Sequence variation and regulatory variation in acetylcholinesterase genes contribute to insecticide resistance in different populations of Leptinotarsa decemlineata

Abstract Although insect herbivores are known to evolve resistance to insecticides through multiple genetic mechanisms, resistance in individual species has been assumed to follow the same mechanism. While both mutations in the target site insensitivity and increased amplification are known to contribute to insecticide resistance, little is known about the degree to which geographic populations of the same species differ at the target site in a response to insecticides. We tested structural (e.g., mutation profiles) and regulatory (e.g., the gene expression of Ldace1 and Ldace2, AChE activity) differences between two populations (Vermont, USA and Belchow, Poland) of the Colorado potato beet…

Can aposematic signals evolve by gradual change?

Aposematic species, which signal conspicuously of their unprofitability to predators, have puzzled evolutionary biologists for over a century1,2. Although conspicuousness of unpalatable prey improves avoidance learning by predators3,4,5, it also involves an evolutionary paradox: with increasing detectability4,6,7,8 the deviant aposematic prey would suffer high predation initially from naive predators. Here we test a neglected idea7,8,9,10,11 that aposematic coloration may evolve by gradual change rather than by major mutations. Weak signals did not suffer high initial predation, but predators (great tits, Parus major) did not learn to separate them from cryptic palatable prey. Furthermore, …

Evolutionary constraints of warning signals: A genetic trade-off between the efficacy of larval and adult warning coloration can maintain variation in signal expression

To predict evolutionary responses of warning signals under selection, we need to determine the inheritance pattern of the signals, and how they are genetically correlated with other traits contributing to fitness. Furthermore, protective coloration often undergoes remarkable changes within an individual's lifecycle, requiring us to quantify the genetic constraints of adaptive coloration across all the relevant life stages. Based on a 12 generation pedigree with > 11,000 individuals of the wood tiger moth (Arctia plantaginis), we show that high primary defense as a larva (large warning signal) results in weaker defenses as adult (less efficient warning color), due to the negative genetic cor…

Can ultraviolet cues function as aposematic signals?

The fact that birds are sensitive to ultraviolet light (UV, 320–400 nm) has been largely ignored by previous studies of aposematism. Therefore, in the present article we investigated whether great tits preferred ultraviolet-reflecting colors compared to colors without UV reflection and whether UV cues alone could function as aposematic signals. We were able to manipulate prey visibility in UV light by changing the UV reflectance of prey items as well as altering the lighting conditions. In order to perform a preference experiment we used three pairs of colors (green UV vs. green, gray UV vs. gray, yellow UV vs. yellow) on a black background. The birds ate both UV types equally for all three…

The importance of pattern similarity between Müllerian mimics in predator avoidance learning

Müllerian mimicry, where unpalatable prey share common warning patterns, has long fascinated evolutionary biologists. It is commonly assumed that Müllerian mimics benefit by sharing the costs of predator education, thus reducing per capita mortality, although there has been no direct test of this assumption. Here, we specifically measure the selection pressure exerted by avian predators on unpalatable prey with different degrees of visual similarity in their warning patterns. Using wild-caught birds foraging on novel patterned prey in the laboratory, we unexpectedly found that pattern similarity did not increase the speed of avoidance learning, and even dissimilar mimics shared the educatio…

Signalling and Reception

Communication, a widespread natural phenomenon, occurs in both animals and plants. Signals are evolved traits that transfer information from one individual (the signaller) to another (the receiver); they can occur in any sensory modality. Keywords: cost and benefits; honesty; mimicry; predation; sexual selection

Underutilized resources for studying the evolution of invasive species during their introduction, establishment, and lag phases

The early phases of biological invasions are poorly understood. In particular, during the introduction, establishment, and possible lag phases, it is unclear to what extent evolution must take place for an introduced species to transition from established to expanding. In this study, we highlight three disparate data sources that can provide insights into evolutionary processes associated with invasion success: biological control organisms, horticultural introductions, and natural history collections. All three data sources potentially provide introduction dates, information about source populations, and genetic and morphological samples at different time points along the invasion trajector…

Inheritance patterns of photoperiodic diapause induction in Leptinotarsa decemlineata

Photoperiod is a reliable indicator of season and an important cue that many insects use for phenological synchronization. Undergoing range expansion insects can face a change in the local photoperiod to which they need to resynchronize. Rapid range expansion can be associated with rapid photoperiodic adaptation, which can be associated with intense selection on strongly heritable polygenic traits. Alternatively, it is proposed that, in insects with an XO sex-determination system, genes with large effect residing on the sex chromosome could drive photoperiodic adaptation because the gene or genes are exposed to selection in the sex carrying only a single X-chromosome. The present study seek…

How Did the Cuckoo Get Its Polymorphic Plumage?

One hundred and fifty years ago, the English naturalist Henry Walter Bates first developed the theory of mimicry ( 1 ). Based on his field observations in the Amazon, he argued that the uncanny likeness of unrelated butterflies is an evolutionary adaptation whereby edible butterflies avoid predation by imitating the coloration of venomous butterfly species without paying the cost of arming themselves. Such “Batesian mimicry” is a dynamic parasitic game between three players, in which a harmless species (the mimic) escapes predation by imitating the warning signals of harmful species (the model) that a shared predator (the dupe) has learned to avoid. On page 578 of this issue, Thorogood and …

Investigating Müllerian mimicry: predator learning and variation in prey defences

Inexperienced predators are assumed to select for similarity of warning signals in aposematic species (Mullerian mimicry) when learning to avoid them. Recent theoretical work predicts that if co-mimic species have unequal defences, predators attack them according to their average unpalatability and mimicry may not be beneficial for the better defended co-mimic. In this study, we tested in a laboratory environment whether a uniform warning signal is superior to a variable one in promoting predator learning, and simultaneously whether co-mimics are preyed upon according to their average unpalatability. There was an interaction of signal variation and unpalatability but inexperienced birds did…

Imperfect Batesian mimicry—the effects of the frequency and the distastefulness of the model

Batesian mimicry is the resemblance between unpalatable models and palatable mimics. The widely accepted idea is that the frequency and the unprofitability of the model are crucial for the introduction of a Batesian mimic into the prey population. However, experimental evidence is limited and furthermore, previous studies have considered mainly perfect mimicry (automimicry). We investigated imperfect Batesian mimicry by varying the frequency of an aposematic model at two levels of distastefulness. The predator encountered prey in a random order, one prey item at a time. The prey were thus presented realistically in a sequential way. Great tits (Parus major) were used as predators. This expe…

Resting metabolic rate can vary with age independently from body mass changes in the Colorado potato beetle, Leptinotarsa decemlineata.

