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.     

Cowpea resistance and bioactivity of two diatomaceous earths against Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae) on three cowpea varieties

The bruchid, Callosobruchus maculatus (F.) is the most important pest of stored cowpea in tropical climates. Experiments were conducted to evaluate the susceptibility of three cowpea varieties and the bioactivity of SilicoSec® and FossilShield® against this insect pest under ambient laboratory conditions at the contents 0, 0.5, 1, 1.5 and 2 g/kg of cowpea. The Dobie susceptibility index for cowpea genotypes was estimated to compare their relative tolerance to this insect pest. Adult mortality was assessed after 1, 2, 4 and 6 days of exposure followed by the evaluation of the F₁ progeny. Population increase, percentage seed damage and weight loss were assessed after three months of storage. The results showed that the CRPS and TN5-78 varieties were highly susceptible to C. maculatus, while Lori variety was moderately susceptible to the weevil. SilicoSec® and FossilShield® caused a significant and increasing bruchid mortality according to the dose rates and periods of exposure. Malathion 5% caused complete mortality after one day exposure time. SilicoSec® was more effective than FossilShield® on the three varieties of cowpea. The greatest dose of SilicoSec® and FossilShield® (2 g/kg) caused 100% mortality on all three varieties; 4 days of exposure in the Lori variety was the shortest exposure time leading to complete mortality. Regarding LD₅₀ and LD₉₅ values, it was observed that C. maculatus adults are more susceptible to SilicoSec® than to FossilShield®.The two formulations of diatomaceous earth (1.5 g/kg) reduced the F₁ progeny by more than 70%, and produced less than 10% of seed damage and weight loss for the three varieties of cowpea. The varietal resistance and the use of diatomaceous earth against C. maculatus in storage could represent an alternative to Malagrain® for the protection of the cowpea.     

Effects of chemical- and bio-fertilizers on cowpea resistance to cowpea weevil,Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae)

The cowpea weevil, Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae) is a destructive pest of cowpea grains in the field and storage. Effects of two chemical fertilizers (triple superphosphate (TSP) and urea) and three bio-fertilizers (Bradyrhizobium japonicum, Pseudomonas putida and mycorrhizal fungi) compared to control were studied on resistance of cowpea pod and grain to C. maculatus. Pod resistance was evaluated as oviposition preference, egg-to-adult survival and developmental time, and grain resistance was measured as life history and life table parameters of the pest. Cowpea plants treated with tested fertilizers were grown under field condition. Then, the full-size green pods and their immature grains were harvested and used in the experiments. The highest oviposition preference and percentage of adults emergence were on control (untreated) pods as compared to treated ones. The longest developmental times were recorded on the pods obtained from TSP and B. japonicum treatments. In most cases, significant differences were observed for the life history and life table parameters of C. maculatus on the grains treated with examined fertilizers. Delayed developmental time was recorded when C. maculatus was fed with P. putida- and B. japonicum-treated grains. The highest and lowest number of eggs laid by each female (fecundity) was on the grains obtained from TSP and control treatments, respectively. The net reproductive rate (R₀), intrinsic rate of increase (r_m) and finite rate of increase (λ) were the lowest on cowpea grains fertilized with B. japonicum and highest on the grains treated with urea. Our results showed that B. japonicum can be suggested to utilize, as an alternative for chemical fertilizers, to minimize cowpea infestation by C. maculatus. These findings could be helpful in developing integrated management of the pest on cowpea.     

Effects of cowpea varietal susceptibility and low pressure on the mortality of life stages of Callosobruchus maculatus (Coleoptera: Bruchidae)

