Melanic moths and charges in epiphytic vegetation in north-west England and north Wales

In 1973 the appearance and epiphyte cover of tree trunks was examined along a 125 km cline of melanic frequency in the peppered moth in NW England and N Wales. The sampled sites were resurveyed in 1986. There has been a marked increase in the crustose lichen Lecanora conizaeoides at the eastern end of the transect, an increase in lichen diversity in the middle of the transect and a possible decline in large foliose forms at the western end. These changes are discussed in relation to the eastward shift of the cline in melanic frequencies in the moth.     

Fitness of insularia morphs of the peppered moth Biston betularia

In recent years the industrial melanic carbonaria morph in the moth Biston betularia (L.) has decreased rapidly in frequency in Britain as air pollution has decreased. The intermediate melanic insularia has shown a variable response. We have estimated the fitness of insularia, compared with the other two morphs, for several data sets. As a rule its fitness lies between that of carbonaria and typical, but nearer to typical and sometimes very close to it. The intermediate position is expected if fitness relates directly to phenotype. The results suggest that insularia may continue polymorphic while carbonaria is likely to disappear. The past high frequency of insularia in South Wales may have been due to an initial increase in insularia frequency before carbonaria reached the region. Differences in dynamics of frequency change in insularia and carbonaria are evidence against induction, which has sometimes been invoked to explain the spread of melanism in this species.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 82 , 359–366.     

Peppered moth revisited: analysis of recent decreases in melanic frequency and predictions for the future

In a recent review article by Mani published in 1990, it was shown that the changes in the frequency of the morph carbonaria can be explained through a migration-selection model and the model is capable of reproducing the decrease in the carbonaria frequency since the enactment of Clean Air Acts. In this paper new data both for carbonaria and for insularia up to 1990 for West Kirby in NW England and for Cambridge are presented. In the earlier review, the data for West Kirby extended to 1987 and for Cambridge to 1985. Data not previously published for the carbonaria and the insularia frequencies at sites in Northwood in Middlesex, Egham in Surrey and Ringwood in Hampshire are also presented. A migration-selection model was used to predict the change in the frequencies of the melanic morphs up to the year 2010. All the data are compared with the predictions of the model. It is encouraging that the data follow the theoretical predictions.     

The understanding of industrial melanism in the peppered moth (Biston betularia) (Lepidoptera: Geometridae)

Melanic polymorphism in B. betularia has been extensively studied. Correlations between high melanic frequency and high levels of air pollution have been demonstrated. Kettlewell and others have shown that differential bird predation has an important effect on the maintenance of the polymorphism, and coefficients of visual selection have been obtained on the assumption that the moth habitually rests on tree trunks. Computer models based on these selective coefficients show that they are not sufficient accurately to explain observed melanic frequencies. Other non-visual selective factors and weak frequency-dependent selection have been invoked to improve fits. Analysis of the resting positions of moths recorded in the wild demonstrates that B. betularia does not usually rest in exposed positions on tree trunks, but rather rests on the underside of branches, on trunks in shaded positions just below major branch joints or on foliate twigs. The results of a pilot selection experiment, while agreeing qualitatively with Kettlewell's results, suggest that fitness estimates that assume trunk-resting are quantitively incorrect. The error is greatest for melanic moths in rural areas. It is suggested that visual selective coefficients based on a true assessment of the resting behaviour of the moths may considerably improve the fit between computer predictions and observed phenotype frequency distributions.     

Non‐morph specific predation of peppered moths (Biston betularia) by bats

Abstract 1. Industrial melanism in the peppered moth, Biston betularia, is one of the foremost examples of natural selection in action. 2. Differential bird predation was suggested as the main agent for the evolution of melanism in the peppered moth by Tutt in the 1890s, with empirical support being published by Kettlewell in the 1950s. 3. Some recent critiques that have attempted to undermine Kettlewell’s work have lacked objectivity, and have been answered previously. 4. One criticism that has not previously been addressed is that of the role of bat predation in the case. 5. The difficulty of using non‐visual differential predation by bats to explain the increase and decrease in melanism in the peppered moth, correlated as it is to pollution levels, is outlined. 6. Predation experiments, in which moths of the typica and carbonaria forms of the peppered moth were released and observed at night, were used to determine whether bats differentially predate these forms. 7. Results of experiments at three sites showed no significant differences in the level of bat predation of the two forms of peppered moth.     

