Juvenile hormone and the control of adult diapause in the Colorado potato beetle,Leptinotarsa decemlineata

One of the major hormonal events preceding environmentally controlled diapause in the Colorado potato beetle, Leptinotarsa decemlineata, is the inhibition of the corpora allata. This paper reviews briefly the data developed by our laboratory on the role of the neuroendocrine system in the control of the corpus allatum. In addition, we demonstrate that during transport in the hemolymph JH is bound to a JH-III specific binding protein. The JH-binding sites of this protein have a high affinity for JH-III and display stereoselectivity. New data are presented showing that, in beetle hemolymph, JH-III specific binding occurs with the lipophorin fraction.     

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.     

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.     

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.     

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.     

Juvenile hormone and the ultrastructural properties of the fat body of the adult Colorado beetle,Leptinotarsa decemlineata say

Summary In the fat body of ovipositing female Colorado beetles, two types of lobes occur. The first type, the internal fat body, is highly specialised for protein synthesis. A lobe of the second type, the peripheral fat body, contains two types of cells, oenocytes and glycogen cells. Ovariectomy, performed at adult moult results in hypertrophy of the glycogen cells of the peripheral fat body. The lobes are characterized by the storage of lipid bodies and glycogen and by numerous mitochondria. Short-day conditions ab ovo, which induce diapause in adults, also result in hypertrophy of glycogen cells of the peripheral fat body. Furthermore, only few mitochondria occur but many proteinaceous bodies may be observed, which conditions are in contrast to the observed effects of castration. The fat body of allatectomized long-day females, has the same structure as that of short day beetles. Consequently a lack of juvenile hormone induces the proteinaceous bodies.Dr. A. De Loof gratefully acknowledges a scholarship as Aspirant of the National Foundation of Scientific Research in Belgium. We wish to thank Prof. Dr. h. C. J. de Wilde for his suggestions and helpfull criticism. We also thank Mr. W. Bohijn for his help in operating the EM and Mr. G. Maes for photography.     

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.     

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.     

A proctolin-like peptide and its immunocytochemical localization in the Colorado potato beetle,Leptinotarsa decemlineata

Summary Evidence is presented that neurons in the adult Colorado potato beetle contain a proctolin-like substance. By use of immunocytochemical methods the location of immunoreactive neurons in the central and stomatogastric nervous systems is described. No such neurons were found in the proto- and deutocerebrum or optic lobe. Few immunoreactive neurons are present in the tritocerebrum and numerous proctolin-immunoreactive neurons occur in all ventral ganglia and in the frontal ganglion. Two groups of neurosecretory cells in the suboesophageal ganglion contain a proctolin-immunoreactive substance. In these cells this material is co-localized with a bovine pancreatic polypeptide/FMRF amide-like substance and with a vasopressin/vasotocin/oxytocin-like substance. Proctolin-immunoreactive axon terminals were found on the musculature of the fore- and hindgut and of the vas deferens, and on some segmental muscles. Furthermore, proctolin-immunoreactive neurosecretory axon terminals were found in the corpus cardiacum. The proctolin-like substance may therefore function both as a neurotransmitter/neuromodulator and as a neurohormone. The presence of a proctolin-like substance was also demonstrated with a sensitive bioassay. On fractionation of extracts of the nervous systems of Leptinotarsa decemlineata with high performance liquid chromatography most of the proctolin-like bioactive material comigrated with authentic proctolin. This shows that a proctolin-like substance in this insect is very similar to, if not identical with, the known pentapeptide proctolin.     

Host-plant discrimination and evolution of feeding preference in the Colorado potato beetle Leptinotarsa decemlineata

ABSTRACT. The behaviour of newly emerged adult Colorado potato beetles on preferred hosts follows a stereotyped pattern of sampling, feeding, grooming and rest. Reduced meal sizes on less-preferred hosts is accompanied by increased sampling and frequent interruptions in feeding. A systematic increase in pre-ingestive sampling on less-preferred foodplants indicates that beetles discriminate among closely related species within the Solanaceae. This ability may depend primarily on stimuli perceived at, and near, the leaf surface. Three geographic populations of beetles have adapted to different local host plants, but have not lost their preference for feeding on an ancestral host species. Host shifts by oligophagous insects to related plant species may evolve through selection for feeding generalists in isolated populations, and may not require genetic changes affecting the perception of a particular novel host.     

Biological control potential of Turkish entomopathogenic nematodes against the Colorado potato beetle,Leptinotarsa decemlineata

Three Turkish isolates of the entomopathogenic nematodes Steinernema carpocapsae, Steinernema feltiae and Heterorhabditis bacteriophora were evaluated under laboratory conditions (in different temperatures and doses) for their biocontrol efficiency against last instar Leptinotarsa decemlineata. Herein, the effects of infective juveniles in aqueous suspension against L. decemlineata were evaluated. S. feltiae appeared to be the most pathogenic nematode among the tested species.     

Ultrastructure of the galeal sensory complex in adults of the Colorado potato beetle, Leptinotarsa decemlineata

ABSTRACT. The structure of galeal sensilla of the Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae), is described using electron microscopical methods. Previous electro-physiological studies indicate that these sensilla respond to amino acids, sucrose and plant saps. One physiological type is particularly sensitive to L-alanine and gamma amino butyric acid (GABA).
Three morphologically different types of sensilla occur on the galeal tip. The more numerous apical pegs are not distinguishable from one another on the basis of external structure, although they differ physiologically. Five sensory cells are associated with most apical pegs. One apical peg, the α-sensillum, contains only four cells. All apical pegs have one cell with a tubular body. The remaining cells have unbranched dendrites and are associated with a single apical pore.
Apical hairs differ from the apical pegs by having double innervation. Within the hair shaft, a dendritic sheath is lacking and the sensillar sinus extends to the base of the hair. The function of this hair type is not known.
Numerous mechanosensory hairs which surround the other sensilla are singly innervated and contain a tubular body at the level of the outer dendritic segments.     

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).     

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.