Fruit removal and postdispersal survivorship in the tropical dry forest shrub Erythroxylum havanense: ecological and evolutionary implications

We studied the relationship between the removal rate and the spatiotemporal availability of ripe fruits of the tropical deciduous shrub Erythroxylum havanense in western Mexico. We also evaluated the effects of dispersal on seed survival during the first stages of establishment. Fast and early dispersal should be favored in E. havanense, since propagules have more time to grow and accumulate resources before the beginning of the severe dry season. In general, high rates of fruit removal imply faster and earlier dispersal. Thus, plants producing large crops should benefit from high removal rates, which will increase the probability of successful establishment by their progeny. To characterize both individual and population fruiting patterns, we made daily counts of fruits on 51 plants arranged in six clumps of different sizes. The daily number of fruits removed per plant was higher for plants with larger initial crop sizes and larger numbers of ripe fruits on a given day, but decreased as clump size increased. Additionally, we monitored postdispersal survival and germination in an experiment manipulating seed density, distance from adult plants, and seed predation. Early establishment was independent of density or distance, and vertebrate seed predation was the main agent of seed mortality. Our results indicate that the critical variable with respect to fruit removal is the number of fruits a plant produces, large plants having higher dispersal rates. Large plants are also more likely to have more seeds escaping postdispersal seed predation.     

Phenology and phenotypic natural selection on the flowering time of a deceit-pollinated tropical orchid, Myrmecophila christinae

BACKGROUND AND AIMS: Flowering phenology is described and the effect of flowering time on pollination success is evaluated in the deceit-pollinated tropical orchid, Myrmecophila christinae. It was expected that, due to this species' deceit pollination strategy and low observed pollinator visit rate, there would be a higher probability of natural selection events favouring individuals flowering away from the population flowering peak. METHODS: The study covers two consecutive years and four populations of M. christinae located along the north coast of the Yucatán Peninsula. For phenological and pollination success data, a total of 110 individuals were monitored weekly in 1998, and 83 individuals in 1999, during all the flowering and fruiting season. KEY RESULTS: The results showed significant differences in the probability of donating and receiving pollen throughout the flowering season. The probability of receiving or donating pollen increased the further an individual flowering was from the flowering peak. Regression analysis showed directional and disruptive phenotypic natural selection gradients, suggesting the presence of selection events unfavourable to flowering during flowering peak, for both male success (pollen removal) and female success (fruit production). However, the intensity and significance of the natural selection events varied between populations from year to year. The variation between seasons and populations was apparently due to variations in the density of reproductive individuals in each population and each season. CONCLUSIONS: As in other deceit-pollinated orchids, natural selection in M. christinae favours individuals flowering early or late in relation to population peak flowering. However, results also suggested a fluctuating regime of selective events act on flowering time of M. christinae.     

Phenotypic natural selection on flower biomass allocation in the tropical treeIpomoea wolcottiana Rose (Convolvulaceae)

Intra-population patterns of phenotypic variation in biomass allocation to flower parts are analyzed for the treeIpomoea wolcottiana, and the effects of this variation on fruit production are evaluated in two consecutive years. Significant variation among individuals was found for biomass of both sexual and attractive structures. Repeatability values showed that variation in these structures could be heritable. Analyses of phenotypic natural selection indicated a positive directional selection for both, gynoecium and petals mass in both years. These results suggest that floral resource allocation can be modified by phenotypic natural selection. To our knowledge this is the first paper that reports phenotypic natural selection in floral characteristics of a tropical tree species.     

Variation of development time until flowering in natural populations ofCapsella bursa-pastoris (Cruciferae)

Seed samples were collected from wild populations ofCapsella bursa-pastoris along a transsect from Northern to Southern Europe. Progeny was grown in (a) open-field random block experiments (47 populations) and (b) in growth chambers under five to seven controlled temperature regimes (18 populations). Beginning of flowering was recorded, and great differences between and also within populations are documented. Some populations are extremely heterogenous whereas others are homogenous in this respect. Some biotypes react positively when exposed to lower temperatures, others are inhibited. In many cases specific effects of day- and/or night-temperatures can be inferred. In some progenies begin of flowering is independent of temperature as long as this exceeds the 5:10°C regimen. Altogether,Capsella bursa-pastoris displays definite intraspecific variation in time required until flowering. Adaptations to local ecological conditions are obvious. In addition to a genotypic component pronounced environmental interactions provide the plants with a component of phenotypic plasticity. The degree of modificability apparently varies itself and seems to be controlled by selection; the phenotypic plasticity, therefore, displays adaptive variation patterns, too.Adaptation in Life History Traits of Colonizing Plant Species; Part of a doctoral thesis by the first author.     

