Carry-Over Niches for Lepidopteran Maize Stemborers and Associated Parasitoids during Non-Cropping Season.

Sources of infestation are the key elements to be considered in the development of habitat management techniques for the control of maize stemborers. Several wild plants, grasses mostly, have been identified that serve as hosts for stemborers and their parasitoids during the off-season when maize is not present in the field. However, their abundance is much lower in wild plants compared to cultivated fields. Thus, the role of wild plants as a reservoir for cereal stemborers and their parasitoids is still controversial, particularly in agro-ecosystems with reduced wild habitat. We studied the occurrence of different maize stemborers and associated parasitoids in maize stem residues and wild grasses during non-cropping seasons as potential carry-over populations to subsequent early season maize plants. Surveys were conducted in the central region of Kenya during long and short dry seasons in maize residues and wild grasses as well as during the two rainy seasons in maize plants at earlier and late whorl stages during the years of 2017 and 2018. Wild habitat had a higher species diversity than maize residues habitat, but maize residues had a higher abundance of maize stemborer species, such as Busseola fusca, Sesamia calamistis, and Chilo partellus, and of associated parasitoid species (i.e., Cotesia flavipes and Cotesia sesamiae) than wild plants. Our surveys, complemented by field parasitoid releases of C. flavipes and C. sesamiae, indicated that maize residues constitute a better refugia reservoir not only of the maize stemborers but also of C. flavipes and C. sesamiae during non-cropping seasons as compared to wild plants and, thus, might constitute in this region the main source of both stemborers and C. flavipes/C. sesamiae carry-over in maize plants during the subsequent cropping season. Thus, systematic destruction of maize residues would not help the biological control of lepidopteran stemborers. This is particularly true in areas with reduced wild habitat.

Spatio-Temporal Interactions Between Maize Lepidopteran Stemborer Communities and Possible Implications From the Recent Invasion of Spodoptera frugiperda (Lepidoptera: Noctuidae) in Sub-Saharan Africa.

Spatio-temporal dynamics of multi-species pest communities and the interactions between them influence the structure of pest complex that attack crops. In East and Southern Africa, cereal crops, especially maize, is attacked by a complex of lepidopteran stemborer species made up of Busseola fusca (Fuller) and Sesamia calamistis Hampson (Lepidoptera: Noctuidae), and Chilo partellus (Swinhoe) (Lepidoptera: Crambidae). There is inadequate information on the extent of single- and multi-species infestations by this pest complex, their current spatio-temporal variations, and the primary abiotic factors that influence these. Furthermore, the recent invasion of the fall armyworm, Spodoptera frugiperda J.E. Smith, in sub-Saharan Africa will likely influence this stemborer community structure. Sampling was conducted in maize fields to record stemborer species and larval numbers from infested plants, in 28 localities found in six agro-ecological zones (AEZs) of Kenya and parts of Tanzania, as well as in one locality in the mid-altitudes where the three stemborer species occurred together. Both single- and multi-species stemborer communities characterized infestation of maize at field and plant levels, but varied in proportions between the AEZs. Infestation patterns and larval densities varied between seasons at mid-locality stemborer communities followed a clustered distribution pattern. Temperature was the most significant abiotic factor influencing the composition of stemborer communities at all spatial scales. Rainfall was significant only at the local scale. Results are discussed in relation to current stemborer community structures in maize fields and what the likely potential implications are, in the light of climate change and the recent establishment of the fall armyworm in Africa.

Phylogeography and Population Structure of the Mediterranean Corn Borer, Sesamia nonagrioides (Lepidoptera: Noctuidae), Across Its Geographic Range.

Sesamia nonagrioides (Lefèbvre) (Lepidoptera: Noctuidae), is a widespread insect pest in Africa, the Middle East, and Europe. However, its pest status varies across its distribution range. It is a major pest of maize in Europe and of sugarcane in Iran. In Africa, it is a major pest of maize in West Africa but not considered as a pest in East Africa. Recent surveys conducted in 2015 recorded S. nonagrioides to be a major pest of sugarcane in Ethiopia and reported the species for the first time in Botswana, outside its known geographic range. The genetic relationship of these records with the previously recorded population of S. nonagrioides was investigated using the cytochrome oxidase subunit I region of the mitochondrial genome. In total, 113 individuals across the geographic range of the species were analyzed and 63 haplotypes were identified. Phylogenetic analysis separated the populations into two clades with no distinct geographic distribution pattern. The genetic differentiation was also not associated with host plants and geographic distances. Results of the molecular analysis revealed the long-time establishment of S. nonagrioides population in Botswana and identified the newly recorded sugarcane population from Ethiopia as part of the wild host population in the country. The phylogeographic patterns observed among population of S. nonagrioides have probably been shaped by Pleistocene's climatic oscillations and geographic range expansions from different refugia with secondary contact and admixture. Possible reasons for the host-plant expansion by the Ethiopian population are discussed.

