Population structure of the boll weevil in cotton fields and subtropical forests of South America: a bayesian approach

The main goal of this contribution is to investigate the genetic structure of boll weevil populations from South America (Argentina and Brazil) and to make further comparisons with a putative source population from USA. Samples were collected in a Paranaense forest under reserve protection, cotton fields and non-cultivated areas. Data from anonymous molecular markers were analysed using both traditional methods of population genetics and Bayesian approaches. Results help to support a previous hypothesis on the presence of two lineages of boll weevil populations in South America: one with characteristics of recent invaders and the other with characteristics of ancient populations. The sample from Urugua-í Provincial Park (Misiones, Argentina) shows the highest percentage of polymorphic loci, the highest values of mean heterozigosity, and the largest number of population-specific alleles, all being typical features of ancient populations. Furthermore, the Urugua-í sample shows two gene pools occurring in sympatry, probably as a consequence of a secondary contact. The remaining samples reveal not only lower percentages of polymorphic loci and heterozygosity values, but also an almost negligible presence of specific alleles. Bayesian methods also suggest the occasional migration of some individuals of ancient lineages from their natural habitats in fragments of the Paranaense forest into cotton fields, and vice versa.     

Effects of soybean Kunitz trypsin inhibitor on the cotton boll weevil (Anthonomus grandis)

The cotton boll weevil, Anthonomus grandis, is an economically important pest of cotton in tropical and subtropical areas of several countries in the Americas, causing severe losses due to their damage in cotton floral buds. Enzymatic assays using gut extracts from larval and adult boll weevil have demonstrated the presence of digestive serine proteinase-like activities. Furthermore, in vitro assays showed that soybean Kunitz trypsin inhibitor (SKTI) was able to inhibit these enzymes. Previously, in vivo effects of black-eyed pea trypsin chymotrypsin inhibitor (BTCI) have been demonstrated towards the boll weevil pest. Here, when neonate larvae were reared on an artificial diet containing SKTI at three different concentrations, a reduction of larval weight of up to 64% was observed for highest SKTI concentration 500 microM. The presence of SKTI caused an increase in mortality and severe deformities of larvae, pupae and adult insects. This work therefore represents the first observation of a Kunitz trypsin inhibitor active in vivo and in vitro against A. grandis. Bioassays suggested that SKTI could be used as a tool in engineering crop plants, which might exhibit increased resistance against cotton boll weevil.     

Isolation and characterization of polymorphic microsatellite loci in the boll weevil,Anthonomus grandis Boheman (Coleoptera: Curculionidae)

The boll weevil (Anthonomus grandis Boheman) is a major insect pest of cotton in North America. Dispersal activity poses a threat to ongoing eradication efforts in the US, but little is known about the frequency of long‐distance migration. Nuclear molecular markers are needed to assess gene flow in relation to geographical distance. A biotin‐enrichment strategy was employed to develop microsatellite markers for the boll weevil. Of 23 loci isolated, 14 were polymorphic with three to 10 alleles per locus. Twelve of the polymorphic loci showed Mendelian inheritance and are likely to be useful in population genetics studies.     

Susceptibility of the boll weevil to Steinernema riobrave and other entomopathogenic nematodes

