Bionomics and polymorphism in Callosobruchus subinnotatus (Coleoptera: Bruchidae)

Callosobruchus subinnotatus (Pic) is the major insect pest of stored bambara groundnuts, Vigna subterranea (L.) Verdcourt, in sub-Saharan West Africa, but little is currently known about its biology or how it may be controlled. A series of laboratory studies was performed to investigate the bionomics of and differences between two apparently different morphs of adult of each sex of this species, here termed 'active' and 'normal'. Major differences in their morphology, physiology and behaviour were identified and are described in detail for the first time. They provide clear evidence of the existence of an adult polymorphism among populations of this species, which is comparable in certain respects to that previously described for C. maculatus (Fabricius) and C. chinensis Linnaeus. Adults can be separated into the correct morph based on characteristic differences in elytral and pygidial colour and pattern. 'Normal' adults are characterized by having high fecundity, short adult life and are relatively sedentary while 'active' adults exhibit reproductive diapause (suspension of reproductive activity), are long lived, and show (at least in females) increased dispersal tendencies. These characteristics suggest adaptation of the 'active' and 'normal' morphs respectively to the different environments of field and seed stores, and the significance of the polymorphism in the life history of C. subinnotatus is discussed in this context. The design of any effective control regime for this bruchid needs to take account of and could potentially be based upon the existence of polymorphism in C. subinnotatus.     

Sources of semiochemicals mediating host finding in Callosobruchus chinensis (Coleoptera: Bruchidae)

Bruchid pests such as Callosobruchus chinensis (Linnaeus) endanger stored legume seeds throughout the tropical belt. The chemical composition of the headspace volatiles from healthy and fourth instar larvae-infested cowpea seeds were identified, characterized, and compared using gas chromatography-mass spectrometry. Y-tube olfactometer bioassays were performed to evaluate the effect of these chemicals on the orientation of conspecific adult females. Analysis of volatiles released from healthy and infested seeds revealed qualitative differences for three out of the 17 compounds identified. Dimethyl disulphide, isobutenyl methyl ketone and methyl trisulphide were found only in the blend emitted from infested but not from healthy seeds. Quantitative differences were apparent for tridecane which was released in larger amounts from infested seeds. While volatiles collected from healthy seeds were attractive to female bruchids, volatiles collected from infested seeds were repellent. To test the hypothesis that the qualitative differences in the chemical composition found may be due to insect-derived components, the volatiles from frass and fourth instar larvae combined were analysed. These volatiles contained both of the sulphides emitted from infested seed but not from healthy seeds. Although a limited induction of volatiles from cowpea seeds cannot be excluded, it is postulated that behavioural differences of the female weevils are largely due to insect-derived semiochemicals. The potential use of such semiochemicals as part of an integrated pest management strategy is discussed.     

Efficacy of natural products against Callosobruchus chinensis (Coleoptera: Bruchidae) in Nepal

This study was conducted to determine the efficacy of local natural products against the beetle, Callosobruchus chinensis L. (Coleoptera: Bruchidae), in stored chickpea Cicer arietinum L. (Fabaceae) in Chitwan, Nepal. Five natural products and one synthetic pesticide (Malathion) and two storage regimes (aluminum sheet bin vs. jute bags with plastic lining) were tested for their effect on stored pulse with respect to damage by C. chinensis. The five natural products included Xanthoylum armatum DC (Rutaceae) fruit powder, Acorus calamus L. (Araceae) rhizome powder, Cinnamom camphora L. (Lauraceae) balls, oil of Sesamum indicum L. (Pedaliaceae), and leaf powder of Azadirachta indica A. Juss. (Meliaceae). Treatments of stored pulses with natural products or with Malathion all caused significantly higher mortality of C. chinensis at 15 d after treatment (DAT) than stored pulses receiving no treatments. The balls of C. camphora, rhizome powder of A. calamus and sesame oil outperformed all other treatments, including the Malathion at 45 and 75 DAT and resulted in significantly reduced damage or deterioration of stored pulses compared with other treatments. Storage regimes performed similarly, although the jute bags did protect seed integrity for some treatments. Our results indicate that incorporating these natural products into stored pulses can protect the seeds from C. chinensis for up to two generations, something that Malathion cannot do. These products are readily available to most farmers in the region and their use will lead to 1) reduction of losses to significant stored product pests, and 2) a reduction of contamination of foodstuffs and the environment by synthetic pesticides like Malathion.     