Theoretical models of melanism in Biston betularia—a review

The various theoretical models that have been constructed to explain the spatial and temporal changes in the frequency of the melanic morph of the moth Biston betularia are reviewed. The assumptions that are made in these models are discussed. It is shown that these models do explain the gross features of the spatial and temporal distribution of melanic variety in England and Wales. The models have been fairly successful in quantitative prediction of the observed decline in the frequencies of the melanic forms by relating selective differentials to the sulphur dioxide levels. Thus these models do yield a reasonable and consistent picture of the gross pattern of changes in melanic frequencies within the limitation of the available data. The models are robust and can thus accommodate changes to the data on the biology and behaviour of the moth. It is shown that neither heterozygote advantage nor non-Darwinian mechanism need to be invoked to understand the observed evolution of melanism in B. betularia.     

A decline of melanism in the peppered moth Biston betularia in The Netherlands

Melanic forms of the peppered moth Biston betularia were well established in The Netherlands by the end of the 19th century, indeed the first records of the black carbonaria form in 1867 are only about 20 years later than in England. Analysis of extensive sampling data collected by B. J. Lempke for a period of several years beginning in 1969 shows that carbonaria was at a frequency of about 60 to 70% in most of the country where epiphyte communities on trees were reduced due to the effects of air pollution. The pale typica and the three intermediate insularia forms were each at similar, low frequencies. Only in the extreme north and south-east of The Netherlands where epiphyte floras were richer was carbonaria at a lower frequency of less than 40%. Samples collected from seven localities in 1988 show that carbonaria has dramatically declined to a frequency of less than 10%. In contrast to England, the fully black form is being replaced not only by typica but also by the darkest of the insularia phenotypes. The decline in melanism coincides with a period of decreasing levels of sulphur dioxide and of increasing species diversity of lichens on trees.     

Evolution of resting behaviour in polymorphic 'industrial melanic' moth species

The behaviour of individually marked melanic and typical forms of Allophyes oxyacanthae was followed in four successive tests in an apparatus comprising bark of three different reflectances. In samples of wild populations, melanics showed a moderate preference for dark bark, while typicals did not seem to prefer or avoid this substrate. However, in four of the eight families of known parentage tested, both melanics and typicals preferred to rest on dark bark. These four families were the progeny of dark typicals which had shown a preference for dark bark and were the families containing the darkest typicals. In A. oxyacanthae the variation in resting behaviour is not therefore closely associated with the melanic allele but may be linked to a 'dark typical' allele which, together with other loci, produces polygenic darkening of the normal phenotype. The results with A. oxyacanthae are compared with those obtained with Biston betularia and other moth species.     

Background selection by the peppered moth (Biston betularia Linn.): individual differences

The hypothesis that dimorphically coloured, cryptic moths select appropriate rest sites by comparing their body scales to substrate reflectance was tested using typical and melanic morphs of the peppered moth, Biston betularia (L.). Experiments designed to block the individual's inspection of its inherited colour phenotype do not support Kettlewell's contrast/conflict (self-inspection) hypothesis. Instead, tracking of marked moths over successive days revealed individual differences in rest-site selection which were not related to treatments, experience (imprinting), nor closely to a moth's inherited colour pattern. Differences between family broods indicate that some genetic bias in background selection exists. The production of artificially selected lines with consistent but opposing preferences will allow us to investigate the co-evolution of pattern and behaviour.     

The reliability of estimates of migration in the peppered moth Biston betularia and some implications for selection-migration models

Migration of adult males is one of the important variables involved in the mathematical models of industrial melanism in Biston betularia. Values for this variable are based on data from a capture-recapture performed by Bishop (1972) using both local and bred moths which were at least one night old at release. We carried out an experiment to compare the rate of recapture close to the point of release for moths allowed to fly away immediately after their emergence around dusk and those which were at least one night old at release. Unheld moths were less likely to be recaptured suggesting that males have an initial dispersal phase on their first night which results in a higher rate of emigration than on subsequent nights. Such a phase would have been largely missed in Bishop's experiment. The implications of this type of behaviour pattern for the models of spatial variation based on a selection-migration balance are discussed.     

