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Dispersal of boll weevils (Coleoptera: Curculionidae) from cotton modules before ginning.

We characterized the level of risk of boll weevil, Anthonomus grandis grandis Boheman, reintroduction to an eradication zone posed by dispersal from cotton modules during and after transport to the gin. Mark-release-recapture experiments in August and September in Texas indicated that most weevils disperse rapidly from the module surface, temperature permitting, unless confined under a module tarp, where most died. Nevertheless, 1-5% of released weevils were recovered alive after 24 h on the side and top surfaces of modules, representing potential dispersants. Mortality of boll weevils caged on the top surface of a module was 95-100% after 1-4 d when maximum air temperatures were > or = 33 degrees C and 72-100% when minimum temperatures were -7 degrees C or lower, but a few survived even after experiencing a minimum daily temperature of -12 degrees C. Under warm (daily maximum temperatures > or = 25 degrees C) and cold (daily minimum temperatures < or = 0 degrees C) weather conditions, survival was higher under the tarp than on the open surface of the module (20 versus 7% and 42 versus 26%, respectively), but mortality was 100% in both locations when temperatures reached 34 degrees C. Our results indicate that although the threat to an eradication zone posed by boll weevil dispersal from an infested module is very low under most environmental conditions, it is probably greatest when 1) a module is constructed and transported from an infested zone during weather too cool for flight, followed by warm weather favorable for flight at the gin yard; or 2) such a module is transported immediately after construction in moderate-to-warm weather.

Agriculture↗

Size-dependent feeding and reproduction by boll weevil (Coleoptera: Curculionidae).

The considerable variation in adult size of the boll weevil, Anthonomus grandis grandis Boheman, has been well documented, but the influences of adult size on reproductive rate are not known. We examined the relationship between the size of boll weevils and their feeding and oviposition. Weevils weighed to the nearest milligram were grouped into five categories based on pupal weight: < or =5, 6-10, 11-15, 16-20, and >20 mg. Numbers of lifetime punctures produced in flower buds (squares) of cotton, Gossypium hirsutum L., by both sexes of adults tended to increase with pupal weight. Boll weevil females with pupal weights >10 mg produced progeny with significantly higher survival to adulthood and also produced a higher percentage of female progeny than those with pupal weights < or =10 mg. The population growth indices for females having pupal weights >10 mg averaged 1.8-fold higher than those of females weighing < or =10 mg. Survivorship of adults of both sexes also tended to increase with pupal weight. The percentage of females laying eggs on any given day averaged 2.1 times higher when their pupal weights were >10 mg than when their pupal weights were < or =10 mg. Although small size negatively affected female reproductive potential, even extremely small females produced some viable offspring. However, the penalties of small adult size, in terms of longevity and reproductive potential, suggest that cultural practices that result in the production of small adults may be used to impact weevil populations.

Animals↗

Proactive spraying against boll weevils (Coleoptera: Curculionidae) reduces insecticide applications and increases cotton yield and economic return.

The current standard practice of two to three preemptive insecticide applications at the start of pinhead (1-2-mm-diameter) squaring followed by threshold-triggered (whenever 10% of randomly selected squares have oviposition punctures) insecticide applications for boll weevil, Anthonomus grandis grandis Boheman, control does not provide a reliably positive impact on cotton, Gossypium hirsutum L., yields in subtropical conditions. This study showed that four fewer spray applications in a "proactive" approach, where spraying began at the start of large (5.5- 8-mm-diameter) square formation and continued at 7- to 8-d intervals while large squares were abundant, resulted in fewer infested squares and 46-56% more yield than the standard treatment at two locations during 2004. The combination of fewer sprays and increased yield made the proactive approach 115-130% more profitable than the standard. The proactive approach entails protection only at the crop's most vulnerable stage (large squares) that, as a source of food, accelerates boll weevil reproduction. In contrast, the standard approach protects early season small squares and later season bolls, both of which contribute less to boll weevil reproduction than large squares. Proaction is an in-season crop protection approach that can be used to increase yield in individual fields during the same season and that could be incorporated into boll weevil eradication strategy that involves later diapause sprays. Because proaction is based on an important relationship between the cotton plant and boll weevil reproduction, the tactic will probably be effective regardless of climate or region.

Animals↗

Short-range dispersal and overwintering habitats of boll weevils (Coleoptera: Curculionidae) during and after harvest in the subtropics.

