the expectation of increased adaptive phenotypic plasticity of European invasive populations of H. axyridis, and stress the importance of the choice of the environmental parameters to be manipulated for assessing phenotypic plasticity variation among populations.     

Analysis of tissue chimerism in nude mouse brain and abdominal xenograft models of human glioblastoma multiforme: what does it tell us about the models and about glioblastoma biology and therapy?

In situ hybridization coupled to immunohistochemistry for antigens of interest allows unequivocal identification of tumor cells from reactive stroma cells and normal adjacent structures in human glioblastoma multiforme grafts transplanted into nude mice. With this methodology, we have explored the development of glioblastoma multiforme solid grafts transplanted into nude mouse brains or flanks. The brain transplants closely resembled the human situation, particularly in relation to differentiation and growth patterns. The morphological features of peritumoral reactive gliosis were similar to those observed in humans. A mouse glial stroma within the main tumor masses was also demonstrated. Kinetic studies showed that the compartment of isolated tumor cells that infiltrated host brains and the reactive gliosis constituted two cycling cell populations. Despite VEGF protein expression by tumor cells and some reactive astrocytes, the abnormally permeable microvascular beds were not hyperplastic. The observation of a non-infiltrative pattern of growth when grafts were established in host flanks demonstrated that the organ-specific environment plays a determining role in the growth and invasive properties of glioblastoma. The phylogenetic distance between man and mouse and the recipient immunoincompetence should not impose serious limitations on the use of this model for studying malignant glioma biology and therapy in vivo.     

Jack of all trades,master of some? On the role of phenotypic plasticity in plant invasions

Invasion biologists often suggest that phenotypic plasticity plays an important role in successful plant invasions. Assuming that plasticity enhances ecological niche breadth and therefore confers a fitness advantage, recent studies have posed two main hypotheses: (1) invasive species are more plastic than non-invasive or native ones; (2) populations in the introduced range of an invasive species have evolved greater plasticity than populations in the native range. These two hypotheses largely reflect the disparate interests of ecologists and evolutionary biologists. Because these sciences are typically interested in different temporal and spatial scales, we describe what is required to assess phenotypic plasticity at different levels. We explore the inevitable tradeoffs of experiments conducted at the genotype vs. species level, outline components of experimental design required to identify plasticity at different levels, and review some examples from the recent literature. Moreover, we suggest that a successful invader may benefit from plasticity as either (1) a Jack-of-all-trades, better able to maintain fitness in unfavourable environments; (2) a Master-of-some, better able to increase fitness in favourable environments; or (3) a Jack-and-master that combines some level of both abilities. This new framework can be applied when testing both ecological or evolutionary oriented hypotheses, and therefore promises to bridge the gap between the two perspectives.     

Phenotypic divergence during the invasion of Phyla canescens in Australia and France: evidence for selection-driven evolution

Rapid adaptive evolution has been advocated as a mechanism that promotes invasion. Demonstrating adaptive evolution in invasive species requires rigorous analysis of phenotypic shifts driven by selection. Here, we document selection-driven evolution of Phyla canescens , an Argentine weed, in two invaded regions (Australia and France). Invasive populations possessed similar or higher diversity than native populations, and displayed mixed lineages from different sources, suggesting that genetic bottlenecks in both countries might have been alleviated by multiple introductions. Compared to native populations, Australian populations displayed more investment in sexual reproduction, whereas French populations possessed enhanced vegetative reproduction and growth. We partitioned evolutionary forces (selection vs. stochastic events) using two independent methods. Results of both analyses suggest that the pattern of molecular and phenotypic variability among regions was consistent with selection-driven evolution, rather than stochastic events. Our findings indicate that selection has shaped the evolution of P . canescens in two different invaded regions.
Ecology Letters (2010) 13: 32–44     