Disruptive selection on female reproductive characters in a hybrid zone of Littorina saxatilis

We studied natural selection in a hybrid zone of the intertidal marine snail Littorina saxatilis located in Galicia (NW Spain) by measuring the number and average size of the embryos carried by females. We related these characters with the females' position on the phenotypic and environmental gradients between the two pure morphs of the hybrid zone. In contrast with previous interpretations of studies made in this hybrid zone, we found a depression in both embryo number and size for phenotypically intermediate females. This disruptive natural selection could play an important role in the maintenance of the population's bimodal phenotypic distribution and in that of the assortative mating between the pure morphs. We found also that intermediate environments tended to be unfavorable for all phenotypes. Although the precise causes for the found depression in female reproductive characters remain to be determined, these results serve to emphasize the importance of studying whole fitness surfaces in hybrid zones, across the complete phenotypic and environmental ranges, instead of merely comparing the fitness averages of the two pure morphs and the intermediate individuals, taken as three discrete phenotypic classes.     

The influence of animal sex hormones on the induction of flowering in Arabidopsis thaliana: comparison with the effect of 24-epibrassinolide

The influence of selected steroids on the in vitro generative development of Arabidopsis thalianawas investigated. The activity of the animal steroids androsterone, androstenedione, progesterone, estrone, estriol, and 17-estradiol was compared to 24-epibrassinolide, a member of the regulatory family of brassinosteroids. A. thaliana plants were cultured in vitro in media containing these steroids. The stimulatory effect of the tested substances was evaluated by measurement of the percentage of generative plants versus vegetative plants in the experimental group. It was established that androstenedione, the main testosterone precursor, and androsterone, a typical male hormone, were more effective in stimulating flowering in A. thaliana than the female hormones, estrogens and progesterone. Androsterone at a concentration of 0.1 M increased the percentage of generative plants up to 96% (control 41%). Estrogens at the same concentration decreased the number of generative plants and 24-epibrassinolide did not stimulate A. thalianagenerative development.     

Pedigree analysis on small laboratory populations of the butterfly Bicyclus anynana: The effects of selection on inbreeding and fitness

The effect of small population size and gene flow on the rate ofinbreeding and loss in fitness in Bicyclus anynana populationswas quantified by means of a pedigree analysis. Laboratorymetapopulations each consisted of four subpopulations with breeding sizeof N = 6 or N = 12 and migration rate of m = 0 or m= 0.33. Pedigrees were established by individually marking about35,000 butterflies. The increase in inbreeding coefficients(F-coefficients) over time was compared to that of simulated populationswith similar N and m. In the seventh generation, the level of inbreedingin larger subpopulations did not deviate significantly from the expectedvalues, but smaller subpopulations were less inbred than expected.Individuals in the small populations still showed considerableinbreeding depression, indicating that only a small proportion of therecessive deleterious alleles had been purged by selection. Two opposingprocesses potentially affected the rate of inbreeding and fitness: (1)Inbreeding depression increased the variance in family size and reducedthe effective population size. This will accelerate the rate ofinbreeding and is expected to selectively purge deleterious recessivealleles. (2) Variance in reproductive success of families was reducedbecause individuals which had a large number of siblings in thepopulation were more likely to mate with a full-sib than individualswith a smaller number of siblings. Subsequent inbreeding depressionreduced the number of viable offspring produced by these full-sibmatings. As a consequence, natural selection purged only some of thedeleterious alleles from the butterfly populations during sevengenerations with inbreeding. These findings emphasise the potentialproblems of using only small numbers of breeding individuals (N10) incaptive populations for conservation purposes.     

