Bookkeeping of insect herbivory trends in herbarium specimens of purple loosestrife (Lythrum salicaria).

The potential use of herbarium specimens to detect herbivory trends is enormous but largely untapped. The objective of this study was to reconstruct the long-term herbivory pressure on the Eurasian invasive plant, purple loosestrife (Lythrum salicaria), by evaluating leaf damage over 1323 specimens from southern Québec (Canada). The hypothesis tested is that that the prevalence of herbivory damage on purple loosestrife is low during the invasion phase and increases throughout the saturation phase. Historical trends suggest a gradual increase in hole feeding and margin feeding damage from 1883 to around 1940, followed by a period of relative stability. The percentage of specimens with window feeding damage did not begin to increase until the end of the twentieth century, from 3% (2-6%) in 1990 to 45% (14-81%) in 2015. Temporal changes in the frequency of window feeding damage support the hypothesis of an increasing herbivory pressure by recently introduced insects. This study shows that leaf damage made by insects introduced for the biocontrol of purple loosestrife, such as coleopterans of the Neogalerucella genus, can be assessed from voucher specimens. Herbaria are a rich source in information that can be used to answer questions related to plant-insect interactions in the context of biological invasions and biodiversity changes.This article is part of the theme issue 'Biological collections for understanding biodiversity in the Anthropocene'.

Experimental insights on Darwin's cross-promotion hypothesis in tristylous purple loosestrife (Lythrum salicaria).

PREMISE OF THE STUDY: Darwin proposed that the reciprocal arrangement of anthers and stigmas in heterostylous plants promotes cross-pollination through pollen segregation on pollinators' bodies. The floral tube in most heterostylous species constrains the feeding posture of pollinators determining the site of contact with sex organs located at different heights within a flower. Here, we evaluate Darwin's hypothesis in tristylous Lythrum salicaria, a species with a partially tubular corolla, and examine the extent to which the location of sex organs within a flower influence compatible and incompatible pollination. We predicted that the proficiency of cross-pollination would increase for more inserted sex organs due to the restrictions imposed by the floral tube on pollinator positioning. METHODS: We used experimental trimorphic and monomorphic arrays and emasculated flowers to quantify intermorph pollen transfer and capture among all sex-organ heights, and estimated the contribution of intraflower self-pollination, geitonogamous self-pollination, and intramorph outcross pollination to total intramorph pollination. KEY RESULTS: As predicted, disassortative pollination varied significantly with sex-organ height and was highest for short-level organs and lowest for long-level organs. In monomorphic arrays, most intramorph pollination resulted from outcross pollination followed by intraflower and geitonogamous self-pollination. CONCLUSIONS: Our results provide evidence for the Darwinian hypothesis. Reciprocal herkogamy promoted varying degrees of disassortative pollination with the magnitude strongly influenced by sex-organ height within a flower.

Variation in style morph frequencies in tristylous Lythrum salicaria in the Iberian Peninsula: the role of geographical and demographic factors.

BACKGROUND AND AIMS: The balance between stochastic forces and negative frequency-dependent selection largely determines style morph frequencies in heterostylous populations. Investigation of morph frequencies at geographical range limits can provide insights into the forces maintaining the floral polymorphism, and the factors causing biased morph ratios. Here, we investigate style morph frequencies in populations at the south-western European range limit of tristylous Lythrum salicaria, to explore the role of demographic and geographical factors influencing morph ratios in its native range. METHODS: We measured morph composition and evenness, and the size of 96 populations, along a north to south latitudinal transect from Galicia to Andalucia, Iberian Peninsula, traversing a steep climatic gradient. To examine the potential influence of morph-specific fitness components on morph ratios, we examined reproductive traits in 19 populations. KEY RESULTS: Most populations of L. salicaria were trimorphic (94·79 %), the majority exhibiting 1 : 1 : 1 morph ratios (68·75 %). Populations with biased morph ratios had a deficiency of the short-styled morph. Population size and morph evenness were positively associated with latitude, with smaller populations and those with less even morph ratios occurring towards the south. Greater variance in morph evenness was evident at the southern range margin. There were no consistent differences in components of reproductive fitness among style morphs, but southern populations produced less fruit and seed than more northerly populations. CONCLUSIONS: Our results demonstrate the influence of finite population size on morph frequencies in L. salicaria. However, they also illustrate the resilience of Iberian populations to the factors causing deviations from isoplethy and morph loss, especially at the southern range limit where populations are smaller. The maintenance of tristyly in small populations of L. salicaria may be aided by the genetic connectivity of populations in agricultural landscapes resulting from gene flow through pollen and seed dispersal.