Simulation Predicts IGFBP2-HIF1α Interaction Drives Glioblastoma Growth

Tremendous strides have been made in improving patients’ survival from cancer with one glaring exception: brain cancer. Glioblastoma is the most common, aggressive and highly malignant type of primary brain tumor. The average overall survival remains less than 1 year. Notably, cancer patients with obesity and diabetes have worse outcomes and accelerated progression of glioblastoma. The root cause of this accelerated progression has been hypothesized to involve the insulin signaling pathway. However, while the process of invasive glioblastoma progression has been extensively studied macroscopically, it has not yet been well characterized with regards to intracellular insulin signaling. In this study we connect for the first time microscale insulin signaling activity with macroscale glioblastoma growth through the use of computational modeling. Results of the model suggest a novel observation: feedback from IGFBP2 to HIF1α is integral to the sustained growth of glioblastoma. Our study suggests that downstream signaling from IGFI to HIF1α, which has been the target of many insulin signaling drugs in clinical trials, plays a smaller role in overall tumor growth. These predictions strongly suggest redirecting the focus of glioma drug candidates on controlling the feedback between IGFBP2 and HIF1α.     

Regulation of Phenotypic Switching and Heterogeneity in Photorhabdus luminescens Cell Populations

Phenotypic heterogeneity in bacterial cell populations allows genetically identical organisms to different behavior under similar environmental conditions. The Gram‐negative bacterium Photorhabdus luminescens is an excellent organism to study phenotypic heterogeneity since their life cycle involves a symbiotic interaction with soil nematodes as well as a pathogenic association with insect larvae. Phenotypic heterogeneity is highly distinct in P. luminescens. The bacteria exist in two phenotypic forms that differ in various morphologic and phenotypic traits and are therefore distinguished as primary (1°) and secondary (2°) cells. The 1 cells are bioluminescent, pigmented, produce several secondary metabolites and exo-enzymes, and support nematode growth and development. The 2° cells lack all these 1°-specific phenotypes. The entomopathogenic nematodes carry 1° cells in their upper gut and release them into an insect's body after slipping inside. During insect infection, up to the half number of 1° cells undergo phenotypic switching and convert to 2° cells. Since the 2° cells are not able to live in nematode symbiosis any more, they cannot re-associate with their symbiosis partners after the infection and remain in the soil. Phenotypic switching in P. luminescens has to be tightly regulated since a high switching frequency would lead to a complete break-down of the nematode-bacteria life cycle. Here, we present the main regulatory mechanisms known to-date that are important for phenotypic switching in P. luminescens cell populations and discuss the biological reason as well as the fate of the 2° cells in the soil.     

Pre-adaptation or genetic shift after introduction in the invasive species Impatiens glandulifera?

Invasive exotic plants often grow fast, reproduce rapidly and display considerable phenotypic plasticity in their invasive range, which may be essential characteristics for successful invasion. However, it remains unclear whether these characteristics are already present in native populations (pre-adaptation hypothesis) or evolve after introduction (genetic shift hypothesis).To test these hypotheses we compared means and phenotypic plasticity of vegetative and reproductive traits between populations of Impatiens glandulifera collected from either the invasive (Norway) or native range (India). Seeds were sown and the resulting plants were exposed to different experimental environments in a glasshouse. We also tested whether trait means and reaction norms harbored genetic variation, as this may promote fitness in the novel environment.We did not find evidence that invasive populations of I. glandulifera grew more vigorously or produced more seeds than native populations. Phenotypic plasticity did not differ between the native and invasive range, except for the number of nodes which was more plastic in the invasive range. Genetic variation in the slope of reaction norms was absent, suggesting that the lack of change in phenotypic plasticity between native and invasive populations resulted from low genetic variation in phenotypic plasticity initially harbored by this species. Post-introduction evolution of traits thus probably did not boost the invasiveness of I. glandulifera. Instead, the species seems to be pre-adapted for invasion.We suggest that differences in habitat between the native and invasive range, more specifically the higher nutrient availability observed in the new environment, are the main factor driving the invasion of this species. Indeed, plants in the more nutrient-rich invasive range had greater seed mass, likely conferring a competitive advantage, while seed mass also responded strongly to nutrients in the glasshouse. Interactions between habitat productivity and herbivore defense may explain the lack of more vigorous growth in the new range.     

