Ultrastructure of the neurosecretory system of the Colorado potato beetle,Leptinotarsa decemlineata (Say)

Summary The ultrastructure of seven types of neurosecretory cells (NSC) in the medial and lateral groups of the protocerebrum is described. The differences among cell types established earlier by light microscopy parallel differences in size and appearance of the neurosecretory particles observed in electron micrographs. No relationship was found between the affinity for Gomori's paraldehyde fuchsin stain and the nature of the particles.The secretions of the A-, A₁-, and C-types of NSC of the medial group are characterized by electron-dense neurosecretory granules of 1250 Å dia., medium-dense granules of 2100 Å, and electron-lucent vesicles of 1700 Å, respectively. The L-type NSCof the lateral group contain smaller (1300 Å) or larger (1700 Å) neurosecretory granules. The medial B- and E-types of NSC and the lateral L_B-type contain granulated vesicles (1200 Å) of the same appearance. These cell types differ in other respects and most likely have separate functions.The author wishes to thank the Laboratory of Virology of the Agricultural University for the use of the electron microscope, Mr. J. Groenewegen and Miss J. van Rinsum for technical assistance, and Professor J. Lattin for correcting the English text. Part of the work has been done while the author was in the service of the Netherlands Organization for the Advancement of Pure Research (ZWO, grant 942-48), and the National Council for Agricultural Research (TNO).     

Resistance of Colorado potato beetle (Leptinotarsa decemlineata Say) to commonly used insecticides in Iran

The Colorado potato beetle (CPB, Leptinotarsa decemlineata Say), is a major pest of potatoes in Iran and many other parts of the world. Injury is caused when adults and larvae feed on the foliage and stems of potato plants, resulting in poor yields and/or plant death. Adult beetles can also vector plant diseases. Historically, the CPB been controlled using different insecticides, but it is currently resistant to nearly all classes of insecticides and remains a serious pest in many parts of the world. All of the resistance mechanisms reported in insects have been demonstrated in CPB. L. decemlineata invaded Iran in the early 1980s, probably through the importation of infested potatoes. It has caused significant damage to potato crops in affected areas, and it accordingly remains a major threat to Iranian potato production. Regrettably, no IPM programs have been developed for managing CPB infestations in Iran. Furthermore, there are no organized CPB resistance monitoring programs in Iran, and the recommended insecticides for CPB control, endosulfan and phosalon, have not changed in over 22 years. Anecdotal evidence from local farmers suggests a reduction in the efficacy of control of CPB by commonly used insecticides, probably due to the reduced susceptibility to these insecticides. Given the economic significance of L. decemlineata infestations, the increasing prevalence of resistance in this species, the rate of spread of infestations, and the extent of the area infested, there is an urgent need to develop effective and sustainable integrated pest management programs for CPB in Iran.     

Quantitative trait locus mapping of pyrethroid resistance in Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae)

Quantitative trait locus (QTL) mapping is a valuable new tool for locating genomic regions that underlie variation in important traits such as insecticide resistance. Because QTL mapping complements a candidate gene strategy for understanding the genetic architecture of important traits, it may also facilitate the identification of genes causing important variation. After mapping the QTL locations, markers closely linked to QTL can be used for genetic analysis of population structure and to measure the spread and increase of resistance-causing QTL alleles. In this study, QTL influencing resistance to the pyrethroid insecticide esfenvalerate were mapped in the Colorado potato beetle Leptinotarsa decemlineata (Say) (CPB). Three QTL contributing to esfenvalerate resistance were identified from a mapping population of 79 individuals analyzed at 90 marker loci. One QTL had a large effect and two QTL had smaller effects. The major QTL occurs on the X chromosome, overlapping the position of a candidate gene (Leptinotarsa decemlineata Voltage sensitive sodium channel [LdVssc1]) previously implicated in pyrethroid resistance. Resistance-increasing alleles at the two minor-effect QTL originated with the susceptible parent, suggesting that alleles of small effect may be segregating in susceptible populations. Comparison of the New York population from which the susceptible parent originated with a more-susceptible population from North Carolina suggests that the minor-effect loci identified here may explain some of the variation in tolerance observed among susceptible populations. DNA sequencing of a portion of LdVssc1 shows that the resistance-conferring allele from the resistant parent does not contain the kdr mutation previously found in CPB and typically observed in other insects that are resistant to pyrethroid insecticides because of changes in this gene.     

