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.     

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.     

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.     

Regulation of vitellogenin synthesis and uptake in the boll weevil, Anthonomus grandis

Abstract. Hormonal factors influencing reproductive development were examined in adult boll weevils, Anthonomus grandis (Boheman) (Coleoptera: Curculionidae). Long-day, high-temperature rearing conditions promote reproduction whereas short-day, low-temperature conditions do not. Implants of corpora allata (CA), brains, or brains plus retrocerebral complexes taken from long-day donors, or hormone analogue treatments were used to examine onset of vitellogenin synthesis and uptake in decapitated bodies of adult weevils reared in short-day, low-temperature conditions. Weevils decapitated within 2 days after eclosion and reared in short-day, low-temperature conditions never initiated vitellogenin production or ovarian development. Females and males decapitated on day 2 showed haemolymph vitellogenin within 5 days following treatment with Juvenile Hormone (JH) analogue or implantation of CA, but not after implantation of brain alone or implantation of muscle (sham). Uptake of vitellogenin into the oocytes did not occur unless both JH analogue and brain were given as replacement therapy. These experiments indicated that JH is necessary and sufficient to stimulate vitellogenin synthesis in this species but that a brain factor must be present for vitellogenin uptake.     

Rates of Juvenile Hormone biosynthesis and degradation during reproductive development and diapause in the boll weevil, Anthonomus grandis

Abstract. The role of Juvenile Hormone (JH) during reproductive development and diapause was investigated in the boll weevil, Anthonomus grandis Boheman (Coleoptera: Curculionidae). JH sythesized by corpora allata (CA) in vitro of A.grandis was identified as JH-UJ by high-performance liquid chromatography and by conversion to the methoxyhydrin. Optimal conditions for the use of a short-term assay in vitro were established to examine profiles of CA activity. In addition, rates of JH degradation by JH-specific esterase were determined. Patterns of CA and JH-esterase activity during reproductive development and the diapause state were established with laboratory-reared reproductive weevils and diapausing weevils collected as larvae and pupae in the field after the cotton-growing season. The results indicate that JH production is elevated in reproductive females whereas males and winter field-collected females show no CA activity. Vitellogenin concentrations in haemolymph and rates of oviposition were studied in relation to CA activity and JH degradation. An attempt to induce diapause in the laboratory failed.     

Forensic pollen geolocation techniques used to identify the origin of boll weevil re-infestation

The boll weevil, Anthonomus grandis, entered the United States of America in the early twentieth century and became a major pest in cotton, Gossypium spp. Shortly after the passage of Tropical Storm Erin on 16 August 2007 through the South Texas/Winter Garden boll weevil eradication zone, over 150 boll weevils were captured in the Southern Rolling Plains (SRP) eradication zone that was essentially weevil-free since 2003. Pollen analyses were made of the SRP weevils and weevils collected in two suspected source zones, Cameron (Southern Blacklands eradication zone) and Uvalde (Winter Garden eradication zone). An additional examination of the palynological evidence and examination of additional pollen residue shed new light on this event and strengthens the conclusion that the Uvalde area was the source of the SRP weevils. A total of 192 pollen grains from 39 taxa were found in the SRP weevils: 1904 pollen grains from ten taxa from the Cameron weevils and 148 grains from 28 taxa in the Uvalde weevils. The SRP weevils shared 16 taxa, including Phermeranthus sp. (flameflower) with the Uvalde weevils and only five taxa with the Cameron weevils. Common taxa between SRP and Uvalde weevils and the lack of the dominant ‘low spine’ Asteraceae that occurred in all Cameron samples confirm that the SRP weevils originated from the South Texas/Winter Garden zone. Problems associated with this type of research are similar to those in forensic palynology. These problems include the unknown origin of the weevils, pollen contamination and care and storage of the samples.     

Functions of the spermathecal muscle of the boll weevil,Anthonomus grandis

In female boll weevils, Anthonomus grandis, spermathecal filling was not affected by severing the spermathecal muscles. Females whose spermathecal muscles were severed 2 to 4 weeks after mating laid infertile eggs and resumed virginal ovipositional behaviour indicating the importance of this muscle in supplying sperm for egg fertilization. The presence of normal active sperm within the spermatheca in no way influenced ovipositional behaviour. In females whose spermathecal muscles had been severed, 22 per cent sperm displacement occurred after a second mating compared with 66 per cent for normal females. The physical displacement of sperm was thus largely dependent on a functional spermathecal muscle.     