On the selective forces acting in the industrial melanism of Biston and Oligia moths (Lepidoptera: Geometridae and Noctuidae)

Melanic and typical morphs of Biston betularius (L.), Oligia latruncula (D. & S.) and 0. strigilis (L.) made choices between vertical trunks and horizontal branches, sprayed with white and black paints, in a transparent plastic cylinder in natural illumination. The moths settled in exposed positions. In neither Biston nor Oligia did the choice for white/black backgrounds differ between the morphs. Biston moths settle on narrow branches (not on twigs) with the body at right angles to the longitudinal axis of the branch. The Oligias showed an asymmetrical light reaction: one eye is kept in shadow so that they settle as a continuation of an irregularity of the surface, often of a lichen.
In nature, Biston betularius probably rests high up in the canopies, on the under surfaces of horizontal branches. The visual selection acting on the morphs is expected to be less intensive than that measured on tree trunks. The mark-release-recapture results of Kettlewell (1955a, 1956) do not show any qualitative change during the self-determination of the moths but the material is too limited for firm conclusions.
Newly-hatched Biston males take off straight from the trunk where they have expanded their wings but the females may climb higher in the tree. A hypothesis is presented to explain the black-and-white coloration of f. carbonaria : the short-winged moths climbing up the trunks might deter bird predation.     

Behavioural studies on the peppered moth Biston betularia and a discussion of the role of pollution and lichens in industrial melanism

We examined the resting behaviour during pairing and egg-laying of bred female Biston betularia (L.) after they were set free at wooded sites in Somerset and Cardiff. Moths were not released immediately on emergence but were held for three nights to overcome an initial period of flight activity. Females (N= 257) were placed on trunks, and on large and small branches of trees, especially oaks, over periods of several weeks in 1984, 1985 and 1986. Releases were not made within 50 m of any known survivor. One hundred and thirty-one pairings were observed. We also examined oviposition behaviour in a cage experiment using different types of branches. Our observations support Mikkola's earlier conclusion from cage experiments with male moths that the species rests predominantly on branches and shows an appropriately specialized resting attitude, demonstrated here in a series of photographs. Many moths will rest underneath, or on the side of, narrow branches in the canopy. Once released females had settled and paired they only moved quite short distances, but positioning and egg-laying were profoundly influenced by the presence of foliose lichens. Females of all phenotypes show a strong preference to oviposit beneath a thallus of foliose lichens. Their tendency to rest against or close to the lichen during the day could markedly influence their crypsis. In the absence of foliose lichens, eggs are laid in cracks in the bark. The survivorship of released females both in different pairing combinations and during their whole reproductive life-span is analysed. These preliminary data suggest that bird predation can give rise to differential mortality of different pairing combinations and of the phenotypes. This technique provides a powerful method of measuring the relative crypsis and differential mortality of phenotypes with living females in natural resting sites. The significance of our observations on resting behaviour is discussed with regard to the role of air pollution and epiphytes in industrial melanism. Particular attention is given to the causal mechanisms underlying recent declines in the frequency of carbonaria melanics in parts of Britain. One important factor is the new growth made by trees since the marked reduction in particulate air pollution in the 1960s. It is emphasized that we need to know much more about the interactions between pollution, epiphytes and resting backgrounds, especially in the canopy, before we can be confident of our understanding of the evolution of industrial melanism.     

A proposed selective mechanism based on metal chelation in industrial melanic moths

Industrial melanism, a phenomenon observed in some moths and especially in the case of the peppered moth (Biston betularia), has received much attention as an example of Darwinian evolution in action. The rapid rise in the proportion of the darker melanic form of the adult moth coincided with the advent of atmospheric pollution resulting from industrialization, and was ascribed to the improved camouflage of the melanotic insects against a background blackened by soot, which conferred a selective advantage in the avoidance of predation by birds. The topic of the increase in melanization during the initial period of industrial expansion and the reversal of the process after the introduction of the Clean Air Act has received much attention. Although there is sound experimental evidence to support selective avian predation as a major mechanism to account for the changes in the relative frequency of melanics, it is not clear that this is the only selective factor involved in industrial melanism. It is possible that other processes may have made a contribution to the preponderance of melanic variants. In the present study, the hypothesis is advanced that melanization may have conferred a selective advantage by protecting the insects from the toxic effects of metals by virtue of the strong metal chelating action of melanin. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 298–301.     

