Fractionation of stable zinc isotopes in the zinc hyperaccumulator Arabidopsis halleri and nonaccumulator Arabidopsis petraea.

Zn isotope fractionation may provide new insights into Zn uptake, transport and storage mechanisms in plants. It was investigated here in the Zn hyperaccumulator Arabidopsis halleri and the nonaccumulator A. petraea. Plant growth on hydroponic solution allowed us to measure the isotope fractionation between source Zn (with Zn(2+) as dominant form), shoot and root. Zn isotope mass balance yields mean isotope fractionation between plant and source Zn Δ(66)Zn(in-source) of -0.19 ± 0.20‰ in the nonaccumulator and of -0.05 ± 0.12‰ in the hyperaccumulator. The isotope fractionation between shoot Zn and bulk Zn incorporated (Δ(66)Zn(shoot-in)) differs between the nonaccumulator and the hyperaccumulator and is function of root-shoot translocation (as given by mass ratio between shoot Zn and bulk plant Zn). The large isotope fractionation associated with sequestration in the root (0.37‰) points to the binding of Zn(2+) with a high affinity ligand in the root cell. We conclude that Zn stable isotopes may help to estimate underground and aerial Zn storage in plants and be useful in studying extracellular and cellular mechanisms of sequestration in the root.

Arsenite efflux is not enhanced in the arsenate-tolerant phenotype of Holcus lanatus.

Arsenate tolerance in Holcus lanatus is achieved mainly through suppressed arsenate uptake. We recently showed that plant roots can rapidly efflux arsenite to the external medium. Here, we tested whether arsenite efflux is a component of the adaptive arsenate tolerance in H. lanatus. Tolerant and nontolerant phenotypes were exposed to different arsenate concentrations with or without phosphate for 24 h, and arsenic (As) speciation was determined in nutrient solutions, roots and xylem sap. At the same arsenate exposure concentration, the nontolerant phenotype took up more arsenate and effluxed more arsenite than the tolerant phenotype. However, arsenite efflux was proportional to arsenate uptake and was not enhanced in the tolerant phenotype. Within 2-24 h, most (80-100%) of the arsenate taken up was effluxed to the medium as arsenite. About 86-95% of the As in the roots and majority of the As in xylem sap (c. 66%) was present as arsenite, and there were no significant differences between phenotypes. Arsenite efflux is not adaptively enhanced in the tolerant phenotype H. lanatus, but it could be a basal tolerance mechanism to greatly decrease cellular As burden in both phenotypes. Tolerant and nontolerant phenotypes had a similar capacity to reduce arsenate in roots.

The distribution of arsenic in the body tissues of wood mice and bank voles.

Arsenic is accumulated by free-living small mammals, but there is little information on the resultant concentrations in different tissues other than liver and kidney. Such information is important because the severity of toxicological effects may be related to the amount of arsenic accumulated in specific organs, and the availability of arsenic to predators is, in part, dependent on which tissues accumulate arsenic. The objective of this study was to quantify the arsenic concentrations and the percentage of the total body burden (%TBB) accumulated in different body tissues of free-living small mammals and to determine how these factors varied with severity of habitat contamination. Arsenic concentrations were measured in various tissues of wood mice (Apodemus sylvaticus) and bank voles (Clethrionomys glareolus) from a range of arsenic-contaminated sites in southwest Britain. Arsenic concentrations in the gastrointestinal (GI) tract (including contents), liver, kidneys, spleen, lung, femur, and fur of both species varied significantly between sites and were higher in mice and voles from heavily contaminated areas. Heart and brain arsenic concentrations did not vary with degree of environmental contamination. The GI tract and excised carcass contained roughly equal amounts of arsenic and, in sum, comprised 75-85% of the TBB on uncontaminated sites and 90-99% on contaminated sites. Although the excised carcass contains about half of the TBB, its importance in food-chain transfer of arsenic to predators may depend on the bioavailability of arsenic sequestered in fur. In contrast, the GI tract and its contents, provided that it is consumed, will always be a major transfer pathway for arsenic to predators, regardless of the severity of habitat contamination.

QTL mapping for a trade-off between leaf and bud production in a recombinant inbred population of Microseris douglasii and M. bigelovii (Asteraceae, Lactuceae): a potential preadaptation for the colonization of serpentine soils.