Low pressure creates a low-oxygen controlled atmosphere that can kill all developmental stages of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). This study investigated the interaction of low pressure with seed variety to determine the susceptibility of different cowpea varieties to developmental stages of C. maculatus. Laboratory studies were performed to determine exposure periods to low pressure (32.5 ± 1.0 mm Hg) necessary to kill all life stages of C. maculatus on different cowpea varieties. The results indicated that with the exception of eggs, differences in exposure periods required to achieve 100% mortality varied with life stage and variety. The larval stages were the most susceptible, and larvae required shorter exposure periods to low pressure when reared on C. maculatus resistant varieties (24-125B-3 and IT89KD-288) than on susceptible varieties (CB-3, CB-5, CB-46 and UC-27). Analysis of variance (ANOVA) showed a significant interaction between the life stages of C. maculatus, exposure time and cowpea varieties on mortality. Integration of low pressure applications with storage of cowpea varieties resistant to C. maculatus has the potential of providing an alternative pest management tool to fumigants, which are sometimes used in protecting cowpea in storage from infestation by C. maculatus.     

Resistance screening of 14 Cajanus cajan (L.) Millsp. cultivars to Callosobruchus maculatus (F.) (Coleoptera:Bruchidae)

Pigeonpea, Cajanus cajan, is a rich source of human dietary protein and carbohydrate as well as an income generator for farmers especially in developing countries. However, a major limiting factor to the viable production and post-harvest preservation of pigeonpea is infestation by the cowpea weevil, Callosobruchus maculatus. An experiment to screen for resistance in 14 newly developed C. cajan cultivars from The University of the West Indies (UWI) was conducted. The experiment compared oviposition preference based on eight parameters, namely, testa thickness, testa hardness, seed sphericity, surface area, total adult emergence, larval + pupal mortality, seed hardness and surface roughness. Four of the pigeon pea cultivars screened were resistant and could be ranked in the order of decreasing resistance as: A16 = A66PL1> A112 > A196PL2. Seed surface roughness, seed testa hardness and seed grain hardness were all found to play a major role in oviposition deterrence with seed roughness being the most effective deterrent. The results suggest that breeding for pigeonpea cultivars with rougher seed coats, harder seed testa and harder grains could enhance pigeonpea seed resistance to C. maculatus by reducing infestation and damage while in storage.     

Resistance of landrace and commercial cowpea genotypes to Callosobruchus maculatus attack

In the northern and north-eastern regions of Brazil, the cultivation of cowpea (Vigna unguiculata L.) is socially and economically important. However, attack by weevils makes storage difficult. Landrace genotypes exhibit high genetic variability, which can confer resistance to pest attacks. In this study, the susceptibility of landrace genotypes of cowpea to the attack of the weevil (Callosobruchus maculatus (Fabr., 1775)) was assessed. A total of 13 genotypes were used (11 landraces and two commercial genotypes). Biological parameters (number of eggs; daily, accumulated, and final emergence; the instantaneous rate of population increase) of C. maculatus, and loss of cowpea seed mass were analysed. The treatments were composed of 30g of each genotype and 20 adult insects in an entirely randomised design with ten replicates. A principal component analysis and grouping analysis were performed due to the similarity between the genotypes, and three groups were described. Group 3 (‘BRS Pujante’, ‘Ligeiro’, and ‘Tardão’ genotypes) had the highest oviposition/number of eggs, daily emergence, number of emerging insects, and instantaneous rate of population increase. Group 2 (‘Azulão’, ‘Costela de Vaca’, and ‘Rabo de Tatu’ genotypes) showed intermediate values, and group 1 (‘Bala’, ‘Canapú’, ‘Corujinha’, ‘Manteiguinha’, ‘Moita’, ‘Seridó’, and ‘Sempre Verde’ genotypes) had the lowest values for all those variables. The commercial genotypes were grouped in groups 1 (‘Sempre Verde’) and 3 (‘BRS Pujante’) and showed similar behaviour to the landrace genotypes of those groups. This study demonstrated that the genotypes in group 3 are susceptible to the attack of C. maculatus, while those in group 1 showed antibiosis resistance. Utilization of insect-resistant genotypes can reduce cowpea infestation and synthetic insecticide application, leading to a positive economic impact.     

