Azuki Bean Weevil Research Articles

Tandemly duplicated genes encoding polygalacturonase inhibitors are associated with bruchid (Callosobruchus chinensis) resistance in moth bean (Vigna aconitifolia)

Bruchids are stored-grain insect pests responsible for serious seed loss in legume crops. A previous study using an F2 population (F2OA) derived from a cross between wild moth-bean (Vigna aconitifolia [Jacq.] Maréchal) accession TN67 (resistant) and cultivated moth-bean accession ICPMO056 (susceptible) revealed that resistance to the azuki bean weevil (Callosobruchus chinensis L.) in TN67 was regulated by a single gene located in the major quantitative trait locus—qVacBrc2.1. In this study, qVacBrc2.1 was finely mapped and candidate genes in this locus were identified using F2OA and another large F2 population (F2NB) derived from the cross mentioned previously. In contrast to the previous study, segregation analysis in the F2NB population revealed that resistance against this pest was controlled by two genes. Furthermore, the addition of novel markers to qVacBrc2.1 and reanalysis of the QTL in the F2OA population demonstrated that qVacBrc2.1 constituted two linked QTLs—qVacBrc2.1-A and qVacBrc2.1-B. The presence of qVacBrc2.1-B was verified using the population F2NB. Comparative genomics using three Vigna spp. strongly suggested the presence of two tandemly duplicated genes, VacPGIP1 and VacPGIP2, which encoded polygalacturonase inhibitors (polygalacturonase-inhibiting proteins) as the candidates for conferring resistance, but only VacPGIP1 could be successfully cloned and sequenced. The alignment of VacPGIP1 coding sequences of TN67 and ICPMO056 revealed eight single nucleotide polymorphisms, three of which altered the amino-acid sequence of the predicted domains of polygalacturonase inhibitors in ICPMO056. Overall, these findings indicate that VacPGIP1 and VacPGIP2 regulated C. chinensis resistance in TN67.

Bruchid a Serious Pest on Pulse Crops: Its Control Measures and Breeding Advancements: A Review

Bruchid (Callosobruchus spp.) are the most destructive, notorious storage pest of pulses in the tropical and sub-tropical region. The yield losses are higher than half of the expected yield with in the short period of time. Bruchid initial infestation started in the field and shortly builds up during storage time and cause severe seed damage up to 100 per cent. Bruchid infestation ranges from 60 to 100% within two to three months of storage period. Among the different bruchid species, cowpea weevils (C. maculatus F.) and azuki bean weevils (Callosobruchus chinensis L.) are the most destructive storage pest of pulses. Though several options are now available to identify the elite genotypes against bruchid infestation, still the development of genotypes with sufficient level of host plant resistance is not achieved. In this pursuit, the present article has given a detailed review of the major species of bruchid, insect life style, management practices, screening technique, sources of resistance, novel breeding strategies and recent advancements including use of molecular markers in marker aided selection and QTL studies for bruchid resistance.

Genetic analysis of seed resistance to Callosobruchus chinensis and Callosobruchus maculatus in cowpea

Callosobruchus chinensis (azuki bean weevil) and Callosobruchus maculatus (cowpea weevil) are the most destructive pests that cause significant losses to cowpea (Vigna unguiculata) seeds during storage. Development of cultivar(s) resistance to bruchids is a major goal in cowpea breeding program. Cowpea accession “TVu 2027” has been identified as moderately resistant to C. maculatus. Genetic studies of the seed resistance in this accession using strains of C. maculatus from Africa and America have demonstrated that the resistance is controlled by one or two recessive genes. However, there is no reports on genetics of the resistance to C. chinensis and to strain(s) of C. maculatus from Asia. Therefore, the objective of this study was to investigate genetics of the resistance to C. chinensis and C. maculatus in TVu 2027. TVu 2027 (P2) and PK2015VTN001 (P1; susceptible cowpea) were used to develop six basic generations (populations), viz. P1, P2, F1 (P1 × P2), F2 (P1 × P2), BC1P1 (P1 × F1) and BC1P2 (P2 × F1). The population were evaluated for percentage of damaged seeds (PDS) and area under the disease progress stairs (AUDPS; indicating infestation severity) by C. chinensis and C. maculatus. The results showed that TVu 2027 was moderately resistant to both bruchid species. Broad-sense heritability for PDS and AUDPS was moderate, being 70% and 73% for C. chinensis, respectively, and 64% and 61% for C. maculatus, respectively. Number of genes controlling C. chinensis resistance and C. maculatus resistance was two genes and one gene, respectively. Generation mean analysis revealed that genes with additive effect and additive × dominance gene interaction are involved in the resistance to both bruchid species. Correlation analysis suggested that the genes controlling resistance to C. chinensis and those conferring resistance to C. maculatus are unlinked.

