Quiescence in the Colorado potato beetle, Leptinotarsa decemlineata

Quiescence (death feigning) was induced in the Colorado potato beetle, Leptinotarsa decemlineata by mechanical disturbance of live and artificial plants on which beetles were walking, feeding or grooming. The response consisted of a sudden cessation of activity lasting 12 to 500 s. Recovery from this immobile state was usually preceded by either antennal or leg movement. The time taken for beetles to come out of the immobile state (i.e. recovery time) was not dependent on age or sex of beetles or on stimulus duration, but was dependent on degree of displacement and on satiation level. Repeated stimulations led to longer recovery times and to a smaller proportion of responding beetles. Stimulation within the immobile state also prolonged recovery time. The most effective frequency of mechanical disturbance for startling beetles was 30 Hz. This response is presumed to be part of the extensive defense repertoire of this beetle, but we have so far been unable to discover a natural enemy that will elicit it.     

Effect of food deprivation on the ambulatory movement of the Colorado potato beetle,Leptinotarsa decemlineata

Modification of dispersal behaviour is a common response of insects to food and water deprivation. The literature suggests that different insects respond with different strategies: changing walking parameters, switching dispersal mode (walking to flight or vice versa), or changing the host searching path. The goal of this study was to add to the limited literature on the subject by investigating, whether the walking parameters of adult male Colorado potato beetles, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), change in response to food and water deprivation. Observations on the distance walked, the travel speed, and the frequency of walking bouts were carried out in laboratory arenas using motion monitoring equipment. Summer and overwintered beetles were exposed to short starvation periods (2, 4, 8, 24 h) and two ranges of long starvation periods (1, 2, 4, 8 days and 1, 2, 4, 8, 16, and 32 days). Only the longest food deprivation periods of 16 and 32 days significantly reduced the walking distance, speed, and frequency of walking bouts of summer beetles. No changes were observed with overwintered beetles. The tolerance of the beetles without access to water to the different periods of food deprivation was similar to that for beetles with water except after a starvation period of 32 days, when the travel speed of summer beetles was significantly reduced by 33%. The absence of increased walking parameters found in this study and earlier observations of increased flight frequency suggest that the strategy of summer beetles will be to change the dispersal mode from walking to flight and/or to change the walking host searching path. The same results of this study and earlier observations of a decrease in the mean frequency of daily flights suggest that the strategy of overwintered L. decemlineata, exposed to food deprivation, will be to change the host search walking path rather than the walking parameters themselves.     

Photosynthetic responses of potato to Colorado potato beetle injury and differences in injury between adult males and females

Establishing rates of injury to plants and the physiological impact of this injury provides essential data in the development of economic injury levels, but variation of sex effects is not often considered. Here, we examined injury by the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), larvae and adult males and females on potato, Solanum tuberosum L. (Solanaceae). Specifically, we looked for adult sex differences between males and females in injury rates (= leaf consumption rates), and examined the impact of all types of injury (larval, adult male, and adult female) on gas exchange parameters of remaining potato leaf tissue. Experiments were conducted in the field and in growth chambers on Frito‐Lay proprietary and Pike chipping‐potato varieties at pre‐blooming and blooming stages. We found no change in photosynthetic rates on remaining (uninjured) leaf tissue infested with male, female, or fourth‐stage larva of Colorado potato beetle. However, when the midrib was cut in trials with male beetles, the remaining tissue above the injury exhibited photosynthetic rate reductions as a result of stomatal limitations. These findings are consistent with the pattern that we and other researchers have observed with gross tissue removal by various insects on other plant species. Adult females consumed more tissue than males, and temperature was positively correlated with feeding rates for both sexes. Sex‐related differences in feeding rate are most important to studies quantifying consumption rates for economically important species because of its potential impact on resulting economic injury level calculations.     

