Factors limiting the recruitment of Quercus schottkyana, a dominant evergreen oak in SW China.

Quercus schottkyana is a dominant species of oak in the Asian evergreen broad-leaved forests in southwestern China but seedlings are uncommon and recruitment is rare. Annual acorn production by Q. schottkyana is variable and the acorns are exposed to a series of mortality risks. Understanding the factors that limit recruitment of the oak requires knowledge of the oak's life cycle from acorn production to germination and seedling establishment. In this study, we first tested the effects of acorn density on establishment of seedlings by placing batches of acorns at different densities throughout the study area. Second, we tested the effects of herbivores on seedling survival by erecting fences around both natural and transplanted seedling populations. Our results show that even though the rate of seedling establishment increases as acorn density increases (for 32-8000 acorns∙m-2), survival rates of seedlings in the field were generally low (0-0.6%). We show that seedling recruitment of Q. schottkyana is mainly limited to the acorn stage where 88% of the acorns died from the combined effects of desiccation and predation by weevils (Curculio) and bark beetles (Coccotrypes sp.). Herbivory results in the death of some seedlings and consequently also affects the recruitment of seedlings of Q. schottkyana.

Temperate grassland songbird species accumulate incrementally along a gradient of primary productivity.

Global analyses of bird communities along elevation gradients suggest that bird diversity on arid mountains is primarily limited by water availability, not temperature or altitude. However, the mechanism by which water availability, and subsequently primary productivity, increases bird diversity is still unclear. Here we evaluate two possible mechanisms from species-energy theory. The more individuals hypothesis proposes that a higher availability of resources increases the total number of individuals that can be supported, and therefore the greater number of species that will be sampled. By contrast, the more specialization hypothesis proposes that increasing resource availability will permit specialists to exploit otherwise rare resources, thus increasing total diversity. We used 5 years of surveys of grassland songbird communities along an elevational gradient in British Columbia, Canada, to distinguish between these hypotheses. Vegetation changed markedly in composition along the gradient and contrary to the expectations of the more specialization hypothesis, bird community composition was remarkably constant. However, both total abundance and species richness of birds increased with increasing water availability to plants. When we used rarefaction to correct species richness for differences in total abundance, much of the increase in bird diversity was lost, consistent with the expectations of the more individuals hypothesis. Furthermore, high species richness was associated with reductions in territory size of common bird species, rather than the fine-scale spatial partitioning of the landscape. This suggests that bird diversity increases when greater resource availability allows higher densities rather than greater habitat specialization. These results help explain a pervasive global pattern in bird diversity on arid mountains, and suggest that in such landscapes conservation of grassland birds is strongly linked to climate and hydrology.

Pre-dispersal strategies by Quercus schottkyana to mitigate the effects of weevil infestation of acorns.

We investigated how pre-dispersal strategies may mitigate the effects of weevil infestation of acorns in a population of Quercus schottkyana, a dominant oak in Asian evergreen broad-leaved forests, and assess if weevil infestation contributes to low seedling recruitment. We counted the number of acorns produced, daily from the end of August to mid-late November for 9 years from 2006-2014. We also recorded the rate of acorn infestation by weevils and acorn germination rates of weekly collections. Annual acorn production was variable, but particularly low in 2011 and 2013. There was no trade-off between acorn production and acorn dry mass. However, acorns produced later in the season were significantly heavier. For most years: (i) the rate of weevil infestation was negatively density dependent (a greater proportion of acorns died with increased acorn density), (ii) the percentage germination of acorns was positively density dependent (proportionately more acorns germinated with increased density), and (iii) as the season progressed, the percentage of infested acorns declined while germination rates increased. Finally, (iv) maximum acorn production, percentage infestation and percentage germination were asynchronous. Although pre-dispersal mortality is important it is unlikely to be the primary factor leading to low recruitment of oak seedlings.

An experimental approach to addressing ecological questions related to the conservation of plant biodiversity in China.