A second VrPGIP1 allele is associated with bruchid resistance (Callosobruchus spp.) in wild mungbean (Vigna radiata var. sublobata) accession ACC41.

Two bruchid species, azuki bean weevil (Callosobruchus chinensis L.) and cowpea weevil (Callosobruchus maculatus F.), are the most important insect pests of mungbean [Vigna radiata (L.) Wilczek] after harvest. Improving bruchid resistance is a major goal for mungbean breeders. Bruchid resistance in mungbean is controlled by a single major locus, Br. The tightly linked VrPGIP1 and VrPGIP2, which encode polygalacturonase-inhibiting proteins (PGIPs), are the candidategenes at the Br locus associated with bruchid resistance. One VrPGIP1 resistance allele and two VrPGIP2 resistance alleles have been identified. In this study, we fine-mapped the bruchid-resistance genes in wild mungbean (V. radiata var. sublobata) accession ACC41 using the F2 population (574 individuals) derived from the 'Kamphaeng Saen 2' (susceptible) × ACC41 (resistant) cross. A QTL analysis indicated that the resistance to the azuki bean weevil and cowpea weevil in ACC41 is controlled by a major QTL (qBr5.1) and a minor QTL (qBr5.2), which are only 0.3cM apart. qBr5.1 and qBr5.2 accounted for about 82% and 2% of the resistance variation in the F2 population, respectively. qBr5.1 was mapped to a 237.35-kb region on mungbean chromosome 5 containing eight annotated genes, including VrPGIP1 and VrPGIP2. An examination of the ACC41 VrPGIP1 and VrPGIP2 sequences revealed a new allele for VrPGIP1 (i.e., VrPGIP1-2). Compared with the wild-type sequence, VrPGIP1-2 has five SNPs, of which four cause amino acid changes (residues 125, 129, 188, and 336). A protein sequence analysis indicated that residues 125 and 129 in VrPGIP1-2 are in a β-sheet B1 region, whereas residues 188 and 336 are in a C10-helix region and at the end of the C-terminal region, respectively. Because the β-sheet B1 region is important for interactions with polygalacturonase (PG), residues 125 and 129 in VrPGIP1-2 likely contribute to bruchid resistance by inhibiting PG. Our results imply that VrPGIP1-2 is associated with the bruchid resistance of wild mungbean accession ACC41. This new resistance allele may be useful for breeding mungbean varieties exhibiting durable bruchid resistance.

Genetic Dissection of Azuki Bean Weevil (Callosobruchus chinensis L.) Resistance in Moth Bean (Vigna aconitifolia [Jaqc.] Maréchal).