Temperature and mass dependency of insect metabolic rates are well known, while less attention has been given to other factors, such as age. Among insect species that experience seasonal variation in environmental conditions, such as in temperate latitudes, age may also have indirect effects on the metabolic rate. We examined the effect of age on the resting metabolic rate of Leptinotarsa decemlineata during 11 days after adult emergence by using flow-through respirometry. Age had a significant mass-independent effect on metabolic rate of beetles. A twofold increase in metabolic rate occurred during the first 2 days of adult life after which metabolic rate decreased with age relatively slow…

Preconditioning of the generalist herbivoreTrialeurodes vaporariorumto greenhouse monocultures and its subsequent performance on wild polycultures

Generalist herbivores can face many challenges when choosing their host plant. This can be particularly difficult if their choice and performance are affected by host experience. Greenhouse whitefly, Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae), is an invasive generalist herbivore, which has established in year-round greenhouses at northern latitudes where it cannot overwinter outdoors. It mainly uses crops such as cucumber (Cucumis sativus L.), tomato (Solanum lycopersicum L.), and ornamentals as host plants. However, every summer the insect escapes greenhouses and is exposed to natural vegetation. We evaluated the performance of T. vaporariorum on diverse vegetation outside…

Evolutionary constraints of warning signals : a genetic trade-off between the efficacy of larval and adult warning coloration can maintain variation in signal expression

To predict evolutionary responses of warning signals under selection, we need to determine the inheritance pattern of the signals, and how they are genetically correlated with other traits contributing to fitness. Furthermore, protective coloration often undergoes remarkable changes within an individual's lifecycle, requiring us to quantify the genetic constraints of adaptive coloration across all the relevant life stages. Based on a 12 generation pedigree with > 11,000 individuals of the wood tiger moth (Arctia plantaginis), we show that high primary defense as a larva (large warning signal) results in weaker defenses as adult (less efficient warning color), due to the negative genetic cor…

Environmental Fluctuations Drive Species' Competitive Success in Experimental Invasions

Climate change is presumed to increase both the number and frequency of fluctuations in environmental conditions. Fluctuations can affect the ecological and evolutionary processes that make species more successful competitors. For example, fluctuating conditions can create selection pressures for traits that are profitable in adaptation to fast climate change. On an ecological timescale, environmental fluctuations can facilitate species competitive success by reducing other species’ population sizes. Climate change could then enhance species invasions into new areas if fluctuation-adapted invaders displace their native competitors in chancing environments. We tested experimentally whether f…

Photoperiodic effects on diapause-associated gene expression trajectories in EuropeanLeptinotarsa decemlineatapopulations

Behavioural and physiological changes during diapause, an important strategy of insects for surviving harsh seasonal conditions, have been intensively studied. The genetic and molecular mechanisms underpinning diapause development are less well known. We took a candidate gene approach to study prediapause gene expression patterns in the Colorado potato beetle (Leptinotarsa decemlineata), an invasive insect that has rapidly spread northwards to high seasonality environments. Newly eclosed beetles originating from southern (Italy) and northern (Russia) Europe were reared under short- [12 h light (L):12 h dark (D)] and long-day (18L:6D) photoperiods for 10 days. This time period includes the s…

Invasion triple trouble : environmental fluctuations, fluctuation-adapted invaders and fluctuation-mal-adapted communities all govern invasion success

AbstractIt has been suggested that climate change will lead to increased environmental fluctuations, which will undoubtedly have evolutionary consequences for all biota. For instance, fluctuations can directly increase the risk of invasions of alien species into new areas, as these species have repeatedly been proposed to benefit from disturbances. At the same time increased environmental fluctuations may also select for better invaders. However, selection by fluctuations may also influence the resistance of communities to invasions, which has rarely been tested. We tested eco-evolutionary dynamics of invasion with bacterial clones, evolved either in constant or fluctuating temperatures, an…

Variation in mortality among populations is higher for pymetrozine than for imidacloprid and spiromesifen inTrialeurodes vaporariorumin greenhouses in Finland

BACKGROUND Insecticide resistance in Trialeurodes vaporariorum W. is unknown in the species' northern distribution range where it inhabits mainly commercial greenhouses. Resistance development in whiteflies feeding on year-round crops in greenhouses is possible owing to the use of chemical treatments to back up biocontrol. The authors tested the response levels to spiromesifen, pymetrozine and imidacloprid in whiteflies collected from seven greenhouses within a 35 km radius in western Finland. RESULTS All except one (PR) population had LC50 values below the recommended concentrations for the tested compounds. However, some populations showed reduced susceptibility to pymetrozine in comparis…

Glyphosate-based herbicide has soil-mediated effects on potato glycoalkaloids and oxidative status of a potato pest

Glyphosate is the most used herbicide worldwide, targeting physiological pathways in plants. Recent studies have shown that glyphosate can also cause toxic effects in animals. We investigated the glyphosate-based herbicide (GBH)-induced changes in potato (Solanum tuberosum) plant chemistry and the effects of a GBH on the survival rate and oxidative status of the Colorado potato beetle (Leptinotarsa decemlineata). The beetles were reared on potato plants grown in pots containing soil treated with a GBH (Roundup Gold, 450 g/l) or untreated soil (water control). The 2nd instar larvae were introduced to the potato plants and then collected in 2 phases: as 4th instar larvae and as adults. The ma…