Effect of diet and feeding history on flight of Colorado potato beetle, Leptinotarsa decemlineata

We evaluated the hypothesis that Colorado potato beetle (Leptinotarsa decemlineata Say) (CPB) flight frequency is related to diet, and that it changes with duration of food unavailability or exposure to poor quality food by exposing adult overwintered and summer CPB populations to an acceptable host plant (conventional foliage), a poor host (insect resistant transgenic foliage expressing Bacillus thuringiensis tenebrionis[Btt] Cry3a toxin) and no host. Exposure to poor host and no host treatments (with or without water) decreased mean daily flight frequencies and the overall number of overwintered CPB flying, but increased the mean daily flight frequency and number of summer population CPB that flew. Overwintered CPB did not react to an absence of plants at emergence whereas summer CPB increased mean daily flight frequencies when plants and water were not available. The flight response to insect resistant foliage was similar to that for starvation treatments in both populations indicating that flight may not be triggered by Btt toxins but by starvation brought on by feeding on poor quality food. Flight was observed in all treatments for the duration of the test with two exceptions; overwintered beetles fed insect resistant foliage ceased flying after day 17 and summer beetles starved without water ceased after day 8 of a 29‐day study.     

Farm‐scale dispersal of Bactericera cockerelli in potato crops measured using Bt mark‐capture techniques

Natural populations of Bactericera cockerelli (Sulc) (Hemiptera: Triozidae), also known as tomato/potato psyllid, were marked in potato [Solanum tuberosum L. (Solanaceae)] crops using Bacillus thuringiensis Berliner (Bt) to investigate the impact of dispersal on crop infestation and management of potential insecticide resistance in New Zealand. The technique was adapted from previous studies that used conventional spray applications of Bt to mark Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), and identified marked individuals with selective microbiological assays and identification of characteristic crystal inclusions. Initially, marking rates of B. cockerelli were improved by using ultra‐low volume applications of undiluted Bt, but this result was not consistent. Several other pests and natural enemies were also marked. In mark‐capture studies, marked B. cockerelli were captured over 3 days on yellow sticky traps in small trap plots of potatoes at 60, 120, 180, 250, and 350 m from the sprayed crop. Bactericera cockerelli flight activity occurred throughout daylight hours with evidence of bimodal diurnal peaks. Significantly greater numbers of B. cockerelli were captured in downwind traps. The combined dispersal curve derived from two mark‐capture experiments estimated a mean dispersal distance for B. cockerelli of 100 m in 3 days and indicated that 10% of the population dispersed further than ca. 250 m. Over the period of a growing season, this level of dispersal suggests that B. cockerelli can disperse throughout a vegetable‐growing region, with implications for crop infestation and management of potential insecticide resistance.     

Insecticidal activity of Cry3Bb1 expressed in Bt maize on larvae of the Colorado potato beetle, Leptinotarsa decemlineata

Environmental risk assessment for genetically modified crops producing insecticidal Cry proteins derived from Bacillus thuringiensis (Bt) includes the evaluation of adverse effects on non-target organisms. Although ELISA concentration measurements indicate the presence of Cry proteins, sensitive insect bioassays determine whether there is biological activity. The insecticidal activity of the coleopteran-active Cry3Bb1 expressed in different tissues of Bt maize, contained in maize-fed herbivores, and in spiked soil was measured in sensitive insect bioassays using larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). Biological activity was confirmed of Cry3Bb1 contained in pulverized Bt maize pollen, roots, leaves, silk, and Bt maize-fed spider mites and western corn rootworm adults. When test substances were incorporated into artificial diet at the same concentrations of Cry3Bb1 (measured by ELISA), maize pollen and leaf litter exhibited lower toxicity than fresh plant material and maize-fed arthropods. This suggests that nutritional quality of food and degradation of Cry proteins may influence toxicity to insects. When soil was spiked with Cry3Bb1, the Bt protein was highly adsorbed and retained its full biological activity. Because toxicity of Cry proteins contained in different matrices cannot always be determined from ELISA values alone, sensitive insect bioassays can improve hazard and exposure assessments in environmental risk assessment of Bt crops.     