Morph frequencies in the snail Cepaea nemoralis: changes with time and their interpretation

Colonies of the polymorphic snail Cepaea nemoralis on the sand dune at Point of Air, north Wales, first sampled in 1962 by A.J. Cain and P.M. Sheppard, have been re-sampled after 35 years. The frequency of the mid-banded morph has increased. Overall, yellow frequency has decreased at the expense of pink, but the change is in different directions in different parts of the dune. Yellow mid-banded and pink five-banded, phenotype combinations controlled by alleles at unlinked loci, are in excess within colonies, which suggests an effect of selection. It is argued that changes in morph frequency are due, at least in part, to movement between colonies. Cepaea populations are slow to respond to environmental changes and displacement of individuals may be greater than is often assumed. It is therefore difficult to interpret frequency changes over time. Correlated changes in groups of colonies may not always be interpretable as indicating selection.     

Localized industrial melanism in the spittlebug Philaenus spumarius (L.) (Homoptera: Aphrophoridae) in Cardiff docks, south Wales

Twenty-four large adult samples of the meadow spittlebug, Philaenus spumarius , were collected from within a 4 times 3 km area containing Cardiff docks, south Wales. All samples are highly polymorphic for colour and pattern and show marked geographic variation in the combined frequency of the eight melanic morphs (from <15% to > 75%). Maximum melanic phenotype frequencies in both sexes coincide with dock wharfs currently containing large accumulations of pulverized coal and these samples are heavily contaminated with small coal particles. However, interpretation of this association is complicated by the existence, until 1976, of a smokeless fuel factory causing local air pollution immediately adjacent to the area of highest melanic frequency. It is suggested that the association of high levels of melanism in P. spumarius with areas heavily contaminated with particulate pollutants is due to direct selective effects of small particles favouring melanies.     

Allele frequencies, genetic relationships and heterozygosity in Poly gala vulgaris populations from contrasting habitats in southern Britain

Polygala vulgaris is a widespread perennial plant species of basic to neutral grassland throughout Britain. It is variable in flower colour and some vegetative characters in most populations. Gene flow is restricted in all populations since the plants are normally self-pollinating (up to 20% outcrossing, calculated from figures of heterozygosity) and seeds are normally dispersed only up to 2 m from the parent plant. Examination of several enzyme systems, using starch gel electrophoresis, showed that all enzyme loci appeared to have been duplicated, probably owing to the fact that P. vulgaris is a tctraploid in a genus which has primarily diploid members in Europe. Polymorphisms were found at both of the duplicated loci in four enzyme systems and at one locus in each of two further systems. Fifty-one populations from a diversity of habitats were sampled in southern Britain. In most populations from the typical habitat of lowland calcareous grassland, polymorphisms occurred at five or more loci, and the alleles were distributed throughout the populations. In all these sites there was a diversity of microhabitat and, in one area, marked differences in microhabitat distribution between different enzyme morphs. It is likely that differential selection is an important factor in the maintenance of the polymorphisms in all these areas. These populations were divided, genetically, into two main groups, those from the Gower peninsula, South Wales, and those from the English chalk. Within each group the populations were quite closely related genetically, though some large differences at particular loci occurred even between adjacent populations. Diffuse clumps of alleles occurred within all these populations, and it is likely that limited gene flow has affected the genetic structure of the populations, particularly since cycles of local extinction and recolonization are likely to be recurring. The genetic structure of each population, at the allozyme loci examined, is likely to be the consequence of a balance between differential selection and restricted gene flow. The number of heterozygotes was small, though higher than expected from an analysis of experimental progeny, suggesting a hcterozygotc advantage. Few strong multilocus associations were found despite the predominance of self-pollination. In the populations from sand dunes, roadsides, submontane grassland and water meadows there was, in general, much less polymorphism than in those from lowland calcareous grassland and some populations were entirely monomorphic; in most others there were large, entirely monomorphic groups of plants. Genetic distances between these populations were variable, even within one habitat and area, and they did not appear to show any particular relationship to each other. It is likely that the genetic structure of these populations, at the loci studied, has arisen largely through chance colonizations and genetic drift.     