A history of phenotypic plasticity accelerates adaptation to a new environment

Can a history of phenotypic plasticity increase the rate of adaptation to a new environment? Theory suggests it can be through two different mechanisms. Phenotypically plastic organisms can adapt rapidly to new environments through genetic assimilation, or the fluctuating environments that result in phenotypic plasticity can produce evolvable genetic architectures. In this article, I studied a model of a gene regulatory network that determined a phenotypic character in one population selected for phenotypic plasticity and a second population in a constant environment. A history of phenotypic plasticity increased the rate of adaptation in a new environment, but the amount of this increase was dependent on the strength of selection in the original environment. Phenotypic variance in the original environment predicted the adaptive capacity of the trait within, but not between, plastic and nonplastic populations. These results have implications for invasive species and ecological studies of rapid adaptation.     

Genetic variability in Irish populations of the invasive zebra mussel, Dreissena polymorpha: discordant estimates of population differentiation from allozymes and microsatellites

1. The recent arrival and explosive spread of the zebra mussel, Dreissena polymorpha (Pallas), in Ireland provided a rare opportunity to study the population genetics of an invasive species.
2. Eight polymorphic allozyme loci ( ACO-1, ACO-2 , EST-D, GPI, IDH-2, MDH, OPDH and PGM ) were used to investigate genetic diversity and population structure in five Irish populations, and the results were compared with those from a previous microsatellite study on the same samples.
3. The mean number of alleles per locus (2.7 ± 0.1) was similar to the mean for the same loci in European populations, suggesting that Irish founder populations were large and/or multiple colonization events took place after foundation. A deficiency of heterozygotes was observed in all populations, but was uneven across loci.
4. Pairwise comparisons, using Fisher's exact tests and F _ST values, revealed significant genetic differentiation among populations. The overall multilocus F _ST estimate was 0.118 ± 0.045, which contrasted with an estimate of 0.015 ± 0.007 from five microsatellite loci on the same samples in a previous study.
5. Assuming that microsatellites can be used as a neutral baseline, the discordant results from allozymes and microsatellites suggest that selection may be acting on some allozyme loci, specifically ACO-1, ACO-2 , IDH-2 and MDH, which contributed most to the significant differentiation between samples.     

NICHE DIMENSIONALITY AND THE GENETICS OF ECOLOGICAL SPECIATION

Niche dimensionality is suggested to be a key determinant of ecological speciation (“multifarious selection” hypothesis), but genetic aspects of this process have not been investigated theoretically. We use Fisher's geometrical model to study how niche dimensionality influences the mean fitness of hybrids formed upon secondary contact between populations adapting in allopatry. Gaussian selection for an optimum generates two forms of reproductive isolation (RI): an extrinsic component due to maladaptation of the mean phenotype, and an intrinsic variance load resulting from what we term transgressive incompatibilities between mutations fixed in different populations. We show that after adaptation to a new environment, RI increases with (1) the mean initial maladaptation of diverging population, and (2) niche dimensionality, which increases the phenotypic variability of fixed mutations. Under mutation selection drift equilibrium in a constant environment, RI accumulates steadily with time, at a rate that also increases with niche dimensionality. A similar pattern can be produced by successive shifts in the optimum phenotype. Niche dimensionality thus has an effect per se on postzygotic isolation, beyond putative indirect effects (stronger selection, more genes). Our mechanism is consistent with empirical evidence about transgressive segregation in crosses between divergent populations, and with patterns of accumulation of RI with time in many taxa.     

CAN REINFORCEMENT OCCUR WITH A LEARNED TRAIT?