Reproductive fitness and artificial selection in animal breeding: culling on fitness prevents a decline in reproductive fitness in lines of Drosophila melanogaster selected for increased inebriation time

Summary The maintenance of reproductive fitness in lines subjected to artificial selection is one of the major problems in animal breeding. The decline in reproductive performance has neither been predictable from heritabilities and genetic correlations, nor have conventional selection indices been adequate to avoid the problem. Gowe (1983) has suggested that the heritabilities of reproductive traits are non-linear, with heritabilities being higher on the lower fitness side. Consequently, he has predicted that culling on reproductive fitness in artificial selection lines will be effective in preventing the usual declines in fitness. An experimental evaluation of Gowe's prediction has been carried out by comparing fitnesses of replicated lines of three treatments: selection for increased inebriation time without culling on fitness (HO), selection for inebriation time with culling of 20% (4/20) of selected females on reproductive fitness (HS), and unselected controls (C). Response to selection for inebriation time in the two selection treatments was similar. After 25 generations, the competitive index, a measure of reproductive fitness, was significantly lower in the HO treatment than the HS treatment, while the HS treatment did not differ from the control lines or the base population. These results demonstrate for the first time that culling on reproductive fitness in selection lines can be used to prevent the usual decline in reproductive performance.     

Non-linear selection response in Drosophila: a strategy for testing the rare-alleles model of quantitative genetic variability

Quantitative genetic theory predicts that variation due to rare alleles at many loci will generate a transient acceleration in the response to directional selection. We have tested this prediction by constructing experimental lines ofDrosophila melanogaster that carry positively selected ethanol resistance alleles at low frequencies, and then subjecting the lines to directional selection for ethanol resistance. Approximately 468,000 files were subjected to artificial selection over 30 generations. The predicted non-linear selection responses were observed in all experimental lines and replicates, on three genetic backgrounds. In contrast, un-selected controls and lines carrying random alleles at low frequencies on the same genetic backgrounds exhibited linear selection responses. These results demonstrate that non-linearities due to rare alleles are detectable and repeatable, provided that experiments are done on a sufficiently large scale. The results suggest that it may be possible to test for rare-alleles as a component of naturally occurring genetic variation by careful examination of selection response curves.     

Ecological and selective effects of stigma-anther separation in the self-incompatible tropical tree Ipomoea wolcottiana (Convolvulaceae)

We evaluated the effect of stigma-anther separation (SAS) in flowers of the self-incompatible tropical tree Ipomoea wolcottiana (Convolvulaceae). We measured the deposition of self pollen on the stigma and the growth of pollen tubes. We also evaluated phenotypic selection on female fertility using univariate regressions of SAS with fruit production. SAS varied among individuals in two years, with a range of 0–6 mm. There was a negative correlation between SAS and the number of self pollen grains on the stigma; trees with SAS of 0–2 mm received averages of ca 80–130 self grains. The effect of loading the stigma with self-pollen was tested by allowing isolated flowers to self-pollinate, then cross-pollinating by hand: there was a negative correlation between the pollen load and the proportion of grains forming pollen tubes. The highest proportion of grains forming tubes was found with loads of <10 grains, while the minimum frequency of tubes occurred with loads of >50 grains. The results suggest that more self pollen deposition leads to fewer tubes from outcross pollen. There was a significant effect of SAS on fruit production in both years. The analyses suggested stabilizing selection; very small SAS is disadvantageous due to the high load of self pollen, while very large SAS probably led to damage to the style during pollinator visits. We suggest that herkogamy may be a key character for determining the success of fruit production in self-incompatible hermaphroditic species.     

Effect of balancing selection on spatial genetic structure within populations: theoretical investigations on the self-incompatibility locus and empirical studies in Arabidopsis halleri

The effect of selection on patterns of genetic structure within and between populations may be studied by contrasting observed patterns at the genes targeted by selection with those of unlinked neutral marker loci. Local directional selection on target genes will produce stronger population genetic structure than at neutral loci, whereas the reverse is expected for balancing selection. However, theoretical predictions on the intensity of this signal under precise models of balancing selection are still lacking. Using negative frequency-dependent selection acting on self-incompatibility systems in plants as a model of balancing selection, we investigated the effect of such selection on patterns of spatial genetic structure within a continuous population. Using numerical simulations, we tested the effect of the type of self-incompatibility system, the number of alleles at the self-incompatibility locus and the dominance interactions among them, the extent of gene dispersal, and the immigration rate on spatial genetic structure at the selected locus and at unlinked neutral loci. We confirm that frequency-dependent selection is expected to reduce the extent of spatial genetic structure as compared to neutral loci, particularly in situations with low number of alleles at the self-incompatibility locus, high frequency of codominant interactions among alleles, restricted gene dispersal and restricted immigration from outside populations. Hence the signature of selection on spatial genetic structure is expected to vary across species and populations, and we show that empirical data from the literature as well as data reported here on three natural populations of the herb Arabidopsis halleri confirm these theoretical results.     

