Identification of sources of resistance in cowpea mini core accessions to Aphis craccivora Koch (Homoptera: Aphididae) and their biochemical characterization.

Cowpea (Vigna unguiculata (L. Walp) is an important grain legume for human and livestock nutrition, especially in sub-Saharan Africa. Aphid, Aphis craccivora Koch (Homoptera: Aphididae), is one of the most widespread and destructive insect pests of cowpea and host-plant resistance is an effective approach to minimize the pest damage at seedling stage. This study was aimed at identifying resistant sources to A. craccivora within the cowpea mini core collection, a set of accessions from the largest world cowpea germplasm collection maintained at the International Institute of Tropical Agriculture (IITA). A total of 375 lines including 373 from IITA mini core collection, one resistant (TVu-801) and one susceptible (TVx-3236) checks were evaluated through artificial infestation in screening cages during the seedling stage. In cages, genotypes were planted in single rows containing four plants. They were arranged in an augmented design in which the two checks were sown in individual cages. Scoring for aphid population and damage levels were carried out on individual plants at 7, 14, and 21 days after planting. Advanced bioassays and biochemical analyses were conducted to investigate the mechanism of resistance to A. craccivora. Overall, three genotypes TVu-6464, TVu-1583, and TVu-15445 showed good levels of resistance comparable to the resistant check TVu-801. The HPLC analyses proved that both low sucrose levels in the plant, as well as high levels of kaempferol and quercetin, aglycones of phenolic compounds, were related with high resistance to aphids. The above genotypes with promising levels of resistance to A. craccivora will be used in cowpea breeding programs to develop improved resistant lines against this pest.

Functional Response of the Predators Alloeocranum biannulipes (Hemiptera: Reduviidae) and Teretrius nigrescens (Coleoptera: Histeridae) Feeding on Dinoderus porcellus (Coleoptera: Bostrichidae) Infesting Yam Chips.

The functional response and some predation parameters of the predators Alloeocranum biannulipes Montrouzier & Signoret (Hemiptera: Reduviidea) and Teretrius nigrescens Lewis (Coleoptera: Histeridae) were evaluated at five different densities of larvae and pupae of Dinoderus porcellus Lesne (Coleoptera: Bostrichidae) with the aim to understand their roles in the biological control of this major pest of stored yam chips. Experiments were performed in petri dishes at 25 ± 1 °C, 60 ± 10% RH, and a photoperiod of 12:12 (L:D) h in a controlled temperature room. Both predators showed Type II of functional response with respect to larvae, determined by a logistic regression model. However, T. nigrescens significantly killed more larvae of D. porcellus compared with A. biannulipes. This behavior, however, changed to a linear functional response (Type I), when pupae of D. porcellus were offered to both predators, possibly because of their immobility. In addition, there was no significant difference between T. nigrescens and A. biannulipes in terms of the killed pupae. Parameters of the Holling disc equation for both predators were estimated. Estimated handling time on larvae of D. porcellus for T. nigrescens and A. biannulipes was 0.254 and 0.677 h and the rate of searching efficiency was 0.289 and 0.348 h-1, respectively. Results indicated that T. nigrescens was a more suitable candidate for augmentative release for D. porcellus control than A. biannulipes. However, semifield studies are required to draw firm conclusions.

Insect fauna associated with Anacardium occidentale (Sapindales: Anacardiaceae) in Benin, West Africa.