We briefly introduce and describe seven questions related to community structure and biodiversity conservation that can be addressed using field experiments, and provide the context for using the vast geographic diversity, biodiversity, and network of Nature Reserves in China to perform these experiments. China is the world's third largest country, has a diverse topography, covers five climatic zones from cold-temperate to tropical, has 18 vegetation biomes ranging from Arctic/alpine tundra and desert to Tropical rain forest, and supports the richest biodiversity in the temperate northern hemisphere (>10% of the world total). But this tremendous natural resource is under relentless assault that threatens to destroy biodiversity and negatively impact the services ecosystems provide. In an attempt to prevent the loss of biodiversity, China has established 2729 nature reserves which cover 14.84% of the nation's area. Unfortunately underfunding, mismanagement, illegal activities, invasive species and global climate change threaten the effectiveness of these protected areas. Attention has focused on protecting species and their habitats before degradation and loss of either species or habitats occur. Here we argue that we must move beyond the simple protection of ecosystems, beyond their description, and by using experiments, try to understand how ecosystems work. This new understanding will allow us to design conservation programs, perform restoration of damaged or degraded areas, and address resource management concerns (e.g., agriculture, logging, mining, hunting) more effectively than with the current approach of ad hoc reactions to ecological and environmental problems. We argue that improving our understanding of nature can best be done using well designed, replicated, and typically manipulative field experiments.

A new positive relationship between pCO2 and stomatal frequency in Quercus guyavifolia (Fagaceae): a potential proxy for palaeo-CO2 levels.

BACKGROUND AND AIMS: The inverse relationship between atmospheric CO2 partial pressure (pCO2) and stomatal frequency in many species of plants has been widely used to estimate palaeoatmospheric CO2 (palaeo-CO2) levels; however, the results obtained have been quite variable. This study attempts to find a potential new proxy for palaeo-CO2 levels by analysing stomatal frequency in Quercus guyavifolia (Q. guajavifolia, Fagaceae), an extant dominant species of sclerophyllous forests in the Himalayas with abundant fossil relatives. METHODS: Stomatal frequency was analysed for extant samples of Q. guyavifolia collected from17 field sites at altitudes ranging between 2493 and 4497 m. Herbarium specimens collected between 1926 and 2011 were also examined. Correlations of pCO2-stomatal frequency were determined using samples from both sources, and these were then applied to Q. preguyavaefolia fossils in order to estimate palaeo-CO2 concentrations for two late-Pliocene floras in south-western China. KEY RESULTS: In contrast to the negative correlations detected for most other species that have been studied, a positive correlation between pCO2 and stomatal frequency was determined in Q. guyavifolia sampled from both extant field collections and historical herbarium specimens. Palaeo-CO2 concentrations were estimated to be approx. 180-240 ppm in the late Pliocene, which is consistent with most other previous estimates. CONCLUSIONS: A new positive relationship between pCO2 and stomatal frequency in Q. guyavifolia is presented, which can be applied to the fossils closely related to this species that are widely distributed in the late-Cenozoic strata in order to estimate palaeo-CO2 concentrations. The results show that it is valid to use a positive relationship to estimate palaeo-CO2 concentrations, and the study adds to the variety of stomatal density/index relationships that available for estimating pCO2. The physiological mechanisms underlying this positive response are unclear, however, and require further research.

Species-specific responses to community density in an unproductive perennial plant community.

Most studies of density dependent regulation in plants consider a single target species, but regulation may also occur at the level of the entire community. Knowing whether a community is at carrying capacity is essential for understanding its behaviour because low density plant communities may behave quite differently than their high density counterparts. Also, because the intensity of density dependence may differ considerably between species and physical environments, generalizations about its effects on community structure requires comparisons under a range of conditions. We tested if: (1) density dependent regulation occurs at the level of an entire plant community as well as within individual species; (2) the intensity (effect of increasing community density on mean plant mass) and importance (the effect of increasing density, relative to other factors, on mean plant mass) of competition increases, decreases or remains unchanged with increasing fertilization; (3) there are species-specific responses to changes in community density and productivity. In 63 1 m2 plots, we manipulated the abundance of the nine most common species by transplanting or removing them to create a series of Initial Community Densities above and below the average natural field density, such that the relative proportion of species was consistent for all densities. Plots were randomly assigned to one of three fertilizer levels. At the community level, negative density dependence of mean plant size was observed for each of the 4 years of the study and both the intensity and importance of competition increased each year. At the species level, most species' mean plant mass were negatively density dependent. Fertilizer had a significant effect only in the final year when it had a negative effect on mean plant mass. Our data demonstrate a yield-density response at the entire community-level using perennial plant species in a multi-year experiment.

Response of grassland biomass production to simulated climate change and clipping along an elevation gradient.