Ecdysteroids and adult diapause in the Colorado potato beetle, Leptinotarsa decemlineata

ABSTRACT. The difference in ecdysteroid activity in short-day (10-h photo-phase) and long-day (16-h photophase) Leptinotarsa decemlineata Say adults was characterized by high performance liquid chromatography, which revealed that for short-day beetles ecdysterone and ecdysone are the major constituents of the free ecdysteroids, whereas in long-day beetles products co-migrating with 2-deoxyecdysone and 2-deoxyecdysterone are just as abundant. Ecdysteroids were detected (15 ng/ml) only in the haemolymph of females. Ligation experiments showed that the induction of the diapause syndrome by inactivation of the corpora allata or the brain not only resulted in the formation of the typical diapause proteins but also in an increase of the ecdysteroid activity. It is suggested that adult diapause in L.decemlineata is regulated by a multifactorial system.     

Genetic relationships of introduced Colorado potato beetle Leptinotarsa decemlineata populations in Xinjiang,China

The Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), is an infamous invasive species worldwide that aggressively attacks potato and other Solanaceae crops. CPB was first found in China in 1993 and has since spread across 2.77 × 10⁵ km² in Xinjiang Uygur Autonomous Region. To better understand genetic variation and migration patterns, we used seven polymorphic microsatellite loci to elucidate the genetic relationships and gene flow among 10 CPB populations across Xinjiang. (i) Overall low levels of genetic diversity were detected on the entire population in Xinjiang but most of the diversity was retained among populations during invasion. (ii) The mean pairwise F_ST was low (0.071 ± 0.043) among populations. The genetic differentiation was little (pairwise F_ST 0.038 ± 0.016) between the five interior populations (Wusu, Urumqi, Jimsar, Qitai and Mulei) and Tacheng population. The six populations might come from the same genetic group via Bayesian clustering and were closely related on a neighbor‐joining tree. Combining the history data, the five interior populations may have originated from Tacheng. (iii) Gene flow was frequent, especially among the five interior populations. Individuals from the interior populations could be assigned to Tacheng at higher probabilities (means 0.518 ± 0.127) than vice versa (means 0.328 ± 0.074), suggesting that the beetle population has spread from the border to the interior in Xinjiang.     

Respiration rhythms and heartbeats of diapausing Colorado potato beetles, Leptinotarsa decemlineata, at low temperatures

Discontinuous gas exchange cycles (DGCs), active muscular ventilation, microcycles of repetitive openings, and heartbeats of diapausing adult Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae), were studied at low temperatures (0, 5, and 10 °C) using an electrolytic respirometer combined with an infrared actograph. The DGC of the adult constriction-flutter-open type was the main respiration mode in fully quiescent beetles at temperatures from 5 to 10 °C. The CO₂ bursts were actively ventilated at temperatures above 5 °C. During the flutter period, a series of microcycles appeared, but no muscular contractions associated with the microcycles were detected. We identified this respiration mode as discontinuous suction ventilation.
The hydration condition of the beetles did not influence the frequency of the gas exchange cycles, but dehydrated beetles showed significantly longer flutter periods and shorter ventilation periods than hydrated beetles. The heartbeat frequencies were influenced by both temperature and hydration status.
We conclude from the results that DGCs are used at rest in adult L. decemlineata under various environmental conditions and also at low temperatures. Our results showed that DGCs are the main respiration mode of resting adult Colorado potato beetle irrespective of its hydration state and temperature. Our method resolves O₂ uptake and subsequent CO₂ release in flutter and ventilation periods and shows that diffusion is replaced by convection to reduce water loss in adult beetles.     

Sperm utilization and post-copulatory female-guarding in the Colorado potato beetle, Leptinotarsa decemlineata

Crosses between white and black color morphs of the Colorado potato beetle, Leptinotarsa decemlineata Say, used as genetic markers, showed that male beetles ride and guard females to achieve the three copulations required to fill the spermatheca. Multiple matings are necessary for the females to realize their full reproductive potential. Sperm mixing occurs in the spermatheca providing partial sperm precedence. The data support the competitive mate searching theory.