Effect of different legume seeds on life table parameters of cowpea weevil,Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae)

The cowpea weevil, Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae) is one of the most important storage pests of legume seeds. The effect of various legume species including chickpea (Cicer arietinum L.) (cultivars Hashem and Mansour), cowpea (Vigna unguiculata L.) (cultivars Mashhad and 1057), green gram (Vigna radiata L.) (cultivar Parto), lentil (Lens culinaris Medikus) (cultivar Bilehsavar) was studied on the life history and life table parameters of C. maculatus at 30 ± 1 °C, relative humidity of 65 ± 5% and complete darkness. The developmental time was longest on lentil and shortest on cowpea 1057. The fecundity (number of eggs laid per reproductive period) of the pest was the lowest on lentil. The longest oviposition period was observed on chickpea Hashem. The gross and net reproductive rates were the highest on chickpea Mansour and lowest on lentil. The intrinsic rate of increase and finite rate of increase were the lowest when C. maculatus was reared on lentil and highest when it was reared on other hosts. The longest and shortest mean generation times were observed on lentil and cowpea 1057, respectively. According to the obtained results, lentil was relatively resistant and the other tested legumes were more susceptible hosts for feeding and population increase of C. maculatus. It is concluded that the resistant host cultivar can be proposed to be incorporated into breeding programs to minimize the economic losses incurred by C. maculatus.     

Automatic, rapid screening of seed resistance in cowpea, Vigna unguiculata (L.) Walpers, to the seed beetle Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) using acoustic monitoring

The biomonitor technique was investigated as a rapid and automatic method for measuring the resistance of cowpea varieties to the seed beetle Callosobruchus maculatus. This technique measures the activity of internally feeding insect larvae by counting ultrasonic emissions produced as they feed. Activity throughout the development of C. maculatus larvae in known susceptible and resistant cowpea varieties was recorded. This showed details of the development of each larval instar, and showed clear differences between the resistant and susceptible cowpeas. A rapid method for comparing cowpeas was proposed in which the activity of larvae was recorded for 24 h starting 14 days after oviposition. Using this method, significant differences in activity were apparent between one susceptible and two resistant cowpea varieties. Further comparisons using a randomised block experimental design also showed a clear difference in activity between one susceptible and one known resistant cowpea variety. This experimental protocol took 21 days. It is suggested that the biomonitor offers a promising method for screening seeds for resistance to insect pests with a reduction in the time and effort required over conventional bioassay methods.     

Correlation between biophysical seed characteristics of rice bean,Vigna umbellata (Fabaceae: Faboideae: Phaseoleae) and the development of Callosobruchus maculatus (Coleoptera: Chrysomelidae: Bruchinae)

The bruchid, Callosobruchus maculatus is a serious pest of stored pulses. Four biophysical seed characteristics viz. Seed size, texture of seed surface, thickness of seed coat and seed hardness of 10 genotypes of rice bean, a wild pulse crop were investigated for assessing mechanism of resistance against C. maculatus. Oviposition preference and larval penetration in seed coat did not differ significantly among different test genotypes. Highest number of larvae which developed up to last larval/pupal instar were observed in susceptible check variety of mungbean, PAU 911 (30.00) and lowest in rice bean genotype, LRB 535 (0.83). Highest and lowest seed weight was noticed in rice bean genotype, LRB 529 (6.81 g) and PAU 911 (3.88 g), respectively. Almost similar seed coat thickness was recorded among different test genotypes. Maximum seed hardness was noted in genotype LRB 507 (6.45 kg) whereas minimum in PAU 911 (2.00 kg) among all test genotypes. Any remarkable correlation of biophysical seed characteristics of test genotypes with ovipositon, larval penetration and development of C. maculatus was not observed. Thus, biophysical seed characteristics of rice bean did not suggest to play a vital role in imparting resistance against C. maculatus.     

Triple bag hermetic technology for post-harvest preservation of Bambara groundnut (Vigna subterranea (L.) Verdc.)