Field experiments in the subtropical Lower Rio Grande Valley of Texas were conducted to determine the extent of adult boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), dispersal from cotton, Gossypium hirsutum L., fields during harvest operations and the noncotton-growing ("overwinter") period between 1 September and 1 February. Using unbaited large capacity boll weevil traps placed at intervals extending outward from commercial field edges, boll weevils did not move in substantial numbers during harvest much beyond 30 m, primarily in the direction of prevailing winds. From traps placed in fallow cotton; citrus; lake edge; pasture; treeline; sorghum, Sorghum bicolor (L.) Moench, and sugarcane, Saccharum spp., habitats during the overwinter period, the most boll weevils were collected in the fallow cotton fields and adjacent treelines during the fall. However, the greatest abundances of boll weevils were found in citrus orchards in the spring, before newly planted cotton fields began to square. One of the three lake edges also harbored substantial populations in the spring. Egg development in females was not detected between November and April, but in cotton fields most females were gravid between May and August when cotton fruiting bodies were available. Mated females, as determined by discoloration of the spermatheca, made up 80-100% of the female population during November and December but declined to approximately 50% in February. The lower incidence of mating indicates a reduction in physical activity, regardless of overwinter habitat, until percentages increased in March and April after cotton fields had been planted and squares were forming.

Animals↗

The boll weevil vitellogenin gene: nucleotide sequence, structure, and evolutionary relationship to nematode and vertebrate vitellogenin genes.

Boll weevil (Anthonomus grandis) eggs contain two yolk proteins, YP47 and YP160. Using anti-YP160 antiserum as probe, a partial-length complementary DNA (cDNA) was isolated from a lambda gt11 adult female cDNA library. A second partial-length cDNA was isolated from a lambda gt10 adult female cDNA library by differential screening with male vs. female cDNAs. Northern blot analysis showed that each cloned cDNA hybridized to a 6-kb female-specific transcript. These cDNAs were used to probe a genomic library, and two overlapping genomic clones were obtained that span the boll weevil vitellogenin gene. The entire transcription unit was sequenced, and introns were mapped by a combination of primer extension experiments, S1 nuclease protection experiments, and polymerase chain reaction-mediated synthesis of two additional cDNA clones. Based on these data, the vitellogenin mRNA is 5511 nucleotides [plus a poly(A) tail of undetermined length] and specifies a provitellogenin of 1790 amino acids. The deduced protein has a Glu+Gln content of 16.3%, which is a relatively high value that is typical of most vitellogenins. Protein sequence similarities including Cys clusters conserved between boll weevil vitellogenin and Xenopus laevis A2 or Caenorhabditis elegans vit-5 vitellogenins indicated that the boll weevil protein is a member of the ancient nematode-vertebrate vitellogenin family. Moreover, the six introns in the boll weevil vitellogenin gene interrupt the coding region at positions closely or exactly corresponding to a subset of the positions of the 34 vertebrate vitellogenin introns, further supporting the argument for a common evolutionary relationship. This report represents the first complete nucleotide sequence and structural analysis of a nondipteran insect vitellogenin gene.

Amino Acid Sequence↗

Replication of Chilo iridescent virus in the cotton boll weevil, Anthonomus grandis, and development of an infectivity assay.

The boll weevil, Anthonomus grandis Boheman, is a devastating pest of cotton. Chemical pesticides are problematic due to relative lack of target specificity and resistance. Microbial pesticides may provide viable alternatives because of their narrow host range. Chilo iridescent virus (CIV) is the type species for genus Iridovirus, family Iridoviridae: large, icosahedral cytoplasmic viruses containing a double-stranded DNA genome. Earlier work suggested that CIV replicated in the boll weevil; however, efficiency or production of infectious virus was not established. We showed that CIV undergoes a productive cycle in A. grandis. CIV DNA levels in boll weevil pupae increased significantly from 0 to 3 days post infection. Moreover, virogenic stromata and complete virus particles were observed in the cytoplasm by 7 days. An endpoint dilution assay using viral DNA replication as indicator suggested a 10(5)-fold increase in infectious virus titer over 7 days. This is the first such demonstration in larval infections with genus Iridovirus. Our study establishes that CIV undergoes a productive cycle in the boll weevil and provides an important and useful model system for replication at the organismal level. These results have important implications for the potential of CIV and its components in boll weevil control.