Herbivory Differentially Affects Plant Fitness in Three Populations of the Perennial Herb Lythrum salicaria along a Latitudinal Gradient.

Herbivory can negatively and selectively affect plant fitness by reducing growth, survival and reproductive output, thereby influencing plant population dynamics and evolution. Latitudinal variation in intensity of herbivory is common, but the extent to which it translates into corresponding variation in effects on plant performance is still poorly known. We tested the hypothesis that variation in the fitness-consequences of herbivory mirror differences in intensity of herbivory among three natural populations of the perennial herb Lythrum salicaria along a latitudinal gradient from southern to northernmost Sweden. We documented intensity of herbivory and examined its effect on survival, growth and reproductive output over two years by experimentally removing herbivores with insecticide. The intensity of herbivory and the effects of herbivory on plant fitness were strongest in the southern population, intermediate in the central population and weakest in the northern population. The mean proportion of the leaf area removed ranged from 11% in the southern to 3% in the northern population. Herbivore removal increased plant height 1.5-fold in the southern and 1.2-fold in the central population, the proportion plants flowering 4-fold in the southern and 2-fold in the central population, and seed production per flower 1.6-fold in the southern and 1.2-fold in the central population, but did not affect plant fitness in the northern population. Herbivore removal thus affected the relative fecundity of plants in the three populations: In the control, seed output per plant was 8.6 times higher in the northern population compared to the southern population, whereas after herbivore removal it was 2.5 times higher in the southern population. The results demonstrate that native herbivores may strongly affect the demographic structure of L. salicaria populations and thereby shape geographic patterns of seed production. They further suggest that the strength of herbivore-mediated selection varies among populations and decreases towards the north.

Indirect effects of herbivory on plant-pollinator interactions in invasive Lythrum salicaria.

PREMISE OF THE STUDY: Herbivory can affect a plant's fitness in a variety of ways, including modifying the biotic interactions of the plant. In particular, when herbivory influences floral display, we hypothesize that pollinator visitation will be altered accordingly. Here we studied the indirect effects of feeding by two beetles, Neogalerucella calmariensis and N. pusilla, released as a biological control, on plant-pollinator interactions and fitness in the invasive plant, purple loosestrife (Lythrum salicaria). METHODS: Two herbivory treatments (ambient and simulated) were applied to plants in a naturally occurring population of purple loosestrife. During flowering, traits of plants in the treatment and control groups were recorded. Data on pollinator visitation behavior was then collected after intense larval herbivory had ended. KEY RESULTS: Plants exposed to herbivory treatments produced more flowers and inflorescences but flowered significantly later than those in the control group. Moreover, we found a significant, positive association of herbivory with the number of flowers probed by bumblebees and with the number of times a foraging pollinator moved among inflorescences on a single plant. No differences in female fitness (fruit or seed production) were detected. CONCLUSIONS: We conclude that herbivore-mediated differences in floral display traits impacted pollinator visitation behavior. However, as we discuss, differences in pollinator visitation did not translate into detectable differences in female reproductive success. We postulate that herbivory could influence other unmeasured aspects of fitness, such as seed quality or the number of seeds sired.

Latitudinal variation in resistance and tolerance to herbivory in the perennial herb Lythrum salicaria is related to intensity of herbivory and plant phenology.

Both the length of the growing season and the intensity of herbivory often vary along climatic gradients, which may result in divergent selection on plant phenology, and on resistance and tolerance to herbivory. In Sweden, the length of the growing season and the number of insect herbivore species feeding on the perennial herb Lythrum salicaria decrease from south to north. Previous common-garden experiments have shown that northern L. salicaria populations develop aboveground shoots earlier in the summer and finish growth before southern populations do. We tested the hypotheses that resistance and tolerance to damage vary with latitude in L. salicaria and are positively related to the intensity of herbivory in natural populations. We quantified resistance and tolerance of populations sampled along a latitudinal gradient by scoring damage from natural herbivores and fitness in a common-garden experiment in the field and by documenting oviposition and feeding preference by specialist leaf beetles in a glasshouse experiment. Plant resistance decreased with latitude of origin, whereas plant tolerance increased. Oviposition and feeding preference in the glasshouse and leaf damage in the common-garden experiment were negatively related to damage in the source populations. The latitudinal variation in resistance was thus consistent with reduced selection from herbivores towards the northern range margin of L. salicaria. Variation in tolerance may be related to differences in the timing of damage in relation to the seasonal pattern of plant growth, as northern genotypes have developed further than southern have when herbivores emerge in early summer.