Oviposition deterrents of Callosobruchus maculatus (Coleoptera: Bruchidae)

Abstract Many bruchids distribute their eggs uniformly among the host seeds available to them and tend to avoid adding eggs to seeds bearing any eggs if pristine seeds remain. Several authors have postulated that the adult cowpea seed beetles, Callosobruchus maculatus (F.) (Coleoptera: Bruchidae), produce pheromones inhibiting oviposition by females exposed to them. It is shown that both sexes produce compounds which can be washed from glass surfaces in a variety of organic solvents which do not deter oviposition as such, but seeds coated with such washings are avoided by ovipositing females when they are selecting seeds for egg-laying and uncoated seeds are available. Some components, at least, of the products persist for at least 30 days, longer than the normal development time of insects in the seeds. The compounds affect the distribution of eggs among the available seeds. At least three different biotypes of the species produce, recognize, and respond in the same way to each other's products. It is emphasized that the eggs themselves may not be the site of the product produced by females but that the physical presence of eggs cannot be overlooked in any discussion of the possible role of pheromones in oviposition and egg distribution by bruchids.     

Evolutionary diversification of the bean beetle genus Callosobruchus (Coleoptera: Bruchidae): traits associated with stored-product pest status

Despite the fact that many plant-feeding insects are pests, little effort has been made to identify key evolutionary trait transitions that allow taxa to acquire or lose pest status. A large proportion of species in the genus Callosobruchus are economically important pests of stored, dry postharvest beans of the tribe Phaseoleae. However, the evolution of this feeding habit is poorly understood. Here, we present a reconstruction of the phylogeny of the Asian and African Callosobruchus based on three mitochondrial genes, and assess which traits have been associated with the evolutionary origin or loss of ability to reproduce on dry beans. Our phylogenetic analysis showed that species group into the chinensis and the maculatus clades, which are also supported by genital morphology, and an additional paraphyletic group. Ancestral ability to use dry beans has been lost in the chinensis clade but acquired again in C. chinensis. Dry-bean use and host-plant use were both phylogenetically constrained and transitions in the two were significantly correlated. Host shifts from the subtribe Phaseolinae to Cajaninae were more common than the reverse and were more likely in species using young beans. The ability to use dry beans was more likely gained when using Phaseolinae hosts and promoted habitat shifts from tropical to temperate regions. Adaptation to arid climate was also associated with the ability to reproduce on dry beans and on Phaseolinae. Thus, our analysis suggests that physiological adaptations to an arid climate and to Phaseolinae hosts both render beetles predisposed to become pests of cultivated beans.     

Conspecific sperm precedence in Callosobruchus subinnotatus (Coleoptera: Bruchidae): mechanisms and consequences

Conspecific sperm precedence (CSP) has been identified as an important post-copulatory, pre-zygotic mechanism that can act to reduce gene flow between populations. The evolution of CSP is thought to have arisen as a by-product of male and female coevolution in response to intraspecific post-copulatory sexual selection. However, little is known about the mechanisms that generate CSP. When Callosobruchus subinnotatus females copulate with both C. subinnotatus and Callosobruchus maculatus males, regardless of mating order, the majority of eggs are fertilized by conspecific sperm. The low number of heterospecific fertilizations does not result from general differences in the viability of sperm in the female reproductive tract, as heterospecific sperm fertilized equivalent numbers of eggs as conspecific sperm in the absence of sperm competition. Instead, CSP results from disadvantages to heterospecific sperm that are manifest only when in competition with conspecific sperm. CSP in C. subinnotatus appears to result from two, not mutually exclusive, mechanisms. First, conspecific sperm are better able to displace heterospecific sperm from female storage. Second, conspecific sperm achieve disproportionately higher numbers of fertilizations relative to their proportional representation in the fertilization set. Thus, we provide evidence of differential sperm use from the female spermatheca.     