We use birdsong as a case study to ask whether reinforcement can occur via the spread of a genetically determined female preference for a socially inherited (learned) male trait. We envision secondary contact between two neighboring populations with different song dialects. An individual's ability to learn song is confined by a genetic predisposition: if predispositions are strong, there will be no phenotypic overlap in song between populations, whereas weak predispositions allow phenotypic overlap, or "mixed" song. To determine if reinforcement has occurred, we consider if an allele for within-population female mating preference, based on song, can spread, and whether population specific songs can concurrently be maintained at equilibrium. We model several scenarios, including costs to mating preferences, mating preferences in hybrids, and hybrids having the ability to learn pure songs. We find that when weak predispositions are fixed within a population reinforcement based on song cannot occur. However, when some individuals have strong predispositions, restricting phenotypic overlap between populations in the trait, reinforcement is only slightly inhibited from a purely genetic model. Generalizing beyond the example of song, we conclude that socially learned signals will tend to prohibit reinforcement, but it may still occur if some individuals acquire trait phenotypes genetically.     

Gene expression and drought response in an invasive thistle

Though rapid phenotypic evolution has been observed in many invasive plant species, less is known about the associated genetic mechanisms. Some hypotheses invoke the evolution of trade-offs in resource allocation to explain phenotypic differences between the native and invaded ranges of a species. Alternately, invasive species may benefit from a generalist strategy and perform well in many environments. Identification of the molecular changes associated with successful invasions can offer clues regarding the mechanistic basis of such hypotheses, even in non-model organisms. To complement studies of phenotypic variation, we investigate gene expression during drought response that might underlie variation in drought tolerance between native and introduced populations of diffuse knapweed (Centaurea diffusa), and possibly contribute to invasion success. Using species-specific microarrays and tissue sampled under drought and control conditions at three time points, we identified genes whose expression either varied constitutively or responded to drought stress differently between ranges. Further, we functionally investigate these genes. Based on these data, invasive populations have constitutively higher levels of expression relating to energy production and lower levels of signal transduction expression relative to native populations. Under drought conditions, invasive populations may maintain energy production and react less strongly to drought than native populations, which may allow them to maintain fitness across moderate environmental variation. This supports the expectation of invasive populations consisting of generalist genotypes capable of fitness homeostasis, which may have facilitated the successful invasion of many environments in North America.     

Environmental fluctuations promote intraspecific diversity and population persistence via inflationary effects

The impact of temporal variation in the environment, specifically the amount of temporal autocorrelation, on population processes is of growing interest in ecology and evolutionary biology. It was recently discovered that temporal autocorrelation in the environment can significantly increase the abundance of populations that would otherwise have low, or even negative long‐term growth rates (via so‐called ‘inflationary effects’), provided that immigration from another source prevents extinction. Here we use a mathematical model to ask whether inflationary effects can also increase population persistence without immigration if different phenotypes within that population partition growth over time and buffer each other from extinction via mutation. Using a combination of analytical and numerical methods, we find that environmental autocorrelation can inflate the abundance of phenotypes that would otherwise be excluded from the population, provided that phenotypes are sufficiently different in their use of the environment. This inflation of abundance at the phenotypic level also generates an inflation of abundance at the population level. Remarkably, intraspecific inflationary effects can increase both phenotypic and whole population abundance even if one or all phenotypes are maladapted to the environment, as long as mutations prevent phenotypic extinction during periods of poor environmental conditions. Given the prevalence of temporally autocorrelated environmental variables in nature, intraspecific inflationary effects have the potential to be of widespread importance for population persistence as well as the maintenance of intraspecific diversity.     

Exploring the Competition between Proliferative and Invasive Cancer Phenotypes in a Continuous Spatial Model

Tumor is characterized by extensive heterogeneity with respect to its microenvironment and its genetic composition. We extend a previously developed monoclonal continuous spatial model of tumor growth to account for polyclonal cell populations and investigate the interplay between a more proliferative and a more invasive phenotype under different conditions. The model simulations demonstrate a transition from the dominance of the proliferative to the dominance of the invasive phenotype resembling malignant tumor progression and show a time period where both subpopulations are abundant. As the dominant phenotype switches from proliferative to invasive, the geometry of tumor changes from a compact and almost spherical shape to a more diffusive and fingered morphology with the proliferative phenotype to be restricted in the tumor bulk and the invasive to dominate at tumor edges. Different micro-environmental conditions and different phenotypic properties can promote or inhibit invasion demonstrating their mutual importance. The model provides a computational framework to investigate tumor heterogeneity and the constant interplay between the environment and the specific characteristics of phenotypes that should be taken into account for the prediction of tumor evolution, morphology and effective treatment.     