The azuki bean weevil (Callosobruchus chinensis L.) is an insect pest responsible for serious postharvest seed loss in leguminous crops. In this study, we performed quantitative trait locus (QTL) mapping of seed resistance to C. chinensis in moth bean (Vigna aconitifolia [Jaqc.] Maréchal). An F2 population of 188 plants developed by crossing resistant accession ‘TN67’ (wild type from India; male parent) and susceptible accession ‘IPCMO056’ (cultivated type from India; female parent) was used for mapping. Seeds of the F2 population from 2014 and F2:3 populations from 2016 and 2017 were bioassayed with C. chinensis, and the percentage of damaged seeds and progress of infestation severity were measured. Segregation analysis suggested that C. chinensis resistance in TN176 is controlled by a single dominant gene, designated as Rcc. QTL analysis revealed one principal and one modifying QTL for the resistance, named qVacBrc2.1 and qVacBrc5.1, respectively. qVacBrc2.1 was located on linkage group 2 between simple sequence repeat markers CEDG261 and DMB-SSR160 and accounted for 50.41% to 64.23% of resistance-related traits, depending on the trait and population. Comparative genomic analysis suggested that qVacBrc2.1 is the same as QTL Brc2.1 conferring C. chinensis resistance in wild azuki bean (V. nepalensis Tateishi and Maxted). Markers CEDG261 and DMB-SSR160 should be useful for marker-assisted selection for C. chinensis resistance in moth bean.

Mung bean (Vigna radiata) cultivars mediated oviposition preference and development of Callosobruchus chinensis (Coleoptera: Chrysomelidae: Bruchinae)

The Azuki bean weevil, Callosobruchus chinensis (L.), is a destructive pest of stored mung bean [Vigna radiata (L.) Wilczek] as well as other leguminous seeds. The development of resistant seeds to manage this pest is of current great interest to plant breeders. In this study, we investigated the oviposition preference and development of C. chinensis on two susceptible mung bean cultivars (Seonhwa and Gyeongseon) and one previously reported resistant cultivar (Jangan), compared to the susceptible cowpea (Vigna unguiculata L.), cultivar (Yeonbun) using both multiple-choice and no-choice tests. In addition, the development of C. chinensis was also examined at four constant temperatures (20, 25, 30, and 35 °C). Both tests found cowpea to be the most suitable seed for oviposition. Total developmental time from oviposition to adult emergence ranged from 27.01 to 38.2 days, being shortest on cowpea and longest on the mung bean, cv. Jangan. However, no successful development of C. chinensis larvae on mung bean, cv. Jangan, occurred at any temperature. The highest rate of adult emergence and the longest adult longevity both occurred on cowpea and certain mung bean cultivars (Seonhwa and Gyeongseon), with the dramatic exception of cv. Jangan. These results suggest that the higher preference and performance of C. chinensis on cowpea (3.3 egg/seed) and least on mung bean, cv. Jangan (0.4 egg/seed). This information may facilitate the exploration of resistant genetic materials and chemicals associated with seeds for successful breeding. Further studies should examine the chemicals associated with mung bean cultivars and its resistant mechanism to develop a control method against bruchines.

Effects of low temperatures on the survival and development of Callosobruchus chinensis (L.) (Coleoptera: Bruchidae) under different storage durations

Azuki bean weevil, Callosobruchus chinensis L. is a cosmopolitan pest of stored grain legumes. Larvae cause huge losses of energy and seeds in the storage. This study assessed the effects of short-term exposure to low-temperatures (4, 0, and −4°C) under seven durations of storage (1, 5, 10, 15, 20, 25, and 30days) on the survival, development, and adult performance of C. chinensis in laboratory. The effects were studied on three life stages (egg, larva [2–3 instar], and pupa) of C. chinensis. The results showed that low-temperature treatments have significant effects on the life variables of C. chinensis. Among the stages, egg and pupa stages were most sensitive to low-temperature. The very low survival rates of eggs at −4°C, no pupa stages of C. chinensis survived after 10days storage. Cold exposure at −4°C for <10days not only increased egg and pupa mortality, but the negative effects also transmitted to the larvae and pupa developed from those eggs (64.3days) and least adults were emerged with minimum longevity. Increasing the duration of cold exposure further reduced survival rates, e.g. about 20% of the eggs survived after 15days exposure at −4°C, but <30% of them survived after 25days treatment at 4 and 0°C. Low-temperature treatments have also affected on the adult performances such as adult weight. However, there were not any negative effects of low-temperatures on the seed germination. The life variables of C. chinensis are discussed in terms of targeting particular susceptibilities to low-temperatures in different durations of storage as an alternative to chemical treatments.