The susceptibility of the boll weevil (BW), Anthonomus grandis Boheman, to Steinernema riobrave and other nematode species in petri dishes, soil (Hidalgo sandy clay loam), and cotton bolls and squares was investigated. Third instar weevils were susceptible to entomopathogenic nematode (EN) species and strains in petri dish bioassays at 30 degrees C. Lower LC(50)'s occurred with S. riobrave TX- 355 (2 nematodes per weevil), S. glaseri NC (3), Heterorhabditis indicus HOM-1 (5), and H. bacteriophora HbL (7) than H. bacteriophora IN (13), S. riobrave TX (14), and H. bacteriophora HP88 (21). When infective juveniles (IJs) of S. riobrave were applied to weevils on filter paper at 25 degrees C, the LC(50) of S. riobrave TX for first, second, and third instars, pupae, and 1-day-old and 10-days-old adult weevils were 4, 5, 4, 12, 13, and 11IJs per weevil, respectively. The mean time to death, from lowest to highest concentration, for the first instar (2.07 and 1.27days) and second instar (2.55 and 1.39days) weevils were faster than older weevil stages. But, at concentrations of 50 and 100IJs/weevil, the mean time to death for the third instar, pupa and adult weevils were similar (1.84 and 2.67days). One hundred percent weevil mortality (all weevil stages) occurred 3days after exposure to 100IJs per weevil. Invasion efficiency rankings for nematode concentration were inconsistent and changed with weevil stage from 15 to 100% when weevils were exposed to 100 and 1IJs/weevil, respectively. However, there was a consistent relationship between male:female nematode sex ratio (1:1.6) and nematode concentration in all infected weevil stages. Nematode production per weevil cadaver increased with increased nematode concentrations. The overall mean yield of nematodes per weevil was 7680IJs. In potted soil experiments (30 degrees C), nematode concentration and soil moisture greatly influenced the nematode efficacy. At the most effective concentrations of 200,000 and 400,000IJs/m(2) in buried bolls or squares, higher insect mortalities resulted in pots with 20% soil moisture either in bolls (94 and 97% parasitism) or squares (92 and 100% parasitism) than those of 10% soil moisture in bolls (44 and 58% parasitism) or squares (0 and 13% parasitism). Similar results were obtained when nematodes were sprayed on the bolls and squares on the soil surface. This paper presents the first data on the efficacy of S. riobrave against the boll weevil, establishes the potential of EN to control the BW inside abscised squares and bolls that lay on the ground or buried in the soil.     

Suppression of Boll Weevil Infestations (Coleoptera: Curculionidae) Occurring on Fallow-Season Cotton in Southern Texas by Augmentative Releases ofCatolaccus grandis(Hymenoptera: Pteromalidae)

Augmentative releases ofCatolaccus grandis(Burks) were conducted in the Lower Rio Grande Valley of Texas in an attempt to suppress infestations of boll weevil,Anthonomus grandisBoheman, occurring in stands of “fallow-season” cotton (i.e., fields in violation of the 1 September stalk destruction deadline mandated by Texas law). In each of five release sites monitored during the study period (October, 1994–March 1995), augmentative releases ofC. grandiswere accompanied by an appreciable increase in the incidence of parasitized boll weevils (primarily third-stage larvae and pupae infesting abcised cotton squares) within a relatively brief time period. The relatively high incidence of host mortality caused byC. grandisin each release site was largely indispensible (i.e., would not have occurred in the absence of the parasite) and served to destroy significant numbers of immature boll weevils that appear to have been predisposed to successfully overwinter. The potential role of parasite augmentation in the management of the overwintering boll weevil population in southern Texas is discussed.     

Distribution and probable physiological role of esterases in reproductive,digestive, and fat-body tissues of the adult cotton boll weevil,Anthonomus grandis Boh

Polyacrylamide gel electrophoresis was used to examine gut, Malpighian tube, fat-body, testes, and ovariole tissues of the adult cotton boll weevil, Anthonomus grandis Boh. Esterases for which the inheritance has been reported previously by Terranova using whole-body homogenates were detected in dissected tissues and the probable physiological function of each allozyme is suggested. EST-1 occurs most frequently in ovarioles and female fat bodies. EST-2 is most often found in fat bodies and may be important in lipid turnover. No sex difference was observed. EST-3^S is found in fat bodies and reproductive tissue, while EST-3^F is always located in gut tissues, indicating that EST-3 is not controlled by a single autosomal locus with two codominant alleles as previously reported. EST-4, the most abundant esterase, can be detected in gut tissue at any age and is probably involved in digestion. EST-5 contains four allozymes which appear most frequently in testes and may be important during reproduction.     

Karyotypic analysis of the cotton boll weevil, Anthonomus grandis Boheman

The diploid chromosome number of the cotton boll weevil, Anthonomus grandis Boheman, is 44. Both C‐ and N‐banding techniques of mitotic cells demonstrated constitutive heterochromatin in the p arm of the eight largest chromosomes, the p arm of the X chromosome, and the centromeric region of autosomal groups A–D. Neither the y nor the group E autosomes appeared to contain constitutive heterochromatin. Supernumerary chromosomes were not found in the boll weevil. Restriction endonuclease banding of primary spermatocytes revealed a rod‐shaped Xy tetrad in which the X and y were terminally associated. The p arm of the large, submetacentric X was C‐band positive. While two of the autosomal tetrads were typically ring‐shaped in primary spermatocytes, the remaining 19 autosomal tetrads were rod‐shaped. This revised version was published online in July 2006 with corrections to the Cover Date.     