Cashew, Anacardium occidentale L. (Sapindales: Anacardiaceae), is an important cash crop in Benin. However, its production is threatened by several biotic factors, especially insects. In Benin, very few studies have focused on insects and just listed species commonly found on cashew worldwide. The present investigation fills this gap by presenting an exhaustive inventory of insect species associated with this crop in the country. The survey was carried out from September 2009 to August 2010 in 22 cashew orchards (5 young and 17 mature) distributed over three major agroecological zones where cashew is most produced in the country. Insects were collected using chemical knock-down technique and visual observation followed by capture with sweep net. In addition, infested plant organs were sampled and incubated to collect emerging insects. In total, 262 insect species were recorded and identified. Among them, the wood borer Apate terebrans Pallas, the leafminer Eteoryctis gemoniella Stainton, and the mirid bugs Helopeltis schoutedeni Reuter., and Helopeltis anacardii Miller., appeared as the most important insect species attacking cashew in Benin. Beneficial insects encountered included some predators, parasitoids, and pollinators. Few vertebrate predators were also recorded on the trees. Differences in agroecological conditions or in field cleanliness did not affect the number of insect species encountered in the cashew orchards. The results of this study represent an important baseline data for the design and implementation of strategies for cashew protection in Benin.

Storage insects on yam chips and their traditional management in Northern Benin.

Twenty-five villages of Northern Benin were surveyed to identify the constraints of yam chips production, assess the diversity of storage insects on yam chips, and document farmers' perception of their impacts on the stocks and their traditional management practices. Damages due to storage insects (63.9% of responses) and insufficiency of insect-resistant varieties (16.7% of responses) were the major constraints of yam chips production. Twelve insect pest species were identified among which Dinoderus porcellus Lesne (Coleoptera, Bostrichidae) was by far the most important and the most distributed (97.44% of the samples). Three predators (Teretrius nigrescens Lewis, Xylocoris flavipes Reuter, and Alloeocranum biannulipes Montrouzier & Signoret) and one parasitoid (Dinarmus basalis Rondani) all Coleoptera, Bostrichidae were also identified. The most important traditional practices used to control or prevent insect attack in yam chips were documented and the producers' preference criteria for yam cultivars used to produce chips were identified and prioritized. To further promote the production of yam chips, diversification of insect-resistant yam varieties, conception, and use of health-protective natural insecticides and popularization of modern storage structures were proposed.

Modeling evolution of resistance by Maruca vitrata (Lepidoptera: Crambidae) to transgenic insecticidal cowpea in Africa.

We created a detailed model of the Maruca vitrata (F.) and cowpea [Vigna unguiculata (L.) Walp] system to study the possible evolution of resistance by the insect to transgenic insecticidal cowpea, which is under development. We focused on population dynamics and genetics in a region of west Africa. We simulated single-toxin and pyramided (two-toxin) cowpea and emphasized conservative, worst-case scenarios in our analysis. The results indicate that as long as a pyramided, transgenic cowpea can be developed, seed saving by farmers and reliance on natural refuge are not major problems for resistance management. Furthermore, it is possible that one or both toxins in the pyramid may not need to be high dose for evolution to be delayed significantly (>20 yr or 80 generations for resistance to become a concern if transgenic cowpea is deployed in areas where M. vitrata is endemic). If efforts are made to deploy transgenic cowpea only into the regions where M. vitrata is not endemic, then there is little to no concern with resistance emerging in the M. vitrata population.

The spatial genetic differentiation of the legume pod borer, Maruca vitrata F. (Lepidoptera: Crambidae) populations in West Africa.

The legume pod borer, Maruca vitrata, is an endemic insect pest that causes significant yield loss to the cowpea crop in West Africa. The application of population genetic tools is important in the management of insect pests but such data on M. vitrata is lacking. We applied a set of six microsatellite markers to assess the population structure of M. vitrata collected at five sites from Burkina Faso, Niger and Nigeria. Observed polymorphisms ranged from one (marker 3393) to eight (marker 32008) alleles per locus. Observed and expected heterozygosities ranged from 0.0 to 0.8 and 0.0 to 0.6, respectively. Three of the loci in samples from Nigeria and Burkina Faso deviated significantly from Hardy-Weinberg Equilibrium (HWE), whereas no loci deviated significantly in samples from Niger. Analysis of molecular variance (AMOVA) indicated that 67.3% level of the genetic variation was within individuals compared to 17.3% among populations. A global estimate of F ST=0.1 (ENA corrected F ST=0.1) was significant (P⩽0.05) and corroborated by pairwise F ST values that were significant among all possible comparisons. A significant correlation was predicted between genetic divergence and geographic distance between subpopulations (R2=0.6, P=0.04), and cluster analysis by the program STRUCTURE predicted that co-ancestry of genotypes were indicative of three distinct populations. The spatial genetic variance among M. vitrata in West Africa may be due to limited gene flow, south-north seasonal movement pattern or other reproductive barriers. This information is important for the cultural, chemical and biological control strategies for managing M. vitrata.

