I.B. Baoua

Mitochondrial genome sequence and expression profiling for the legume pod borer Maruca vitrata (Lepidoptera: Crambidae).

We report the assembly of the 14,054 bp near complete sequencing of the mitochondrial genome of the legume pod borer (LPB), Maruca vitrata (Lepidoptera: Crambidae), which we subsequently used to estimate divergence and relationships within the lepidopteran lineage. The arrangement and orientation of the 13 protein-coding, 2 rRNA, and 19 tRNA genes sequenced was typical of insect mitochondrial DNA sequences described to date. The sequence contained a high A+T content of 80.1% and a bias for the use of codons with A or T nucleotides in the 3rd position. Transcript mapping with midgut and salivary gland ESTs for mitochondrial genome annotation showed that translation from protein-coding genes initiates and terminates at standard mitochondrial codons, except for the coxI gene, which may start from an arginine CGA codon. The genomic copy of coxII terminates at a T nucleotide, and a proposed polyadenylation mechanism for completion of the TAA stop codon was confirmed by comparisons to EST data. EST contig data further showed that mature M. vitrata mitochondrial transcripts are monocistronic, except for bicistronic transcripts for overlapping genes nd4/nd4L and nd6/cytb, and a tricistronic transcript for atp8/atp6/coxIII. This processing of polycistronic mitochondrial transcripts adheres to the tRNA punctuated cleavage mechanism, whereby mature transcripts are cleaved only at intervening tRNA gene sequences. In contrast, the tricistronic atp8/atp6/coxIII in Drosophila is present as separate atp8/atp6 and coxIII transcripts despite the lack of an intervening tRNA. Our results indicate that mitochondrial processing mechanisms vary between arthropod species, and that it is crucial to use transcriptional information to obtain full annotation of mitochondrial genomes.

Transcriptome Sequencing, and Rapid Development and Application of SNP Markers for the Legume Pod Borer Maruca vitrata (Lepidoptera: Crambidae)

The legume pod borer, Maruca vitrata (Lepidoptera: Crambidae), is an insect pest species of crops grown by subsistence farmers in tropical regions of Africa. We present the de novo assembly of 3729 contigs from 454- and Sanger-derived sequencing reads for midgut, salivary, and whole adult tissues of this non-model species. Functional annotation predicted that 1320 M. vitrata protein coding genes are present, of which 631 have orthologs within the Bombyx mori gene model. A homology-based analysis assigned M. vitrata genes into a group of paralogs, but these were subsequently partitioned into putative orthologs following phylogenetic analyses. Following sequence quality filtering, a total of 1542 putative single nucleotide polymorphisms (SNPs) were predicted within M. vitrata contig assemblies. Seventy one of 1078 designed molecular genetic markers were used to screen M. vitrata samples from five collection sites in West Africa. Population substructure may be present with significant implications in the insect resistance management recommendations pertaining to the release of biological control agents or transgenic cowpea that express Bacillus thuringiensis crystal toxins. Mutation data derived from transcriptome sequencing is an expeditious and economical source for genetic markers that allow evaluation of ecological differentiation.

Geographic distribution of phylogenetically-distinct legume pod borer, Maruca vitrata (Lepidoptera: Pyraloidea: Crambidae)

Maruca vitrata Fabricius is a pantropical lepidopteran pest of legumes. Phylogenetic analysis of a mitochondrial cytochrome c oxidase-I gene (cox1) fragment indicates that three Maruca sp. mitochondrial lineages have unique geographic distributions [lineages 1 and 2: Australia, Taiwan, and West Africa (Niger, Nigeria, and Burkina Faso), and lineage 3: Puerto Rico]. The haplotype (T30, T114) is specific to lineages 1&2 and was assayed by NsiI and SacI polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) within population samples; it was not observed in the Puerto Rican samples, but was nearly fixed among samples from West Africa, Australia and Taiwan (85.5–100%). Re-sequencing and phylogenetic analyses of PCR-RFLP defined cox1 haplotypes indicate that nucleotide diversity is highest among samples from West Africa. Phylogenetic reconstruction based upon ribosomal DNA (rDNA) internal transcribed spacer-2 (ITS-2) sequences provided additional evidence for three Maruca sp. clades. These data suggest that multiple unique Maruca species or subspecies are present worldwide, which has implications for the management of this pest species-complex.

ANIMATED VIDEOS AS A LEARNING TOOL IN DEVELOPING NATIONS: A PILOT STUDY OF THREE ANIMATIONS IN MARADI AND SURROUNDING AREAS IN NIGER

Access to information and knowledge has always been a challenge for illiterate or low literate learners in developing nations. As a consequence, pedagogical advances, for low literate learners, have lagged behind, as compared to educational strategies for literate or highly literate learners. The technological revolution based on access to Internet and cell‐phones has brought new opportunities to one billion low literate learners in the planet. There is evidence that cell‐phones are becoming easily available for users in developing nations. “Scientific Animations Without Borders” (SAWBO) creates educational animations that can be watched on cell‐phones and other devices and can be transmitted from mobile phone to mobile phone through Bluetooth® technology. In this article we report the results of a short survey conducted by students from Maradi (Niger) with different groups; farmers, teachers in rural areas, informal entrepreneurs and women in which several 3 D animations were showed on mobile phones: cholera prevention, neem seed extract for insects pest control and triple bagging for storage to prevent post harvest looses. The objectives were to find out the perception of usefulness of the content and easiness of the technology used as a mode of transmission of information and knowledge.