Experiments were conducted to evaluate the performance of hermetic triple bagging using Purdue Improved Crop Storage (PICS) bags for storage of Bambara groundnut (Vigna subterranea (L.) Verdc.). One set of experiments used grain heavily infested by Callosobruchus maculatus (F.) while a second set began with a low level infestation. Each experiment consisted of V. subterranea grain kept in four replicate 50 kg PICS bags or two replicate woven bags as controls. Two to five days after the beginning of the experiments, oxygen level inside the bags averaged about 21% (v/v) in the controls but decreased significantly in PICS bags, reaching 10% (v/v) with the heavily infested grain but falling only slightly in the lightly infested grain. After 7 months of storage, (i) the number of C. maculatus adults found between and within the grains, (ii) the 100 seed weight, (iii) the number of seed with holes, and (iv) the percentage germination of grain stored in PICS bags did not differ from what had been measured on the day that the experiment was set up. In woven bags, by contrast, there was a massive increase in C. maculatus numbers with means of 309 and 251 adults per 500 g in heavily and lightly infested grain, respectively. Grain weight losses in the woven bag controls ranged from 8 to 19% and the percentage of C. maculatus emergence holes per 100 seed increased from 51 to 135%.     

Toxicity of some citrus peels to Dermestes maculatus Deg. and Callosobruchus maculatus (F)

The toxicity of powdered sun-dried orange and grapefruit peels to Callosobruchus maculatus and Dermestes maculatus was evaluated in the laboratory. LD₅₀ values of orange and grapefruit peels admixed with cowpea grains on adult C. maculatus were 4.00 g (peel)/100 g (cowpea) and 5.62 g (peel/100 g (cowpea) respectively. LD₅₀ values of the ground peels on adult D. maculatus were much higher at 14.13 g (orange peel)/100 g (fish chips) and 14.29 g (grapefruit peel/100 g (fish chips). Orange and grapefruit peels deterred adult test insects from admixed cowpea and dried fish chips respectively. Orange peel at high dosages was also shown to depress progeny development of D. maculatus. The possible use of ground orange peel as a protectant for at least cowpea grains from C. maculatus infestation in small-scale storage in Nigeria is discussed.     

Semiochemical modulation of host preference of Callosobruchus maculatus on legume seeds

Host preference of Callosobruchus maculatus (F.) on seeds of three legume cultivars, Ife-brown and black-eyed cowpeas [Vigna unguiculata L. (Walp)], and soybean (Glycine max L.), was investigated. Mated female C. maculatus showed high (90–95%) attraction to the three legume cultivars in Y-tube bioassays. However, the weevils discriminated among the cultivars in four-choice tests and showed greater attraction to Ife-brown cowpea (50%) than to soybean (30%) and black-eyed cowpea (15%). Coupled gas chromatography-electroantennography (GC-EAD) and GC–MS analyses of the headspace volatile organic compounds (VOCs) emitted by the legume seeds identified 2-ethyl hexanol as the principal EAD active component. Emission of 2-ethyl hexanol was two-fold greater in Ife-brown cowpea (∼0.54 μg g⁻¹ seeds) compared with black-eyed cowpea (∼0.23 μg g⁻¹ seeds) and soybean (∼0.21 μg g⁻¹ seeds). Synthetic 2-ethyl hexanol attracted 68% of female C. maculatus at 0.01 μg dose in Y-tube bioassays. These results demonstrated that host preference in C. maculatus is odor-mediated, and identified 2-ethyl hexanol as a potential attractant for C. maculatus.     

Variation in the response of Callosobruchus maculatus (F.) to a resistant veriety of cowpea

A Nigerian cultivar of the cowpea has previously been identified as being less suitable than conventional varieties for the development of a local strain of C. maculatus (F.). The survival and duration of development of the immature stages of three strains of C. maculatus in the dried seeds of this cultivar were compared with the performance of the same strains of beetle on a susceptible variety. In all beetle strains, fewer larvae survived to produce adults, and the duration of development was significantly extended, in the resistant cowpea cultivar. A Brazilian strain produced fewest adults and a Nigerian strain had the longest mean development period. Combining all the data, it appeared that a strain from the Yemen Arab Republic was least influenced and that from Brazil most adversely affected by the resistant cowpea. Evidence is presented indicating that factors in both the cowpea seeds and the beetles themselves may contribute to the poorer performance of bruchids in the resistant as compared to the susceptible variety.     