Animals↗

Influence of cotton fruit stages as food sources on boll weevil (Coleoptera: Curculionidae) fecundity and oviposition.

Boll weevil, Anthonomus grandis grandis Boheman (Coloeoptera: Curculionidae), populations in the Lower Rio Grande Valley of Texas increase strongly during the squaring stage, and preemptive insecticide sprays at the pinhead square stage were designed to capitalize on that association. Laboratory assays showed that cotton plant volatiles, or leaves as a food source, do not elicit egg production in wild weevils. Boll weevils fed on large (5.5-8-mm diameter) squares for 7 d resulted in > or = 3.8-fold more gravid females that developed 4.8-fold more chorionated eggs per female than weevils fed on match-head-sized squares, or postbloom, young (5-10-d-old), or old (3-5-wk-old) bolls. When presented with a choice, nongravid females preferred to feed on young and old bolls 4.7- and 8.4-fold more, respectively, than on large squares. In the field, large squares had 7.8- and 25-fold more feeding punctures than match-head-sized squares and bolls, respectively. Oviposition increased > or = 2.7-fold when females fed on large squares compared with match-head-sized squares and bolls. Greater feeding on large squares during squaring and the associated greater fecundity explain rapid weevil population buildups shortly after large squares become well established.

Animals↗

Changes in volatile terpene and diterpene resin acid composition of resistant and susceptible white spruce leaders exposed to simulated white pine weevil damage.

Induced (traumatic) resin in white spruce (Picea glauca (Moench) Voss) leaders resistant or susceptible to the white pine weevil (Pissodes strobi Peck) was analyzed for volatile terpenes and diterpene resin acids after simulated white pine weevil damage. Leaders from 331 trees were wounded just below the apical bud with a 1-mm diameter drill, coinciding with the natural time of weevil oviposition in the spring. Leaders were removed in the fall, and the bark and xylem from the upper and lower regions of the leader extracted and analyzed by gas chromatography. Unwounded trees had low amounts of resin in xylem compared with bark. In response to wounding, volatile terpenes and diterpene resin acids increased in the upper xylem (area of wounding), with resistant trees showing a greater increase than susceptible trees. Wounding caused monoterpenes in particular to decrease in the lower region of the leader (away from the drilled area) in greater amounts in susceptible trees than in resistant trees. In response to wounding, the proportion of monoterpene to resin acid increased in the upper and lower xylem of resistant trees, and slightly increased in the upper xylem of susceptible trees. Monoterpene-enriched resin is more fluid than constitutive resin, and probably flows more readily into oviposition cavities and larval mines, where it may kill immature weevils. Loss of resin components in the lower xylem suggested catabolism and transport of these materials to the site of wounding; however, energetic and regulatory data are necessary to confirm this hypothesis. This study provides a basis for measuring the ability of a tree to undergo traumatic resinosis that could be used to screen for resistance to white pine weevil.

Animals↗

Effect of nitrogen on resistance of sweet potato to sweetpotato weevil (Coleoptera: Curculionidae) and on storage root chemistry.

The effects of nitrogen fertilizer on sweet potato, Ipomoea batatas (L.) Poir., resistance to the sweetpotato weevil, Cylas formicarius elegantulus (Summers), was studied. Adult weevil feeding and oviposition preference, larval survival, and pupal weight were used as measures of sweet potato resistance. Sweet potato resin glycosides and caffeic acid concentrations in the periderm tissue of storage roots also were measured. Sweet potato genotypes (Beauregard, Excel, W-244, W-250, and Sumor) with varying levels of resistance to sweetpotato weevil were grown in the field under three nitrogen regimes (0, 45, and 135 kg N/ha). Harvested storage roots were evaluated in the laboratory for feeding and oviposition activity of sweetpotato weevil female adults under no-choice and choice test conditions. Larval survival rate and pupal weight were determined by rearing the insects individually on storage root sections. Nitrogen level had a significant effect on the number of eggs deposited, but not on the number of feeding punctures. Sweetpotato weevils laid fewer eggs on plants with the highest level of nitrogen. Nitrogen levels did not significantly affect larval survival and pupal weight. Genotype had a significant effect on feeding, oviposition, and larval survival. Beauregard had higher levels of feeding, oviposition, and larval survival compared with the other genotypes. No interaction effects between nitrogen and genotype were significant. Resin glycosides and caffeic acid concentrations were significantly different among genotypes and between years. Nitrogen levels significantly affected the concentrations of caffeic acid in 1997.