Development of Reference Transcriptomes for the Major Field Insect Pests of Cowpea: A Toolbox for Insect Pest Management Approaches in West Africa

Cowpea is a widely cultivated and major nutritional source of protein for many people that live in West Africa. Annual yields and longevity of grain storage is greatly reduced by feeding damage caused by a complex of insect pests that include the pod sucking bugs, Anoplocnemis curvipes Fabricius (Hemiptera: Coreidae) and Clavigralla tomentosicollis Stål (Hemiptera: Coreidae); as well as phloem-feeding cowpea aphids, Aphis craccivora Koch (Hemiptera: Aphididae) and flower thrips, Megalurothrips sjostedti Trybom (Thysanoptera: Thripidae). Efforts to control these pests remain a challenge and there is a need to understand the structure and movement of these pest populations in order to facilitate the development of integrated pest management strategies (IPM). Molecular tools have the potential to help facilitate a better understanding of pest populations. Towards this goal, we used 454 pyrosequencing technology to generate 319,126, 176,262, 320,722 and 227,882 raw reads from A. curvipes, A. craccivora, C. tomentosicollis and M. sjostedti, respectively. The reads were de novo assembled into 11,687, 7,647, 10,652 and 7,348 transcripts for A. curvipes, A. craccivora, C. tomentosicollis and M. sjostedti, respectively. Functional annotation of the resulting transcripts identified genes putatively involved in insecticide resistance, pathogen defense and immunity. Additionally, sequences that matched the primary aphid endosymbiont, Buchnera aphidicola, were identified among A. craccivora transcripts. Furthermore, 742, 97, 607 and 180 single nucleotide polymorphisms (SNPs) were respectively predicted among A. curvipes, A. craccivora, C. tomentosicollis and M. sjostedti transcripts, and will likely be valuable tools for future molecular genetic marker development. These results demonstrate that Roche 454-based transcriptome sequencing could be useful for the development of genomic resources for cowpea pest insects in West Africa.

Biological control of the millet head miner Heliocheilus albipunctella in the Sahelian region by augmentative releases of the parasitoid wasp Habrobracon hebetor: effectiveness and farmers’ perceptions

On-farm augmentative releases of the parasitoid Habrobracon hebetor (Say) for controlling the millet head miner (MHM) Heliocheilus albipunctella (de Joannis) were tested in Burkina Faso, Mali and Niger from 2007 to 2009. In addition, a survey of farmers’ perceptions of insect pests, with particular focus on MHM, and the biological control program (BCP) was carried out. There was a significant increase of MHM parasitization rate after the releases, with up to 97% mortality. The survey on farmers’ perceptions revealed a fair knowledge of the MHM and the ability of farmers to describe the pest and the damage it caused. Farmers claimed that the biocontrol agent H. hebetor is effective and perceived a significant gain in grain yield due to this control strategy. Implications of these findings for a large extension of the MHM biocontrol program are discussed.

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.

Potential use of Sesbania pachycarpa (Fabace ae: Papilionoideae) as a refugia for the legume pod borer Maruca vitrata (Lepidoptera: Crambidae)

Deployment of cowpea with Bacillus thuringiensis (Bt-cowpea), to control Maruca vitrata Fab., must be preceded by the development of an insect resistance management (IRM) plan to ensure a sustainable use of the in-plant protection offered by the transgenic variety. One of the components of a resistance management plan involves the use of wild or cultivated host plants as refugia. In West Africa, wild refugia have the potential to be a major component of such an IRM strategy. In the current study, we examined the occurrence of M. vitrata on three cultivated cowpea varieties and one wild alternative host, Sesbania pachycarpa D.C. Our results indicate that M. vitrata population overlapped on the wild host and cowpea. The overall insect population on S. pachycarpa represents 9–13% of the population on cultivated cowpea. Based on these findings, we suggest that S. pachycarpa may contribute as a refuge for M. vitrata population in case of deployment of Bt-cowpea.

Triple bag hermetic technology for controlling a bruchid (Spermophagus sp.) (Coleoptera, Chrysomelidae) in stored Hibiscus sabdariffa grain

We assessed the performance of hermetic triple layer Purdue Improved Crop Storage (PICS) bags for protecting Hibiscus sabdariffa grain against storage insects. The major storage pest in the grain was a bruchid, Spermophagus sp.. When we stored infested H. sabdariffa grain for six months in the woven polypropylene bags typically used by farmers, the Spermophagus population increased 33-fold over that initially present. The mean number of emergence holes per 100 seeds increased from 3.3 holes to 35.4 holes during this time period, while grain held for the same length of time in PICS bags experienced no increase in the numbers of holes. Grain weight loss in the woven control bags was 8.6% while no weight loss was observed in the PICS bags. Seed germination rates of grain held in woven bags for six months dropped significantly while germination of grain held in PICS bags did not change from the initial value. PICS bags can be used to safely store Hibiscus grain after harvest to protect against a major insect pest.

Wild host plants of legume pod borer Maruca vitrata (Lepidoptera: Pyraloidea: Crambidae) in southern Niger and northern Nigeria

We surveyed three locations in West Africa, two in northern Nigeria and one in southern Niger, for the presence of wild alternative hosts of the legume pod borer (LPB) Maruca vitrata Fabricius, which could serve as natural unstructured refugia for transgenic Bacillus thuringiensis (Bt)-cowpea. We surveyed a total of 67 sites in proximity to cowpea fields in the Sudan Savannah (Kano, Nigeria), the Northern Guinea Savannah (Zaria, Nigeria) and the Sahel Savannah (Maradi, Niger) ecological zones. The results indicate that alternative host plants for LPB are scarce or absent during the cowpea-growing season in these areas.