Effects of temperature on development of the azuki bean weevil, Callosobruchus chinensis (Coleoptera: Bruchidae) on two leguminous seeds

The azuki bean weevil, Callosobruchus chinensis L., is one of the most serious stored product pests in the world. The temperature driven stage-specific development of C. chinensis was examined at eight constant temperatures of 15.97, 18.31, 22.09, 26.15, 30.50, 34.39, 38.00, and 40.00 °C on azuki bean (Vigna angularis [Wild.] Ohwi & Ohashi) and cowpea (Vigna unguiculata L.) seeds. Parameters for stage-specific development such as threshold temperatures and thermal constants were determined by linear and nonlinear models. The total developmental time (egg - adult) decreased with increasing temperature from 14 to 38 °C on both seeds. However, total developmental time varied from 127.93 to 122.25 days at 15.97 °C to 19.31 and 19.17 days at 38.00 °C on azuki bean and cowpea seeds, respectively. The nonlinear Logan-6 and Performance models estimated optimal temperatures as 37.3, 37.0, 36.5 and 37.1 °C on azuki bean, and as 36.7, 36.4, 34.5 and 36.6 °C on cowpea for egg, larva, pupa and egg to adult, respectively. The linear model estimated the lower temperature thresholds (Tmin) of egg, larva, pupa and egg to adult as 16.97, 10.47, 1.36, and 10.28 °C on azuki bean, respectively, and 16.84, 10.33, 10.92, and 9.83 °C on cowpea, respectively. The thermal constants (K) for each stage completion was 35.13, 255.21, 233.11, and 503.41° days (DD) for egg, larva, pupa, and egg to adult on azuki bean, respectively and 32.45, 246.50, 107.04, and 490.02 DD for egg, larva, pupa, and egg to adult on cowpea, respectively. Temperature also influenced the adult longevity. These findings on thermal requirements and temperature thresholds can be used to predict the occurrence, number of generations and population dynamics of C. chinensis.

A gene encoding a polygalacturonase-inhibiting protein (PGIP) is a candidate gene for bruchid (Coleoptera: bruchidae) resistance in mungbean (Vigna radiata).

The Br locus confers bruchid resistance in mungbean; VrPGIP2 (encoding a polygalacturonase inhibitor) is a strong candidate gene for this resistance. The VrPGIP2 sequence differs between resistant and susceptible lines. Azuki bean weevil (Callosobruchus chinensis) and cowpea weevil (Callosobruchus maculatus) are serious insect pests of mungbean during storage. Bruchid resistance in mungbean is controlled by a single dominant locus, Br. Although the Br locus has been located on a genetic map, molecular basis and function of the gene remain unknown. In this study, high-resolution mapping using a BC11F2 population of 418 plants derived from a cross between 'Kamphaeng Saen 1' (KPS1; susceptible) and 'V2802' (resistant) using simple sequence repeat (SSR) markers delimited the Br locus to a genomic region of 38Kb of chromosome 5 containing two annotated genes. EST-SSR marker DMB-SSR158 co-segregated perfectly with the Br locus. Bioinformatics analyses revealed that DMB-SSR158 corresponds to a gene encoding a polygalacturonase inhibitor (polygalacturonase-inhibiting protein PGIP) and was designated as VrPGIP2. Comparison of VrPGIP2 coding sequences between four bruchid-resistant (V2802, V1128, V2817 and TC1966) and four bruchid-susceptible (KPS1, Sulu-1, CM and an unknown accession) mungbean lines revealed six single nucleotide polymorphisms (SNPs) between the resistant and susceptible groups. Three of the six SNPs resulted in amino acid changes; namely, alanine (A) to serine (S) at position 320, leucine (L) to proline (P) at position 332, and threonine (T) to P at position 335 of the VrPGIP2 sequence in resistant lines, compared with that in susceptible lines. The A to S change at position 320 may affect the interaction between PGIP and polygalacuronase. These results indicate that VrPGIP2 is very likely the gene at the Br locus responsible for bruchid resistance in mungbean.