Internal Spatiotemporal Population Dynamics of Infection with Three Wolbachia Strains in the Adzuki Bean Beetle, Callosobruchus chinensis (Coleoptera: Bruchidae)

The adzuki bean beetle, Callosobruchus chinensis, is infected with three distinct lineages of endosymbiotic bacteria belonging to the genus Wolbachia, which were designated wBruCon, wBruOri, and wBruAus. In an attempt to understand the mechanisms underlying the infection with these three organisms, the spatiotemporal infection dynamics of the three Wolbachia strains was investigated in detail by using a quantitative PCR technique. During the development of C. chinensis, the wBruCon, wBruOri, and wBruAus infection levels consistently increased but the growth patterns were different. The levels of infection plateaued at the pupal stage at approximately 3 × 10⁸, 2 × 10⁸, and 5 × 10⁷ wsp copy equivalents per insect for wBruCon, wBruOri, and wBruAus, respectively. At the whole-insect level, the population densities of the three Wolbachia types did not show remarkable differences between adult males and females. At the tissue level, however, the total densities and relative levels of the three Wolbachia types varied significantly when different tissues and organs were compared and when the same tissues derived from males and females were compared. The histological data obtained by in situ hybridization and electron microscopy were concordant with the results of quantitative PCR analyses. Based on the histological data and the peculiar Wolbachia composition commonly found in nurse tissues and oocytes, we suggest that the Wolbachia strains are vertically transmitted to oocytes not directly, but by way of nurse tissue. On the basis of our results, we discuss interactions among the three coinfecting Wolbachia types, reproductive strategies of Wolbachia, and factors involved in the different cytoplasmic incompatibility phenotypes.     

Internal spatiotemporal population dynamics of infection with three Wolbachia strains in the adzuki bean beetle,Callosobruchus chinensis (Coleoptera: Bruchidae)

The adzuki bean beetle, Callosobruchus chinensis, is infected with three distinct lineages of endosymbiotic bacteria belonging to the genus Wolbachia, which were designated wBruCon, wBruOri, and wBruAus. In an attempt to understand the mechanisms underlying the infection with these three organisms, the spatiotemporal infection dynamics of the three Wolbachia strains was investigated in detail by using a quantitative PCR technique. During the development of C. chinensis, the wBruCon, wBruOri, and wBruAus infection levels consistently increased but the growth patterns were different. The levels of infection plateaued at the pupal stage at approximately 3 x 10(8), 2 x 10(8), and 5 x 10(7) wsp copy equivalents per insect for wBruCon, wBruOri, and wBruAus, respectively. At the whole-insect level, the population densities of the three Wolbachia types did not show remarkable differences between adult males and females. At the tissue level, however, the total densities and relative levels of the three Wolbachia types varied significantly when different tissues and organs were compared and when the same tissues derived from males and females were compared. The histological data obtained by in situ hybridization and electron microscopy were concordant with the results of quantitative PCR analyses. Based on the histological data and the peculiar Wolbachia composition commonly found in nurse tissues and oocytes, we suggest that the Wolbachia strains are vertically transmitted to oocytes not directly, but by way of nurse tissue. On the basis of our results, we discuss interactions among the three coinfecting Wolbachia types, reproductive strategies of Wolbachia, and factors involved in the different cytoplasmic incompatibility phenotypes.     

Consequences of inbreeding for the cowpea seed beetle, Callosobruchus maculatus (F.)(Coleoptera: Bruchidae)

Inbreeding is said to reduce vigour and fitness. It may also determine how a population responds to selection. Local populations of Callosobruchus maculatus , the cowpea seed beetle, are established annually from small numbers of founders and the species has been distributed to many parts of the world where isolated populations may have been founded by very small numbers of individuals. After more than 20 generations of inbreeding, inbred lines have been shown to diverge from a common ancestral stock in similar directions with respect of some variables such as developmental speed, but haphazardly in respect of other parameters such as male weight. The respective roles of drift and of selection as effective evolutionary forces in inbred lines are discussed in the light of these results. It is argued that some intraspecific differences in C. maculatus may be explained as a product of periodic inbreeding, but that the process does not impair the ability to adapt to local conditions so contributing to the status of the species as a pest of international importance.     