Proteolytic processing of the vitellogenin precursor in the boll weevil,Anthonomus grandis

The soluble proteins of the eggs of the coleopteran insect Anthonomus grandis Boheman, the cotton boll weevil, consist almost entirely of two vitellin types with M_rs of 160,000 and 47,000. We sequenced their N-terminal ends and one internal cyanogen bromide fragment of the large vitellin and compared these sequences with the deduced amino acid sequence from the vitellogenin gene. The results suggest that both the boll weevil vitellin proteins are products of the proteolytic cleavage of a single precursor protein. The smaller 47,000 M vitellin protein is derived from the N-terminal portion of the precursor adjacent to an 18 amino acid signal peptide. The cleavage site between the large and small vitellins at amino acid 362 is adjacent to a pentapeptide sequence containing two pairs of arginine residues. Comparison of the boll weevil sequences with limited known sequences from the single 180,000 M_r honey bee protein show that the honey bee vitellin N-terminal exhibits sequence homology to the N-terminal of the 47,000 M_r boll weevil vitellin. Treatment of the vitellins with an N-glycosidase results in a decrease in molecular weight of both proteins, from 47,000 to 39,000 and from 160,000 to 145,000, indicating that about 10–15% of the molecular weight of each vitellin consists of N-linked carbohydrate. The molecular weight of the deglycosylated large vitellin is smaller than that predicted from the gene sequence, indicating possible further proteolytic processing at the C-terminal of that protein. © 1993 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

Endocrine regulation of reproduction and diapause in the boll weevil,Anthonomus grandis Boheman

A review of the literature for the hormonal control of reproduction and diapause in Coleoptera is presented. The role of juvenile hormone and juvenile hormone esterase (JHE) in the control of the different life history strategies of the boll weevil are examined. Elevated levels of hemolymph JHE were found to be positively correlated with survival throughout the winter in South Texas population of weevils. Winter weevils were examined for hemolymph vitellogenin (Vg) and their subsequent survival was monitored. The majority of weevils surviving beyond ten weeks had no hemolymph Vg. We conclude that elevated hemolymph JHE and the absence of Vg are good predictors of survivors in the South Texas population. The hormonal basis of diapause termination was examined using hormone treatment and implant therapy. We were unsuccessful in our attempt to induce post-diapause reproductive development with all of our treatments. Only weevils given access to a food source were capable of reproductive development. Our recent experimental findings in the control of Vg synthesis and uptake in the boll weevil are reviewed. Arch. Insect Biochem. Physiol. 35:455–477, 1997. © 1997 Wiley-Liss, Inc.     

Molecular cloning of alpha-amylases from cotton boll weevil,Anthonomus grandis and structural relations to plant inhibitors: an approach to insect resistance

Anthonomus grandis, the cotton boll weevil, causes severe cotton crop losses in North and South America. Here we demonstrate the presence of starch in the cotton pollen grains and young ovules that are the main A. grandis food source. We further demonstrate the presence of -amylase activity, an essential enzyme of carbohydrate metabolism for many crop pests, in A. grandis midgut. Two -amylase cDNAs from A. grandis larvae were isolated using RT-PCR followed by 5 and 3 RACE techniques. These encode proteins with predicted molecular masses of 50.8 and 52.7 kDa, respectively, which share 58% amino acid identity. Expression of both genes is induced upon feeding and concentrated in the midgut of adult insects. Several -amylase inhibitors from plants were assayed against A. grandis -amylases but, unexpectedly, only the BIII inhibitor from rye kernels proved highly effective, with inhibitors generally active against other insect amylases lacking effect. Structural modeling of Amylag1 and Amylag2 showed that different factors seem to be responsible for the lack of effect of 0.19 and -AI1 inhibitors on A. grandis -amylase activity. This work suggests that genetic engineering of cotton to express -amylase inhibitors may offer a novel route to A. grandis resistance.     

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.     

Oviposition behaviour of Bracon mellitor, a parasitoid of the boll weevil (Anthonomus grandis)

ABSTRACT. The parasitoid, Bracon mellitor , will probe with its ovipositor plant structures containing its host. The identification of methyl parahydroxy-benzoate (methyl parasept), an antimicrobial additive to the semi-synthetic diet used for rearing the host, as a releaser of this behaviour prompted an investigation of the parasitoid's response to this chemical. Its absence from field hosts suggested the response was learned. Female parasitoids reared in host-infested cotton plant buds from the fields or in methyl parasept-free hosts did not respond to methyl parasept alone until exposed to hosts contaminated with it. This is taken to imply associative learning by the females reared from the methyl parasept-free hosts. A few responses to methyl parasept were elicited from inexperienced females reared from methyl parasept-contaminated hosts, but their responsiveness increased after they had been exposed to methyl parasept-contaminated hosts as oviposition stimuli. These results point to the caution that must be exercised in the isolation and identification of kairomones.     