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Résumé Différentes combinaisons d'accouplements entre des mutants noirs et blancs du doryphore de la pomme de terre, Leptinotarsa decemlineata, servirent à étudier le comportement de guarde de la femelle par le mâle après un premier accouplement. Les résultats indiquent que le sperme de deux mâles accouplant la même femelle se mélange dans la spermathèque assurant ainsi un certain niveau de précédence du sperme du dernier mâle.La masse de sperme dans la spermathèque était inférieure à sa capacité chez les femelles ayant été accouplées une ou deux fois. Le doryphore mâle aurait donc avantage à demeurer sur la femelle après un premier accouplement afin d'obtenir le minimum de trois accouplements requis pour remplir la spermathèque avant que les autres mâles puissent accoupler cette femelle. Les autres mâles ont avantage à essayer de dérober cette femelle pour remplir eux mêmes la spermathèque ou tout au moins profiter d'une certaine précédence du sperme du dernier mâle pour fertiliser une partie des oeufs.Sans accouplements multiples, la femelle ne possède pas suffisamment de sperme pour féconder tous ses oeufs. II semble aussi que les accouplements répétés puissent augmenter sa fécondité.Les résultats supportent une stratégie reproductive de compétition plutôt qu'une simple maximisation du nombre d'accouplements.

     

短时低温对马铃薯甲虫种群增长的影响

[目的]马铃薯甲虫Leptinotarsa decemlineata是我国重要的检疫性害虫,对茄科植物危害严重.本研究旨在明确出现倒春寒短时低温对马铃薯甲虫种群增长的影响.[方法]马铃薯甲虫卵在8℃下分别处理1,3和5d,以27℃下饲养的卵作为对照,调查卵孵化率及孵化后幼虫的生长发育和成虫繁殖情况,用种群参数评估短时低...     

Development and characterization of microsatellite markers in the Colorado potato beetle,Leptinotarsa decemlineata

The characterization of 11 Leptinotarsa decemlineata microsatellite loci isolated using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol is reported. Three to 15 alleles per locus were detected in 54 beetles collected from four populations of L. decemlineata. The mean number of alleles was 7.4 ± 3.2 and the level of expected heterozygosity ranged from 0.451 to 0.798. The total exclusionary probabilities using these loci for the first and the second parent were 0.982 and 0.999, respectively. These are the first microsatellite loci characterized from the Colorado potato beetles that can be used for estimating genetic diversity, population structure and parentage analysis.     

Combining genetic engineering and traditional breeding to provide elevated resistance in potatoes to Colorado potato beetle

The sustainable deployment of resistant crop varieties is a critical issue for the implementation of biotechnology in crop pest management. Feeding, biomass accumulation, and mortality were evaluated for susceptible, insecticide‐resistant, and Bacillus thuringiensis (Bt) Cry 3A‐selected Colorado potato beetle (Leptinotarsa decemlineata Say) (Coleoptera, Chrysomelidae) larvae fed on: cultivated potato, a Solanum chacoense line expressing leptine glycoalkaloids, a transformed line expressing Bt toxin, or the leptine line transformed to express Bt toxin. Larvae selected for resistance to Bt‐Cry3A performed better on Bt foliage, but not as well on the leptine foliage, compared to susceptible or insecticide‐resistant larvae. Neither leptine nor Bt toxin completely inhibited the feeding and growth of 3rd and 4th instars of all three strains of Colorado potato beetle. However, for all three strains of Colorado potato beetle on leptine + Bt foliage, feeding was almost zero, growth was zero or negative, and mortality was near 100%.     

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 (Coleoptera: Chrysomelidae). We reared full‐sib families of L. decemlineata from the current northernmost European population at 15 °C (temperature below that in the current range) and 25 °C (optimal temperature) and on three potato varieties. We monitored development time, adult weight and larval‐to‐adult survival, and estimated the amount of heritable variance. The development time and adult weight of progenies were more variable between than within families. Thus, there was genetic variability in traits relevant to the ability to adapt to a colder environment (i.e., accelerate development and become heavier) allowing range expansion further north in Europe, even though low temperature increased beetle mortality. Temperature strongly affected all traits measured. Potato variety, in turn, did not strongly affect beetles’ performance. Beetle ability to adapt to a cool environment was further enhanced by the fact that size was not constrained by fast development. The results showed that beetle populations possess genetic variation allowing a response to temperature and thus have the evolutionary potential to adapt and spread beyond their current range.     