Trehalose and proline failed to enhance cold tolerance of the cowpea weevil,Callosobruchus maculatus (F.) (Col.: Bruchidae)

The cowpea weevil, Callosobruchus maculatus (F.) (Col.: Bruchidae) is a cosmopolitan field-to-store pest ranked as the major post-harvest pest of cowpea in tropical regions. The cold tolerance of an insect species can vary as a result of abiotic features including food resources. In this study, C. maculatus larvae were fed with proline and trehalose (10, 20, and 40 mmol) treated cowpea seeds to determine the effects of these potential cryoprotectants on the supercooling (SCP) and cold hardiness of the upcoming adult beetles. The SCPs of the control, proline-fed and trehalose-fed adults non-significantly changed from −18.2 °C for the control to −17.2 °C for trehalose-fed adults. The cold hardiness (24 h at 0, -5.0, −7.5, −10.0, and −12.5 °C) of the adults was almost the same for control and treatments. Median lethal times (LT₅₀; lethal time for 50% mortality) were 6.3, 6.0, and 5.4 h, respectively. Moreover, feeding the larvae with proline and trehalose-treated seeds did not affect the proline and trehalose contents of the adult beetles. Our results showed that C. maculatus could not tolerate subzero temperatures well above their SCP, indicating that this species might be a chill-susceptible insect.     

Susceptibility of selected maize seed genotypes to Sitophilus zeamais (Coleoptera: Curculionidae)

Studies were carried out under laboratory conditions of 25 ± 2 °C and 70 ± 5% RH in an incubator to determine the relative susceptibility of eighteen maize genotypes to attack by the maize weevil, Sitophilus zeamais Motschulsky. The experiment was laid out in a Completely Randomized Design (CRD) with three replications. Adult mortality, number of F₁ progeny, weevil development time, susceptibility index, percentage seed damage, percentage weight loss, and weight of powder produced were determined after storage period of three months. The susceptibility index was determined using Dobie’s formula and the genotypes were classified into different susceptibility groups. The genotypes exhibited varying degrees of susceptibility to S. zeamais attack. Only Aseda was regarded as resistant and TZE-Y POP STR as moderately resistant to S. zeamais. Kpari-Faako, Tintim, WACCI-M-1215, WACCI-M-1594 and Wang-Dataa were regarded as moderately susceptible to S. zeamais. However, Abontem, Bihilifa, Ewul-Boyu, Sanzal-Sima, TZE-I 17, WACCI-M-1205, WACCI-M-1508 and WACCI-M-1510 were regarded as susceptible genotypes. Furthermore, Aburohemaa, Obaatanpa and Omankwa were regarded as highly susceptible to S. zeamais. Number of F₁ progeny, seed damage, seed weight loss, weight of dust produced and seed moisture content had positive relationship with varietal susceptibility while adult weevil mortality, median development period and seed germination after infestation had an inverse relationship with susceptibility in maize. The use of insect resistant genotypes would offer a sustainable way of minimizing postharvest losses of seeds in storage especially for smallholder farmers who keep harvested grains for future use as food and seed.     

Effect of monoterpenoids on oviposition and mortality of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) under hermetic conditions

Monoterpeniods have been shown to cause mortality in certain stored-product insect pests. The current report investigated the prospects of using monoterpenoids as oviposition deterrents of the cowpea beetle, Callosobruchus maculatus (Fabricius), as well as in the management of populations of the beetle. The monoterpenoids investigated include E-anethole, estragole, S-carvone, linalool, L-fenchone, geraniol, γ-terpinene and DL-camphor, and at the concentrations of 66.7, 33.3, 16.7, 8.33 and 0 μL/L. Exposure of the life stages of the beetle, which included eggs, young larvae (first instar), 4th instar, pupae and adults to different concentrations of the monoterpenoids over 24 h period caused varying levels of mortality. The stages of the beetle that were the least susceptible to the monoterpenoids were the 4th instar, and the pupae, which required high concentrations of the monoterpenoids to achieve 99% mortality. The adults and the eggs exhibited the highest susceptibility to the monoterpenoids. Mated C. maculatus females that were offered cowpea seeds upon treatment with low doses (8.33 μL/L) of the monoterpenoids did not lay eggs, while control female beetles offered untreated seeds laid several eggs. However, mated C. maculatus females laid eggs on cowpea seeds treated with monoterpenoids 3 weeks before to the day of experimentation. The monoterpenoids did not exhibit residual toxicity to the cowpea beetles. These monoterpenoids could be further investigated for the postharvest management of seed beetles of grain legumes.     