Esterases of laboratory-reared and field-collected cotton boll weevils,Anthonomus grandis Boh.: Polymorphism of adult esterases and formal genetics of esterase II

The esterases of the cotton boll weevil were separated by polyacrylamide gel electrophoresis into four major regions. These were named Est I–IV in order of migration from anode to origin. Polymorphism was observed in all regions. The Est II region was shown to consist of no more than two bands (fast and slow). The inheritance of the fast and slow bands of Est II was demonstrated to be controlled by codominant autosomal alleles. Analysis of the gene frequency of the Est II region showed that one field population was consistent with the Hardy-Weinberg law (P=0.995), while a second field population was not at equilibrium (P<0.001).This work was supported in part by a Faculty Research Grant from Memphis State University.     

Survival of boll weevil （Coleoptera： Curculionidae） adults after feeding on pollens from various sources

The survival of overwintering boll weevil, Anthonomus grandis grandis （Boheman）, adults on non-cotton hosts in the Lower Rio Grande Valley （LRGV） of Texas was examined from 2001 to 2006. The success of the Boll Weevil Eradication Program, which was reintroduced into the LRGV in 2005, depends on controlling overwintering boll weevil populations. Laboratory studies were conducted using boll weevil adults that were captured in pheromone traps from September through March. The number of adults captured per trap declined significantly in the field from fall to the beginning of spring （3.5-7.0-fold）. The proportion of trapped males and females did not differ significantly. The mean weight of boll weevil adults captured in September was 13.3 mg, while those of captured adults from November to February were significantly lower and ranged from 6.7 to 7.8 mg. Our results show that boll weevil adults can feed on different plant pollens. The highest longevity occurred when adults were fed almond pollen or mixed pollens （72.6 days and 69.2 days, respectively） and the lowest when they fed on citrus pollen or a non-food source （9.7 days or 7.4 days, respectively）. The highest adult survival occurred on almond and mixed pollens [88.0%-97. 6% after 1st feeding period （10 days）, 78.0%-90.8% after 3rd feeding period （10 days）, 55. 0%-83.6% after 5th feeding period （10 days）, and 15.2%-32.4% after lOth feeding period （10 days）]. The lowest adult survival occurred on citrus pollen [52.0%-56.0% after 1st feeding period （10 days）, 13.3% after 3rd and 5th feeding periods （10 days）, and 0 after 6th feeding period （10 days）]. Pollen feeding is not a behavior restricted to adult boll weevils of a specific sex or physiological state. Understanding how boll weevil adults survive in the absence of cotton is important to ensure ultimate success of eradicating this pest in the subtropics.     

Factors contributing to resistance and susceptibility of cotton and other hosts to the boll weevil, Anthonomus grandis

A large portion of the world's germ plasm of cotton has been screened at the USDA Boll Weevil Research Laboratory, State College, Mississippi during the period 1962–68 utilizing developed techniques.An oviposition suppression factor causing 25–40% reduction in number of eggs laid by the weevil has been found in Gossypium barbadense and successfully moved into upland cotton, G. hirsutum.Two years field data with frego cottons indicate that the frego character contributes a significant degree of non-preference for egg laying by the boll weevil under field conditions. The frego character is currently being combined with selected lines of G. hirsutum carrying the oviposition suppression factor.A seedling screening technique utilizing field cages and greenhouse sand flats has been shown to have significant value in screening for individual resistant plants within a commercial variety. A number of selections with 25% or more resistance to oviposition are under extensive investigation.A number of biologically active materials have been detected in the cotton plant and other hosts of the boll weevil which significantly influences feeding stimulation, feeding deterrency, repellency and attractancy of the weevil to its hosts.A highly active square abscission factor has been detected and extracted successfully from 2nd and 3rd instar larvae. The material injected into cotton squares causes 95% abscission of squares within 48–60 hours.