First Report of Soybean Witches'-Broom Disease Caused by Group 16SrII Phytoplasma in Soybean in Malawi and Mozambique.

Soybean (Glycine max L.) is an important grain legume cultivated on approximately 1.24 million ha in Africa (1). Malawi ranks fourth in area of production in Africa, with 75,000 ha in 2009 (1). Soybean is also gaining importance in Mozambique and several other southern African countries due to diversification programs. During a field survey conducted in March 2010, soybean plants with phyllody and witches'-broom disorders typical of phytoplasma infection were observed in three of five fields surveyed in Lilongwe (Chitedze Research Station) and Salima (Channa, Chitala) districts in Malawi and three of four fields surveyed in Zambezia Province in Mozambique. Symptoms consisted of shoot proliferation, reduced leaflets, shortened internodes, proliferated auxiliary shoots producing witches'-brooms, virescence, and phyllody. Incidence of symptomatic plants was <1% in Malawi and 10 to 15% in Mozambique. Yield loss was 100% in affected plants. Five leaf samples each from symptomatic and asymptomatic plants were collected from six fields; total genomic DNAs were isolated and used as templates in PCR using phytoplasma-universal primer pair P1 and P7 for 16S-23S ribosomal RNA encoding region (3). PCR amplicons (1,709 bp) were produced from only templates derived from symptomatic plants. Amplicons from a symptomatic plant each from Malawi (Channa, Salima District) and Mozambique (Mutequelse, Zambezia Province) were directly sequenced in both directions and submitted to the GenBank (Accession Nos. HQ840717 and HQ845208). Nucleotide sequences of the two African soybean witches'-broom (SoyWB) phytoplasma strains were 100% identical. The virtual restriction fragment length polymorphism (RFLP) pattern derived from these sequences using iPhyClassifier software (4) was similar to the reference pattern of the 16Sr group II, subgroup C (cactus phytoplasma, Accession No. AJ293216), with a pattern similarity coefficient of 0.99. A BLASTn search revealed that the African SoyWB phytoplasma sequences had a nucleotide sequence identity of 99% with those of soybean phytoplasma from Thailand (Accession No. EF193353), cactus phytoplasma from China (Accession No. EU099561), and several other members of 16SrII group. Phylogenetic analysis revealed the clustering of these strains with members of 16SrII group. In 1984, the occurrence of phyllody and witches'-broom symptoms in soybean in Mozambique was reported (2), however, no comprehensive details on the pathogen are available. To our knowledge, this is the first report of phyllody and witches'-broom disease in soybean in Malawi and the first molecular evidence of association of a 16SrII-C group 'Candidatus phytoplasma' with the disease in Malawi and Mozambique. Phyllody and witches'-broom is a destructive disease, and its widespread occurrence can adversely affect soybean production in sub-Saharan Africa. Identification of alternative hosts and vector species would improve our understanding of the disease's epidemiology and contribute to development of appropriate tactics to prevent escalation of this problem into a major disease. References: (1) FAOSTAT. http://faostat.fao.org/site/567/default.aspx . Retrieved 28 December 2010. (2) P. Plumb-Dhindsa and A. M. Mondjane. Trop. Pest Manage. 30:407, 1984. (3) L. B. Sharmila et al. J. Plant Biochem. Biotech. 13:1, 2004. (4) Y. Zhao et al. Int. J. Syst. Evol. Microbiol. 59:2582, 2009.

Minor components in the sex pheromone of legume podborer: Maruca vitrata development of an attractive blend.