A comprehensive test of evolutionarily increased competitive ability in a highly invasive plant species.

BACKGROUND AND AIMS: A common hypothesis to explain plants' invasive success is that release from natural enemies in the introduced range selects for reduced allocation to resistance traits and a subsequent increase in resources available for growth and competitive ability (evolution of increased competitive ability, EICA). However, studies that have investigated this hypothesis have been incomplete as they either did not test for all aspects of competitive ability or did not select appropriate competitors. METHODS: Here, the prediction of increased competitive ability was examined with the invasive plant Lythrum salicaria (purple loosestrife) in a set of common-garden experiments that addressed these aspects by carefully distinguishing between competitive effect and response of invasive and native plants, and by using both intraspecific and interspecific competition settings with a highly vigorous neighbour, Urtica dioica (stinging nettle), which occurs in both ranges. KEY RESULTS: While the intraspecific competition results showed no differences in competitive effect or response between native and invasive plants, the interspecific competition experiment revealed greater competitive response and effect of invasive plants in both biomass and seed production. CONCLUSIONS: The use of both intra- and interspecific competition experiments in this study revealed opposing results. While the first experiment refutes the EICA hypothesis, the second shows strong support for it, suggesting evolutionarily increased competitive ability in invasive populations of L. salicaria. It is suggested that the use of naturally co-occurring heterospecifics, rather than conspecifics, may provide a better evaluation of the possible evolutionary shift towards greater competitive ability.

Rapid adaptation to climate facilitates range expansion of an invasive plant.

Adaptation to climate, evolving over contemporary time scales, could facilitate rapid range expansion across environmental gradients. Here, we examine local adaptation along a climatic gradient in the North American invasive plant Lythrum salicaria. We show that the evolution of earlier flowering is adaptive at the northern invasion front where it increases fitness as much as, or more than, the effects of enemy release and the evolution of increased competitive ability. However, early flowering decreases investment in vegetative growth, which reduces fitness by a factor of 3 in southern environments where the North American invasion commenced. Our results demonstrate that local adaptation can evolve quickly during range expansion, overcoming environmental constraints on propagule production.

Evidence for pollen limitation of a native plant in invaded communities.

Animal-pollinated invasive species have frequently been demonstrated to outcompete native species for pollinator attention, which can have detrimental effects on the reproductive success and population dynamics of native species. Many animal-pollinated invasive species exhibit showy flowers and provide substantial rewards, allowing them to act as pollinator 'magnets', which, at a large scale, can attract more pollinators to an area, but, at a smaller scale, may reduce compatible pollen flow to local native species, possibly explaining why most studies detect competition. By performing pollen limitation experiments of populations in both invaded and uninvaded sites, we demonstrate that the invasive plant Lythrum salicaria appears to facilitate, rather than hinder, the reproductive success of native confamilial Decodon verticillatus, even at a small scale, in a wetland habitat in southeastern Ontario. We found no evidence for a magnet species effect on pollinator attraction to invaded sites. Germination experiments confirmed that seeds from invaded sites had similar germination rates to those from uninvaded sites, making it unlikely that a difference in inbreeding was masking competitive effects. We describe several explanations for our findings. Notably, there were no differences in seed set among populations at invaded and uninvaded sites. Our results underscore the inherent complexity of studying the ecological impacts of invasive species on natives.

Response to enemies in the invasive plant Lythrum salicaria is genetically determined.