Temperature-dependent development and reproduction of the boll weevil (Coleoptera: Curculionidae)

Abstract Effects of temperature on development, survival, and fecundity of boll weevil, Anthonomus grandis grandis Boheman, were assessed at 10, 11, 12, 15,20,25,30,35,45, and 46 °C; 65% relative humidity; and a photoperiod of 13:11 (L: D) h. The mortality of boll weevil immature stages was 100% at 12°C and decreased to 36.4% as the temperature increased to 25°C. When the temperature increased from 30 °C to 45 °C, the mortality of weevils also increased from 50.1% to 100%. From 15°C to 35°C, the bollweevilpreimaginal development rate was linearly related to temperature. The average development time of total boll weevil immature lifestages decreased 3.6-fold and the preovipositional period decreased 3.3-fold when the temperature was increased from 15°C to 30°C. The lower threshold for development was estimated at 10.9, 6.6, 7.0, and 9.0 °C for eggs, larval, pupal, and total immature stages, respectively, with total thermal time requirement to complete immature stages of 281.8 DD (degree day) (15°C) and 247.8 DD (35 °C). At 1LC and 46°C, weevil females did not oviposit. Longevity of adult females decreased 4.6-fold with increasing temperatures from 15°C to 35°C. Fecundity increased with increasing temperatures up to 30°C and significantly decreased thereafter. These findings will be useful in creating a temperature-based degree-day model for predicting the occurrence of key life stages in the field. An accurate predictor of a pest's development can be very important in determining sampling protocols, timing insecticide applications, or implementing an integrated pest management control strategy targeting susceptible life stages.     

Thermal response and reversibility of prolonged larval diapause in the chestnut weevil, Curculio sikkimensis

Curculio sikkimensis undergoes prolonged larval diapause that is terminated by chilling and warming cycles. To examine the effects of warming temperatures and their duration on diapause termination, we exposed diapause larvae that had not been reactivated after chilling at 5 °C to 20 or 25 °C and chilled them again before incubation at 20 °C. With increasing warming duration at 20 °C, diapause termination after chilling increased and shorter chilling durations became effective. In contrast, few or no larvae warmed at 25 °C terminated diapause after chilling, irrespective of the warming duration. To investigate the effect of warming temperature on diapause intensity, larvae with diapause weakened by initial incubation at 20 °C after the first chilling were subsequently incubated at 15, 20, or 25 °C, then chilled at 5 °C before incubation at 20 °C. Diapause termination increased significantly after the larvae were treated at 15 or 20 °C but decreased significantly after they were treated at 25 °C. The intensification of prolonged diapause at 25 °C was reversed when the larvae were transferred to 20 °C. Diapause intensity in C. sikkimensis therefore decreases at 20 °C, increases at 25 °C, and can be reversed by alternately exposing diapause larvae to 20 and 25 °C. In C. sikkimensis, prolonged diapause does not always proceed in one direction, and its intensity fluctuates in response to ambient temperature conditions.     

Repeated cycles of chilling and warming effectively terminate prolonged larval diapause in the chestnut weevil, Curculio sikkimensis

Curculio sikkimensis (Coleoptera: Curculionidae) requires one or more years to complete its life cycle, owing to prolonged larval diapause. To compare the effects of temperature cycles and total periods of chilling on the termination of prolonged diapause, larvae were subjected to different chilling (5 degrees C) and warming (20 degrees C) cycles ranging from 30 to 720 days, and all cycles were repeated until the sum of chilling and warming periods reached 720 days. The prolonged diapause of C. sikkimensis was more effectively terminated by repeated cycles of chilling and warming than by prolonging the continuous chilling period. However, extremely short temperature cycles were not highly effective in enhancing diapause termination, even when such cycles were repeated many times. To examine the role of warming periods on diapause termination, diapause larvae were subjected to a sequence of chilling (120 days at 5 degrees C) and warming (240 days at 20 degrees C) with a warming period (0-120 days at 20 degrees C) inserted in the chilling period. Diapause larvae that were not reactivated in the first chilling period required exposure to a certain period of warming before they were able to complete diapause development in the subsequent chilling. Thus, C. sikkimensis appears to spread its reactivation times over several years in response to seasonal temperature cycles.     