Genetic footprints of adaptive divergence in the bracovirus of Cotesia sesamiae identified by targeted resequencing.

The African parasitoid wasp Cotesia sesamiae is a generalist species structured in locally adapted populations showing differences in host range. The recent discovery of Cotesia typhae, a specialist, sister species to C. sesamiae, provides a good framework to study the genetic determinants of parasitoid host range. To investigate the genomic bases of divergence between these populations and species, we used a targeted sequencing approach on 24 samples. We targeted the bracovirus genomic region encoding virulence genes involved in the interaction with the lepidopteran hosts of the wasps. High sequencing coverage was obtained for all samples, allowing the study of genetic variation between wasp populations and species. By combining population genetic estimations, such as nucleotide diversity (π), relative differentiation (FST ) and absolute divergence (dxy ), with branch-site dN/dS measures, we identified six of 98 bracovirus genes showing significant divergence and evidence of positive selection. These genes, belonging to different gene families, are potentially involved in host adaptation and in the specialization process. Fine-scale analyses of genetic variation also revealed mutations and large deletions in certain genes inducing pseudogenization and loss of function. The image emerging from these results is that adaptation mediated by bracovirus genes happens through selection of particularly adaptive alleles and loss of nonadaptive genes. These results highlight the central role of the bracovirus in the molecular interactions between the wasps and their hosts and in the evolutionary processes of specialization.

Influence of Temperature on Intra- and Interspecific Resource Utilization within a Community of Lepidopteran Maize Stemborers.

Competition or facilitation characterises intra- and interspecific interactions within communities of species that utilize the same resources. Temperature is an important factor influencing those interactions and eventual outcomes. The noctuid stemborers, Busseola fusca and Sesamia calamistis and the crambid Chilo partellus attack maize in sub-Saharan Africa. They often occur as a community of interacting species in the same field and plant at all elevations. The influence of temperature on the intra- and interspecific interactions among larvae of these species, was studied using potted maize plants exposed to varying temperatures in a greenhouse and artificial stems kept at different constant temperatures (15°C, 20°C, 25°C and 30°C) in an incubator. The experiments involved single- and multi-species infestation treatments. Survival and relative growth rates of each species were assessed. Both intra- and interspecific competitions were observed among all three species. Interspecific competition was stronger between the noctuids and the crambid than between the two noctuids. Temperature affected both survival and relative growth rates of the three species. Particularly at high temperatures, C. partellus was superior in interspecific interactions shown by higher larval survival and relative growth rates. In contrast, low temperatures favoured survival of B. fusca and S. calamistis but affected the relative growth rates of all three species. Survival and relative growth rates of B. fusca and S. calamistis in interspecific interactions did not differ significantly across temperatures. Temperature increase caused by future climate change is likely to confer an advantage on C. partellus over the noctuids in the utilization of resources (crops).

Ongoing ecological speciation in Cotesia sesamiae, a biological control agent of cereal stem borers.

To develop efficient and safe biological control, we need to reliably identify natural enemy species, determine their host range, and understand the mechanisms that drive host range evolution. We investigated these points in Cotesia sesamiae, an African parasitic wasp of cereal stem borers. Phylogenetic analyses of 74 individual wasps, based on six mitochondrial and nuclear genes, revealed three lineages. We then investigated the ecological status (host plant and host insect ranges in the field, and host insect suitability tests) and the biological status (cross-mating tests) of the three lineages. We found that one highly supported lineage showed all the hallmarks of a cryptic species. It is associated with one host insect, Sesamia nonagrioides, and is reproductively isolated from the other two lineages by pre- and postmating barriers. The other two lineages had a more variable phylogenetic support, depending on the set of genes; they exhibited an overlapping and diversified range of host species and are not reproductively isolated from one another. We discuss the ecological conditions and mechanisms that likely generated this ongoing speciation and the relevance of this new specialist taxon in the genus Cotesia for biological control.

Predicting the Impact of Temperature Change on the Future Distribution of Maize Stem Borers and Their Natural Enemies along East African Mountain Gradients Using Phenology Models.