Presence of the storage seed protein vicilin in internal organs of larval Callosobruchus maculatus (Coleoptera: Bruchidae)

Variant vicilins (7S storage globulins) of cowpea seeds (Vigna unguiculata) are considered as the main resistance factor present in some African genotypes against the bruchid Callosobruchus maculatus. It has been suggested that the toxic properties of vicilins may be related to their recognition and interaction with glycoproteins and other membrane constituents along the digestive tract of the insect. However, the possibility of a systemic effect has not yet been investigated. The objective of this work was to study the fate of 7S storage globulins of V. unguiculata in several organs of larvae of the cowpea weevil C. maculatus. Results demonstrated binding of vicilins to brush border membrane vesicles, suggesting the existence of specific receptors. Vicilins were detected in the haemolymph, in the midgut, and in internal organs, such as fat body and malpighian tubules. There is evidence of accumulation of vicilins in the fat body of both larvae and adults. The absorption of vicilins and their presence in insect tissues parallels classical sequestration of secondary compounds.     

Talisia esculenta lectin and larval development of Callosobruchus maculatus and Zabrotes subfasciatus (Coleoptera: Bruchidae)

Bruchid larvae cause major losses in grain legume crops throughout the world. Some bruchid species, such as the cowpea weevil and the Mexican bean weevil, are pests that damage stored seeds. Plant lectins have been implicated as antibiosis factors against insects, particularly the cowpea weevil, Callosobruchus maculatus. Talisia esculenta lectin (TEL) was tested for anti-insect activity against C. maculatus and Zabrotes subfasciatus larvae. TEL produced ca. 90% mortality to these bruchids when incorporated in an artificial diet at a level of 2% (w/w). The LD(50) and ED(50) for TEL was ca. 1% (w/w) for both insects. TEL was not digested by midgut preparations of C. maculatus and Z. subfasciatus. The transformation of the genes coding for this lectin could be useful in the development of insect resistance in important agricultural crops.     

Comparative seed resistance in eighty cowpea accessions to the seed bruchid,Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae)

Eighty cowpea accessions comprising both improved and local were assessed for their resistance to Callosobruchus maculatus (F.) attack under laboratory conditions (29 ± 3?°C and 65 ± 5% R.H.). Sixty-nine accessions were derived from the International Institute for Tropical Agriculture breeding programmes, four were local cultivars, while seven were improved cultivars from national research institutes. Results showed that there were significant differences among the accessions in terms of number of initial bruchid eggs laid on them, adult emergence, number of seeds damaged and bruchid-induced seed weight loss at three months after artificial infestation with C. maculatus. On IT04K-334-2, IT04K-343-1, IT06K-141, IT99K-216-48-1, IT99K-494-6 and IT99K-529-2, fewer initial eggs were laid and no adult emergence, seed damage or weight loss and were considered highly resistant to C. maculatus. On the other hand, IT03K-337, IT04K-217-51, IT04K-332-1, IT04K-339-1, IT06K-188-49, IT81D-1137, IT845-2231-15, IT93K-452-1, IT04K-154-1, IT96K-610, IT97K-1069-6, IT97K-390-2, Ife Bpc, IT99K-573-2-1, Ife Modupe, IT845-2246, Drum, Ife Brown and Oloyin were the most susceptible with the highest damage indices values (p?<?0.05). The range of each of the resistance indices measured in the susceptible varieties was: number of initial eggs laid (61–100), adult emergence (40.0–62.10%), number of seeds damaged (53.0–85.5) and seed weight loss (58.0–66.25%).     

Contest and scramble competitions in two bruchid species,Callosobruchus analis andC. phaseoli (Coleoptera: Bruchidae)

Larval competition between contest and scramble strategists was investigated using the two bruchid species, C. analis (contest species) and C. phaseoli (scramble species) with two different sized mung beans (large and small beans). In both sized beans, the adult emergences of each species dependen on total density of the initial larval densities of the two species and the ratio of the two densities. The emergence of one species was suppressed by the existence of the other species when the initial larval density per bean of the former species was less than that of the latter one. There were many cases in which both C. analis and C. phaseoli emerged from one bean in large beans, but such cases were quite rare in small beans. C. analis performed interference behavior only at late larval stages, whereas C. phaseoli was superior in exploitative competition all through their larval stages. These, combined with the niche segregation inside a bean, are throught to be the major factors of observed density- and frequency-dependent competition results. Based on the above experimental results, long-term competition results between the contest and scramble species were predicted.     