The legume podborer, Maruca vitrata (syn. M. testulalis) (F.) (Lepidoptera: Pyralidae) is a pantropical pest of legume crops. Sex pheromone was collected by gland extraction or trapping of volatiles from virgin female moths originating in India, West Africa, or Taiwan. Analysis by GC-EAG and GC-MS confirmed previously published findings that (E,E)-10,12-hexadecadienal is the most abundant EAG-active component with 2-5% of (E,E)-10,12-hexadecadienol also present. At least one other EAG response was detected at retention times typical of monounsaturated hexadecenals or tetradecenyl acetates, but neither could be detected by GC-MS. Laboratory wind-tunnel bioassays and a field bioassay of blends of (E,E)-10,12-hexadecadienal with (E,E )-10,12-hexadecadienol and a range of monounsaturated hexadecenal and tetradecenyl acetate isomers indicated greatest attraction of males was to those including (E,E)-10,12-hexadecadienol and (E)-10-hexadecenal as minor components. In subsequent trapping experiments in cowpea fields in Benin, traps baited with a three-component blend of (E,E)-10,12-hexadecadienal and these two minor components in a 100:5:5 ratio caught significantly more males than traps baited with the major component alone, either two-component blend, or virgin female moths. Further blend optimization experiments did not produce a more attractive blend. No significant differences in catches were found between traps baited with polyethylene vials or rubber septa, or between lures containing 0.01 and 0.1 mg of synthetic pheromone. Significant numbers of female M. vitrata moths, up to 50% of total catches, were trapped with synthetic blends but not with virgin females. At present there is no clear explanation for this almost unprecedented finding, but the phenomenon may improve the predictive power of traps for population monitoring.

Resistance in accessions of cowpea to the coreid pod-bug Clavigralla tomentosicollis (Hemiptera: Coreidae).

Laboratory and screenhouse experiments were conducted to identify antibiosis and tolerance in four wild accessions of cowpea, Vigna unguiculata subsp. dekindtiana, to Clavigralla tomentosicollis Stål, the most damaging pod-sucking bug on cultivated Vigna genotypes in West Africa. The wild accession TVnu 151 showed antibiosis resistance, causing >50% mortality of the nymphs within 3 d of placing them on pods. Nymphs died more quickly on TVnu 151 than on TVnu 72, the wild and resistant control of the V. vexillata species which affected the weights of surviving insects to a much greater degree than TVnu 151. The three other accessions of the subspecies dekindtiana (TVnu 369, TVnu 517, and TVnu 707) did not cause significant mortality to the bugs, but rather extended their developmental time, with surviving adults showing lower weights and slower oviposition rates than those on the susceptible control IT84S-2246. Wild accessions affected male and female bugs differently, resulting in differential survival. This was reflected by the sex ratio which was male biased on the wild accessions (1:0.3-1:0.9), and female biased on IT84S-2246 (1:1.5). No evidence of tolerance was found in the four accessions of V. unguiculata subsp. dekindtiana. Overall, seed traits seemed to be the major resistance component in these wild accessions.

Interaction between pod age and position on damage to cowpea Vigna unguiculata by hemipteran pod-sucking bugs.

Laboratory and screenhouse studies were carried out to assess the relationship between pod age and pod position of cowpea and damage by different pod bug species. The coreids Clavigralla tomentosicollis Stäl and Riptortus dentipes Fabricius caused significant damage to young pods of cultivated genotypes, in contrast to the coreid Anoplocnemis curvipes Fabricius and the pentatomid Aspavia armigera Fabricius which exhibited minor feeding activity. Percent seed damage declined with pod age, the critical stage for pod bug infestation being when pods were about eight days old. Clavigralla tomentosicollis and R. dentipes caused significantly higher damage to pods located within the leaf canopy, thus behaving differently from Anoplocnemis curvipes which showed a distinct preference for pods growing above the leaf canopy. The feeding activity of Aspavia armigera was not affected by the position of pods on the plant. Overall, the study suggests that cowpea genotypes with a short flowering period and pods held above the leaf canopy offer the most promise in the management of pod-sucking pests.