Sublethal Pyrethroid Insecticide Exposure Carries Positive Fitness Effects Over Generations in a Pest Insect

AbstractStress tolerance and adaptation to stress are known to facilitate species invasions. Many invasive species are also pests and insecticides are used to control them, which could shape their overall tolerance to stress. It is well-known that heavy insecticide usage leads to selection of resistant genotypes but less is known about potential effects of mild sublethal insecticide usage. We studied whether stressful, sublethal pyrethroid insecticide exposure has within-generational and/or maternal transgenerational effects on fitness-related traits in the Colorado potato beetle (Leptinotarsa decemlineata) and whether maternal insecticide exposure affects insecticide tolerance of offspring…

The effects of short-term glyphosate-based herbicide exposure on insect gene expression profiles

Glyphosate-based herbicides (GBHs) are the most frequently used herbicides worldwide. The use of GBHs is intended to tackle weeds, but GBHs have been shown to affect the life-history traits and antioxidant defense system of invertebrates found in agroecosystems. Thus far, the effects of GBHs on detoxification pathways among invertebrates have not been sufficiently investigated. We performed two different experiments—1) the direct pure glyphosate and GBH treatment, and 2) the indirect GBH experiment via food—to examine the possible effects of environmentally relevant GBH levels on the survival of the Colorado potato beetle (Leptinotarsa decemlineata) and the expression profiles of their deto…

Seasonal changes in predator community switch the direction of selection for prey defences

Insect communities consist of aposematic species with efficient warning colours against predation, as well as abundant examples of crypsis. To understand such coexistence, we here report results from a field experiment where relative survival of artificial larvae, varying in conspicuousness, was estimated in natural bird communities over an entire season. This takes advantage of natural variation in the proportion of naive predators: naivety peaks when young birds have just fledged. We show that the relative benefit of warning signals and crypsis changes accordingly. When naive birds are rare (early and late in the season), conspicuous warning signals improve survival, but conspicuousness b…

Sublethal effects of deltamethrin exposure of parental generations on physiological traits and overwintering inLeptinotarsa decemlineata

Although the evolution of insecticide resistance has received a lot of attention, less is known about sublethal effects of insecticide stress experienced by the preceding generations on the performance of pest populations. We investigated whether three generations of parental exposure to a deltamethrin insecticide influences physiological traits and overwintering success of offspring in Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). Beetles descending from insecticide-exposed parents had lower adult body mass but higher relative lipid content and resting metabolic rate than those descending from non-insecticide-exposed parents. Also, a higher proportion of beetles descending f…

Effects of a glyphosate-based herbicide on survival and oxidative status of a non-target herbivore, the Colorado potato beetle (Leptinotarsa decemlineata).

Abstract Glyphosate is the globally most used herbicide against a wide range of weeds. Glyphosate has been considered safe to animals as it mainly targets physiological pathways in plants. However, recent toxicological studies have revealed that glyphosate can cause various toxic effects also on animals. In this study, we investigated the direct toxic effects of a glyphosate-based herbicide (GBH, Roundup® Bio) on 1) survival and 2) oxidative status of a non-target herbivore by using Colorado potato beetles (Leptinotarsa decemlineata), originating from Poland and USA, as model species. Larvae were randomly divided into three groups: 1) high concentration (100% Roundup Bio, 360 g/l), 2) low c…

THERMOREGULATION CONSTRAINS EFFECTIVE WARNING SIGNAL EXPRESSION

Evolution of conspicuous signals may be constrained if animal coloration has nonsignaling as well as signaling functions. In aposematic wood tiger moth (Parasemia plantaginis) larvae, the size of a warning signal (orange patch on black body) varies phenotypically and genetically. Although a large warning signal is favored as an antipredator defense, we hypothesized that thermoregulation may constrain the signal size in colder habitats. To test this hypothesis, we conducted a factorial rearing experiment with two selection lines for larval coloration (small and large signal) and with two temperature manipulations (high and low temperature environment). Temperature constrained the size and br…

Pre-invasion history and demography shape the genetic variation in the insecticide resistance-related acetylcholinesterase 2 gene in the invasive Colorado potato beetle

Abstract Background Invasive pest species offers a unique opportunity to study the effects of genetic architecture, demography and selection on patterns of genetic variability. Invasive Colorado potato beetle (Leptinotarsa decemlineata) populations have experienced a rapid range expansion and intense selection by insecticides. By comparing native and invasive beetle populations, we studied the origins of organophosphate (OP) resistance-associated mutations in the acetylcholinesterase 2 (AChE2) gene, and the role of selection and demography on its genetic variability. Results Analysis of three Mexican, two US and five European populations yielded a total of 49 haplotypes. Contrary to the exp…

Does predation maintain eyespot plasticity in Bicyclus anynana?