The different response to growth on serpentine soil is a major autecological difference between the annual asteracean species Microseris douglasii and M. bigelovii, with nearly non-overlapping distribution ranges in California. Early flowering and seed set is regarded as a crucial character contributing to escape drought and thus is strongly correlated with survival and reproductive success on serpentine as naturally toxic soil. M. bigelovii (strain C94) from non-serpentine soil produces more leaves at the expense of bud production in the first growing phase than M. douglasii (B14) from serpentine soil. A QTL mapping study for this trade-off and for other growth-related traits was performed after six generations of inbreeding (F7) from a single interspecific hybrid between B14 and C94 on plants that were grown on serpentine and alternatively on normal potting soil. The trade-off is mainly correlated with markers on one map region on linkage group 03a (lg03a) with major phenotypic effects (phenotypic variance explained [PVE] = 18.8 - 31.7 %). Plants with the M. douglasii allele in QTL-B1 (QTL-NL1) produce more buds but fewer leaves in the first 119 days on both soil types. Three modifier QTL could be mapped for bud and leaf production. In one modifier (QTL-B2 = QTL-NL4) the M. douglasii allele is again associated with more buds but fewer leaves. QTL mapped for bud set in the F6 co-localize with QTL-B1 (major QTL) and QTL-B3. Two additional QTL for leaf length and red coloration of leaves could be mapped to one map region on lg03a. Co-localization of the two QTL loci with major phenotypic effects on bud and leaf production strongly suggests that a major genetic locus controls the trade-off between the two adaptive traits. The importance of mutational changes in major genes for the adaptation to stressful environments is discussed.

The potential for floral mimicry in rewardless orchids: an experimental study.

More than one-third of orchid species do not provide their pollinators with either pollen or nectar rewards. Floral mimicry could explain the maintenance of these rewardless orchid species, but most rewardless orchids do not appear to have a rewarding plant that they mimic specifically. We tested the hypothesis that floral mimicry can occur through similarity based on corolla colour alone, using naive bumble-bees foraging on arrays of plants with one rewarding model species, and one rewardless putative mimic species (Dactylorhiza sambucina) which had two colour morphs. We found that when bees were inexperienced, they visited both rewardless morphs randomly. However, after bees had gained experience with the rewarding model, and it was removed from the experiment, bees resampled preferentially the rewardless morph most similar to it in corolla colour. This is the first clear evidence, to our knowledge, that pollinators could select for floral mimicry. We suggest that floral mimicry can be a selective force acting on rewardless orchids, but only under some ecological conditions. In particular, we argue that selection on early-flowering rewardless orchids that receive visits from a large pool of naive pollinators will be weakly influenced by mimicry.

Negative frequency-dependent selection maintains a dramatic flower color polymorphism in the rewardless orchid Dactylorhiza sambucina (L.) Soo.

The orchid Dactylorhiza sambucina shows a stable and dramatic flower-color polymorphism, with both yellow- and purple-flowered individuals present in natural populations throughout the range of the species in Europe. The evolutionary significance of flower-color polymorphisms found in many rewardless orchid species has been discussed at length, but the mechanisms responsible for their maintenance remain unclear. Laboratory experiments have suggested that behavioral responses by pollinators to lack of reward availability might result in a reproductive advantage for rare-color morphs. Consequently, we performed an experiment varying the relative frequency of the two color morphs of D. sambucina to test whether rare morph advantage acted in the natural habitat of the species. We show here clear evidence from this manipulative experiment that rare-color morphs have reproductive advantage through male and female components. This is the first demonstration, to our knowledge, that negative frequency-dependent selection through pollinator preference for rare morphs can cause the maintenance of a flower-color polymorphism.

Zinc tolerance and accumulation in metallicolous and nonmetallicolous populations of Arabidopsis halleri (Brassicaceae).

Zinc tolerance was investigated in five populations of Arabidopsis halleri (syn.: Cardaminopsis halleri) raised from seeds collected from contaminated and uncontaminated sites. Tolerance was measured by determining the concentration which inhibited root growth (EC100 ). A. halleri populations from contaminated and uncontaminated sites were found to be Zn-tolerant compared with the Zn-nontolerant species Arabidopsis thaliana and A. lyrata subsp. petraea. At very high Zn concentrations, populations of A. halleri from uncontaminated sites were slightly less Zn-tolerant than those from contaminated sites. These observations support the hypothesis that in A. halleri, Zn tolerance is largely a constitutive property. One population from an uncontaminated site and one population from a contaminated site were studied for Zn uptake. Zinc content was measured in shoots and roots using a colorimetric test under laboratory conditions. The results showed that whatever their origin, individuals from both populations are Zn accumulators compared with the nonaccumulator species A. thaliana. Moreover, the population from the uncontaminated area accumulated Zn in its shoots and roots more quickly than the population from the contaminated site. These results suggest that, in A. halleri, Zn accumulation to very high concentration is a constitutive property.

Arsenic contamination in wood mice (Apodemus sylvaticus) and bank voles (Clethrionomys glareolus) on abandoned mine sites in southwest Britain.