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Zusammenfassung Ein großer Teil des Weltsortiments der Baumwolle wurde in den Jahren 1962–68 im USA-Baumwollkäfer-Institut der Staatshochschule von Mississippi mit modernen Methoden getestet.Ein die Eiablage unterdrückender Faktor, der die Anzahl der abgelegten Eier um 25–40% herabsetzt, wurde bei Gossypium barbadense gefunden und erfolgreich in Hochland-Baumwolle, G. hirsutum, eingekreuzt.Zweijährige Erfahrungen mit frego-Baumwolle weisen darauf hin, daß das frego-Merkmal unter Feldbedingungen in einem signifikanten Ausmaße zur Nonpräferenz für die eierablegenden Käfer beiträgt. Das frego-Merkmal wird laufend mit selektierten Linien von G. hirsutum kombiniert, die den Faktor für Eiablage-Unterdrückung tragen.Eine Sämlings-Prüfungstechnik, die Feldkäfige und Gewächshaus-Sandbetee benutzt, hatte nachweislich signifikanten Wert für die Auslese resistenter Einzelpflanzen innerhalb einer Handelssorte. Eine Anzahl Selektionslinien mit 25 oder mehr Prozent Eiablage-Resistenz wird intensiv geprüft.Eine Anzahl biologisch aktiven Materials wurde bei Baumwolle und anderen Wirtspflanzen endeckt, das Fraßstimulation, Fraßabschreckung, Repellenz und Attraktanz der Käfer in bezug auf ihre Wirte signifikant beeinflußt.Ein hochaktiver Kapsel-Abwurf-Faktor wurde entdeckt und erfolgreich aus Larven des 2. und 3. Stadiums extrahiert. In Baumwollkapseln injiziert, verursachte dieses Material innerhalb 48–60 Stunden Abwurf von 95% der Kapseln.

     

Phagocytic and humoral immunity of the adult cotton boll weevil, Anthonomus grandis (Coleoptera: Curculionidae), to Serratia marcescens

This investigation determined phagocytic, lysozymal, and bactericidal defensive responses of adult laboratory-reared cotton boll weevils, Anthonomus grandis. Phagocytosis was first demonstrated in boll weevils at 3 hr following injection of live Serratia marcescens. Maximum phagocytosis was found in 16.4% of plasmatocytes at the end of 16 hr postinjection. Lysozyme activity was demonstrated in both inoculated and uninoculated boll weevils. Peak lysozyme activity of 6.9 μg/ml was found at 48 hr following inoculation of heat-killed Serratia marcescens. Bactericidal activity was demonstrated in inoculated boll weevils but not in uninoculated boll weevils. Peak bactericidal activity occurred at 24 hr following inoculation of heat-killed Serratia marcescens. Lysozymal and bactericidal activities were shown to be separate functions.     

Biosynthesis of grandlure,the pheromone of the boll weevil,Anthonomus grandis,from acetate,mevalonate, and glucose

The steam-distilled faeces of adult male boll weevils, Anthonomus grandis, that had been injected with acetate-1¹⁴C, acetate-2¹⁴C, mevalonic acid-2¹⁴C, or glucose¹⁴C(U) showed by column and gas chromatography that approximately 0·02 per cent of the administered radioactivity was incorporated into the volatile fraction. Also, 4 components of the pheromone comprised 57 to 80 per cent of the radioactivity of the volatiles but only 39 per cent of the total content of volatiles. Thus, evidence was obtained for de novo synthesis of the components. Although the boll weevil is essentially an obligate insect of cotton, this insect does not appear to require any specific component in cotton for biosynthesis of the pheromone.     

Cholesterol oxidase: an oöstatic and larvicidal agent active against the cotton boll weevil,Anthonomus grandis

The enzyme cholesterol oxidase (E.C. 1.1.3.6), purified from Streptomyces culture filtrate was previously found to have oral insecticidal activity on neonate larvae of the boll weevil (Anthonomus grandis grandis Boheman) from a laboratory population. In the present study, second instar larvae were also controlled by the enzyme at diet concentrations similar to those which control neonates (12 day LC₅₀ = 2.4 μg.ml^?1 in diet). Larvae from field-collected adults were similarly susceptible to cholesterol oxidase in the diet. When ingested by adult females during the mating/pre-oviposition period, cholesterol oxidase greatly reduced subsequent oviposition (83% reduction in eggs laid as compared to controls) and larval survival (97% reduction from controls). Dissection of treated adult females revealed poorly developed ovaries and few developing oöcytes. These studies were conducted to further evaluate the utility of cholesterol oxidase in a program to establish boll weevil-resistant transgenic cotton.