Changes in rainfall and temperature regimes are altering plant productivity in grasslands worldwide, and these climate change factors are likely to interact with grassland disturbances, particularly grazing. Understanding how plant production responds to both climate change and defoliation, and how this response varies among grassland types, is important for the long-term sustainability of grasslands. For 4 years, we manipulated temperature [ambient and increased using open-top chambers (OTC)], water (ambient, reduced using rainout shelters and increased using hand watering) and defoliation (clipped, and unclipped) in three grassland types along an elevation gradient. We monitored plant cover and biomass and found that OTC reduced biomass by 15%, but clipping and water treatments interacted with each other and their effects varied in different grassland types. For example, total biomass did not decline in the higher elevation grasslands due to clipping, and water addition mitigated the effects of clipping on subordinate grasses in the lower grasslands. The response of total biomass was driven by dominant plant species while subordinate grasses and forbs showed more variable responses. Overall, our results demonstrate that biomass in the highest elevation grassland was least effected by the treatments and the response of biomass tended to be dependent on interactions between climate change treatments and defoliation. Together, the results suggest that ecosystem function of these grasslands under altered climate patterns will be dependent on site-specific management.

Declining diversity in abandoned grasslands of the carpathian mountains: do dominant species matter?

Traditional haymaking has created exceptionally high levels of plant species diversity in semi-natural grasslands of the Carpathian Mountains (Romania), the maintenance of which is jeopardized by recent abandonment and subsequent vegetation succession. We tested the hypothesis that the different life history strategies of dominant grasses cause different patterns of diversity loss after abandonment of traditional haymaking in two types of meadow. Although diversity loss rate was not significantly different, the mechanism of loss depended on the life history of dominant species. In meadows co-dominated by competitive stress-tolerant ruderals, diversity loss occurred following the suppression of dominant grasses by tall forbs, whereas in meadows dominated by a stress-tolerant competitor, diversity loss resulted from increased abundance and biomass of the dominant grass. We conclude that management for species conservation in abandoned grasslands should manipulate the functional turnover in communities where the dominant species is a weaker competitor, and abundance and biomass of dominant species in communities where the dominant species is the stronger competitor.

Mechanisms for success after long-term nutrient enrichment in a boreal forest understory.

Global levels of reactive nitrogen are predicted to rise in the coming decades as a result of increased deposition from the burning of fossil fuels and the large-scale conversion of nitrogen into a useable form for agriculture. Many plant communities respond strongly to increases in soil nitrogen, particularly in northern ecosystems where nitrogen levels are naturally very low. An experiment in northern Canada that was initiated in 1990 has been investigating the effects of long-term nutrient enrichment (fertilizer added annually) on a boreal forest understory community. We used this experiment to investigate why some species increase in abundance under nutrient enrichment whereas others decline. We focused on four species that differed in their responses to fertilization: Mertensia paniculata and Epilobium angustifolium increased in abundance, Achillea millefolium remained relatively constant and Festuca altaica declined. We hypothesized that the two species that were successful in the new high-nutrient, light-limited environment would be taller, have higher specific leaf area, change phenology by growing earlier in the season and be more morphologically plastic than their less successful counterparts. We compared plant height, specific leaf area, growth spurt date and allocation to leaves in plants grown in control and fertilized plots. We demonstrated that each of the two species that came to dominate fertilized plots has a different combination of traits and responses that likely gave them a competitive advantage; M. paniculata has the highest specific leaf area of the four species whereas E. angustifolium is tallest and exhibits morphological plasticity when fertilized by increasing biomass allocation to leaves. These results indicate that rather than one strategy determining success when nutrients become available, a variety of traits and responses may contribute to a species' ability to persist in a nutrient-enriched boreal forest understory.

Plant identity influences decomposition through more than one mechanism.

Plant litter decomposition is a critical ecosystem process representing a major pathway for carbon flux, but little is known about how it is affected by changes in plant composition and diversity. Single plant functional groups (graminoids, legumes, non-leguminous forbs) were removed from a grassland in northern Canada to examine the impacts of functional group identity on decomposition. Removals were conducted within two different environmental contexts (fertilization and fungicide application) to examine the context-dependency of these identity effects. We examined two different mechanisms by which the loss of plant functional groups may impact decomposition: effects of the living plant community on the decomposition microenvironment, and changes in the species composition of the decomposing litter, as well as the interaction between these mechanisms. We show that the identity of the plant functional group removed affects decomposition through both mechanisms. Removal of both graminoids and forbs slowed decomposition through changes in the decomposition microenvironment. We found non-additive effects of litter mixing, with both the direction and identity of the functional group responsible depending on year; in 2004 graminoids positively influenced decomposition whereas in 2006 forbs negatively influenced decomposition rate. Although these two mechanisms act independently, their effects may be additive if both mechanisms are considered simultaneously. It is essential to understand the variety of mechanisms through which even a single ecosystem property is affected if we are to predict the future consequences of biodiversity loss.