Impact of negative frequency-dependent selection on mating pattern and genetic structure: a comparative analysis of the S-locus and nuclear SSR loci in Prunus lannesiana var. speciosa

Mating processes of local demes and spatial genetic structure of island populations at the self-incompatibility (S-) locus under negative frequency-dependent selection (NFDS) were evaluated in Prunus lannesiana var. speciosa in comparison with nuclear simple sequence repeat (SSR) loci that seemed to be evolutionarily neutral. Our observations of local mating patterns indicated that male-female pair fecundity was influenced by not only self-incompatibility, but also various factors, such as kinship, pollen production and flowering synchrony. In spite of the mating bias caused by these factors, the NFDS effect on changes in allele frequencies from potential mates to mating pollen was detected at the S-locus but not at the SSR loci, although the changes from adult to juvenile cohorts were not apparent at any loci. Genetic differentiation and isolation-by-distance over various spatial scales were smaller at the S-locus than at the SSR loci, as expected under the NFDS. Allele-sharing distributions among the populations also had a unimodal pattern at the S-locus, indicating the NFDS effect except for alleles unique to individual populations probably due to isolation among islands, although this pattern was not exhibited by the SSR loci. Our results suggest that the NFDS at the S-locus has an impact on both the mating patterns and the genetic structure in the P. lannesiana populations studied.     

Fragmentation by weirs in a riverine system: A study of genetic variation in time and space among populations of European grayling (Thymallus thymallus) in a Danish river system

Human induced habitat alterations affect the genetic structure of many fish populations. Weirs in particular have caused fragmented populations previously connected by gene-flow. We studied the effects of weirs on the distribution of genetic variation within European grayling (Thymallus thymallus) populations from the Skjern River, Denmark. We compared microsatellite data from DNA extracted from historic scale samples collected 60 years ago with data from contemporary samples. Pairwise multilocus F _ST estimates between all contemporary population samples were significant as well as exact tests for population differentiation. Assignment tests of individuals to a set of baseline samples showed correct assignment to the population of origin of between 54 and 79%. Assignment of individuals from recent samples to the historic population samples showed highly variable results (3–83%) of correct assignment suggesting different population histories. Pairwise multilocus F _ST estimates were significantly correlated with the number of intervening weirs but not with waterway distance. A simulation procedure was used to estimate differences in relative population sizes, which indicated that the main river population was approximately three times larger than those of the tributaries. There were no signs of any loss of genetic variation for the river system as a whole during these 60 years. The results show that weirs can be an important factor for creating the current distribution patterns of genetic variation among grayling populations, most likely by only allowing passive downstream drift of fry and obstructing active upstream migration. This revised version was published online in July 2006 with corrections to the Cover Date.     

Factors affecting the initiation and growth of aboveground adventitious roots in a tropical cloud forest tree: an experimental approach

Some of the proximate factors that would induce aboveground stems to produce adventitious roots were investigated experimentally on Senecio cooperi, a tropical cloud forest tree. Stem segments were air-layered with different treatments to promote root formation, and the number of roots initiated and rates of root growth were monitored for 20 weeks. Treatments were the application of wet epiphytes or dry epiphytes plus associated humus, sponges wetted with either water or nutrient solutions, or dry sponges. Controls (stem segments with nothing applied) were also monitored. Numbers of adventitious roots formed and rates of subsequent root growth differed among treatments. Wet epiphyte/humus and nutrient solutions were most effective in producing roots, which suggests that epiphytes and the nutrients they intercept and retain within the canopy may cue adjacent host tissue to exploit this resource.     

Effect of variation in photoperiodic response on diapause induction and developmental time in the willow leaf beetle, Plagiodera versicolora

The willow leaf beetle, Plagiodera versicolora (Coleoptera: Chrysomelidae) overwinters in adult diapause. In this study, the photoperiodic responses for diapause induction and developmental time were examined in the Ishikari (Hokkaido, Japan) population of P. versicolora. All females entered reproductive diapause under short daylength (L10:D14), but 31.7% of females did not enter diapause under long daylength (L16:D8). The developmental time from oviposition to adult emergence was significantly longer at L10:D14 than that at L16:D8. Norm of reaction curves illustrated variation among families in the photoperiodic responses for diapause induction and for developmental time. ANOVA indicated significant family × photoperiod interactions in the developmental time. At L16:D8, developmental time was positively correlated with the incidence of diapause in females. This means that a female having a longer developmental time tends to have a longer critical photoperiod. Such variation may be maintained by differences in selection pressures on the growth rate and the critical photoperiod for diapause induction between univoltine and bivoltine genotypes because Ishikari is located in a transitional area between populations with univoltine and bivoltine life cycles.     