BACKGROUND AND AIMS: The enemy release hypothesis assumes that invasive plants lose their co-evolved natural enemies during introduction into the new range. This study tested, as proposed by the evolution of increased competitive ability (EICA) hypothesis, whether escape from enemies results in a decrease in defence ability in plants from the invaded range. Two straightforward aspects of the EICA are examined: (1) if invasives have lost their enemies and their defence, they should be more negatively affected by their full natural pre-invasion herbivore spectrum than their native conspecifics; and (2) the genetic basis of evolutionary change in response to enemy release in the invasive range has not been taken sufficiently into account. METHODS: Lythrum salicaria (purple loosestrife) from several populations in its native (Europe) and invasive range (North America) was exposed to all above-ground herbivores in replicated natural populations in the native range. The experiment was performed both with plants raised from field-collected seeds as well as with offspring of these where maternal effects were removed. KEY RESULTS: Absolute and relative leaf damage was higher for introduced than for native plants. Despite having smaller height growth rate, invasive plants attained a much larger final size than natives irrespective of damage, indicating large tolerance rather than effective defence. Origin effects on response to herbivory and growth were stronger in second-generation plants, suggesting that invasive potential through enemy release has a genetic basis. CONCLUSIONS: The findings support two predictions of the EICA hypothesis - a genetically determined difference between native and invasive plants in plant vigour and response to enemies - and point to the importance of experiments that control for maternal effects and include the entire spectrum of native range enemies.

Variability of total flavonoid, polyphenol and tannin contents in some Lythrum salicaria populations.

Lythrum salicaria L. can vary in some morphological and phytochemical features according to its habitat. The present study investigates and compares the total flavonoid, polyphenol and tannin composition of twelve populations of L. salicaria collected from different ecological habitats in south-west Hungary. We studied the plant heights and soil moistures of the habitats analyzing the potential correlation between the morphological and environmental factors and chemical compositions. Total flavonoid, polyphenol and tannin contents were determined according to the valid spectroscopic methods of the European Pharmacopoeia. Total flavonoid content was higher in the populations collected during the main blooming period in August than at the beginning of flowering in July. Higher values for total polyphenol and tannin were detected in flowering branch tips in August than in July, and higher ones in leaves and shoots in July than in August. The highest flavonoid content was measured in the leaves, followed by the flowering branches and shoots as opposed to the total polyphenol and tannin contents, which were higher in the flowering branch tips than in the other organs.

Increased relative abundance of an invasive competitor for pollination, Lythrum salicaria, reduces seed number in Mimulus ringens.

When exotic plant species share pollinators with native species, competition for pollination may lower the reproductive success of natives by reducing the frequency and/or quality of visits they receive. Exotic species often become numerically dominant in plant communities, and the relative abundance of these potential competitors for pollination may be an important determinant of their effects on the pollination and reproductive success of co-occurring native species. Our study experimentally tests whether the presence and abundance of an invasive exotic, Lythrum salicaria L. (Lythraceae), influences reproductive success of a co-flowering native species, Mimulus ringens L. (Phrymaceae). We also examine the mechanisms of competition for pollination and how they may be altered by changes in competitor abundance. We found that the presence of Lythrum salicaria lowered mean seed number in Mimulus ringens fruits. This effect was most pronounced when the invasive competitor was highly abundant, decreasing the number of seeds per fruit by 40% in 2006 and 33% in 2007. Reductions in the number of seeds per fruit were likely due to reduced visit quality resulting from Mimulus pollen loss when bees foraged on neighboring Lythrum plants. This study suggests that visit quality to natives may be influenced by the presence and abundance of invasive flowering plants.

Evolutionary constraints on adaptive evolution during range expansion in an invasive plant.

Biological invasions may expose populations to strong selection for local adaptation along geographical gradients in climate. However, evolution during contemporary timescales can be constrained by low standing genetic variation and genetic correlations among life-history traits. We examined limits to local adaptation associated with northern migration of the invasive wetland plant purple loosestrife (Lythrum salicaria) using a selection model incorporating a trade-off between flowering time and size at reproduction, and common garden experiments of populations sampled along a latitudinal transect of approximately 1200 km in eastern North America. A strong trade-off between flowering time and size at reproduction caused early-flowering plants to be smaller with reduced seed production in northern populations. Northward spread was associated with a decline in genetic variance within populations and an increase in genetic skew for flowering time and size, with limited genetic variation for small, early-flowering genotypes. These patterns were predicted by our selection model of local adaptation to shorter growing seasons and were not consistent with expectations from non-adaptive processes. Reduced fecundity may limit population growth and rates of spread in northern populations. Identifying genetic constraints on key life-history traits can provide novel insights into invasion dynamics and the causes of range limits in introduced species.

Negative per capita effects of two invasive plants, Lythrum salicaria and Phalaris arundinacea, on the moth diversity of wetland communities.

