Reproductive potential of field-collected overwintering boll weevils (Coleoptera: Curculionidae) fed on pollen in the laboratory

Abstract  The reproductive potential of overwintering boll weevil, Anthonomus grandis grandis (Boheman), females collected from pheromone traps in September, November and January, fed for 1, 3, and 5 weeks on plant pollens, and then provided cotton squares, was determined in the laboratory at 27 ± 1°C, 65% RH, and a photoperiod 13 : 11 (L : D) h. Duration of pollen feeding by overwintering boll weevils did not significantly influence egg and feeding punctures, or puncture ratios (egg to total punctures) for any of the three months of parent weevil collections when provided cotton squares on a daily basis. However, punctures and puncture ratios are significantly different when comparing mean data between months of boll weevil collections. When boll weevils were provided with cotton squares daily, the pre-ovipositional periods of female parents captured in September, November and January were 5, 9 and 14 days, respectively. The rate of eggs by females was significantly lower during November and January than September. Female parents collected in September produced a significantly higher percentage of eggs yielding adult progeny than those collected in November and January. Life table parameters indicated that net reproductive rate ( R _o) of boll weevil females collected in September was 1.2-fold higher than those collected in November and 10.7-fold higher than those collected in January. Except for testes size, no differences in male reproductive parameters were observed during the cotton-free period compared with males captured during mid-cotton (June). The number of oocytes in the ovarioles and the number of oocytes containing yolk were significantly lower during September, November and January compared with June. The reproductive potential of overwintering boll weevil females collected in different months is an important consideration in determining the success of any control strategy.     

Sublethal effects of malathion on boll weevil (Coleoptera: Curculionidae) fecundity when maintained on cotton squares or artificial diet

Mated 3‐day‐old female boll weevils, Anthonomus grandis grandis Boheman, reared from field‐infested cotton (Gossypium hirsutum L.) squares were topically treated with an estimated LD₅₀ of malathion (2 μg) to assess its effects on fecundity, oviposition, and body fat condition. Two different food sources, cotton squares and artificial diet, were assessed in malathion‐treated and nontreated (control) weevils. The LD₅₀ caused ～50% mortality in the square‐fed malathion treatment, but the artificial diet‐fed malathion‐treated weevils were less susceptible. LD₅₀ survivors fed on the squares produced ≥ 9 times more chorionated eggs in the ovaries and oviposited ≥ 19‐fold more than survivors fed artificial diet, regardless of the malathion treatment. Boll weevils that survived a 2 μg LD₅₀ malathion and also fed squares were ～4.5‐fold leaner than diet‐fed weevils. Our findings demonstrate that non‐resistant boll weevils surviving a sublethal dose of malathion will reproduce without any delay or significant loss in fecundity, and the food source for which boll weevils are maintained when conducting these assays will directly affect the results. The significance of these findings and how they are related to the final stages of eradicating the boll weevil from the US are discussed.     

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.     

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.     

Syntheses of Several Compounds Related to Helminthosporol and their Plant Growth-regulating Activities

Several compounds related to helminthosporol (I), a natural plant-growth promoter, were prepared from I and their plant-growth regulating activities were examined together with those of the derivatives reported in the previous paper, using rice and lettuce seedlings. Among the compounds tested, helminthosporic acid (III) exhibited marked elongation effect to the shoot growth of rice and lettuce seedlings, whereas the effect of I was specific to the former. Results of the biological test are reported in details.     

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.     

Plant Growth-regulating Activities of 4-Methoxybenzophenones

4-Methoxybenzophenones showed diverse plant growth-regulating actions; inhibition of shoot and root growth, induction of chlorosis, and a disturbance in phototropism or geotropism. No clear relationship was found between the inhibitory activities and Hammett’s σ of the substituents. A brief discussion of the relationship between the growth-inhibitory activity and the absorption wavelength explains the mode of action.     

Molecular modeling and inhibitory activity of cowpea cystatin against bean bruchid pests

Plant cystatins show great potential as tools to genetically engineer resistance of crop plants against pests. Two important potential targets are the bean weevils Acanthoscelides obtectus and Zabrotes subfasciatus, which display major activities of digestive cysteine proteinases in midguts. In this study a cowpea cystatin, a cysteine proteinase inhibitor found in cowpea (Vigna unguiculata) seeds, was expressed in Escherichia coli and purified with a Ni-NTA agarose column. It strongly inhibited papain and proteinases from midguts of both A. obtectus and Z. subfasciatus bruchids, as seen by in vitro assays. When the protein was incorporated into artificial seeds at concentrations as low as 0.025%, and seeds were consumed by the bruchids larva, dramatic reductions in larval weight, and increases in insect mortality were observed. Molecular modeling studies of cowpea cystatin in complex with papain revealed that five N-terminal residues responsible for a large proportion of the hydrophobic interactions involved in the stabilization of the enzyme-inhibitor complex are absent in the partial N-terminal amino acid sequencing of soybean cystatin. We suggest that this structural difference could be the reason for the much higher effectiveness of cowpea cystatin when compared to that previously tested phytocystatin. The application of this knowledge in plant protein mutation programs aiming at enhancement of plant defenses to pests is discussed.     