Animals↗

Effects of kaolin-based particle film application on boll weevil (Coleoptera: Curculionidae) injury to cotton.

This study examined a non-insecticidal tactic for suppressing boll weevil, Anthonomus grandis grandis Boheman, damage to cotton, Gossypium hirsutum L. In cage assays, kaolin, a reflective white mineral, applied to excised cotton squares or to the cotton foliage, initially resulted in lower levels of boll weevil injury to squares than nontreated squares. Boll weevil oviposition and feeding on kaolin-treated squares and squares on kaolin-treated cotton plants increased when nontreated squares and cotton plants were in short supply. A laboratory assay and field trials suggested that boll weevils distinguished between cotton plots based on color differences caused by kaolin and this appeared to influence levels of damage to squares. Random sampling in small plots indicated that oviposition damage to squares in plots treated with kaolin was reduced (P < 0.05) compared with nontreated controls, except when rain washed the kaolin off the foliage. Lint yield differences were not detected between the small plots, but the kaolin-treated small plots yielded as much as 2.36 times more cotton lint than a large but unreplicated adjacent nontreated control plot, and up to 1.39 times more than another large but unreplicated adjacent plot sprayed twice with preemptive applications of azinphosmethyl when cotton squares were first developing (pinhead stage). Potentially important avenues for future research on boll weevil injury suppression using kaolin are discussed.

Animals↗

Deterrent effects of four neem-based formulations on gravid female boll weevil (Coleoptera: Curculionidae) feeding and oviposition on cotton squares.

Three commercial neem-based insecticides, Agroneem, Ecozin, and Neemix, and a neem seed extract formulation, bitters, containing 1,036, 16,506, 471, and 223 microg/ml azadirachtin, respectively, were assessed for feeding and oviposition deterrence against gravid female boll weevils, Anthonomus grandis grandis Boheman, in the laboratory. In choice assays, excised cotton squares dipped in the separate formulations were first physically contacted by the weevils' tarsi or antennae fewer times than nontreated control squares. In choice and no-choice assays, each formulation repelled the weevils for > or = 90 min. After 24 h in the choice assays, feeding punctures on the squares treated with Agroneem, Ecozin, or bitters were significantly fewer compared with controls. Egg punctures on the Ecozin- and the bitters-treated squares were significantly fewer than on control squares after 24 h. In the no-choice assay, no significant difference was detected. Aging the formulations under outdoor conditions for 24 h before weevils were exposed resulted in 46-60% and 62-82% reductions in feeding and oviposition punctures, respectively, compared with controls. Agroneem- and bitters-treated squares had > 37% fewer feeding punctures after being aged for 48 h. No significant difference was detected after 72 h of aging. Because the deterrence of the gravid female boll weevils was not correlated with amounts of azadirachtin, azadirachtin does not seem to be the only, or the most influential, component of neem that induced the observed deterrence.

Animals↗

IgE-mediated inhalant allergy in inhabitants of a building infested by the rice weevil (Sitophilus oryzae).

The rice weevil (Sitophilus oryzae) is known as a stored product pest. Occupants living in a factory used previously as a granary and known to be contaminated by the rice weevil developed rhinitis and asthma. To study the role of the potential insect allergens, extracts were prepared from whole body and a grain dust of the rice and grain weevil (Sitophilus granarius). The extracts were coupled to cyanogen bromide-activated paper disks. Skin prick tests (SPT) were performed using common inhalant allergens as well as the insect allergen preparations. In cases of a positive SPT, specific serum IgE to the insect products was measured. Histamine release studies with peripheral leukocytes (HR) and studies using immunoblot techniques were performed. Fifteen of 39 subjects living in the infested rooms demonstrated positive SPT to the extracts of insect origin. Histamine equivalent wheal size induced by the whole body extract of the rice weevil required 3-160 micrograms protein/mL. Five of 15 subjects (with positive SPTs) had elevated specific IgE-levels to the insect material. RAST-inhibition studies indicated cross allergenicity between rice and grain weevils, and also between the frass of both beetles. Five of 15 subjects were positive in the HR assay (Ag30, allergen concentration for 30% HR for the whole body extract: range, 10(-3) to 400 micrograms protein/mL, maximum release: 26% to 89%). IgE binding was detected after SDS-PAGE by immunoblot techniques at different locations: 35-38, 54, 67, 70, and > 94 kD.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗

Molecular genetic variation of boll weevil populations in North America estimated with microsatellites: implications for patterns of dispersal.