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.     

马铃薯甲虫空间分布型及抽样技术研究

马铃薯甲虫Leptinotarsa decemlineata(Say)是我国马铃薯恶性入侵害虫.本研究利用聚集度指标检验、Taylor幂法则和Iwao回归法对马铃薯甲虫成虫、幼虫和卵在田间的空间分布型进行了研究.聚集度指标检测各虫态均为聚集分布,且种群聚集主要是由马铃薯甲虫各虫态本身的聚集行为,或由其本身的聚集行为与环...     

Assessment of genetic polymorphism in Colorado potato beetle Leptinotarsa decemlineata 8Say) by RAPD markers

Four decanucleotide primers were chosen to detect polymorphism for randomly amplified polymorphic DNA (RAPD) in the populations of Colorado potato beetle. The variability of RAPD patterns was studied in three oblasts (Kiev, Moscow and Kurgan) that are geographically distant and considerably differ in ecological and climatic conditions affecting the development of this beetle: Kiev, Moscow, and Kurgan oblasts. A high level of intrapopulation polymorphism exceeding interpopulation differences was revealed. RAPD-marker subsets differing in the frequency and distribution throughout the studied populations were revealed.     

Bacillus thuringiensis cry3Ca1 protein is toxic to the Colorado potato beetle, Leptinotarsa decemlineata (Say)

We expressed the wild-type cry3Aa3 and cry3Ca1 Bacillus thuringiensis genes, which code for insecticidal proteins, in an Escherichia coli expression system. Highly purified preparations of the soluble delta-endotoxins were used to perform comparative bioassays with third-instar larvae of the Colorado potato beetle (CPB). Acute mortality data showed that Cry3Ca1 (LD(50) = 320.1 ng) was 2-fold more toxic than Cry3Aa3 (LD(50) = 672.9 ng). We also compared the chronic effects of sublethal doses of these toxins by measuring the consumption of untreated foliage and monitoring survival and development for 6 days after intoxication. No significant additional mortality was recorded, but we found that surviving larvae fed Cry3Ca1 consumed foliage at a slower rate than the larvae fed Cry3Aa3, suggesting more damage to their digestive epithelium. This study, the first assessment of the toxicity of cry3Ca1 in third-instar CPB, suggests cry3Ca1 will prove useful for the control of this important insect pest.     

Diet-related plasticity of the digestive proteolytic system in larvae of the Colorado potato beetle (Leptinotarsa decemlineata Say)

Quantitative and qualitative changes in digestive proteolytic activities were monitored in fourth-instar larvae of the Colorado potato beetle (Leptinotarsa decemlineata Say) subjected to three different leaf diets. Depending on the diet, the larvae exhibited variable growth rates, similar for potato (Solanum tuberosum) and eggplant (Solanum melongena) diets but lower for the tomato (Lycopersicon esculentum) diet. Interestingly, these growth rates were not associated with total protease activity in the midgut. While growth of tomato-fed insects was negligible, midgut protease activity in these insects was 1.5 and 4.2 times higher than that measured for potato- and eggplant-fed insects, respectively. As seen on gelatin-containing polyacrylamide gels, midgut extracts from insects that ingested eggplant leaves contained only a few proteinase forms, while numerous forms were observed in extracts of potato- and tomato-fed larvae. Although several forms were common to the three diets, their relative importance in the insect midgut varied. This diet-related plasticity of the digestive proteolytic system in Colorado potato beetle larvae leads one to question the potential for control approaches based on the inhibition of digestive proteases. Arch. Insect Biochem. Physiol. 36:241–250, 1997. © 1997 Wiley-Liss, Inc.     

A QTL that confers resistance to Colorado potato beetle (Leptinotarsa decemlineata [Say]) in tetraploid potato populations segregating for leptine