Strong habitat selection and the development of population structure in a mosquito

Summary Remarkably simple and regular patterns of genic variation for two electrophoretically assayed loci are presented against a background of strong habitat selection, population isolation, and population structuring in the pitcher-plant mosquitoWyeomyia smithii. The genic variation assessed thus far comes from 29 local populations, all but one of which lie within the glaciated region of North America. Although the conclusions drawn from these patterns are still quite tentative, it appears that the fact that the same higher frequency electromorphs occurred everywhere for both loci may be due more to selection than drift. Drift or selection, acting alone or in concert may be responsible for the almost complete absence of electromorphs of intermediate frequency. A weak association was found between the two-locus genic variation and the well-studied diapause-development time variation in this species. It is suggested that a Wrightian interaction system and fluctuating-stabilizing selection might account for maintenance of the genic level polymorphism. The observations reported are circumstantially in accord with Wright's shifting balance process of evolution. Future studies in the older parts of the species range will put these hypotheses to a strong test.     

The phenology of the San Jose scaleQuadraspidiotus perniciosus [Hom.: Diaspididae] and its association with its natural enemies on almond trees in northern Greece

The phenology of the San Jose scale was studied in a heavily-infested almond orchard in northern Greece. It appears that the scale there completes 3 generations per year. The majority of the population hibernates as 1^st instar larvae. A series of releases were made using the introduced Hymenoptera parasiteProspaltella perniciosi Tower. Sampling results showed a very high mortality of the San Jose scale, Apart from the percentage due to climatic factors, the remainder is a result of the action of natural enemies. A considerable part of this action is attributable to the local entomophagous insects and especially the predators, of whichCybocephalus fodori Endrody-Younga appears to be the key species. The rest is attributable to the action of the released populations ofP. perniciosi. The main part of this paper was presented during the International Symposium held in Budapest (15–20 August 1983).     

Inoculation experimental animals withParacoccidioides brasiliensis strains: An attempt to reestablish the dimorphic process and variation in pathogenicity as a function of time of preservation under mineral oil

In an attempt to reestablish the dimorphic process in strains ofParacoccidioides brasiliensis in the transition phase (Y ⇄ M) and to reisolate them, five strains in the transitional phase due to the long time of preservation under mineral oil and two strains in the yeast-like phase were inoculated into male albino rats. The animals were then studied for the presence of paracoccidioidomycotic granulomata. Of the seven strains inoculated, five caused granulomatous nodules in several organs of the animals and only two of these five strains, which had been preserved for the shortest period of time (9 years) were reisolated in culture. Two strains were unable to provoke infection, with no lesions detected in any organ. It is assumed that the long period of time during which the strains were left under oil favored the alteration of celt wall contents, leading to differences in pathogenicity.     

Photosynthetic oxygen evolution and chlorophyll fluorescence in intact isolated chloroplasts on a solid support: the influence of orthophosphate

We devised recently a method to trap intact isolated chloroplasts on a solid support consisting of membrane filters made of cellulose nitrate (Cerovi et al., 1987, Plant Physiol. 84, 1249–1251). The addition of alkaline phosphatase to the reaction medium enabled continuous photosynthesis by spinach (Spinacia oleracea L.) chloroplasts to be sustained by hydrolysis of newly produced and exported triose phosphates and recycling of orthophosphate. In this system, simultaneous measurements of chlorophyll fluorescence and oxygen evolution were performed and their dependence on orthophosphate concentration was investigated. Optimal photosynthesis was obtained at a much higher initial orthophosphate concentration (2–4 mM) compared to intact chloroplasts in suspension. Secondary kinetics of chlorophyll fluorescence yield were observed and were shown to depend on the initial orthophosphate concentration.Abbreviations Chl chlorophyll - CSS intact isolated chloroplasts on solid support - ICS intact isolated chloroplasts in suspension - P_i orthophosphate - v rate of O₂ evolution - PPFD photosynthetic photon flux density The authors wish to thank Dr. Marijana Plesniar, from the University of Novi Sad, for stimulating discussions. This work was supported by the Fond for Science of the Republic of Serbia. Z.G.C.'s visit to the Robert Hill Laboratory was supported by the British Council and the University of Sheffield.