The butterfly Bicyclus anynana exhibits phenotypic plasticity involving the wet-season phenotype, which possesses marginal eyespots on the ventral surface of the wings, and the dry-season form, which lacks these eyespots. We examined the adaptive value of phenotypic plasticity of B. anynana in relation to the defence mechanisms of crypsis and deflection. We assessed the visibility differences between spotless and spotted butterflies against backgrounds of brown (dry season) or green (wet season) leaves. Spotless butterflies were highly cryptic and less predated by adult bird predators than were spotted ones when presented against brown leaf litter. However, the advantage of crypsis disappea…

Sequencing, De Novo Assembly and Annotation of the Colorado Potato Beetle, Leptinotarsa decemlineata, Transcriptome

Background. The Colorado potato beetle (Leptinotarsa decemlineata) is a major pest and a serious threat to potato cultivation throughout the northern hemisphere. Despite its high importance for invasion biology, phenology and pest management, little is known about L. decemlineata from a genomic perspective. We subjected European L. decemlineata adult and larval transcriptome samples to 454-FLX massively-parallel DNA sequencing to characterize a basal set of genes from this species. We created a combined assembly of the adult and larval datasets including the publicly available midgut larval Roche 454 reads and provided basic annotation. We were particularly interested in diapause-specific g…

Variation in mortality among populations is higher for pymetrozine than for imidacloprid and spiromesifen in Trialeurodes vaporariorum in greenhouses in Finland

Abstract. BACKGROUND: Insecticide resistance in Trialeurodes vaporariorum W. is unknown in the species’ northern distribution range where it inhabits mainly commercial greenhouses. Resistance development in whiteflies feeding on year-round crops in greenhouses is possible due to use of chemical treatments to back up biocontrol. We tested the response levels to spiromesifen, pymetrozine and imidacloprid in whiteflies collected from seven greenhouses within a 35 km radius in Western Finland. RESULTS: All except one (PR) population had LC50 values below the recommended concentrations for the tested compounds. However, some populations showed reduced susceptibility to pymetrozine in comparison …

Stress for invasion success? Temperature stress of preceding generations modifies the response to insecticide stress in an invasive pest insect

Adaptation to stressful environments is one important factor influencing species invasion success. Tolerance to one stress may be complicated by exposure to other stressors experienced by the preceding generations. We studied whether parental temperature stress affects tolerance to insecticide in the invasive Colorado potato beetle Leptinotarsa decemlineata. Field-collected pyrethroid-resistant beetles were reared under either stressful (17°C) or favourable (23°C) insecticide-free environments for three generations. Then, larvae were exposed to pyrethroid insecticides in common garden conditions (23°C). Beetles were in general tolerant to stress. The parental temperature stress alone affect…

Can experienced birds select for Müllerian mimicry?

Field experiments have shown that avian predators in the wild can select for similarity of warning signals in aposematic prey (Müllerian mimicry) because a common signal is better protected than a signal that is novel and rare. The original theory of Müllerian mimicry assumes that the mechanism promoting mimicry is predator learning; by sharing a signal, the comimic species share the mortality that is due to sampling by inexperienced predators. Predation events have not been observed in the wild, and learning experiments with naive bird predators in a laboratory have not unambiguously shown a benefit of a uniform signal compared with different signals. As predators in the field experiments …

Variation in Hsp70 Levels after Cold Shock: Signs of Evolutionary Responses to Thermal Selection among Leptinotarsa decemlineata Populations

Individuals of widely spread species are expected to show local adaption in temperature tolerance as they encounter a range of thermal conditions. We tracked thermal adaptations of the Colorado potato beetle (Leptinotarsa decemlineata) that invaded Europe within the last 100 years. It has occupied various conditions although, like the majority of invasive species, it lost a measurable amount of neutral genetic variation due to bottleneck effect when it invaded Europe. We exposed diapausing beetles originated from three different latitudes (54°N, 59°N, 60°N) to cold shock (−5°C, 1.5 hrs) in order to test if beetles from the northern populations express differential levels of cold-induced and…

Northward range expansion requires synchronization of both overwintering behaviour and physiology with photoperiod in the invasive Colorado potato beetle (Leptinotarsa decemlineata)

Abstract. Photoperiodic phenological adaptations are prevalent in many organisms living in seasonal environments. As both photoperiod and growth season length change with latitude, species undergoing latitudinal range expansion often need to synchronize their life cycle with a changing photoperiod and growth season length. Since adaptive synchronization often involves a large number of time-consuming genetic changes, behavioural plasticity might be a faster way to adjust to novel conditions. We compared behavioural and physiological traits in overwintering (diapause) preparation in three latitudinally different European Colorado potato beetle (Leptinotarsa decemlineata) populations reared u…

Propagule pressure increase and phylogenetic diversity decrease community’s susceptibility to invasion

Background Invasions pose a large threat to native species, but the question of why some species are more invasive, and some communities more prone to invasions than others, is far from solved. Using 10 different three-species bacterial communities, we tested experimentally if the phylogenetic relationships between an invader and a resident community and the propagule pressure affect invasion probability. Results We found that greater diversity in phylogenetic distances between the members of resident community and the invader lowered invasion success, and higher propagule pressure increased invasion success whereas phylogenetic distance had no clear effect. In the later stages of invasion,…

Agroecosystems shape population genetic structure of the greenhouse whitefly in Northern and Southern Europe

International audience; Background: To predict further invasions of pests it is important to understand what factors contribute to the genetic structure of their populations. Cosmopolitan pest species are ideal for studying how different agroecosystems affect population genetic structure within a species at different climatic extremes. We undertook the first population genetic study of the greenhouse whitefly (Trialeurodes vaporariorum), a cosmopolitan invasive herbivore, and examined the genetic structure of this species in Northern and Southern Europe. In Finland, cold temperatures limit whiteflies to greenhouses and prevent them from overwintering in nature, and in Greece, milder tempera…

Co-mimics have a mutualistic relationship despite unequal defences

Defensive mimicry, where species have evolved to resemble others in order to evade predators, is quite common in the animal kingdom. The two extremes of the mimicry spectrum are known as 'batesian' and 'mullerian'. Batesian mimics develop signals — visual cues for instance — that are similar to those of species being mimicked, but stop short of adopting the attribute that makes it unprofitable prey to predators. Mullerian mimics both resemble the model species and share the anti-predation attribute — by being dangerous or unpalatable. These different types of mimic were identified a century ago, but the dynamics of mimicry between unequally defended prey remain unresolved. In an experiment …

Repeated exposure of fluazinam fungicides affects gene expression profiles yet carries no costs on a nontarget pest