Genetic resistance to Colorado potato beetle (Leptinotarsa decemlineata [Say]) from Solanum chacoense has been incorporated in the tetraploid potato selection, ND4382-19, which is highly resistant and contains moderate level of foliar leptines. We recently reported using ND4382-19 progeny, population ND5873 (ND4382-19 × Chipeta), to map two genes that segregated as complementary epistatic genes that allow accumulation of leptinidine (Lep) and acetyl-leptinidine (AL) on chromosomes 2 and 8, respectively. We describe here the characterization of a second half-sib population NDG116 (ND4382-19 × N142-72). In this population, solasodine from parent N142-72, which has Solanum berthaultii in its background, was predominant over solanidine-based alkaloids. Concentrations of solanidine, leptinidine, and acetyl-leptinidine were 15-, 5-, and 14-fold lower than in the ND5873 population. Nevertheless, Lep and AL mapped to the same locations on chromosomes 2 and 8 of parent ND4382-19, respectively. The two populations were evaluated for resistance to Leptinotarsa in field assays, and by detached leaf assay for population NDG116. In both families, QTL analysis identified a major QTL from ND4382-19 on the distal end of chromosome 2, close to the Lep locus. The contribution of this QTL to resistance ranged from 11 to 34% for ND5873 at four field sites. Contribution to resistance from the linkage group that contains the gene AL for the accumulation of leptine was not detected. In family NDG116, the same chromosome 2 QTL was detected for field and detached leaf assays, explaining 26 and 12% of the variance for defoliation and larval development, respectively. These data may indicate another resistance mechanism besides leptine in the Leptinotarsa resistance observed in these populations.     

Effects of four Bacillus spp. of soil origin on the Colorado potato beetle Leptinotarsa decemlineata (Say)

Four species of Bacillus were isolated from soil in an effort to find safe, effective and alternative biological control agents against plant pests. These bacteria were identified as Bacillus pumilus, Bacillus sphaericus, Bacillus megaterium and Bacillus cereus on the basis of fatty acid methyl ester analysis and carbon utilization profiles by using Microbial Identification and Biolog Microplate Systems. Laboratory experiments carried out to determine the insecticidal activities of these isolates showed that B. pumilus caused 95.7 and 26.7% mortality and B. sphaericus caused 74.5 and 23.3% mortality of Leptinotarsa decemlineata larvae and adults, respectively. B. cereus and B. megaterium showed 51.1 and 29.7%, respectively, of L. decemlineata larvae. This study presents at least two Turkish isolates from the genus Bacillus showing high insecticidal activity against L. decemlineata.     

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.     

Effect of vermicompost on nutrition and intermediary metabolism of Colorado potato beetle,Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae)

Colorado potato beetle (CPB) is the important pest of potato throughout the world. The study showed the effects of vermicompost on nutritional indices, digestive enzyme activities and intermediary metabolism of the larvae and the adults of CPB. Vermicompost affected significantly the efficiency of conversion of ingested food (ECI) and efficiency of conversion of digested food (ECD) in addition to activities of carbohydrases and proteases in both larvae and adults. Amount of total phenol compounds increased in the leaves of the potatoes grown on the soil containing 30% of vermicompost compared to control and it were amended by 15% of vermicompost. In case of intermediary metabolism, activities of aspartate aminotransferase and γ-glutamyl transferase showed no significant differences in the control and the treated larvae, but those were fluctuated upper and lower when 15% of vermicompost was added into growth plots. The amount of low-density lipoproteins (LDL) and high-density lipoproteins (HDL) in the larvae of CPB showed no significant differences among treatments. However, the amounts of LDL, HDL, glycogen and protein of the adults significantly increased using 30% of vermicompost. Results of the current study clearly revealed significant effects on some physiological processes in CPB fed on the plants grown in the different vermicompost treatments.     

Seasonal occurrence and relative abundance of aphids on potato plants with classical and transgenic characters of resistance to Colorado potato beetle Leptinotarsa decemlineata (Say)

The seasonal abundance and dynamics of potato-infesting aphids were studied in two seasons, on potato Solanum tuberosum L. with various types of transgenic and classical resistance to primary pests, especially the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say). The lines tested were the NewLeaf potato, a transgenic expressing the Bacillus thuringiensis CryIIIA toxin (BTT); NYL 235-4 a parental line derived from S. berthaultii (NYL) with a high density of glandular trichomes; and a transgenic line expressing oryzacystatln I (OCI), a proteinase inhibitor gene from rice; potatoes from two commercial cultivars, Superior and Kennebec, served as controls. Over the two seasons, Macrosiphum euphorbiae (Thomas) and, to a lesser extent, Myzus persicae (Sulzer) were by far the most abundant aphids observed in the experimental plots. M. persicae was observed relatively late in the season, and was most abundant on the Kennebec-OCI potato. Macrosiphum euphorbiae was more regular over seasons, and its highest densities were observed on the NYL 235-4 line. The population of M. euphorbiae showed markedly different patterns on the five lines studied, which was clearly related to potato plant phenology. Density of M. euphorbiae was observed on the NYL 235-4 line, in late July and early August.     