Invasive plants have been shown to negatively affect the diversity of plant communities. However, little is known about the effect of invasive plants on the diversity at other trophic levels. In this study, we examine the per capita effects of two invasive plants, purple loosestrife (Lythrum salicaria) and reed canary grass (Phalaris arundinacea), on moth diversity in wetland communities at 20 sites in the Pacific Northwest, USA. Prior studies document that increasing abundance of these two plant species decreases the diversity of plant communities. We predicted that this reduction in plant diversity would result in reduced herbivore diversity. Four measurements were used to quantify diversity: species richness (S), community evenness (J), Brillouin's index (H) and Simpson's index (D). We identified 162 plant species and 156 moth species across the 20 wetland sites. The number of moth species was positively correlated with the number of plant species. In addition, invasive plant abundance was negatively correlated with species richness of the moth community (linear relationship), and the effect was similar for both invasive plant species. However, no relationship was found between invasive plant abundance and the three other measures of moth diversity (J, H, D) which included moth abundance in their calculation. We conclude that species richness within, and among, trophic levels is adversely affected by these two invasive wetland plant species.

Re-establishment of clinal variation in flowering time among introduced populations of purple loosestrife (Lythrum salicaria, Lythraceae).

Range expansion during biological invasion requires that invaders adapt to geographical variation in climate, which should yield latitudinal clines in reproductive phenology. We investigated geographic variation in life history among 25 introduced populations of Lythrum salicaria, a widespread European invader of North American wetlands. We detected a strong latitudinal cline in initiation of flowering and size at flowering, which paralleled that reported among native populations. Plants from higher latitudes flowered earlier and at a smaller size than those from lower latitudes, even when raised in a uniform glasshouse. Early flowering was associated with greatly reduced reproductive output, but this was not associated with latitudinal variation in abundance, and probably did not result from a genetic correlation between time to and size at flowering. As introduction to North America c. 200 years ago, L. salicaria has re-established latitudinal clines in life history, probably as an evolutionary response to climatic selection.

Phenotypic plasticity of native vs. invasive purple loosestrife: a two-state multivariate approach.

The differences in phenotypic plasticity between invasive (North American) and native (German) provenances of the invasive plant Lythrum salicaria (purple loosestrife) were examined using a multivariate reaction norm approach testing two important attributes of reaction norms described by multivariate vectors of phenotypic change: the magnitude and direction of mean trait differences between environments. Data were collected for six life history traits from native and invasive plants using a split-plot design with experimentally manipulated water and nutrient levels. We found significant differences between native and invasive plants in multivariate phenotypic plasticity for comparisons between low and high water treatments within low nutrient levels, between low and high nutrient levels within high water treatments, and for comparisons that included both a water and nutrient level change. The significant genotype x environment (G x E) effects support the argument that invasiveness of purple loosestrife is closely associated with the interaction of high levels of soil nutrient and flooding water regime. Our results indicate that native and invasive plants take different strategies for growth and reproduction; native plants flowered earlier and allocated more to flower production, while invasive plants exhibited an extended period of vegetative growth before flowering to increase height and allocation to clonal reproduction, which may contribute to increased fitness and invasiveness in subsequent years.

Relationships among three pathways for resource acquisition and their contribution to plant performance in the emergent aquatic Plant Lythrum salicaria (L.).

Three pathways for resource acquisition exist in the emergent aquatic plant, Lythrum salicaria (L.); a subterranean root system, a free-floating adventitious root system, and arbuscular mycorrhiza (AM) fungal hyphae colonizing subterranean roots. This study examined the relationship(s) among these pathways and their contribution to plant performance. If the free-floating adventitious root system and/or AM fungi contribute to plant growth in wetland habitats, we predicted that their absence would result in a significant reduction in plant performance. Furthermore, if a reduction in resource uptake, effected by an absence of free-floating adventitious roots and/or AM fungi, is compensated for by increased allocation to remaining pathway(s) for resource uptake, we predicted altered patterns of resource allocation among shoots and the remaining pathway(s) for resource uptake. Contrary to our predications, plants experiencing adventitious root removal and/or grown in the absence of AM fungi generally had greater biomass and total shoot height than controls. Similarly, while levels of AM colonization and subterranean root biomass displayed a treatment effect, the observed responses did not correspond with our predictions. This was also true for shoot : subterranean root dry weight ratios. Our results indicate that there is interaction among the 3 pathways for resource acquisition in L. salicaria and an effect on plant performance. The adaptive significance of these characteristics is unclear, highlighting the potential difficulties in extrapolating from terrestrial to aquatic plant species and among aquatic plant species with potentially different life history strategies.