The boll weevil (Anthonomus grandis Boheman) is an insect pest of cotton that underwent a well-documented range expansion across the southeastern U.S. from Mexico beginning about 110 years ago. Eleven microsatellite loci were surveyed to infer the magnitude and pattern of genetic differentiation among boll weevil populations from 18 locations across eight U.S. states and northeast Mexico. Estimates of genetic diversity (allelic diversity and heterozygosity) were greater in Southern than Northern populations, and were greater in the west than the east among Northern populations. Boll weevil populations were genetically structured as a whole across the geographic range sampled, with a global F (ST) of 0.241. South-central populations exhibit classic isolation by distance, but evidence suggests that populations within the Eastern and Western regions have not yet reached genetic equilibrium. Gene flow appears to be relatively high among populations within the Eastern region. Population assignment data and estimates of gene flow indicate that migration between locations separated by < 300 km is frequent. The database of microsatellite genotypes generated in this study now makes it possible, through population assignment techniques, to identify the most likely geographic source of a boll weevil reintroduced to an eradication zone, which will help action agencies decide the most appropriate mitigation response.

Animal Migration↗

Toxicity to cotton boll weevil Anthonomus grandis of a trypsin inhibitor from chickpea seeds.

Cotton (Gossypium hirsutum L.) is an important agricultural commodity, which is attacked by several pests such as the cotton boll weevil Anthonomus grandis. Adult A. grandis feed on fruits and leaf petioles, reducing drastically the crop production. The predominance of boll weevil digestive serine proteinases has motivated inhibitor screenings in order to discover new ones with the capability to reduce the digestion process. The present study describes a novel proteinase inhibitor from chickpea seeds (Cicer arietinum L.) and its effects against A. grandis. This inhibitor, named CaTI, was purified by using affinity Red-Sepharose Cl-6B chromatography, followed by reversed-phase HPLC (Vydac C18-TP). SDS-PAGE and MALDI-TOF analyses, showed a unique monomeric protein with a mass of 12,877 Da. Purified CaTI showed significant inhibitory activity against larval cotton boll weevil serine proteinases (78%) and against bovine pancreatic trypsin (73%), when analyzed by fluorimetric assays. Although the molecular mass of CaTI corresponded to alpha-amylase/trypsin bifunctional inhibitors masses, no inhibitory activity against insect and mammalian alpha-amylases was observed. In order to observe CaTI in vivo effects, an inhibitor rich fraction was added to an artificial diet at different concentrations. At 1.5% (w/w), CaTI caused severe development delay, several deformities and a mortality rate of approximately 45%. These results suggested that CaTI could be useful in the production of transgenic cotton plants with enhanced resistance toward cotton boll weevil.

Agriculture↗

Host effects of glandular-haired alfalfa on alfalfa weevil (Coleoptera: Curculionidae) and potato leafhopper (Homoptera: Cicadellidae) populations in Virginia.

Cultivars of glandular-haired alfalfa, Medicago sativa L., such as '54H69', are currently available and marketed as being resistant to potato leafhopper, Empoasca fabae (Harris). Between 2000 and 2002, studies were conducted to compare the effects of '54H69' and a standard, nonglandular-haired alfalfa cultivar, 'Choice', on alfalfa weevil, Hypera postica (Gyllenhal), and potato leafhopper populations at Campbell and Montgomery counties, Virginia. '54H69' had no effect on alfalfa weevil populations. At each location, densities of alfalfa weevil in '54H69' and 'Choice' were similar, but pest pressure was higher at Campbell Co. than at Montgomery Co. and always exceeded the economic threshold before insecticide was applied. Densities of potato leafhopper also did not differ between '54H69' and 'Choice' in any year at the two locations. Insecticide treatment effectively reduced potato leafhopper densities in the two cultivars, although populations were below the economic threshold at both locations when the insecticides were applied. Overall, postinsecticide treatment comparisons showed that the densities of alfalfa weevil and potato leafhoppers were similar or higher in untreated '54H69' compared with insecticide-treated 'Choice'. In addition, there were no differences in seasonal dry yields between '54H69' and 'Choice' in any year at either location. Our results indicate that the glandular-haired alfalfa '54H69' does not provide acceptable resistance to potato leafhopper and also does not offer a yield advantage to growers in Virginia.