Green leaf volatiles enhance aggregation pheromone of boll weevil, Anthonomus grandis

Enhancement of an insect pheromone response by green leaf volatiles is reported for the first time in the boll weevil, Anthonomus grandis Boh. (Coleoptera: Curculionidae). Single cell recordings from antennal olfactory receptors revealed a class of cells selectively responsive to six carbon alcohols and aldehydes (i.e., green leaf volatiles). Field tests with released weevils demonstrated enhanced trap captures with trans-2-hexen-l-ol, cis-3-hexen-l-ol, or l-hexanol paired with grandlure, the boll weevil aggregation pheromone, when in competition with grandlure alone. Although dose-response curves constructed from electroantennograms were indicative of similar populations of receptor cells for selected six carbon alcohols, one of the compounds tested, cis-2-hexen-l-ol, was inactive in field tests. Trans-2-hexenal was active in single cell recordings, but was also inactive in field tests. In tests in cotton fields with indigenous weevil populations, trans-2-hexen-l-ol not only enhanced pheromone trap captures, but also extended the longevity of attractiveness of pheromone-baited traps. The combined electrophysiological and field data support across-fiber coding of green leaf volatiles by boll weevil olfactory receptors. The results are discussed with regard to the chemistry of the host plant of the boll weevil, cotton (Gossypium hirsutum L.), and potential economic significance for boll weevil eradication/suppression.

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Résumé Pour la première fois, l'augmentation de la puissance de la phéromone d'agrégation sous l'action de l'odeur verte est signalée chez A. grandis. Les enregistrements unitaires de cellules des récepteurs olfactifs des antennes a mis en évidence une catégorie de cellules réagissant sélectivement aux alcools et aux aldéhydes à 6 carbones (c'est-à-dire aux substances volatiles des feuilles vertes). Des essais dans la nature ont montré une augmentation des captures par les pièges de grandlure (phéromone d'agrégation du charançon) renforcés par du trans-2-hexen-l-ol, du cis-3-hexen-l-ol, ou du l-hexanol.Bien que les courbes des réponses en fonction des doses construites à partir des électroantennogrammes indiquent des catégories de cellules réceptrices identiques pour les alcools à 6 carbones examinés, l'une de ces substances, cis-2-hexen-l-ol, s'est révélée inactive dans la nature. Le trans-2-hexenal qui était actif avec des enregistrements unitaires de cellules, était lui aussi inactif dans la nature. Dans des essais dans des champs de coton avec des populations indigènes de charançons, le trans-2-hexen-l-ol a, non seulement augmenté l'efficacité des pièges à phéromone, mais a prolongé leur attractivité. Les résultats d'électrophysiologie combinés aux données de la nature appuient le modelage transversal des substances vertes volatiles des feuilles par les récepteurs olfactifs du charançon. Ces résultats sont interprétés en relation avec la chimie de la plante-hôte de A. grandis, le coton, Gossypium hirsutum L., et leur possibilité d'utilisation économique pour l'éradication du charançon.

     

Parental effects: physiological age, mating pattern, and diapause duration on diapause incidence of progeny in the cabbage beetle, Colaphellus bowringi Baly (Coleoptera: Chrysomelidae)

The cabbage beetle, Colaphellus bowringi Baly, is a short-day species that has both a facultative summer and winter adult diapause. In this study, we systematically investigated the effects of parental physiological age, mating pattern (including mating frequency and delayed mating), and diapause duration on the diapause incidence of progeny in the beetle. As parents aged, progeny diapause incidences showed considerable inter-parental variability and an abnormally varying pattern, but their predicting line was approximate to horizontal when reared under constant conditions. There was significant difference in progeny diapause incidence between multiple-mating and once-mating treatments. Higher diapause incidences in the progeny were observed in delayed-mating treatments. Progeny diapause incidences increased as diapause duration in the parental generation increased, and hibernating individuals produced more diapausing progeny than aestivating ones. Reciprocal cross tests between the post-diapause adults and non-diapause adults with no diapause history showed that the effect of parental diapause duration on the progeny diapause incidence was determined by both mothers and fathers but the mothers appeared to have a stronger effect than the fathers. Our results suggest that the parental age effect in Colaphellus bowringi might not be simply age-dependent, but may be controlled by the interaction of multiple factors, including physiological, environmental, behavioral and genetic factors. We discuss the ecological and physiological significance of these findings.     