Sperm competition in Callosobruchus maculatus (Coleoptera: Bruchidae): a comparison of two methods used to estimate paternity

Abstract.

• 1 This study investigates sperm competition in Cullosobruchus maculatus (F.) (Coleoptera: Bruchidae), and is the first study to make a direct, controlled comparison of the sterile male and genetic marker techniques to estimate sperm precedence.
• 2 P₂ values (the proportion of offspring fathered by the second male) obtained from the two methods were similar: P₂ (sterile male) = 0.82, P₂ (genetic marker) = 0.85. Both methods are therefore suitable for studying sperm precedence if the appropriate correction factors are applied.
• 3 The importance of general fertilization ability, differential fertilizing capacity and differential zygote mortality are examined and discussed.

     

Adaptive plasticity of egg size in response to competition in the cowpea weevil,Callosobruchus maculatus (Coleoptera: Bruchidae)

Life history theory predicts that larger propagules should be produced when the offspring are expected to experience intense competition. This study tested whether female cowpea weevils responded to high larval or adult density by producing larger eggs. In a splitbrood design I measured the effect of density experienced by females at their larval stage (1 vs. 4–6 larvae/cowpea) on the size of eggs produced just after emergence. The females were then kept either at low adult density (1 female+1 male per vial), or at high adult density (10 females+10 males) for 2 days, and tested for the effect of this adult density treatment on the size of eggs laid subsequently. I measured egg length and width, as well as the diameter of the entrance tunnel made by the larva, which can be regarded as a crude measure of larval size. Females that experienced high adult density subsequently laid slightly wider eggs than those kept at low density. This difference, albeit small (about 1–4% after correction for female weight and the effect of family, depending on the statistical model used), was statistically significant and robust to alterations of the statistical model. It may be a remnant of a larger plastic response of egg size to competition that has become eroded during many generations in high-density laboratory cultures. There was no difference in egg length or the diameter of the entrance tunnel. Eggs laid just after emergence by females reared at high larval density also tended to be wider than those produced by females that had no competitors. This effect was only marginally significant, however, and sensitive to the statistical model. Both egg length and width and the diameter of the entrance tunnel increased with female weight and decreased with female age. The tunnel diameter was positively correlated with both egg length and width, but the effect of width was larger.     

Wall-making behavior as a proximate mechanism to generate variation in larval competition in Callosobruchus maculatus (Coleoptera: Bruchidae)

Variation from contest to scramble in larval competition types was observed among laboratory lines derived from a geographic strain of Callosobruchus maculatus. In contest competition, only one adult can emerge from a small bean because the successful larva monopolizes resources. In scramble competition, however, multiple adults can emerge from the bean because larvae share resources. To explain the variation in competition types, we used six lines of the geographic strain to test the hypothesis that the larval competition type is determined by the larval behavior of building walls, which prevent larvae from interfering with each other, allowing multiple adults to emerge from a single bean. We also investigated the proportions of wall-making in contest-scramble hybrid lines to test whether the formation of a wall structure was genetically determined. Results support our hypothesis that wall-making behavior determines the type of larval competition within a geographic strain, and that the behavior is genetically determined. Scramble-type lines exhibited higher frequencies of wall-making than contest-type lines when two larvae of the same line infested a bean. Larval competition type and the tendency towards wall formation in contest-scramble hybrid lines ranged intermediate of parental lines. We concluded that the variation in larval competition type is determined by the variation in larval wall-making behavior among laboratory lines derived from the geographic strain. We will discuss the evolution of scramble-type larvae in C. maculatus based on our results.     