Replication and mutation on neutral networks

Folding of RNA sequences into secondary structures is viewed as a map that assigns a uniquely defined base pairing pattern to every sequence. The mapping is non-invertible since many sequences fold into the same minimum free energy (secondary) structure or shape. The pre-images of this map, called neutral networks, are uniquely associated with the shapes and vice versa. Random graph theory is used to construct networks in sequence space which are suitable models for neutral networks. The theory of molecular quasispecies has been applied to replication and mutation on single-peak fitness landscapes. This concept is extended by considering evolution on degenerate multi-peak landscapes which originate from neutral networks by assuming that one particular shape is fitter than all the others. On such a single-shape landscape the superior fitness value is assigned to all sequences belonging to the master shape. All other shapes are lumped together and their fitness values are averaged in a way that is reminiscent of mean field theory. Replication and mutation on neutral networks are modeled by phenomenological rate equations as well as by a stochastic birth-and-death model. In analogy to the error threshold in sequence space the phenotypic error threshold separates two scenarios: (i) a stationary (fittest) master shape surrounded by closely related shapes and (ii) populations drifting through shape space by a diffusion-like process. The error classes of the quasispecies model are replaced by distance classes between the master shape and the other structures. Analytical results are derived for single-shape landscapes, in particular, simple expressions are obtained for the mean fraction of master shapes in a population and for phenotypic error thresholds. The analytical results are complemented by data obtained from computer simulation of the underlying stochastic processes. The predictions of the phenomenological approach on the single-shape landscape are very well reproduced by replication and mutation kinetics of tRNA(phe). Simulation of the stochastic process at a resolution of individual distance classes yields data which are in excellent agreement with the results derived from the birth-and-death model.     

Comparing indigenous and introduced populations of Melaleuca quinquenervia (Cav.) Blake: response of seedlings to water and pH levels

Melaleuca quinquenervia is a wetland tree species indigenous to eastern Australia. It was separately introduced to east and west Florida as an ornamental, but has since become invasive, dominating several habitat types. We tested the predictions that (1) Australian populations would exhibit more genetic variation than Florida populations, due to founder effect, and (2) high phenotypic plasticity would be found in all populations, due to the wide range of habitats occupied. We compared the phenotypic plasticity and familial variation among three Australian populations, two east Florida, and two west Florida populations in a greenhouse experiment. We grew seedlings collected from different maternal trees in each population under two water levels and three pH levels, reflecting the natural range of water levels and soil pH in Florida and Australian Melaleuca stands. We measured leaf size and shape, growth rate and above-ground biomass of seedlings and determined the components of phenotypic variance (familial, environmental, and their interaction) using univariate and multivariate analysis of variance. All traits showed significant among-population and among-family variation, as well as significant phenotypic plasticity, in response to both water level and pH level changes. Sensitivity to pH was particularly high, presumably because plants were grown under pHs ranging from 4.7 to 7.4, and because pH can influence nutrient availability. Familial variation contains genetic variation, but it may also be confounded with maternal environmental effects. Comparing Australian to Floridian Melaleuca, amounts of familial variation and phenotypic plasticity varied by trait. Overall, Australian Melaleuca had more among-population variation than Floridian Melaleuca, presumably reflecting the wider latitudinal range and longer time for evolutionary change in Australia, but had similar amounts of among-family variation, within any one population. If maternal effects are strong, among-population differences may merely reflect greater environmental differences among Australian sites than Florida sites. Australian Melaleuca had less phenotypic plasticity, possibly due to founder effects in Florida or to subsequent adaptive evolution of phenotypic plasticity in Floridian populations. Floridian Melaleuca shows little loss of familial variation, compared to indigenous Australian populations, and that, in combination with its high phenotypic plasticity, should allow it to continue to colonize new areas successfully.