Effect of temperature on the termination of prolonged larval diapause in the chestnut weevil Curculio sikkimensis (Coleoptera: Curculionidae)

Adults of the chestnut weevil Curculio sikkimensis emerged over a 3-year period under laboratory and quasi-field conditions due to a prolonged diapause that occurred at the mature larval stage. Variable proportions of the larvae remained in diapause after a single cold (5 degrees C) treatment of 120 days. Extension of the chilling period to as long as 540 days did not increase the percentage of diapause termination, and excessively long chilling actually reduced the percentage. Chilling was not indispensable to the termination of larval diapause. Diapause intensity was very high and variable, and more than 1000 days at 20 degrees C was necessary to reactivate all diapause larvae. When the diapause larvae were exposed to cycles of low (5 degrees C for 120 days) and high (20 degrees C for 240 days) temperatures, the percentage of diapause termination reached 100% after two or three such cycles. Thus, the prolonged diapause of C. sikkimensis has characteristics similar to the common short winter diapause in other insects, but has unique characteristics that ensure polymodal reactivation over several years.     

Heterochromatin protein 1 (HP1) is associated with induced gene expression in Drosophila euchromatin

Heterochromatin protein 1 (HP1) is a conserved nonhistone chromosomal protein, which is involved in heterochromatin formation and gene silencing in many organisms. In addition, it has been shown that HP1 is also involved in telomere capping in Drosophila. Here, we show a novel striking feature of this protein demonstrating its involvement in the activation of several euchromatic genes in Drosophila. By immunostaining experiments using an HP1 antibody, we found that HP1 is associated with developmental and heat shock-induced puffs on polytene chromosomes. Because the puffs are the cytological phenotype of intense gene activity, we did a detailed analysis of the heat shock-induced expression of the HSP70 encoding gene in larvae with different doses of HP1 and found that HP1 is positively involved in Hsp70 gene activity. These data significantly broaden the current views of the roles of HP1 in vivo by demonstrating that this protein has multifunctional roles.     

Length variation in a specific region of the period gene correlates with differences in pupal diapause incidence in the flesh fly, Sarcophaga bullata

We report differences in the length of a specific region of the circadian clock gene period (per) that correlate with different capacities for pupal diapause in the flesh fly, Sarcophaga bullata. The conspicuous difference is located in a region we refer to as the putative C-terminal photoperiodic (CP) region. The length of the CP region correlates inversely with the incidence of diapause. A deletion of 33 amino acids in this region correlates with a significant increase in the incidence of diapause, from 78.1% to 93.0%, and an insertion of 9 amino acids in the same area correlates with a drop in the diapause incidence to 4.0%. This correlation suggests a possible functional role for this region of per in photoperiodism.     

Interactions between dietary nitrogen and simulated autumn conditions on diet consumption and reproductive development in the boll weevil, Anthonomus grandis

ABSTRACT. Dietary nitrogen content and short-day, cool scotophase temperature were investigated for effects on diet consumption and reproductive development in the female boll weevil, Anthonomus grandis Boheman (Coleoptera: Curculionidae). Experimental weevils were maintained from the day of adult eclosion on cottonseed flour diets on a LD10:14 h, 27:15C cycle. Control weevils were maintained under long-day conditions (LD13:11) at a constant 27C.
Diets containing 4% and 1.7% N were labelled with a ¹⁴C-amino acid mixture and fed to individual females for 24 h to permit determinations of diet consumption rate, amino acid absorption, and reproductive allocation of the assimilated amino acids.
Females fed the 1.7% N diet under long-day, 27C laid 50% fewer eggs than females fed the 4% N diet under the same regimen. Egg production was depressed further by the 1.7% N diet when combined with short-day, and cool scotophase temperatures. These latter conditions delayed, but did not prevent, attainment of a high rate of egg production by females fed the 4% N diet.
The short-day, cool temperature regimen also stimulated diet consumption but reduced the allocation of absorbed ¹⁴C amino acids towards oogenesis.