Animals↗

Wound-induced terpene synthase gene expression in Sitka spruce that exhibit resistance or susceptibility to attack by the white pine weevil.

We analyzed the expression pattern of various terpene synthase (TPS) genes in response to a wounding injury applied to the apical leader of Sitka spruce (Picea sitchensis Bong. Carr.) genotypes known to be resistant (R) or susceptible (S) to white pine weevil (Pissodes strobi Peck.) attack. The purpose was to test if differences in constitutive or wound-induced TPS expression can be associated with established weevil resistance. All wounding treatments were conducted on 9-year-old R and S trees growing under natural field conditions within the range of variation for weevil R and S genotypes. Representative cDNAs of the monoterpene synthase (mono-TPS), sesquiterpene synthase (sesqui-TPS), and diterpene synthase (di-TPS) classes were isolated from Sitka spruce to assess TPS transcript levels. Based on amino acid sequence similarity, the cDNAs resemble Norway spruce (Picea abies) (-)-linalool synthase (mono-TPS; PsTPS-Linl) and levopimaradiene/abietadiene synthase (di-TPS; PsTPS-LASl), and grand fir (Abies grandis) delta-selinene synthase (sesqui-TPS; PsTPS-Sell). One other mono-TPS was functionally identified as (-)-limonene synthase (PsTPS-Lim). No significant difference in constitutive expression levels for these TPSs was detected between R and S trees. However, over a postwounding period of 16 d, only R trees exhibited significant transcript accumulation for the mono- and sesqui-TPS tested. Both R and S trees exhibited a significant accumulation of PsTPS-LASl transcripts. An assessment of traumatic resin duct formation in wounded leaders showed that both R and S trees responded by forming traumatic resin ducts; however, the magnitude of this response was significantly greater in R trees. Collectively, our data imply that the induced resinosis response is an important aspect of defense in weevil R Sitka spruce trees growing under natural conditions.

Alkyl and Aryl Transferases↗

Phylogeography and the geographic cline in the armament of a seed-predatory weevil: effects of historical events vs. natural selection from the host plant.

Japanese camellia (Camellia japonica) and its seed predator, the camellia weevil (Curculio camelliae), provide a notable example of a geographic mosaic of coevolution. In the species interaction, the offensive trait of the weevil (rostrum length) and the defensive trait of the plant (pericarp thickness) are involved in a geographically-structured arms race, and these traits and selective pressures acting on the plant defence vary greatly across a geographical landscape. To further explore the geographical structure of this interspecific interaction, we tested whether the geographical variation in the weevil rostrum over an 800-km range along latitude is attributed to local natural selection or constrained by historical (phylogeographical) events of local populations. Phylogeographical analyses of the mitochondrial DNA sequences of the camellia weevil revealed that this species has experienced differentiation into two regions, with a population bottleneck and subsequent range and/or population expansion within each region. Although these phylogeographical factors have affected the variation in rostrum length, analyses of competing factors for the geographical variation revealed that this pattern is primarily determined by the defensive trait of the host plant rather than by the effects of historical events of populations and a climatic factor (annual mean temperature). Thus, our study suggests the overwhelming strength of coevolutionary selection against the effect of historical events, which may have limited local adaptation.

Animals↗

Molecular cloning of a cysteine proteinase cDNA from the cotton boll weevil Anthonomus grandis (Coleoptera: Curculionidae).

The cotton boll weevil (Anthonomus grandis) causes severe cotton crop losses in North and South America. This report describes the presence of cysteine proteinase activity in the cotton boll weevil. Cysteine proteinase inhibitors from different sources were assayed against total A. grandis proteinases but, unexpectedly, no inhibitor tested was particularly effective. In order to screen for active inhibitors against the boll weevil, a cysteine proteinase cDNA (Agcys1) was isolated from A. grandis larvae using degenerate primers and rapid amplification of cDNA ends (RACE) techniques. Sequence analysis showed significant homologies with other insect cysteine proteinases. Northern blot analysis indicated that the mRNA encoding the proteinase was transcribed mainly in the gut of larvae. No mRNA was detected in neonatal larvae, pupae, or in the gut of the adult insect, suggesting that Agcys1 is an important cysteine proteinase for larvae digestion. The isolated gene will facilitate the search for highly active inhibitors towards boll weevil larvae that may provide a new opportunity to control this important insect pest.

Animals↗