Oviposition preference and development of azuki bean weevil, Callosobruchus chinensis, on five different leguminous seeds

This study was conducted to determine the oviposition preference and development of azuki bean weevil, Callosobruchus chinensis (L.) (Coleoptera: Bruchidae) on five different leguminous seeds a) cowpea cultivar Yeonbundongbu, b) white kidney bean cultivar Hwanghyubilho, c) soybean cultivar Daepung, d) mung bean cultivar Seonhwanogdu, and e) azuki bean cultivar Hongeon. A multiple-choice test and a no-choice test were conducted to record the number of eggs laid, total developmental time, number of adults emerged, and adult longevity of C. chinensis on the five different leguminous seeds. Both choice and no-choice tests revealed cowpea seed as the most preferred oviposition substrate. Total developmental time from egg to adult stage ranged from 28 to 31 days; developmental time was shortest on the cowpea. Successful development of C. chinensis was not observed on soybean or on kidney bean. Rate of adult emergence was higher on azuki and mung bean seeds with longer adult longevity of those emerged from azuki bean and cowpea. Higher egg laying preference and shorter developmental time with longer adult longevity of the beetle on cowpea probably signifies importance of physical as well as chemical attributes of the seed as an oviposition substrate other than evolutionary history of the insect.

Cause of egg mortality in azuki bean weevils (Callosobruchus chinensis): physical or chemical?

AbstractThe cause of high egg mortality at high adult density in azuki bean weevil is still controversial. It may be physical, as a result of adult trampling, or chemical, as a result of substance(s) secreted by adults and coated on the bean surface. In this study, I first constructed simple mathematical models based on each mechanism, which could predict the change of egg hatchability oviposited on different dates at different adult densities. I then devised an experimental method that could identify the fate of eggs oviposited on different dates. The experimental results showed that egg mortality was most likely caused by adult trampling.

Direct Determination of the Stereoisomeric Composition of Callosobruchusic Acid, the Copulation Release Pheromone of the Azuki Bean Weevil, Callosobruchus chinensis L., by the 2D-Ohrui-Akasaka Method

The stereoisomeric composition of the copulation release pheromone of the azuki bean weevil, Callosobruchus chinensis L., was determined to be R:S=3.3-3.4:1 by the 2D-Ohrui-Akasaka method.

Assessment of the importance of α-amylase inhibitor-2 in bruchid resistance of wild common bean

Both alpha-amylase inhibitor-2 (alphaAI-2) and arcelin have been implicated in resistance of wild common bean (Phaseolus vulgaris L.) to the Mexican bean weevil (Zabrotes subfasciatus Boheman). Near isogenic lines (NILs) for arcelin 1-5 were generated by backcrossing wild common bean accessions with a cultivated variety. Whereas seeds of a wild accession (G12953) containing both alphaAI-2 and arcelin 4 were completely resistant to Z. subfasciatus, those of the corresponding NIL were susceptible to infestation, suggesting that the principal determinant of resistance was lost during backcrossing. Three independent lines of transgenic azuki bean [Vigna angularis (Willd.) Ohwi and Ohashi] expressing alphaAI-2 accumulated high levels of this protein in seeds. The expression of alphaAI-2 in these lines conferred protection against the azuki bean weevil (Callosobruchus chinensis L.), likely through inhibition of larval digestive alpha-amylase. However, although the seed content of alphaAI-2 in these transgenic lines was similar to that in a wild accession of common bean (G12953), it did not confer a level of resistance to Z. subfasciatus similar to that of the wild accession. These results suggest that alphaAI-2 alone does not provide a high level of resistance to Z. subfasciatus. However, alphaAI-2 is an effective insecticidal protein with a spectrum of activity distinct from that of alphaAI-1, and it may prove beneficial in genetic engineering of insect resistance in legumes.

