Testing sex-allocation theory: flowers vs seeds in hermaphrodite plants

Predators are usually thought to be rarer (in the sense of having lower population densities) than non-predators. Recent analyses have suggested that this is not the case because a decline in species richness compensates for the well-known decline in number of individuals with increasing trophic rank. I show that a variety of invertebrate communities contain more species of predators than would be expected from the number of predatory individuals. This is not due to differences in dominance or taxonomic resolution between predatory and non-predatory guilds, and implies that predators are indeed relatively rare. I suggest that patterns of energy flow and body size make it likely that there will be a higher proportion of predatory species than individuals in a community, provided that predators have moderately specialized diets.     

Resource-dependent sex-allocation in a simultaneous hermaphrodite

Most sex allocation theory is based on the relationship between the resource investment into male and female reproduction and the consequent fitness returns (often called fitness-gain curves). Here we investigate the effects of resource availability on the sex allocation of a simultaneously hermaphroditic animal, the free-living flatworm Macrostomum lignano. We kept the worms under different resource levels and determined the size of their testes and ovaries over a period of time. At higher resource levels, worms allocated relatively more into the female function, suggesting a saturating male fitness-gain curve for this species. A large part of the observed effect was due to a correlated increase in body size, showing size-dependent sex allocation in M. lignano. However, a significant part of the overall effect was independent of body size, and therefore likely due to the differences in resource availability. Moreover, in accordance with a saturating male fitness-gain curve, the worms developed the male gonads first. As the group size was kept constant, our results contrast with expectations from sex allocation models that deal with local mate competition alone, and with previous experiments that test these models.     

Predictions of species interactions from consumer-resource theory: experimental tests with grasshoppers and plants

We tested the ability of consumer-resource theory to predict direct and indirect interactions among species, using an experimental system of insect herbivores and herbaceous plants. Specifically, we examined interactions among three species of grasshoppers (Melanoplus femur-rubrum, Spharagemon collare, andPhoetaliotes nebrascensis; Orthoptera, Acrididae) and herbaceous plants in experimental field cages placed over existing fertilized or unfertilized vegetation in a Minnesota old field. For the conditions inside these cages, we addressed whether (1) grasshopper diet predicted the presence of competition among grasshopper species, and (2) direct effects of grasshoppers on plants produced indirect interactions among plants, grasshoppers and soil nitrogen. Overall,M. femur-rubrum ate a greater proportion of forbs in cages, while the other two species ate primarily grasses. As expected, a pair of grasshopper species competed if they had similar diets. However, there were important exceptions that could be explained from observed indirect effects, although alternative explanations were also possible. First, all three grasshopper species significantly shifted their diets in the presence of other species, and these shifts occurred most often when competition was expected or occurred. Second, the two grassfeeding species reduced the biomass of the dominant grass (Schizachyrium scoparium) and increased available soil nitrogen and biomass of forbs. This effect may explain why the grass-feedingP. nebrascensis had a positive effect on the forb-feedingM. femur-rubrum on unfertilized plots. Overall, we show that direct effects of consumers on resources can predict competition and other important indirect interactions within a community.     

A systematic error in tests of ideal free theory

Classical ideal free theory predicts that the distribution of consumers within a patchy environment should correspond to the distribution of resources. Tests of this prediction have inappropriately compared ratios of mean resource levels and mean consumer densities, rather than means of ratios. We show that this error, which has propagated through hundreds of studies, leads to a systematic bias: the theory will appear to underestimate the number of consumers occupying poor patches. We explain the correct way to test ideal free theory and apply it to published data; the classical model is then seen to yield far more accurate predictions than previously thought.     

Experimental tests to assess emotionality in horses

Different tests were used to assess different aspects of the emotionality of 1–3 year-old horses: arena test; a `novel object' test; and a handling test. In reaction to the test situations no important differences were observed according to age or sex in the behaviour patterns, but clear individual differences were observed within these classes. The arena test seemed to reveal the degree of gregariousness of the animals whereas the results in the two other tests were correlated and seemed to reflect an inherent degree of fearfulness in the horse. Indices were developed that enabled to rank the animals, by taking into account all behaviour patterns shown. Such individual characteristics might have some genetic basis: half-siblings tended to behave the same way in most cases.     

Empirical tests of reciprocity theory: Problems in assessment

Observations and experiments designed to test Trivers' (1971) theory of reciprocal altruism face two difficulties. First, in many cases the costs and benefits of behaviors being exchanged cannot be expressed directly in terms of effects on the actors' fitness. This is particularly true when an exchange of cooperative acts involves different types of behavior, such as grooming and alliance formation in non-human primates. Second, in many social groups individuals differ widely in their ability to confer benefits on others. High-ranking animals can, for example, offer greater assistance than low-ranking animals and adults can help offspring more than offspring can help adults. As a result, what appears not to be a reciprocal interaction from the observer's point of view may in fact support Trivers' theory when costs and benefits are calculated from the animals' own perspectives. We discuss these issues with special reference to our own experiments on reciprocal exchanges of grooming and alliance formation in free-ranging vervet monkeys.     