RNA interference in the Colorado potato beetle,Leptinotarsa decemlineata: Identification of key contributors

RNA interference (RNAi) is a useful reverse genetics tool for investigation of gene function as well as for practical applications in many fields including medicine and agriculture. RNAi works very well in coleopteran insects including the Colorado potato beetle (CPB), Leptinotarsa decemlineata. We used a cell line (Lepd-SL1) developed from CPB to identify genes that play key roles in RNAi. We screened 50 genes with potential functions in RNAi by exposing Lepd-SL1 cells to dsRNA targeting one of the potential RNAi pathway genes followed by incubation with dsRNA targeting inhibitor of apoptosis (IAP, silencing of this gene induces apoptosis). Out of 50 genes tested, silencing of 29 genes showed an effect on RNAi. Silencing of five genes (Argonaute-1, Argonaute-2a, Argonaute-2b, Aubergine and V-ATPase 16 kDa subunit 1, Vha16) blocked RNAi suggesting that these genes are essential for functioning of RNAi in Lepd-SL1 cells. Interestingly, Argonaute-1 and Aubergine which are known to function in miRNA and piRNA pathways respectively are also critical to siRNA pathway. Using ³²P labeled dsRNA, we showed that these miRNA and piRNA Argonautes but not Argonaute-2 are required for processing of dsRNA to siRNA. Transfection of pIZT/V5 constructs containing these five genes into Sf9 cells (the cells where RNAi does not work well) showed that expression of all genes tested, except the Argonaute-2a, improved RNAi in these cells. Results from Vha16 gene silencing and bafilomycin-A1 treatment suggest that endosomal escape plays an important role in dsRNA-mediated RNAi in Lepd-SL1 cells.     

Contribution of habitat type to residency and dispersal choices by overwintered and summer adult Colorado potato beetles

The walking and flight dispersal of marked overwintered and summer Colorado potato beetles (CPB), Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), released in field box‐plots was monitored simultaneously in six habitats over a period of 4 days. The emigration out of plots by walking beetles was calculated from the catch in linear pitfall traps completely surrounding each box‐plot and emigration flight was estimated from the number of beetles missing from the plot or captured by the trap. Overwintered beetles dispersed sooner after release than summer beetles. Overall, the mean number of beetles retained by the habitat was significantly higher in the host habitat (potato) than in any non‐host habitat tested (soybean, pasture, bare ground, water, woodland). Unexpectedly, there was no or little difference in overall beetle retention between non‐host habitats except for higher retention in the water habitat. No difference in the ratio of flight over walking could be detected by the study between overwintered and summer CPB except in the water and woodland habitats. Twenty‐four hours after release, the highest ratios were obtained in the water and woodland habitats and the lowest in the bare‐ground habitat, but ratios were similar for all habitats, except water, after 96 h. As a population, under these experimental conditions, 96 h after release, it seems that CPB displayed a slight preference for flight over walking, with walking as a default mode. A fed and starved pre‐release treatment had no effect on dispersal rates or mode of dispersal. Essentially, our results showed that over a 96‐h period, northeastern North American CPB emigrated at similar rates from the various non‐host habitats encountered, except for water, using walking as much as flight. The host habitat retained CPB significantly longer than non‐host habitats but with a mode of dispersal ratio similar to that in non‐host habitats. The impact on dispersal of the various habitats encountered by CPB in the agro‐ecosystem was less important than expected suggesting that the interaction of environmental parameters is likely to have the most significant impact in determining dispersal rates and dispersal modes.     