Development of an interspecific Vigna linkage map between Vigna umbellata (Thunb.) Ohwi &amp; Ohashi and V. nakashimae (Ohwi) Ohwi &amp; Ohashi and its use in analysis of bruchid resistance and comparative genomics

AbstractTo facilitate transfer of bruchid resistance to azuki bean (Vigna angularis) from its relatives an interspecific mapping population was made between rice bean, V. umbellata, and the related wild species V. nakashimae. The V. umbellata parent is completely resistant and V. nakashimae is completely susceptible to the bruchid beetle pests, azuki bean weevil (Callosobruchus chinensis) and cowpea weevil (C. maculatus). There is very low cross compatibility between V. umbellata and azuki bean. Therefore, V. nakashimae, that crosses with both V. umbellata and V. angularis without the need for embryo rescue, is used as a bridging species. A genetic linkage map was constructed based on an interspecific F2 mapping population between V. umbellata and V. nakashimae consisting of 74 plants. A total of 175 DNA marker loci (74 RFLPs and 101 SSRs) were mapped on to 11 linkage groups spanning a total length of 652 cM. Segregation distortion was observed but only three markers were not linked to any linkage group due to severe segregation distortion. This interspecific genome map was compared with the genome map of azuki bean. Of 121 common markers on the two maps, 114 (94.2%) were located on the same linkage groups in both maps. The marker order was highly conserved between the two genome maps. Fifty F2 plants that produced sufficient seeds were used for quantitative trait locus (QTL) analysis and locating gene(s) for C. chinensis and C. maculatus resistance in V. umbellata. The resistance reaction of these F2 plants differed between C. chinensis and C. maculatus. Both resistances were quantitatively inherited with no F2 plants completely susceptible to C. chinensis or C. maculatus. One putative QTL for resistance to each of these bruchid species was located on different linkage groups. Other putative QTLs associated with resistance to both C. chinensis and C. maculatus were localized on the same linkage group 1. Linked markers associated with the bruchid‐resistant QTL will facilitate their transfer to azuki bean breeding lines.

Characterization of Callosobruchus chinensis (L.) resistance in Vigna umbellata (Thunb.) Ohwi & Ohashi

Resistance to azuki bean weevil, Callosobruchus chinensis, was studied in a series of field and laboratory experiments in two accessions of rice bean ( Vigna umbellata (Thunb.) Ohwi & Ohashi), one accession of black gram ( V. mungo (L.) Hepper), and one accession of mungbean, ( V. radiata (L.) Wilczek). Weevil damage to immature pods of the rice bean accessions, ‘Menaga’ and ‘Miyazaki’, was significantly less than to the susceptible mungbean, VC1973A. In mature pods, the pest damage to the pod wall of Menaga was significantly higher than to VC1973A, whereas the damage to Miyazaki was similar to VC1973A. Seeds within the pods of both rice bean accessions were resistant no matter when the pods were harvested. When the insects were exposed directly on dry seeds, both rice bean accessions and a black gram accession VM2164 were resistant to them. In artificial seeds made by mixing flour of the individual resistant Vigna accessions with VC1973A and subsequently exposed to bruchid oviposition, the higher the quantity of resistant Vigna flour the lower the number of bruchids that emerged from such seeds. No bruchids emerged from artificial seeds containing crude starch fraction from the three resistant Vigna accessions when such seeds were exposed to bruchid infestation, whereas many insects emerged from the seeds containing starch of VC1973A or flour of VC1973A alone. In artificial seeds made by mixing crude protein fractions of the three resistant Vigna accessions with flour of VC1973A, as the concentration of protein increased the number of C. chinensis adults that emerged decreased. Fractionation of crude proteins into acetone-precipitable proteins and peptide and amino acid portions resulted in the loss of antibiosis effect. Artificial seeds made from purified starch-polysaccharides fraction, however, exhibited antibiosis effects if prepared from the rice bean seed of Menaga and Miyazaki but not if made from the black gram seed, VM2164.