The Effect of Single, Double, and Triple Matings on the Lifetime Fecundity of Callosobruchus analis and Callosobruchus maculatus (Coleoptera: Bruchidae)

This study examined the effect of single, double, and triple matings on female lifetime fecundity in two closely related species of beetle, Callosobruchus analis and C. maculatus. Multiple mating resulted in elevated lifetime fecundities in both species, although the pattern of elevation differed between the two species. When oviposition resource was plentiful the elevation was apparent after a second but not a third copulation for C. maculatus and after a second and after a third for C. analis. By altering the availability of oviposition sites to C. maculatus females, we were able to alter the pattern of fecundity elevation between matings. When oviposition sites were limited, fecundity increased after a third mating but not a second mating. We suggest that the pattern of fecundity elevation associated with multiple mating in these species is primarily the result of oviposition stimulants delivered by males, rather than a nutritional donation brought about by the metabolism of ejaculates. We also suggest that the intra- and interspecific differences in the relationship between mating frequency and fecundity are the result of differences in egg maturation rates.     

Variation in responses to susceptible and resistant cowpeas among West African populations of Callosobruchus maculatus (Coleoptera: Bruchidae)

The cowpea seed beetle, sometimes also known as the cowpea weevil, Callosobruchus maculatus (F.), is a major pest of stored cowpeas (Vigna unguiculata Walpers) in West Africa. Control methods have included development of 'resistant' varieties as an environmentally benign alternative to insecticides, but there is concern over their effectiveness because of population variation among the insects and the possibility of adaptation overcoming seed resistance. Populations of C. maculatus from Nigeria, Ghana, Benin, and Niger, were used to examine variation in response to resistant and susceptible cowpea varieties at two geographical scales. Among seven Nigerian populations, there were significant differences in development times, the pattern of adult emergence, adult weights, and female fecundity when reared under identical conditions. Development in the resistant variety was retarded, produced higher mortality and lower adult weights. Significant interactions between variety and population were evident in terms of their effects on adult weight and development time; development times in the resistant variety were longer and emergences occurred over a longer period in some populations than in others. Population responses to resistant seeds were therefore unpredictable, but there was no evidence to suggest adaptation to overcome seed resistance within three generations. On a larger geographical scale, variation in performance was much greater and therefore, even less predictable. Mortality in resistant seeds was also higher among populations collected from outside Nigeria and may be explained by significant adaptation among Nigerian populations to previous release of resistant varieties. The findings are discussed in relation to understanding the extent of intraspecific variation in C. maculatus and its implications for future pest management.     

Insecticidal and biological efficacy of two vegetable oils against Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae) under laboratory conditions

Bioefficacy of two vegetable oils, namely ethyl oleate and Acorus calamus, were conducted against the pulse beetle Callosobruchus maculatus (Fabr.) under a constant temperature of 29 ± 1 °C and a relative humidity of 65 ± 5% R.H. Four concentrations (0.125, 0.25, 0.5 and 1.0%) of each oil were used against the adult and tested their latent effect on the immature stages of the beetle. The results obtained show insecticidal effect of both ethyl oleate and A. calamus oil; however, A. calamus oil proved to be more toxic to the adults at all the concentrations tested in comparison to the ethyl oleate oil. As the concentration of the oils decreased, the percentage mortality decreased. The obtained results concentrated on biological efficacy of the oils tested against the adults. Both oils affected the longevities of both male and female C. maculatus adults in comparison to the control. The oviposition periods of the adult female were affected significantly in comparison to the control. A. calamus oil prohibited oviposition completely at high concentrations of 0.5 and 1.0% and caused complete sterility to the females. Lower concentrations decreased the oviposition period greatly and reduced the number of eggs laid in comparison to the control. Ethyl oleate had an inconsiderable latent effect on the incubation period as well as the percentage of hatching of the resulting eggs. A. calamus oil decreased the percentage of egg hatchability with serious increase in the percent sterility. The two tested oils had slight extension in either larval or pupal stage of the resulting progeny of C. maculatus. Both oils show oviposition deterrency towards C. maculatus adults. The oviposition deterrent index was 100% when mung bean seeds were treated with A. calamus oil. The bruchid females laid all eggs on control seeds and no eggs were laid on treated seeds.