Fungicides are used to control pathogenic fungi of crop species, but they have also been shown to alter behavioral, life history and fitness related traits of non-target insects. Here, we tested the fungicide effects on feeding behavior, survival and physiology of the non-target pest insect, the Colorado potato beetle (Leptinotarsa decemlineata). Feeding behavior was studied by a choice test of adult beetles, which were allowed to choose between a control and a fungicide (fluazinam) treated potato leaf. Larval survival was recorded after 24-hour and 72-hour exposure to control and fungicide-treated leaves with two different concentrations. The adults did not show fungicide avoidance behavio…

Characterizing the pigment composition of a variable warning signal of Parasemia plantaginis larvae

Summary 1. Aposematic animals advertise their defences to predators via warning signals that often are bright colours combined with black patterns. Predation is assumed to select for large pattern elements and conspicuousness of warning signals because this enhances avoidance learning of predators. However, conspicuousness of the colour pattern can vary among individuals of aposematic species, suggesting that warning signal expression may be constrained by opposing selection pressures. If effective warning signals are costly to produce, variation in signal expression may be maintained via physiological trade-offs. To understand the costs of signalling that might underlay both physiological …

Multiple benefits of gregariousness cover detectability costs in aposematic aggregations

Understanding the early evolution of aposematic (warning) coloration has been a challenge for scientists, as a new conspicuous morph in a population of cryptic insects would have a high predation risk and would probably die out before local predators learnt to avoid it1,2,3,4. Fisher5 presented the idea of aggregation benefit through the survival of related individuals; however, his theory has been strongly debated6,7,8 as the mechanisms that favour grouping have never been explored experimentally with the incorporation of detectability costs. Here we create a comprehensive ‘novel world’ experiment with the great tit (Parus major) as a predator to explore simultaneously the predation-relate…

Energy use, diapause behaviour and northern range expansion potential in the invasive Colorado potato beetle

Summary 1. As organisms expand their range towards northern latitudes they will encounter selective factors like harsh winter conditions. The ability to cope with and adapt to harsh winters may depend on the variability and evolutionary potential of relevant traits. 2. One adaptation in insects is winter diapause. It is characterized by changes in physiology, behaviour or in both. Physiological changes include lowered metabolic rate that enhances survival by saving limited energy reserves during overwintering. Active behavioural changes like burrowing into the soil allow individuals to escape harsh conditions. 3. We examined variation in overwintering body mass, resting metabolic rate (CO2 …

DIET QUALITY AFFECTS WARNING COLORATION INDIRECTLY: EXCRETION COSTS IN A GENERALIST HERBIVORE

Aposematic herbivores are under selection pressure from their host plants and predators. Although many aposematic herbivores exploit plant toxins in their own secondary defense, dealing with these harmful compounds might underlay costs. We studied whether the allocation of energy to detoxification and/or sequestration of host plant defense chemicals trades off with warning signal expression. We used a generalist aposematic herbivore Parasemia plantaginis (Arctiidae), whose adults and larvae show extensive phenotypic and genetic variation in coloration. We reared larvae from selection lines for small and large larval warning signals on Plantago lanceolata with either low or high concentratio…

Prolonged diapause has sex-specific fertility and fitness costs

AbstractDiapause in seasonal environments allows insects to survive adverse seasons. However, individuals can sometimes enter a prolonged diapause for more than a year, and also skip favourable seasons, which can bring additional costs through e.g. loss of metabolic resources. At the same time, prolonged diapause can be beneficial if it allows individuals to have a risk-spreading strategy to skip potentially suboptimal breeding seasons. We studied if prolonged diapause (2-year diapause) negatively affects the fertility and fitness of female and male Colorado potato beetles (Leptinotarsa decemlineata) compared to control (1-year diapause) beetles. We also tested the parental effects on the s…

Strong antiapostatic selection against novel rare aposematic prey

The evolution of aposematism, a phenomenon where prey species conspicuously advertise their unprofitability to predators, is puzzling. How did conspicuousness evolve, if it simultaneously increased the likelihood of an inexperienced predator to detect the prey and presumably kill it? Antiapostatic selection, where rare prey is predated relatively more often, is considered as another major difficulty for aposematism to evolve. However, the risk of being conspicuous in low frequencies has not been experimentally tested. We designed an experiment to test how frequency (4%, 12%, 32%) of conspicuous aposematic prey and its dispersion type (solitary vs. aggregated) affect an initial predation ri…

Climate Change Effects on Agricultural Insect Pests in Europe

Abstract In this chapter, we will discuss observations of climate change effects on agricultural pests in Europe, the possible mechanisms behind these observed effects and finally delve into more detail through some relatively well studied model species (the Colorado potato beetle and the rape beetle). Direct effects of climate change on agricultural pests in Europe are difficult to dissect from all the human-induced changes that have taken place in parallel with an increased mean annual temperature. During the past decade, agriculture has become more professionalized in terms of land use, crop cultivation techniques and pest management strategies. We review the effect of climate change on …

Relative importance of taste and visual appearance for predator education in Müllerian mimicry

Mullerian mimicry, by definition, is the visual resemblance between two or more aposematic prey species. According to classical Mullerian mimicry theory, comimics draw mutual benefits from the resemblance because predators have to learn to avoid only one colour pattern. In contrast, the relatively untested quasi-Batesian mimicry theory suggests that, because of differences in unpalatablility, the less toxic mimic acts like a parasite on the more defended prey, decreasing its fitness. We tested predation pressures on artificial mimicry complexes in which comimics varied both in visual similarity and in taste. Both signal and taste were important for the survival of comimics. Predators learne…

Evolutionary Considerations in Potato Pest Management

Incorporating our knowledge of fundamental evolutionary processes into pest control practices is essential for maximizing their efficiency. The insect pest complex of potato is characterized by a high degree of plasticity and adaptability. In particular, the Colorado potato beetle quickly expanded its host range to include cultivated potato, and has shown a remarkable ability to evolve resistance to a wide variety of chemicals. Another major pest of potatoes, the green peach aphid, is also very adaptable to insecticides. Both of these insects can also develop resistance to non-chemical methods of their suppression, such as biological control and crop rotation. In addition, understanding ins…