Arsenic can be highly toxic to mammals but there is relatively little information on its transfer to and uptake by free-living small mammals. The aim of this study was to determine whether intake and accumulation of arsenic by wild rodents living in arsenic-contaminated habitats reflected environmental levels of contamination and varied between species, sexes and age classes. Arsenic concentrations were measured in soil, litter, wood mice (Apodemus sylvaticus) and bank voles (Clethrionomys glareolus) from six sites which varied in the extent to which they were contaminated. Arsenic residues on the most contaminated sites were three and two orders of magnitude above background in soil and litter, respectively. Arsenic concentrations in the stomach contents, liver, kidney and whole body of small mammals reflected inter-site differences in environmental contamination. Wood mice and bank voles on the same sites had similar concentrations of arsenic in their stomach contents and accumulated comparable residues in the liver, kidney and whole body. Female bank voles, but not wood mice, had significantly higher stomach content and liver arsenic concentrations than males. Arsenic concentration in the stomach contents and body tissues did not vary with age class. The bioaccumulation factor (ratio of arsenic concentration in whole body to that in the diet) in wood mice was not significantly different to that in bank voles and was 0.69 for the two species combined, indicating that arsenic was not bioconcentrated in these rodents. Overall, this study has demonstrated that adult and juvenile wood mice and bank voles are exposed to and accumulate similar amounts of arsenic on arsenic-contaminated mine sites and that the extent of accumulation depends upon the level of habitat contamination.

Zinc tolerance and hyperaccumulation are genetically independent characters.

The hyperaccumulation of metals by a rare class of plants is a fascinating and little understood phenomenon. No genetic analysis has been possible since no intraspecific variation is known for this character. Here, we report on crosses between the zinc-hyperaccumulating and -tolerant species Arabidopsis halleri and the non-hyperaccumulating, non-tolerant species Arabidopsis petraea. The F2 segregates for both characters and it appears that the two characters are genetically independent. The data for tolerance are consistent with a single major gene for this character (although the number of genes for hyperaccumulation cannot be determined), and is probably not very large.

Arsenic residues in predatory birds from an area of Britain with naturally and anthropogenically elevated arsenic levels.

Birds of prey forage over large areas and so might be expected to accumulate contaminants which are elevated but heterogeneously distributed in the general environment. The aim of this study was to test the hypothesis that arsenic levels in raptors from a region with elevated environmental arsenic concentrations were higher than those in birds from an uncontaminated part of Britain. Arsenic concentrations in the liver, kidney and muscle of kestrels, Falco tinnunculus, sparrowhawks, Accipiter nisus, and barn owls, Tyto alba, from south-west (SW) England, an area with naturally and anthropogenically (through mining) elevated environmental arsenic concentrations, were compared with those in birds from SW Scotland, where no such geochemical anomaly exists. Arsenic residues in kestrels from SW England were approximately three times greater than those in birds from SW Scotland for the three tissue types analysed. This was not the case for the other species in which arsenic residues were similar in birds from both regions. It is suggested that differences between species in both diet and arsenic metabolism could explain why kestrels have elevated arsenic tissue burdens in response to general environmental contamination but sparrowhawks and barn owls do not.

Pollen tube competition as a mechanism of prezygotic reproductive isolation between Mimulus nasutus and its presumed progenitor M. guttatus.

This paper considers the extent to which differences in pollen tube growth rates can provide prezygotic reproductive isolation between Mimulus nasutus and its presumed progenitor, Mimulus guttatus. Mimulus nasutus is partially cleistogamous, but its larger chasmogamous flowers offer appreciable opportunity for outcrossing. Mimulus nasutus was found to have smaller pollen grains and shorter styles than M. guttatus. No differences were observed in pollen grain germination on conspecific and heterospecific stigmas. However, pollen tube growth rates of M. nasutus were found to be much slower than those of M. guttatus in the styles of that species. Consequently, any M. nasutus pollen transferred to an M. guttatus stigma was found to be competitively disadvantaged in an M. gutattus style. By contrast, no difference in pollen tube growth rate was detected between the species when growing in M. nasutus styles, possibly because M. nasutus styles are unable to support fast pollen tube growth. We tested the prediction from the pollen tube studies that a 50∶50 mix of M. guttatus and M. nasutus pollen would produce 50% hybrid seeds when M. nasutus was the maternal parent, and near to 0% hybrid seed when M. guttatus was the maternal parent. The results were found to support this prediction. We conclude that pollen-pistil interactions can effect strong reproductive isolation between these species, as M. guttatus pollen tubes have a competitive advantage over those of M. nasutus in an M. guttatus style, but not in an M. nasutus style.

The genetic architecture of interspecific variation in mimulus.