Dominant species and diversity: linking relative abundance to controls of species establishment.

Ecological theories make divergent predictions about whether extant species inhibit or promote the establishment of new species and which aspects of community composition determine these interactions; diversity, individual dominant species, and neutral interactions have all been argued to be most important. We experimentally tested these predictions by removing plant biomass (0%, 7%, 100%) from boreal forest understory communities. The 7% removals were restricted to the numerically dominant species, the second most dominant species, or many low-abundance species, thereby separating the effects of species composition from those of biomass. We tested the effects of all removal treatments on seedling establishment. Competitive effects were driven by one dominant species and were inconsistent with resource complementarity, neutral, or competition-colonization models. Facilitative effects were apparent only following removal of all vegetation, of which the most dominant species comprised more than 80%. Our results indicate that numerically dominant species in a community can influence the establishment of new species more than species diversity, but the direction of interaction can shift from facilitative to competitive as community density increases.

Serengeti birds maintain forests by inhibiting seed predators.

Of fundamental interest in conservation ecology are the regulatory mechanisms that maintain communities. We document a mechanism that maintains forests in the Serengeti ecosystem, Tanzania, and the destabilization when disturbance opens forest canopy. Forest birds, by consuming seeds, protected them from beetle attack. Consumption increased the germination rate and the density of seedlings and recruits, which was sufficient to maintain the forest. Opening of the canopy resulted in loss of birds, increased beetle attack, and loss of germination. Thus, frugivorous birds are necessary for the maintenance of forests. Their absence could have resulted in the observed forest decline since 1966.

Adaptive phenotypic plasticity of Pseudoroegneria spicata: response of stomatal density, leaf area and biomass to changes in water supply and increased temperature.

BACKGROUND AND AIMS: Changes in rainfall and temperature brought about through climate change may affect plant species distribution and community composition of grasslands. The primary objective of this study was to test how manipulation of water and temperature would influence the plasticity of stomatal density and leaf area of bluebunch wheatgrass, Pseudoroegneria spicata. It was hypothesized that: (1) an increased water supply will increase biomass and leaf area and decrease stomatal density, while a reduced water supply will cause the opposite effect; (2) an increase in temperature will reduce biomass and leaf area and increase stomatal density; and (3) the combinations of water and temperature treatments can be aligned along a stress gradient and that stomatal density will be highest at high stress. Methods The three water supply treatments were (1) ambient, (2) increased approx. 30% more than ambient through weekly watering and (3) decreased approx. 30 % less than ambient by rain shades. The two temperature treatments were (1) ambient and (2) increased approx. 1-3 degrees C by using open-top chambers. At the end of the second experimental growing season, above-ground biomass was harvested, oven-dried and weighed, tillers from bluebunch wheatgrass plants sampled, and the abaxial stomatal density and leaf area of tillers were measured. KEY RESULTS: The first hypothesis was partially supported--reducing water supply increased stomatal density, but increasing water supply reduced leaf area. The second hypothesis was rejected. Finally, the third hypothesis could not be fully supported--rather than a linear response there appears to be a parabolic stomatal density response to stress. CONCLUSIONS: Overall, the abaxial stomatal density and leaf area of bluebunch wheatgrass were plastic in their response to water and temperature manipulations. Although bluebunch wheatgrass has the potential to adapt to changing climate, the grass is limited in its ability to respond to a combination of reduced water and increased temperature.

The mutualism-parasitism continuum in ectomycorrhizas: a quantitative assessment using meta-analysis.