Trophic interactions between two herbivorous insects, Galerucella calmariensis and Myzus lythri, feeding on purple loosestrife, Lythrum salicaria, and two insect predators, Harmonia axyridis and Chrysoperla carnea.

The effects of two herbivorous insects, Galerucella calmariensis Duftschmid and Myzus lythri L. (Coleoptera: Chrysomelidae), feeding on purple loosestrife, Lythrum salicaria L. (Myrtiflorae: Lythraceae), were measured in the presence of two insect predators, Harmonia axyridis Pallas (Coleoptera: Coccinellidae) and Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae). A greenhouse cage experiment examined the direct effects of these predators on these herbivores, and indirect effects of predation on aboveground biomass, defoliation, number of leaves, and internode length. Eight treatment combinations with G. calmariensis, M. lythri, H. axyridis and C. carnea were applied to caged L. salicaria. The experiment ended when G. calmariensis adults were observed, 11 to 13 days after release of first instar G. calmariensis. G. calmariensis larvae alone removed significant amounts of leaf tissue and reduced the number of L. salicaria leaves. Predators did not reduce levels of defoliation by G. calmariensis. C. carnea had no effect on G. calmariensis survival, but H. axyridis reduced G. calmariensis survival in the presence of M. lythri. Both predators reduced the survival of M. lythri. This short duration greenhouse study did not demonstrate that predator-prey interactions altered herbivore effects on L. salicaria.

Rapid growth and early flowering in an invasive plant, purple loosestrife ( Lythrum salicaria L.) during an El Niño spring.

Phenological shifts may play a role in the success of invasive species, especially in association with climatic variability. We studied the response of a North American population of the invasive plant, Lythrum salicaria L., to changes in local climate associated with the El Niño/Southern Oscillation Event (ENSO) of 1997-1998. For L. salicaria plants at two wetland sites near North Bay, Ontario, Canada, we made weekly observations of flowering phenology and monthly measurements of aboveground biomass during the 1997 and 1998 growing seasons (April-October). Reproductive output was measured as cumulative length and biomass of inflorescences at the end of the growing season. Temperature and precipitation during the 1997 growing season were typical for the region and provided good baseline data for comparison to the full effects of the ENSO event in 1998, which increased spring temperatures and reduced precipitation in the study area. In response to these conditions, populations of L. salicaria began to flower 14 days earlier (Julian day = 181 +/- 10) in 1998 than in 1997 (Julian day = 195 +/- 12), and accumulated more aboveground biomass early in the growing season (P < 0.05). However, by the end of the growing season, there were no significant differences between years in aboveground biomass or total inflorescence lengths, and senescence of plants occurred at similar times for both growing seasons. Advances in spring phenology during ENSO events offer several potential advantages to L. salicaria, and could have a significant impact on biological control programs initiated for this species in North America.

Phosphorus, arbuscular mycorrhizal fungi and performance of the wetland plant Lythrum salicaria L. under inundated conditions.

The role of arbuscular mycorrhizal (AM) fungi in aquatic and semi-aquatic environments is poorly understood, although they may play a significant role in the establishment and maintenance of wetland plant communities. We tested the hypothesis that AM fungi have little effect on plant response to phosphorus (P) supply in inundated soils as evidenced by an absence of increased plant performance in inoculated (AM+) versus non-inoculated (AM-) Lythrum salicaria plants grown under a range of P availabilities (0-40 mg/l P). We also assessed the relationship between P supply and levels of AM colonization under inundated conditions. The presence of AM fungi had no detectable benefit for any measures of plant performance (total shoot height, shoot dry weight, shoot fresh weight, root fresh weight, total root length or total root surface area). AM+ plants displayed reduced shoot height at 10 mg/l P. Overall, shoot fresh to dry weight ratios were higher in AM+ plants although the biological significance of this was not determined. AM colonization levels were significantly reduced at P concentrations of 5 mg/l and higher. The results support the hypothesis that AM fungi have little effect on plant response to P supply in inundated conditions and suggest that the AM association can become uncoupled at relatively high levels of P supply.