Preconditioning of the generalist herbivore Trialeurodes vaporariorum to greenhouse monocultures and its subsequent performance on wild polycultures

Generalist herbivores can face many challenges when choosing their host plant. This can be particularly difficult if their choice and performance are affected by host experience. Greenhouse whitefly, Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae), is an invasive generalist herbivore, which has established in year-round greenhouses at northern latitudes where it cannot overwinter outdoors. It mainly uses crops such as cucumber (Cucumis sativus L.), tomato (Solanum lycopersicum L.), and ornamentals as host plants. However, every summer the insect escapes greenhouses and is exposed to natural vegetation. We evaluated the performance of T. vaporariorum on diverse vegetation outside…

Hairiness and warning colours as components of antipredator defence: additive or interactive benefits?

To deter predator attack, aposematic prey species advertise their unprofitability with one or more conspicuous warning signals that, in turn, enhance the avoidance learning of predators. We studied the costs and benefits of multicomponent signalling in Parasemia plantaginis moths. The hairy moth larvae have an orange patch on their otherwise black bodies. The patch varies phenotypically and genetically in size. We studied whether the detection risk associated with patch size varied against two backgrounds (green or brown) with two different predators: naive chicks, Gallus gallus domesticus, and experienced great tits, Parus major. We also evaluated the signal value of different defence trai…

The voyage of an invasive species across continents: genetic diversity of North American and European Colorado potato beetle populations

The paradox of successful invading species is that they are likely to be genetically depauperate compared to their source population. This study on Colorado potato beetles is one of the few studies of the genetic consequences of continent-scale invasion in an insect pest. Understanding gene flow, population structure and the potential for rapid evolution in native and invasive populations offers insights both into the dynamics of small populations that become successful invaders and for their management as pests. We used this approach to investigate the invasion of the Colorado potato beetle (Leptinotarsa decemlineata) from North America to Europe. The beetles invaded Europe at the beginnin…

Inheritance patterns of photoperiodic diapause induction inLeptinotarsa decemlineata

Photoperiod is a reliable indicator of season and an important cue that many insects use for phenological synchronization. Undergoing range expansion insects can face a change in the local photoperiod to which they need to resynchronize. Rapid range expansion can be associated with rapid photoperiodic adaptation, which can be associated with intense selection on strongly heritable polygenic traits. Alternatively, it is proposed that, in insects with an XO sex-determination system, genes with large effect residing on the sex chromosome could drive photoperiodic adaptation because the gene or genes are exposed to selection in the sex carrying only a single X-chromosome. The present study seek…

Is a change in juvenile hormone sensitivity involved in range expansion in an invasive beetle?

Introduction: It has been suggested that rapid range expansion could proceed through evolution in the endocrinological machinery controlling life-history switches. Based on this we tested whether the Colorado potato beetle, Leptinotarsa decemlineata, which has rapidly expanded its range across latitudinal regions in Europe, and shows photoperiodic adaptation in overwintering initiation, has different sensitivities to juvenile hormone (JH) manipulation along a latitudinal gradient. Results: A factorial experiment where beetles were reared either under a long or short day photoperiod was performed. Hormone levels were manipulated by topical applications. An allatostatin mimic, H17, was used t…

Effects of a glyphosate-based herbicide on survival and oxidative status of a non-target herbivore, the Colorado potato beetle (Leptinotarsa decemlineata)

Glyphosate is the globally most used herbicide against a wide range of weeds. Glyphosate has been considered safe to animals as it mainly targets physiological pathways in plants. However, recent toxicological studies have revealed that glyphosate can cause various toxic effects also on animals. In this study, we investigated the direct toxic effects of a glyphosate-based herbicide (GBH, Roundup® Bio) on 1) survival and 2) oxidative status of a non-target herbivore by using Colorado potato beetles (Leptinotarsa decemlineata), originating from Poland and USA, as model species. Larvae were randomly divided into three groups: 1) high concentration (100% Roundup Bio, 360 g/l), 2) low concentrat…

Life-history constraints and warning signal expression in an arctiid moth

Summary 1 Traditionally, large pattern elements in conspicuous warning signals are assumed to be selected by predation because increasing signal strength enhances education and avoidance of predators. However, variation in the colour pattern of a warningly coloured species often exists even within a location, and many warning signals have relatively small pattern elements. This suggests that there must be opposing selection pressures that can constrain warning signal expression, resulting in variation in warning signals as well as suboptimal signals for predator education. 2 We quantified the variation in the warning signal of Parasemia plantaginis-moth larvae. With a full-sib rearing exper…

The Diapause Lipidomes of Three Closely Related Beetle Species Reveal Mechanisms for Tolerating Energetic and Cold Stress in High-Latitude Seasonal Environments

During winter insects face energetic stress driven by lack of food, and thermal stress due to sub-optimal and even lethal temperatures. To survive, most insects living in seasonal environments such as high latitudes, enter diapause, a deep resting stage characterized by a cessation of development, metabolic suppression and increased stress tolerance. The current study explores physiological adaptations related to diapause in three beetle species at high latitudes in Europe. From an ecological perspective, the comparison is interesting since one species (Leptinotarsa decemlineata) is an invasive pest that has recently expanded its range into northern Europe, where a retardation in range expa…

Transgenerational effects of insecticides - implications for rapid pest evolution in agroecosystems

Although pesticides are a major selective force in driving the evolution of insect pests, the evolutionary processes that give rise to insecticide resistance remain poorly understood. Insecticide resistance has been widely observed to increase with frequent and intense insecticide exposure, but can be lost following the relaxation of insecticide use. One possible but rarely explored explanation is that insecticide resistance may be associated with epigenetic modifications, which influence the patterning of gene expression without changing underlying DNA sequence. Epigenetic modifications such as DNA methylation, histone modifications, and small RNAs have been observed to be heritable in art…

Population dependent effects of photoperiod on diapause related physiological traits in an invasive beetle (Leptinotarsa decemlineata).