Context dependency is deemed to position the outcomes of species interactions along a continuum of mutualism to parasitism. Thus, it is imperative to understand which factors determine where a particular interspecific interaction falls along the continuum. Over the past 20 years research on the ectomycorrhizal symbiosis has resulted in sufficient independent studies to now generalize about the factors and mechanisms that affect host response to ectomycorrhizas. Using meta-analysis we quantitatively evaluated the role of biotic (partner identity and colonization levels of ectomycorrhizal fungi) and abiotic (phosphorus levels) factors in determining host biomass, height, and shoot:root responses to ectomycorrhizal associations. On average, seedlings across multiple host genera increased in total biomass when inoculated with ectomycorrhizal fungi regardless of the identity of the fungal associate; host genera differed in the magnitude of response for both total biomass and shoot:root ratio. Association with different fungal genera modified only host allocation of biomass to shoots and roots. Neither level of colonization on inoculated seedlings nor the level of contamination on control seedlings relative to colonization levels by target fungi on inoculated seedlings was important in explaining variation in effect sizes for any growth response. None of our proposed factors (identity of partners, colonization level, magnitude of contamination, or duration of association) explained variation in effect sizes for shoot height, although in general seedlings were taller when inoculated with ectomycorrhizal fungi. Phosphorus additions did not influence effect sizes. Although the general trend across studies was for a positive response of hosts to ectomycorrhizal inoculation, publication bias and methodological issues effectively reduce and distort the spectrum on which we evaluate host responses to ectomycorrhizal inoculation. Our results indicate that the variation in ectomycorrhizal fungi perceived by the host may be of a discrete (presence/absence of ectomycorrhizal fungi) rather than continuous nature (variation in identity or abundance of ectomycorrhizal fungi).

Quantitative partitioning of regional and local processes shaping regional diversity patterns.

Much recent research explaining plant community diversity has focused on comparing the relative impacts of regional and local processes. We employed a novel analysis to quantify the effects of multiple regional and local processes on species richness, and to make quantitative comparisons of those effects across two sites that differ in plot-level species richness, productivity and environmental conditions. While abiotic stress and competition limited richness within the communities at both sites, only differences in the overall pool of species at the site, likely resulting from long-distance dispersal and climate fluctuations, explained the differences in plot-level richness between sites. Patterns in local richness may be driven by a temporal storage effect, with greater richness in the site with greater climatic variability. By identifying both the factors that impact diversity within communities and those that vary systematically across communities, our integrated approach provides a better understanding of regional diversity patterns.

Dispersal, competition, and shifting patterns of diversity in a degraded oak savanna.

Diversity is a balance between processes that add and limit species (e.g., dispersal vs. competition), but reconciling their contributions remains a challenge. Recruit-ment-based models predict that dispersal barriers are most limiting for diversity, while competition-based models predict that dispersal matters only when competition is minimized. Testing these models is difficult because their influence varies with scale and site productivity. In a degraded oak savanna, we used plot-level (seed additions, burning) and site-level (proportions of regional functional groups found locally) analyses in areas with variable soil depth to examine how dispersal and competition influence diversity. At the plot level, added species persisted where they were formerly absent, but few established naturally despite fire-induced resource enrichment and nearby populations, revealing the importance of dispersal limitation for diversity. This result did not vary with soil depth or standing crop. Although competition could not prevent establishment in unburned plots, it significantly lowered survival, indicating that resource limitations exacerbate dispersal inefficiencies. At the site level, the concordance between regional and local diversity for native species was associated with soil depth heterogeneity, not dispersal or competition. This suggests that persistence is determined primarily by the influence of the environment on population demographics. Given that the formation of new populations is unlikely, those remaining appear to be confined to optimal habitat where they resist competitive or stochastic displacement, possibly explaining why species loss is rare despite substantial habitat loss and invasion. For exotics, there was no relationship between diversity and soil depth heterogeneity. Annuals with presumed dispersal capabilities were significantly overrepresented in all sites while perennial forbs, the largest regional functional group, were significantly underrepresented. We interpret the native-exotic discrepancies as reflecting the recent arrival of exotics (150 years ago), suggesting that local establishment occurs slowly even for species with regional prevalence. The accumulation lag may be explained by the need for founder populations to be demographically stable; otherwise persistence requires continual immigration favoring overrepresentation by dispersers. Our findings support the view that dispersal limitation restricts diversity within plant communities, but suggests that the impacts of environment on demographic performance ultimately determine the pattern and rate of community assembly.

Morphological plasticity of white clover (Trifolium repens L.) in response to spatial and temporal resource heterogeneity.