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.

     

Influence of fertilization on the Colorado potato beetle, Leptinotarsa decemlineata, in organic potato production

The abundance of the Colorado potato beetle, Leptinotarsa decemlineata (Say), in organically grown potato did not change significantly in response to increasing rates of dehydrated poultry manure. However, peaks of abundance of larvae were shifted forward in time in response to the high rate of organic fertilizer. Tests using excised foliage showed that the shift was not caused by differential larval mortality or longer developmental times. Time allocation to resting, walking, and feeding by adults was similar regardless of fertilizer rate. Adult foliage consumption was unaffected by organic fertilizer rates in no choice tests and significantly affected in few choice tests. A 22% longer larval development time on plants treated with low fertilizer rate than on plants with high rate was the most significant effect. Even though maximum plant height, canopy, biomass, and yield were significantly smaller in the organic than in conventional plots, the suitability of the plants was not affected except for reduced feeding by summer beetles. Summer adults spent less time feeding and consumed two to five times less foliage on organic potato than on inorganically fertilized and conventionally produced plants. The overall influence of fertilizer on Colorado potato beetle populations was limited and therefore can only play a secondary role in management strategies for organic potato. Avoidance of excessive organic fertilizer that promotes short larval development time and extension of the period over which large Colorado potato beetle larvae are present should be recommended.     

Behavioural and physical reactions of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) walking on a slanted surface

 A natural reaction of the adult Colorado potato beetle [Leptinotarsa decemlineata (Say)] is to walk uphill on sloped surfaces. The geotaxis reaction of the beetle was observed on slopes of 20° or steeper. It was demonstrated that the uphill orientation behaviour was not a consequence of physical limitation for across-slope locomotion. The walking speed of insects deviating from the fall line did not change within the range of slope angles tested. The speed of adult beetles decreased with an increase in the slope of the substrate as a reaction to the increased gravitational force vector opposing uphill movement. The larger size of the hind legs might make uphill locomotion more efficient than traversing a sloped surface. As the angle of the slope increased, the gait changed from a 3/3 to a 5/1, as did the posterior and anterior extreme position of the legs. This behaviour might be triggered by the need to maintain balance on slanted surfaces as the vertical projection of the centre of mass on the substrate moved outside the support base pattern at the steeper angles. In one experiment beetles were made to pull a load when walking over a horizontal surface. The loads pulled were equivalent to the gravitational loads opposing forward motion when walking up a slope. No differences in forward speed or gait were observed at the lower-angle equivalent compared to beetles walking on slopes. Differences in speed were noted at slope angles higher than 40° indicating that adaptation of the walking strategy might be needed on steeply slanted surfaces. Received: 2 May 2000 / Accepted in revised form: 11 September 2000     

Intraspecific mitochondrial DNA variation in the Colorado potato beetle,Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

Restriction endonuclease analyses of mitochondrial DNA (mtDNA) were used to examine genetic variability and population structure inLeptinotarsa decemlineata (Say). A group of three enzymes, EcoRI, HpaI, and PstI, was used to reveal polymorphism both within and among some of the 10 populations tested, yielding 16 haplotypes in combination. The frequencies of these 16 haplotypes differed significantly across geographic regions, indicating some partitioning of mtDNA haplotypes. Estimates of mtDNA sequence divergence (δ) between haplotypes ranged from 0.016 to 0.135%, suggesting local differentiation of mtDNA in some populations. Analysis of these data suggests that Texas was colonized by more than one mtDNA lineage, most likely originating in Mexico. We hypothesize that a larger founder size for the initial introductions or high levels of variability in the parent population at the edge of the CPB expanding range led to the initial partitioning of haplotypes observed in samples from Texas.     

Genetic structure of populations of the Colorado potato beetle Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

The genetic structure of a Colorado potato beetle population from Kiev oblast was examined by cluster analysis of individual RAPD patterns. The obtained clustering indicates that the population is structured. This may be explained by adaptation to pyrethroid insecticides used for controlling the population size of this pest. Micro-evolutionary factors affecting the genetic structure of local populations of Colorado potato beetle are discussed.