The genetic architecture of various floral and morphological differences between Mimulus cupriphilus and Mimulus guttatus is investigated. M. cupriphilus is believed to have speciated from M. guttatus in the recent past. The two parent species, the F(1) and F(2), and two backcrosses were grown and scored for 23 different characters. The analysis of means revealed significant epistasis for a number of the floral characters, particularly those involving the length of parts. Dominance was generally toward M. guttatus, except for the characters related to flowering time. Analysis of the genetic correlations between characters revealed that there were at least four different polygenic genetic systems, governing flowering time, size of flower, number of spots on the corolla, and general size. An analysis of minimum gene number suggested that there were at least 3-7 genes controlling floral size, and a different three controlling floral spot number. Two other characters, corolla lobe shape and stem color, were produced by independent major gene differences. Annuality was also shown to be heritable. The two species appear to utilize the same gene for copper tolerance. The results are discussed in the light of current theories of speciation.

Heavy metal tolerance in plants: A model evolutionary system.

Evolved tolerance to toxic concentrations of heavy metals in plants inhabiting spoil heaps of mines is a well known phenomenon that has been the subject of much research in the last two decades. These plants are useful models for studying processes involved in the early stages of the speciation of edaphic endemics. Recent work has revealed the importance of several phenomena in the differentiation of tolerant populations, including natural selection, founder effects and 'hitch-hiking', and has demonstrated the early evolution of morphological differentiation and reproductive isolating mechanisms. Further studies of the biochemistry and molecular biology of heavy metal tolerance will help to show why some plant groups, such as Agrostis, are far more prone to evolve tolerance than others.

The number of genes on the second chromosome of Drosophila melanogaster and a comment on the genetic structure of eukaryotes.

Recessive lethals on the second chromosome were extracted from genetically isolated populations in Australia and the U.K. The frequency of allelism, used in a manner analogous to capture-recapture of animal populations, indicated that the number of genes capable of mutating to lethal had a 95 per cent probability of being in the range 247 to 1140, although excluding possible heterotic and synthetic lethals altered this to 309 to 3568. Possible sources of bias are discussed. The disagreement between these values and those obtained the direct measurements of DNA (more than 10 times greater) is clear. It is suggested that many of the genes of eukaryotes have been duplicated by unequal exchange during recombination and occur in functionally related groups or in supergenes. Some consequences of this hypothesis to population genetics are noted.

The Genetics of Copper Tolerance in the Yellow Monkey Flower, MIMULUS GUTTATUS. I. Crosses to Nontolerants.

The biometrical genetics of copper tolerance has been investigated in two Californian populations of Mimulus guttatus by crosses to a nontolerant British population. A simple biometrical model involving only additive and dominance effects is not sufficient. When the first order interactions are included, the model is shown to fit the data. Interactions between the dominance effects of different loci, and between dominance and additive effects, are the most important. These interactions can be explained either by a threshold model, or by postulating dominance modification.

Models of parent-offspring conflict. II. Promiscuity.

The population genetics of Trivers (1974) concept of parent-offspring is examined for species in which the effects of the conflict are felt by future half-sibs, as in promiscuous mating systems in which the male shows no parental care. Whether or not a rare conflictor gene will spread in a non-conflictor population depends on f(m) greater than (m + 1)/(0.5m + 1.5) for a dominant gene, and on f(m) greater than 1/4(7 + 3) for a recessive gene; f(m) is the fitness gained by a conflictor relative to a non-conflictor offspring [f(m) greater than 1], and m is the amount of parental investment taken by a conflictor relative to m = 1 for a non-conflictor. The ESS value for conflict (mo) in promiscuous species with zero male parental care has mo = f(mo)/4[df(mo)/dmo]. However, where the male maintains the same harem for several breeding seasons, or where there is promiscuity but both sexes contribute equally to parental care, conditions for conflict are equivalent to monogamy.

Models of parent-offspring conflict. I. Monogamy.

Theoretical models for Trivers (1974) concept of parent-offspring conflict are examined for species in which the effects of the conflict are felt by full sibs. A rare conflictor gene will spread if (f(m) greater than 1/2(m + 1), where f(m) is the fitness gained by a conflictor relative to a non-conflictor offspring (f(m) greater than 1), and m is the amount of parental investment taken by a conflictor relative to m = 1 for a non-conflictor. The range of m alleles which can spread against the parent optimum decreases as the cost to the parent increases until a point is reached where there is no conflict of evolutionary interests. There would be no polymorphism for conflictor: non-conflictor alleles unless special conditions prevail. The conflictor allele which spreads most rapidly as a rare mutant against the parental optimum is not an evolutionarily stable strategy (ESS). The ESS for parent-offspring conflict in monogamous species has mo = f(mo)/2[df(mo)/dmo]. The analytical solutions are confirmed throughout by simulations.