Identification,biochemical characterization and biological role of defense proteins from common bean genotypes seeds in response to Callosobruchus maculatus infestation

Common bean is a legume of significant socioeconomic importance and is cultivated worldwide. This crop is affected by several pests and diseases, which cause considerable economic losses and reduce yield. In recent years, several studies have demonstrated the role of proteins and peptides with activity against a wide range of insects and pathogens. The objective of this work was to identify defense proteins, such as antimicrobial peptides, protease and amylases inhibitors in common bean genotypes and evaluate the relationship of these proteins with Phaseolus vulgaris seed resistance to Callosobruchus maculatus infestation. Nineteen common bean genotypes were subjected to protein extraction, pH 5.4, and precipitation with ammonium sulfate at 70% saturation. The obtained extracts were separated by tricine gel electrophoresis. Experiments were carried out with natural seeds of common beans and artificial seeds (Vigna unguiculata seeds covered with seed coats of common beans) to evaluate the rate of oviposition and development of the insect species Callosobruchus maculatus. Lipid-transfer proteins were identified in nine genotypes whereas defensins were present in all genotypes. The inhibitory activity of α-amylases and trypsin and fungal development were determined in crude extracts (50 μg mL⁻¹). The results also indicated that the extracts from all bean genotypes inhibited the activity of human salivary α-amylase and C. maculatus larval α-amylase. Except for the extracts of four genotypes, all other extracts inhibited trypsin activity. None of the extracts from the evaluated bean genotypes inhibited the growth of tested fungi. Natural seeds from all genotypes did not inhibit insect oviposition, however, the larvae did not survive after feeding on these seeds. Artificial seeds containing seed coat flour of all genotypes inhibited the oviposition of C. maculatus, indicating that the seed coat was also repellent to insect.     

Efficacy of ozone for Callosobruchus maculatus and Callosobruchus chinensis control in cowpea seeds and its impact on seed quality

The insecticidal efficacy of ozone was evaluated against the adults of Callosobruchus maculatus (F.) and C. chinensis L. on stored cowpea seeds under laboratory conditions. Ozone was assessed at concentrations of 0.25, 0.5, 1.0, 1.5, and 2.0 g/m³. The effect of ozone treatments on the adult mortality after 1, 3, 5 and 7 days of treatment, progeny production after 45 days, cowpea seed weight loss, seed germination and chemical constituents of cowpea seeds were determined. Adult mortality of C. maculatus and C. chinensis was improved with the increase in ozone concentration. Thus, all tested concentrations caused complete adult mortality of C. chinensis after 7 days of treatment, while the concentrations of 1.0, 1.5, and 2.0 g/m³ caused complete adult mortality of C. maculatus after the same periods. Progeny of both species was significantly decreased in all ozone concentrations after 45 days of treatment. Nevertheless, strong suppression in progeny production was achieved at the highest concentration of ozone (2.0 g/m³). Also, the ozone treatment at the highest concentration protected the cowpea seeds from damage caused by C. maculatus and C. chinensis for 45 days. In addition, there was no significant effect of ozone treatments on the cowpea seed germination compared with untreated seeds. Moreover, the chemical analysis of treated cowpea seeds showed a slight decrease in protein, fat, carbohydrate, moisture, total phenolics, total flavonoids and tannins contents, and a slight increase in fiber and ash contents compared with untreated cowpea seeds. Our findings suggest the ozone can be effectively used for the control C. maculatus and C. chinensis and can provide sufficient protection of stored cowpea seeds.     