Organisms undergoing latitudinal range expansion face a change in the photoperiod which can lead to a mismatch between the timing of seasonal changes in physiological and life history traits with seasonal environmental changes. This mismatch can lead to lowered survival, for example, due to unsynchronized diapause timing. Successful range expansion even in recent introductions requires that organisms which use the photoperiod for seasonal predictions should show interpopulational differences in photoperiodic responses at different latitudes, as the photoperiod is a function of latitude. We investigated among population differences in photoperiodic responses of life history and physiological…

Adaptation to fluctuations in temperature by nine species of bacteria

Rapid environmental fluctuations are ubiquitous in the wild, yet majority of experimental studies mostly consider effects of slow fluctuations on organism. To test the evolutionary consequences of fast fluctuations, we conducted nine independent experimental evolution experiments with bacteria. Experimental conditions were same for all species, and we allowed them to evolve either in fluctuating temperature alternating rapidly between 20°C and 40°C or at constant 30°C temperature. After experimental evolution, we tested the performance of the clones in both rapid fluctuation and in constant environments (20°C, 30°C and 40°C). Results from experiments on these nine species were combined meta…

Ultraviolet reflection and predation risk in diurnal and nocturnal Lepidoptera

According to our extensive data on Lepidoptera (883 species), UV wing patterns are almost three times more common in nocturnal than in diurnal Lepidoptera. This might be due to predation, because the primary diurnal predators, birds, utilize UV light in foraging and even prefer UV-reflecting prey. To test this hypothesis, we conducted a field experiment with tethered living moths whose wings were artificially manipulated to reflect (UV+, reflection at UV wavelength: 15%) or absorb (UV - ) UV light, keeping longer wavelengths identical. Thus, any difference found in survival rates would be the result of the difference in wing patterns in UV spectrum. Significantly more UV+ moths th…

Responses in metabolic rate to changes in temperature in diapausing Colorado potato beetleLeptinotarsa decemlineatafrom three European populations

Many insects survive adverse periods in seasonal environments by entering diapause, a deep resting stage, during which energy consumption is typically low and gas exchange is in the form of a discontinuous gas exchange cycle (DGC). Because insects in high-latitude environments are severely time constrained during summer, an effective diapause termination with careful regulation of metabolic rate is important. The present study examines whether diapausing Colorado potato beetles Leptinotarsa decemlineataSay originating from three latitudinally different regions in Europe differ in their quantitative or qualitative gas exchange patterns in response to an increasing temperature. Overall produc…

The art of animal colouring

Predator experience on cryptic prey affects the survival of conspicuous aposematic prey.

Initially, aposematism, which is an unprofitable trait, e.g. noxiousness conspicuously advertised to predators, appears to be a paradox since conspicuousness should increase predation by naive predators. However, reluctance of predators for eating novel prey (e.g. neophobia) might balance the initial predation caused by inexperienced predators. We tested the novelty effects on initial predation and avoidance learning in two separate conspicuousness levels of aposematic prey by using a 'novel world' method. Half of the wild great tits (Parus major) were trained to eat cryptic prey prior to the introduction of an aposematic prey, which potentially creates a bias against the aposematic morph. …

Can Indirect Herbicide Exposure Modify the Response of the Colorado Potato Beetle to an Organophosphate Insecticide?

AbstractOrganisms live in complex multivariate environments. In agroecosystems, this complexity is often human-induced as pest individuals can be exposed to many xenobiotics simultaneously. Predicting the effects of multiple stressors can be problematic, as two or more stressors can have interactive effects. Our objective was to investigate whether indirect glyphosate-based herbicide (GBH) exposure of the host plant has interactive effects in combination with an insecticide (azinphos-methyl) on an invasive pest Colorado potato beetle (Leptinotarsa decemlineata Say). We tested the effects of GBH and insecticide on the survival, insecticide target genes expression (acetylcholinesterase genes)…

Responses of a native plant species from invaded and uninvaded areas to allelopathic effects of an invader

Invaders exert new selection pressures on the resident species, for example, through competition for resources or by using novel weapons. It has been shown that novel weapons aid invasion but it is unclear whether native species co‐occurring with in‐ vaders have adapted to tolerate these novel weapons. Those resident species which are able to adapt to new selective agents can co‐occur with an invader while others face a risk of local extinction. We ran a factorial common garden experiment to study whether a native plant species, Anthriscus sylvestris, has been able to evolve a greater tolerance to the allelochemicals exerted by the invader, Lupinus polyphyllus. Lupinus polyphyllus produces …

Genetic variation in growth and development time under two selection regimes in Leptinotarsa decemlineata

It is possible to predict the potential range of a species on the basis of its ecological characteristics and those of the invaded ecosystem. The existence of genetic variation indicates a species' potential to respond to new environmental conditions, thus facilitating its success as an invader. Accordingly, evolutionary and ecological approaches are needed to identify the factors explaining both species' range and their potential to invade new areas. We combined these two approaches and studied whether genetic variation in life-history traits under abiotic (temperature) and biotic (host plant) selection pressures contributes to the potential range expansion of Leptinotarsa decemlineata Say…

THE EFFECT OF ALTERNATIVE PREY ON THE DYNAMICS OF IMPERFECT BATESIAN AND MÜLLERIAN MIMICRIES

Both Batesian and Müllerian mimicries are considered classical evidence of natural selection where predation pressure has, at times, created a striking similarity between unrelated prey species. Batesian mimicry, in which palatable mimics resemble unpalatable aposematic species, is parasitic and only beneficial to the mimics. By contrast, in classical Müllerian mimicry the cost of predators' avoidance learning is shared between similar unpalatable co-mimics, and therefore mimicry benefits all parties. Recent studies using mathematical modeling have questioned the dynamics of Müllerian mimicry, suggesting that fitness benefits should be calculated in a way similar to Batesian mimicry; that i…