Glandular trichomes of Solanum berthaultii and resistance to the Colorado potato beetle

The defensive mechanisms of the wild potato, solanum berthaultii Hawkes, to larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say), were studied by selective removal of glandular trichomes and trichome exudates from leaflets, and by comparing performance on S. berthaultii and on the cultivated potato, S. tuberosum L., which lacks defensively active type A and B glandular trichomes. Removal of type A trichomes increased the proportion of larvae that fed on S. berthaultii. Removal of the exudate from type B trichomes increased the proportion of larvae that fed and led to a decrease in mortality. The predominant active compounds in type B exudate, i.e. fatty acid esters of sucrose, were only effective in the presence of type A trichomes. Sucrose esters did not affect larval feeding on S. tuberosum leaflets or on S. berthaultii leaf discs from which the type A trichomes had been removed. Growth of surviving larvae was not significantly affected by removing type A trichomes or type B exudate. Growth of larvae was significantly increased when S. berthaultii leaflets were presented in artificial diet which eliminated the physical barrier of the type B stalks. Growth was no different on artificial diet containing either S. berthaultii or S. tuberosum leaf material (fresh or lyophilized powder) but was poorer on these diets than on S. tuberosum leaflets. The presence of type A trichomes is a fundamental requirement for expression of S. berthaultii resistance to L1 L. decemlineata. Type B droplets containing sucrose esters increase the expression of resistance in the presence of defensively-active type A trichomes.

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Résumé Les mécanismes de défense de la pomme de terre sauvage, S. berthaultii Hawkes, aux larves de Leptinotarsa decemlineata Say, ont été étudiés par ablation sélective des trichomes glandulaires et par l'élimination de leur exsudat des folioles, et par comparaison avec S. tuberosum L. qui a perdu les trichomes glandulaires défensifs A et B. L'ablation des trichomes A a augmenté la proportion de larves ayant consommé S. berthaultii. L'élimination de l'exsudat des trichomes B a augmenté la proportion de consommatrices et réduit la mortalité. Les principaux composés actifs de l'exsudat B, c'est-à-dire des esters d'acides gras de sucrose, n'étaient actifs qu'en présence de trichomes A. Les esters de sucrose n'ont pas modifié la consommation larvaire sur folioles de S. tuberosum, ou sur disques de feuilles de S. berthaultii dont les trichomes A avaient été enlevés. La croissance des larves survivantes n'a pas été modifiée significativement par l'ablation des trichomes A ou l'élimination de l'exsudat de B. La croissance des larves a été significativement augmentée quand les folioles de S. berthaultii ont été incorporés dans l'aliment artificiel après élimination de la barrière physique due aux pédoncules B. La croissance a été de même importance sur aliments artificiels contenant des feuilles (fraiches ou en poudre lyophylisée) de S. berthaultii ou de S. tuberosum, mais plus faible que sur folioles de S. tuberosum. La présence de trichomes A est indispensable à la résistance de S. berthaultii aux L, de L. decemlineata. Les gouttelettes de type B contenant des esters de sucrose augmentent l'expression de la résistance en présence d'une défense active par trichomes A.

     

The structure of feeding behavior in the colorado potato beetle,Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

Feeding behavior, in an ad libitum situation on potato plants in the laboratory, was continuously observed for approximately 7 h/day on 2 successive days for 18 adult femaleLeptinotarsa decemlineata. Additional behaviors were also recorded including resting, walking, biting, local movements, grooming, defecating, and regurgitating. These data were used to calculate a time budget for the various behaviors. The feeding data were analyzed to describe the structure of feeding for young adult females on their normal host plant. The criterion for a meal (minimum intermeal interval) was determined to be 286 s. This criterion was used to distinguish between intra- and intermeal interruptions in feeding for all subsequent analyses. Meals taken from leaves that were young, medium aged, or old did not differ, but on average beetles took 60% of their meals from young leaves. Meal size and meal duration were equally good predictors of when a meal would end. Feeding from stems was a prominent feature for most beetles. The stem meals had much longer durations than leaf meals, but stem feeding did not affect subsequent leaf feeding. The structure of feeding by these beetles is compared with that found in other insects, especiallyLocusta migratoria.