Physiological Effects of Nondialyzable Melanoidin in Rats

The absorption and excretion of melanoidins, a mixture of the low- and high-molecular weight components (LMM and HMM), and the LMM component prepared from a l-lysine^d-glucose system, at pH 7.4 and 9.0, respectively, and the effects of these melanoidins on cholesterol metabolism were examined in rats. Weanling male rats of the Wistar strain weighing about 50 g were fed diets containing 10% casein (10C) with 3% of a melanoidin or 25% casein (25C) with 4% of the melanoidin for 12 weeks, and the 25C diet including 3% of the melanoidin or LMM for 8 weeks. The growth and organ weights of the melanoidin-supplied groups were not different from those of the control ones. In rats given the melanoidin diets with both protein levels, the kidneys became dark brownish due to the accumulation of the melanoidin, though the accumulated amount was extremely small (nearly 0.5 mg/g wet kidney), and gel filtration chromatography of a water extract of the kidneys on Sephadex G-75 showed that the deposited melanoidin was the LMM component. Most of the ingested melanoidin, however, was excreted in the feces, and on comparison of the gel chromatographic patterns, the melanoidin groups were found to have more fecal LMM than the control ones. When rats were provided with LMM, HMM increased in their feces. The addition of melanoidin suppressed the level of plasma total cholesterol and elevated the fecal excretion of total lipids and total cholesterol. The urinary contents of protein and total creatinine did not differ from those in the control groups.     

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.     

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.     

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.     

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.     

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.     

Production of Ribotides of 4-Hydroxy- and 4-Aminopyrazolo[3, 4-d]pyrimidine by Brevibacterium ammoniagenes

Brevibacterium ammoniagenes ATCC 6872 was previously reported to accumulate large amounts of IMP, AMP, ADP, ATP, GMP, GDP and GTP from the corresponding purine bases. The organism was also reported to convert various derivatives of purine and 8-azapurine to the corresponding ribotides.

Using the similar process, ribotidation of pyrazolo[3, 4-d]pyrimidines was attempted, and it was found that the same organism was able to produce remarkable amounts of 4-hydroxy-1-β-d-ribofuranosylpyrazolo[3, 4-d]pyrimidine 5′-monophosphate (HPP-RP) from 4-hydroxypyrazolo[3, 4-d]pyrimidine (HPP, allopurinol) and 4-amino-1-β-d-ribofuranosylpyrazolo[3, 4-d]pyrimidine 5′-monophosphate and 5′-diphosphate from 4-amino-pyrazolo[3, 4-d]pyrimidine.

The crystals of HPP-RP (Na-salt) were isolated from the cultured broth of Br. ammoniagenes incubated with HPP, and characterized based on UV-spectra, IR-spectrum, NMR and others.

It was also found that HPP-RP was converted to the corresponding riboside by hydrolysis in aqueous solution (pH 4.0 ~ 9.0) for 6 hr at 140°C. The hydrolysis of HPP-RP was also accomplished with various organisms.     

Isolation and characterization of farnesyl diphosphate synthase from the cotton boll weevil,Anthonomus grandis

Farnesyl diphosphate synthase (FPPS) catalyzes the consecutive condensation of two molecules of isopentenyl diphosphate with dimethylallyl diphosphate to form farnesyl diphosphate (FPP). In insects, FPP is used for the synthesis of ubiquinones, dolicols, protein prenyl groups, and juvenile hormone. A full‐length cDNA of FPPS was cloned from the cotton boll weevil, Anthonomus grandis (AgFPPS). AgFPPS cDNA consists of 1,835 nucleotides and encodes a protein of 438 amino acids. The deduced amino acid sequence has high similarity to previously isolated insect FPPSs and other known FPPSs. Recombinant AgFPPS expressed in E. coli converted labeled isopentenyl diphosphate in the presence of dimethylallyl diphosphate to FPP. Southern blot analysis indicated the presence of a single copy gene. Using molecular modeling, the three‐dimensional structure of coleopteran FPPS was determined and compared to the X‐ray crystal structure of avian FPPS. The α‐helical fold is conserved in AgFPPS and the size of the active site cavity is consistent with the enzyme being a FPPS. © 2009 Wiley Periodicals, Inc.     

Inhibitory Activities of 3-Thiophenecarboxylic Acid and Related Compounds on Plant Growth

3-Thiophenecarboxylic acid (1) showed strong growth-inhibitory activity toward the following plants but not Glycine max Merrill; Brassica campestris subsp. rapa Hook. fil. et Anders, Sesamum indicum L., Lactuca sativa L. var. longifolia Lam, Echinochloa utilis Ohwi et Yabuno and Allium tuberosum Rottler. Compound 1 strongly inhibited the growth of roots of S. indicum and L. sativa even at the low concentration of 5.0 × 10^?5 m. The growth-inhibitory activity of 1-related compounds (2–6) on S. indicum was also studied. Among the compounds, 3-thiopheneacetic acid (6) showed the strongest inhibitory activity, but 3-thiophenecarboxaldehyde (2), 3-thiophenemethanol (3), and 3-thiophenecarboxamide (5) showed no activity. The radicles of plants treated with these active compounds showed negative geotropism.     

植物蛋白酶抑制剂基因结构、调控及其控制害虫的策略

各种不同类型的植物蛋白酶抑制剂基因已被分离,它们的特异产物(单基因或多基因组合),对昆虫体内各种生化和生理过程会产生不同程度的影响,在对昆虫和病原体防御体系中起重要作用。多种蛋白酶抑制剂重组,协同保护植物的方法,已成为害虫综合防治计划的一部分。尽管它们近期内尚不能代替化学杀虫剂,但可作为有效的替补。目前,大多数抑制剂的作用和机理正在详尽地研究中,该文综述了植物蛋白酶抑制剂的基因结构、调控与表达并讨论了培育转基因作物控制害虫的策略。