Alternative prey can change model-mimic dynamics between parasitism and mutualism

Classical (conventional) Mullerian mimicry theory predicts that two (or more) defended prey sharing the same signal always benefit each other despite the fact that one species can be more toxic than the other. The quasi-Batesian (unconventional) mimicry theory, instead, predicts that the less defended partner of the mimetic relationship may act as a parasite of the signal, causing a fitness loss to the model. Here we clarify the conditions for parasitic or mutualistic relationships between aposematic prey, and build a model to examine the hypothesis that the availability of alternative prey is crucial to Mullerian and quasi-Batesian mimicry. Our model is based on optimal behaviour of the pr…

Negatively condition dependent predation cost of a positively condition dependent sexual signalling.

Predation is considered as an important factor constraining the expression of sexual signals. Nevertheless, direct quantitative evidence for predation provoking significant viability costs on individuals signalling at high rates is scarce. Moreover, it is unclear whether high rate signallers are able to balance presumably increased predation costs. We examined whether a condition dependent audible sexual signal, drumming, makes Hygrolycosa rubrofasciata male spiders more prone to predation by pied flycatchers (Ficedula hypoleuca), and whether sexual signalling rate is related to escaping ability once attacked. When birds were given a choice between two spider males manipulated to drum eithe…

Cold tolerance during larval development: effects on the thermal distribution limits of Leptinotarsa decemlineata

Insects' cold tolerance during their development is a surprisingly understudied subject in ecology, despite the fact that subzero temperatures during the growing season are common at high altitudes and latitudes. Subzero temperatures can have detrimental effects on organisms, restricting a species' range. This study addresses the question whether night frosts during the growing season have an instant or delayed negative impact on larval mortality of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). We also tested whether populations from the centre (Poland) and margins (Russia) of the distribution range of L. decemlineata differ in their responses to s…

PERSPECTIVE: Underutilized resources for studying the evolution of invasive species during their introduction, establishment, and lag phases

The early phases of biological invasions are poorly understood. In particular, during the introduction, establishment, and possible lag phases, it is unclear to what extent evolution must take place for an introduced species to transition from established to expanding. In this study, we highlight three disparate data sources that can provide insights into evolutionary processes associated with invasion success: biological control organisms, horticultural introductions, and natural history collections. All three data sources potentially provide introduction dates, information about source populations, and genetic and morphological samples at different time points along the invasion trajector…

Data from: Propagule pressure increase and phylogenetic diversity decrease community’s susceptibility to invasion

Invasions pose a large threat to native species, but the question of why some species are more invasive, and some communities more prone to invasions than others, is far from solved. Using ten different three-species bacterial communities, we tested experimentally if the phylogenetic relationships between an invader and a resident community and propagule pressure affect invasion probability. We found that greater diversity in phylogenetic distances between the resident community members and the invader lowered invasion success, and higher propagule pressure increased invasion success whereas phylogenetic distance had no clear effect. In the later stages of invasion phylogenetic diversity ha…

Data from: Stress for invasion success? Temperature stress of preceding generations modifies the response to insecticide stress in an invasive pest insect

Adaptation to stressful environments is one important factor influencing species invasion success. Tolerance to one stress may be complicated by exposure to other stressors experienced by the preceding generations. We studied whether parental temperature stress affects tolerance to insecticide in the invasive Colorado potato beetle Leptinotarsa decemlineata. Field-collected pyrethroid-resistant beetles were reared under either stressful (17°C) or favourable (23°C) insecticide-free environments for three generations. Then, larvae were exposed to pyrethroid insecticides in common garden conditions (23°C). Beetles were in general tolerant to stress. The parental temperature stress alone affect…

The effect of environmental fluctuations – Could climate change promote species’ invasion success?

The global climate change is presumed to increase the amount of fluctuations in the environmental conditions. This could increase the amount of species invasion into new areas if fluctuations affect the ecological and evolutionary processes that make species successful as invaders, and native communities and their environments more susceptible to invasions1. Disturbed environments are assumed to be more prone to invasions and the fluctuations in invasive species’ home range could pre-adapt them to tolerate similar conditions elsewhere. Under fluctuating conditions, natural selection could potentially favor traits like generalism, which is profitable in adaptation to wide range of conditions…

Supplementary data to: Sequence and Regulatory Variation in Acetylcholinesterase Genes Contribute to Insecticide Resistance in Different populations of Leptinotarsa decemlineata

This is data used in Aigi Margus, Saija Piiroinen, Philipp Lehmann, et al. Sequence and Regulatory Variation in Acetylcholinesterase Genes Contribute to Insecticide Resistance in Different populations of Leptinotarsa decemlineata. Authorea. September 26, 2021. Data contains bioassay data, enzyme activity data, gene expression data and gene sequence data used to research insect herbivores resistance to insecticides. See the attached data description file for more information.

Original data for manuscript: Can indirect herbicide exposure modify the response of the colorado potato beetle to an organophosphate insectiside

Organisms live in complex multivariate environments. In agroecosystems, this complexity is often human-induced as pest individuals can be exposed to many xenobiotics simultaneously. Predicting the effects of multiple stressors can be problematic, as two or more stressors can have interactive effects. Our objective was to investigate whether indirect glyphosate-based herbicide (GBH) exposure of the host plant has interactive effects in combination with an insecticide (azinphos-methyl) on an invasive pest Colorado potato beetle (Leptinotarsa decemlineata Say). We tested the effects of GBH and insecticide on the survival, insecticide target genes expression (acetylcholinesterase genes) and oxi…