Growth rate change driven by external perturbation in the azuki bean weevil

In laboratory experiments we obtain that the apparent growth rate of the population becomes larger than one under the normal condition, triggered by the external perturbation as the removal of individuals. The changed growth rate is stable for a while. We also propose a simple model of population dynamics allowing both matching and mis-matching the trend of the external perturbation, and show that the growth rate of the model population is changeable and stable to some extent.

Host-recognizing kairomones for parasitic wasp, Anisopteromalus calandrae, from larvae of azuki bean weevil, Callosobruchus chinensis.

Host-recognizing kairomones for the stinging behavior of the parasitic wasp, Anisopteromalus calandrae, were identified on host azuki bean weevil larvae, Callosobruchus chinensis (L.). The kairomones were extracted with acetone from Chinese green beans, from which emerged wasps and host weevils had been removed. The kairomones are a mixture of triacylglycerols and fatty acids, each of which is separately active, and with no observable synergistic effect between them. These compounds are known to be constituents of an oviposition-marking pheromone of host azuki bean weevils. However, they differ from the previously reported saturated hydrocarbons and diacylglycerols of the kairomone that another parasitic wasp, Dinarmus basalis, uses for the host recognition of C. chinensis. Thus, A. calandrae and D. basalis selectively utilize different constituents of the oviposition-marking pheromone of C. chinensis as host-recognizing kairomones.

Realized heritability of behavioral responsiveness to an oviposition deterring pheromone in the azuki bean weevil Callosobruchus chinensis

Artificial selection was conducted to reduce the behavioral responsiveness of female bruchid beetles, Callosobruchus chinensis, to the oviposition deterring pheromone excreted by conspecific females. Significant responses to selection were observed after two generations of selection. Realized heritability was estimated as 0.052 ± 0.017 from cumulative responses regressed on cumulative selection differentials. These results indicate that this pheromone communication system has significant additive genetic variance needed for its evolution.

Parasitic wasp, Dinarmus basalis, utilizes oviposition-marking pheromone of host azuki bean weevils as host-recognizing kairomone.

A host-recognizing kairomone responsible for the stinging behavior of the parasitic wasp, Dinarmus basalis, was studied. Fresh azuki beans coated with an acetone extract of the azuki beans, from which both emerged wasps and their host weevils were removed, elicited stinging behavior from female wasps. The kairomone is a mixture of saturated hydrocarbons and diacylglycerols, both of which are required for activity. The kairomone is composed of normal and methyl-branched hydrocarbons with carbon numbers ranging from 25 to 35, most of which are known as the hydrocarbon constituents of an oviposition-marking pheromone of the host azuki bean weevils, Callosobruchus chinensis. This indicates that D. basalis utilizes the oviposition-marking pheromone of its host weevils as a host-recognizing kairomone.

Insecticidal activity of an α-amylase inhibitor-like protein resembling a putative precursor of α-amylase inhibitor in the common bean, Phaseolus vulgaris L.

α-Amylase inhibitor (αAI) in the common bean, Phaseolus vulgaris L., protects seeds from insect pests such as the cowpea weevil ( Callosobruchus maculatus) and the azuki bean weevil ( C. chinensis). Cultivars which lack αAI still show resistance to both bruchids. These cultivars have a glycoprotein that reacts with anti-αAI-1 antibodies. The glycoprotein with a molecular mass of 29 kDa (Gp29) was purified and the encoding gene was isolated. The primary structure of Gp29 is the same as α-amylase inhibitor-like protein (AIL) from which the encoding gene has already been isolated. AIL resembles a putative precursor of αAI, even though it does not form the active inhibitor. However, AIL has some inhibitory effect on the growth of C. maculatus but not C. chinensis. The presence of AIL alone is insufficient to explain the bruchid resistance of common bean cultivars lacking α-AI. Common bean seeds appear to contain several factors responsible for the bruchid resistance.