Heterogeneity in resource distribution has been an important selective force shaping morphological plasticity in plants. When resources are patchily distributed, changes in morphology are assumed to affect placement of the resource-acquiring structures (roots and leaves) such that they enhance the plant's capacity for resource uptake. Morphological development of four white clover (Trifolium repens) genets was studied in two glasshouse experiments. In the spatial experiment, two substrates (potting soil and sand) were used to create the following discrete patch combinations, sand-sand, soil-sand, sand-soil, and soil-soil. Stolons grew across each combination and consecutive ramets from a given stolon permitted rooting in each substrate pair. In the temporal experiment, the two ramets were first rooted in sand only. After a predetermined period, the sand was replaced and the same substrate combinations created as in the spatial experiment. In each experiment, total developmental time within a given substrate combination was held constant. All measurements were conducted on the second (i.e., younger) of the ramet pairs. In the spatial experiment, ramets rooted in soil had significantly greater branching frequencies than ramets rooted in the sand substrate, regardless of genotype or the preceding substrate type. Ramets occupying the sand-sand combination had the lowest branching frequencies but branch production for the ramet rooted in sand was higher if the preceding ramet was rooted in soil. The substrate occupied by a preceding ramet had no influence upon branching propensity if a ramet was rooted in soil. There were no significant differences in branching frequencies between the sand and soil substrates in the temporal experiment. The relationship between branching and substrate thus depended upon whether a ramet was exposed to a given substrate type during its early development. In both experiments, branched ramets in the soil-soil combinations had significantly greater shoot mass than corresponding ramets in the sand-sand combinations. Internode length was significantly shorter in the soil versus sand combinations of the spatial experiment but was unaffected by substrate in the temporal experiment. Leaf area and stolon width showed significant genotype×treatment interactions in both experiments but no consistent trends were evident; petiole length was unaffected by substrate.

Differential effects of light quality, provided by different grass neighbours, on the growth and morphology of Trifolium repens L. (white clover).

The ability to respond in a specific manner to different light conditions imposed by different species of grass is a major factor contributing to white clover persistence in pastures. Gaps in a pasture provide light with a higher red:far-red ratio (R:FR) and higher photosynthetic photon flux density (PPFD) than the light filtered through neighbours. White clover (Trifolium repens L.) was grown under different light conditions in ways that tried to simulate as closely as possible some of the light conditions experienced in a natural field situation, being partially shaded and receiving light reflected from neighbouring grasses. The objective was to determine specifically if the mere presence of neighbouring grasses could influence the growth and morphology of white clover individuals without physically contacting them, and thereby send a signal of impending competition. In the first experiment, white clover was subjected to shading cast from three different grass species. There were differences in both the quantity and quality of light received under the various grass canopies. The canopies reduced overall growth and branching of clones, while increasing the length of and biomass allocation to petioles. Lolium perenne L. canopy shade had different effects compared to Holcus lanatus L. or Dactylis glomerata L., but between the latter two species, no differences were detected. In the second experiment, light reflected from grass neighbours was provided simultaneously with direct light. There was a strong increase in FR and a resulting decrease in the R:FR due to neighbouring D. glomerata, but few consistent effects on white clover growth and morphology; there was evidence of phototropic movement by the leaves. We show that plants must experience partial shading, and not just reflected light, in order to alter their morphology in response to the presence of different species of grass neighbours.

Within-population variation in localized and integrated responses of Trifolium repens to biotically patchy environments.

Genets of Trifolium repens (white clover) were collected from three patches of old permanent pasture dominated by Agrostis capillaris, Holcus lanatus or Lolium perenne. Plants derived from the genets were grown with plants of one grass species present on one side of each T. repens, and a different grass species on the other side, in all combinations of two of the three grasses. Different modules (a node with its associated internode, leaf, and axillary bud) on the same clover plant responded independently to the microenvironment provided by their own neighbouring grasses. In contrast, all apical meristems on the plant reacted similarly, showing a unified response and integrating the effects of the different microenvironments experienced by the whole clover plant. This is consistent with what is known both physiologically about the nutrition of meristems and modules, and ecologically about the exploratory growth habit of the species. Averaged over all associated grasses, there was no significant variation in the final dry weight of the different clover genets but these differed in their growth habit response to different grasses. In response to Agrostis as a neighbour, each meristem of T. repens rapidly produced many small modules. New modules were produced more slowly and were larger when Holcus or Lolium was the neighbour. The same pattern of differences occurred among clovers sampled from different backgrounds. Either genetic differences paralleled plastic responses, or plastic changes in phenotype that developed in response to different neighbours in the field persisted in the greenhouse. Plants taken from backgrounds of different grass species showed different responses to growing with those grass species. The differences were manifest primarily in a "positive leading diagonal" effect of Holcus or not-Holcus. They were the result primarily of differences in the dry weight per module and the probability of development of the axillary bud into a branch. This confirms earlier results, and implicates the central importance of branching as a means of local response to the microenvironment.

The analysis of contact sampling data.

Use of the chi-square statistic for investigating interspecific association has been incorrectly applied to the contact sampling method because of an invalid probability argument. The usual method of calculating expected frequencies leads to a large overestimation and hence bias towards negative association. A correct method of analysis is presented and discussed.