Primary and secondary metabolite compounds in cowpea seeds resistant to the cowpea bruchid [Callosobruchus maculatus (F.)] in postharvest storage

Cowpea is a one important legume crop in Sub-Saharan Africa due to its high content of protein and other nutrients. However, seeds of cowpea varieties are destroyed by Callosobruchus maculatus in storage. This study investigated various biochemical compounds of susceptible and resistant cowpea genotypes, to determine the biochemical compounds underlying cowpea resistance to bruchid. Six cowpea genotypes were analyzed in three replicates. One-way analysis of variance, Pearson correlation and path analysis were used to determine the influence of the biochemical compounds on the cowpea status. Amongst the cowpea genotypes assayed the biochemical compounds analyzed (Vicilin; α-amylase Inhibitor; Phenols; Condensed Tannin; Tannin; Carbohydrate; Flavonoid and Protein), were present in amounts. Resistant genotypes exhibited higher contents of Phenols (22.29 mg/g for WC66*5T and 19.74 mg/g for WC36); Tannin (2.45CEmg/g for WC36 and 2.52CEmg/g for TVU13677); and Carbohydrate (72.82% for TVU13677 and 71.09% for WC36). The most susceptible genotypes had the lowest content of Phenols (13.5 mg/g for TVU946 and 12.72 mg/g for Glessissaffodo); Tannin (0.74CEmg/g for Akounado and 0.97CEmg/g for Glessissaffodo) and Carbohydrate (60.95% for Akounado and 61.39% for TVU946), while Condensed Tannin was associated to their seed coat colour but not to their resistance status against bruchid. The resistant genotypes TVU13677; WC36 and WC66*5T were found to contain a higher amount of a subset of biochemical compounds as defensive compounds against cowpea bruchid. The susceptible cowpea genotypes (Akounado; Glessissaffodo; TVU946) screened for their biochemical content presented low amounts of these defensive compounds. The path analysis implemented highlighted the existence of causal relationships among biochemical compounds analyzed and resistance parameters, thus revealing the resistance basis of cowpea seed biochemical compounds. These latter results point out the possibilities to consider the presence of these biochemical compounds underlying the resistance of cowpea to bruchid in the breeding program toward the resistance to bruchid.     

Characterization of host response to bruchids (Callosobruchus chinensis and C. maculatus) in 39 genotypes belongs to 12 Cajanus spp. And assessment of molecular diversity inter se

Thirty nine genotypes belong to 12 Cajanus species were characterized for host response to bruchids (C. chinensis and C. maculatus) using no-choice based in vivo and in vitro antibiosis assay, and these assays revealed seven genotypes of primary gene pool (C. cajan) were more susceptible to bruchid infestation as compared to the genotypes of secondary and tertiary gene pools. Among all the genotypes, C. cajanifolius acc. ICPW-31 showed higher degree of resistance to both the bruchids. The molecular diversity using CDDP and SCoT markers showed wide range of genetic variations among the 12 species of Cajanus, and was supported by estimates of Nei's genetic distance and fixation index (F_ST) based haplotype matrix. The genetic structure showed clustering of 39 genotypes into eight distinct groups (K = 8) on the basis of their allelic composition, and among them C. cajanifolius acc. ICPW 31 and ICPW 30 showed close affinity with the cultivars of C. cajan by sharing several alleles. AMOVA analyses showed the existence of higher extent of genetic variation both at the genotype (37%) and species (63%) level in the genus Cajanus. The dendrogram and the principal coordinate analysis (PCoA) placed the 39 genotypes into six major clusters at par with their sectional classification. MCheza and ARLEQUIN based outlier analysis revealed 13 common loci under balancing selection, which are supposed to be involved in maintenance of genetic polymorphism either at species or genotype level, and are assigned to putative gene families such as KNOX, WRKY, ERF, MYB, ABP1 and MADS. The in vivo and in vitro bruchid assay vis-à-vis the DNA marker based molecular diversity analysis affirmed the possible use of C. cajanifolius acc. ICPW-31 as donor genotype for the introgression of bruchid resistance allele(s) into cultivated genetic background.