Conservation of microsatellites in three species of salmonids

The conservation of flanking regions of ten microsatellites has been investigated in three salmonid species. The proportion of conserved microsatellites and the level of polymorphism are high in the three species. This confirms the potential interest of microsatellite loci for interspecific comparisons.     

Allele and genotype frequencies of polymorphic FMO3 gene in two genetically distinct populations

The aims of this study were to analyze flavin-containing monooxygenase 3 (FMO3) polymorphisms and allele and genotype frequencies in 256 Han Chinese and 50 African-American individuals, to compare the allele and genotype frequencies of these populations with those of other world populations. For Han Chinese, genotyping of three common single nucleotide polymorphisms, E158K, V257M and E308G was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). For African-Americans, genotyping of all coding exons was performed by modified PCR-single strand conformational polymorphism (SSCP). Evolutionary rates of FMO3 were estimated computationally. We found that there were significant differences in allele and genotype frequencies among Han Chinese, African-Americans and other world populations. In Han Chinese, the minor allele frequencies (MAFs) were 0.229 (E158K), 0.203 (V257M) and 0.148 (E308G), respectively. In African-Americans, MAFs were 0.48 (E158K), 0.05 (V257M) and 0 (E308G), respectively. There was rapid evolution during the divergence of primate FMO3. This is the first report comparing FMO alleles and genotypes between Han Chinese and African-Americans. A Han Chinese population database has been established for three gene polymorphisms. The data presented here justify further pharmacogenetic studies for potentially optimizing recommended drug dosages and evaluating relationships with disease processes.     

Global‐scale species distributions predict temperature‐related changes in species composition of rocky shore communities in Britain

Changes in rocky shore community composition as responses to climatic fluctuations and anthropogenic warming can be shown by changes in average species thermal affinities. In this study, we derived thermal affinities for European Atlantic rocky intertidal species by matching their known distributions to patterns in average annual sea surface temperature. Average thermal affinities (the Community Temperature Index, CTI) tracked patterns in sea surface temperature from Portugal to Norway, but CTI for communities of macroalgae and plant species changed less than those composed of animal species. This reduced response was in line with the expectation that communities with a smaller range of thermal affinities among species would change less in composition along thermal gradients and over time. Local‐scale patterns in CTI over wave exposure gradients suggested that canopy macroalgae allow species with ranges centred in cooler than local temperatures (‘cold‐affinity’) to persist in otherwise too‐warm conditions. In annual surveys of rocky shores, communities of animal species in Shetland showed a shift in dominance towards warm‐affinity species (‘thermophilization’) with local warming from 1980 to 2018 but the community of plant and macroalgal species did not. From 2002 to 2018, communities in southwest Britain showed the reverse trend in CTI: declining average thermal affinities over a period of modest temperature decline. Despite the cooling, trends in species abundance were in line with the general mechanism of direction and magnitude of long‐term trends depending on the difference between species thermal affinities and local temperatures. Cold‐affinity species increased during cooling and warm‐affinity ones decreased. The consistency of responses across different communities and with general expectations based on species thermal characteristics suggests strong predictive accuracy of responses of community composition to anthropogenic warming.     

工业生物技术中DNA合成发展现状及展望

DNA合成是生命科学领域的共性支撑技术和合成生物学的关键使能技术。以合成生物学为基础的工业生物技术持续快速发展,迫切需要更加便捷、经济、安全的DNA来源以满足其日益增长的大规模DNA合成需求。工业化DNA合成在通量、成本、速度等方面的优势日益凸显,有力推动了工业生物技术研发效率的提升和研发成本的下降。但是现有技术在生产过程中还存在着使用大量有机试剂、资源浪费等问题。随着DNA合成规模的持续快速提升,有毒化学品危害、成本负担、环境负担等问题日益突出。本文结合我们的工作实践,对工业生物技术中DNA合成需求、合成策略以及可持续发展面临的问题和解决方案研究进展进行探讨。