Experimental tests of nutrient limitation in freshwater picoplankton

On the basis of correlative studies, picoplankton in Calder Lake, New York, are apparently unaffected by seasonal fluxes in nutrient (N and P) levels. In this small eutrophic lake, picoplankton (<2.0- to 0.2-mum size) and nanoplankton (<20 to >2 mum) predominate. Microplankton (>20 mum) are typically least important. Experiments were conducted in situ to test whether N, P or N/P ratios affect the predominance of these smaller organisms. Manipulations were run in 4-liter microcosms during June, July, and August 1988, corresponding to periods of increasing stratification and nutrient depletion. Following nutrient additions, phytoplankton were harvested and fractionated into three size classes. Microplankton and nanoplankton were significantly stimulated by both N (2.5 to 50 muM) and P (1 to 20 muM) additions. The severity of nutrient limitation was greatest during July. Picoplankton responded less strongly to N additions and were never P limited. These field data support laboratory studies which indicate that bacterium-sized phytoplankton use nutrients more efficiently and are superior competitors within mixed communities.     

Experimental tests of charge conservation in macromolecular interactions

A combination of equilibrium dialysis and ultrafiltration has been used to demonstrate the conservation of charge in the interaction between bovine serum albumin and methyl orange in Tris-HCl buffer, pH 7.4, I = 0.05 M; and also in the dimerization of alpha-chymotrypsin in acetate/chloride buffer, pH 3.9, I = 0.11 M, containing various concentrations of indole (0-10 mM) in order to displace the equilibrium position towards monomer. In the former study the magnitude of the negative charge on the albumin was shown to increase linearly with the number of molecules of methyl orange bound to the protein, the observed slope (0.96 +/- 0.08) of this relationship being in excellent agreement with that predicted on the basis of charge conservation for attachment of the univalent, negatively charged methyl orange ligand. In the study of alpha-chymotrypsin, the net charge (expressed per monomeric enzyme unit) was +10 in solutions in which the mole fraction of monomer varied between 0.47 and 0.88, the extent of this range having been established by means of constituent association equilibrium constants obtained from sedimentation equilibrium studies.     

Field tests on managing resistance to Bt-engineered plants

Several important crops have been engineered to express toxins of Bacillus thuringiensis (Bt) for insect control. In 1999, US farmers planted nearly 8 million hectares (nearly 20 million acres) of transgenic Bt crops approved by the EPA. Bt-transgenic plants can greatly reduce the use of broader spectrum insecticides, but insect resistance may hinder this technology. Present resistance management strategies rely on a "refuge" composed of non-Bt plants to conserve susceptible alleles. We have used Bt-transgenic broccoli plants and the diamondback moth as a model system to examine resistance management strategies. The higher number of larvae on refuge plants in our field tests indicate that a "separate refuge" will be more effective at conserving susceptible larvae than a "mixed refuge" and would thereby reduce the number of homozygous resistant (RR) offspring. Our field tests also examined the strategy of spraying the refuge to prevent economic loss to the crop while maintaining susceptible alleles in the population. Results indicate that great care must be taken to ensure that refuges, particularly those sprayed with efficacious insecticides, produce adequate numbers of susceptible alleles. Each insect/Bt crop system may have unique management requirements because of the biology of the insect, but our studies validate the need for a refuge. As we learn more about how to refine our present resistance management strategies, it is important to also develop the next generation of technology and implementation strategies.     

The effect of sex-allocation biasing on the evolution of worker policing in hymenopteran societies

Mutual policing is thought to be important in conflict suppression at all levels of biological organization. In hymenopteran societies (bees, ants, and wasps), multiple mating by queens favors mutual policing of male production among workers (worker policing). However, worker policing of male production is proving to be more widespread than predicted by relatedness patterns, occurring in societies headed by single-mated queens in which, paradoxically, workers are more related to the workers' sons that they kill than the queen's sons that they spare. Here we develop an inclusive-fitness model to show that a second reproductive conflict, the conflict over sex allocation, can explain the evolution of worker policing contrary to relatedness predictions. Among ants, and probably other social Hymenoptera, workers kill males to favor their more related sisters. Importantly, males are killed at the larval stage, presumably because workers cannot determine the sex of queen-laid eggs. Sex-allocation biasing favors worker policing because policing removes some males (the workers' sons) at low cost at the egg stage rather than at higher cost at the larval stage. Our model reveals an important interaction between two reproductive conflicts in which the presence of one conflict (sex allocation) favors the suppression of the other (male production by workers).     

DNA tests of neutral theory: applications in marine genetics

The principal methods of using DNA sequence information to test the neutral theory of evolution and polymorphism are described. These include the use of synonymous and nonsynonymous substitutions for detecting purifying and positive selection, the analysis of nucleotide diversity, mismatch analysis and the HKA, McDonald-Kreitman, Tajima and Ewens-Watterson tests. Analysis of the covariation of different kinds of molecular markers and the relationship between genetic variation and fitness is also considered. Examples of the use of these approaches in a wide variety of marine organisms are described. It is emphasised that tests of neutral theory, in addition to providing important fundamental knowledge about the action of evolutionary forces, provide valuable information about the influence of environmental and demographic factors.     

Experimental tests of villin subdomain folding simulations

We have used laser temperature-jump to investigate the kinetics and mechanism of folding the 35 residue subdomain of the villin headpiece. The relaxation kinetics are biphasic with a sub-microsecond phase corresponding to a helix-coil transition and a slower microsecond phase corresponding to overall unfolding/refolding. At 300 K, the folding time is 4.3(+/-0.6) micros, making it the fastest folding, naturally occurring protein, with a rate close to the theoretical speed limit. This time is in remarkable agreement with the prediction of 5 (+11,-3) micros by Zagrovic et al. from atomistic molecular dynamics simulations using an implicit solvent model. We test their prediction that replacement of the C-terminal phenylalanine residue with alanine will increase the folding rate by removing a transient non-native interaction. We find that the alanine substitution has no effect on the folding rate or on the equilibrium constant. Implications of this result for the validity of the simulated folding mechanism are discussed.     

Experimental tests of the cellular tensegrity hypothesis

The tensegrity model depicts the cytoskeleton (CSK) as a prestressed network of interconnected filaments. The prestress is generated by the CSK contractile apparatus and is partly balanced by traction at the cell-substrate interface and partly by CSK internal compression elements such as microtubules (MTs). A key feature of tensegrity is that the shear modulus (G) must increase in proportion with the prestress. Here we have tested that prediction as well as the idea that compression of MTs balance a portion of the cell prestress. Airway smooth muscle cells were studied. Traction microscopy was used to calculate traction. Because traction must be balanced by the stress within the cell, the prestress could be computed. Cell G was measured by oscillatory magnetic cytometry. The prestress was modulated using graded concentrations of contracting (histamine) or relaxing (isoproterenol) agonists and by disrupting MTs by colchicine. It was found that G increased in proportion with the prestress and that compression of MTs balanced a significant, but a relatively small fraction of the prestress. Taken together, these results do not disprove other models of cell deformability, nor they prove tensegrity. However, they do support a priori predictions of tensegrity. As such, it may not be necessary to invoke more complex mechanisms to explain these central features of cell deformability.     

Overview of QTL detection in plants and tests for synergistic epistatic interactions

Improvements in the usefulness of QTL analysis arise from better statistical methods applied to the problem, ability to analyze more complex mating designs, and the fitting of less simplified genetic models. Here we review the advantages of different plant mating designs in QTL analysis and conclude that diallel designs have several favorable properties. We then turn to the detection of systematic genome-wide synergistic epistasis. This form of epistasis has important implications from evolutionary (maintenance of sexual reproduction and concealment of cryptic genetic variation) and practical perspectives (response to pyramided favorable alleles). We develop two methods for detecting systematic synergistic epistasis, one based on analyzing interactions between locus effects and predicted individual genotypic values and one based on analyzing pairwise locus interactions. Using the first method we detect synergistic epistasis in a barley and a wheat dataset but not in a maize dataset. We fail to detect synergistic epistasis with the second method. We discuss our results in the light of theoretical questions concerning the mechanisms of synergistic epistasis.     

Diagnosis of Phytophthora infections in raspberry and strawberry plants by ELISA tests

Plant diseases caused by Phytophthora spp. may be reproducibly diagnosed by DAS ELISA techniques, but this type of analysis has long been hampered by the presence of phenolic and related compounds in plants to be tested, not least in strawberry and raspberry plants. The compounds will interfere with the ELISA test procedure, leading to high non-specific optical density readings. To overcome this, a series of experiments was performed. Phenolic and related compounds in the samples were first absorbed to polymers during antigen extraction and thereafter separated by filtration at slow rate. To inhibit non-specific binding of enzyme-conjugated antibodies, the plastic wells were preincubated with non-sensitized wells, equally high background values of optical density were seen under untreated conditions and after the use of polymeric adsorbents. A marked reduction in optical density was, however, seen after blocking with non-fat dry milk, but the optimal conditions for all concentrations ofantigen were seen first after combined pre-treatment with polymeric adsorbents and non-fat dry milk. When the threshold absorbance for positive detection was calculated, the low optical density values from healthy plants, at all antigen concentrations studied, contributed to an excellent discrimination between samples from diseased and healthy plants.     

Experimental tests of the Münch–Horwitz theory of phloem transport: effects of loading rates*

Abstract Extended square-wave tracer kinetics using ¹¹CO₂ were used to measure the speed of transport and activity level (proportional to concentration) in the phloem at high and low loading rates in six species of plants. In all cases, increased loading rates resulted in increased concentration. In most cases speed also increased, however, in two cases speed was lower and tracer activity was much higher at the higher loading rate. All the responses are consistent with the Münch Horwitz theory of phloem transport, depending upon the equation used to represent the unloading mechanism as described in a previous paper (Goeschl & Magnuson, 1986). For example, the latter two cases are consistent with the assumption that the unloading rate was limited by a process with saturable kinetics (enzyme-like).