Lepidopteran stem borers are among the most important pests of maize in East Africa. The objective of the present study was to predict the impact of temperature change on the distribution and abundance of the crambid Chilo partellus, the noctuid Busseola fusca, and their larval parasitoids Cotesia flavipes and Cotesia sesamiae at local scale along Kilimanjaro and Taita Hills gradients in Tanzania and Kenya, respectively. Temperature-dependent phenology models of pests and parasitoids were used in a geographic information system for mapping. The three risk indices namely establishment, generation, and activity indices were computed using current temperature data record from local weather stations and future (i.e., 2055) climatic condition based on downscaled climate change data from the AFRICLIM database. The calculations were carried out using index interpolator, a sub-module of the Insect Life Cycle Modeling (ILCYM) software. Thin plate algorithm was used for interpolation of the indices. Our study confirmed that temperature was a key factor explaining the distribution of stem borers and their natural enemies but other climatic factors and factors related to the top-down regulation of pests by parasitoids (host-parasitoid synchrony) also played a role. Results based on temperature only indicated a worsening of stem borer impact on maize production along the two East African mountain gradients studied. This was attributed to three main changes occurring simultaneously: (1) range expansion of the lowland species C. partellus in areas above 1200 m.a.s.l.; (2) increase of the number of pest generations across all altitudes, thus by 2055 damage by both pests will increase in the most productive maize zones of both transects; (3) disruption of the geographical distribution of pests and their larval parasitoids will cause an improvement of biological control at altitude below 1200 m.a.s.l. and a deterioration above 1200 m.a.s.l. The predicted increase in pest activity will significantly increase maize yield losses in all agro-ecological zones across both transects but to a much greater extent in lower areas.

Adaptive selection on bracovirus genomes drives the specialization of Cotesia parasitoid wasps.

The geographic mosaic of coevolution predicts parasite virulence should be locally adapted to the host community. Cotesia parasitoid wasps adapt to local lepidopteran species possibly through their symbiotic bracovirus. The virus, essential for the parasitism success, is at the heart of the complex coevolutionary relationship linking the wasps and their hosts. The large segmented genome contained in the virus particles encodes virulence genes involved in host immune and developmental suppression. Coevolutionary arms race should result in the positive selection of particular beneficial alleles. To understand the global role of bracoviruses in the local adaptation or specialization of parasitoid wasps to their hosts, we studied the molecular evolution of four bracoviruses associated with wasps of the genus Cotesia, including C congregata, C vestalis and new data and annotation on two ecologically differentiated populations of C sesamie, Kitale and Mombasa. Paired orthologs analyses revealed more genes under positive selection when comparing the two C sesamiae bracoviruses belonging to the same species, and more genes under strong evolutionary constraint between species. Furthermore branch-site evolutionary models showed that 17 genes, out of the 54 currently available shared by the four bracoviruses, harboured sites under positive selection including: the histone H4-like, a C-type lectin, two ep1-like, ep2, a viral ankyrin, CrV1, a ben-domain, a Serine-rich, and eight unknown genes. Lastly the phylogenetic analyses of the histone, ep2 and CrV1 genes in different African C sesamiae populations showed that each gene described differently the individual relationships. In particular we found recombination had happened between the ep2 and CrV1 genes, which are localized 37.5 kb apart on the wasp chromosomes. Involved in multidirectional coevolutionary interactions, C sesamiae wasps rely on different bracovirus mediated molecular pathways to overcome local host resistance.

Parasitism of lepidopterous stem borers in cultivated and natural habitats.

Plant infestation, stem borer density, parasitism, and parasitoid abundance were assessed during two years in two host plants, Zea mays (L.) (Cyperales: Poaceae) and Sorghum bicolor (L.) (Cyperales: Poaceae), in cultivated habitats. The four major host plants (Cyperus spp., Panicum spp., Pennisetum spp., and Sorghum spp.) found in natural habitats were also assessed, and both the cultivated and natural habitat species occurred in four agroecological zones in Kenya. Across habitats, plant infestation (23.2%), stem borer density (2.2 per plant), and larval parasitism (15.0%) were highest in maize in cultivated habitats. Pupal parasitism was not higher than 4.7% in both habitats, and did not vary with locality during each season or with host plant between each season. Cotesia sesamiae (Cameron) and C. flavipes Cameron (Hymenoptera: Braconidae) were the key parasitoids in cultivated habitats (both species accounted for 76.4% of parasitized stem borers in cereal crops), but not in natural habitats (the two Cotesia species accounted for 14.5% of parasitized stem borers in wild host plants). No single parasitoid species exerted high parasitism rates on stem borer populations in wild host plants. Low stem borer densities across seasons in natural habitats indicate that cereal stem borer pests do not necessarily survive the non-cropping season feeding actively in wild host plants. Although natural habitats provided refuges for some parasitoid species, stem borer parasitism was generally low in wild host plants. Overall, because parasitoids contribute little in reducing cereal stem borer pest populations in cultivated habitats, there is need to further enhance their effectiveness in the field to regulate these pests.

Intraspecific specialization of the generalist parasitoid Cotesia sesamiae revealed by polyDNAvirus polymorphism and associated with different Wolbachia infection.

As a result of an intense host-parasite evolutionary arms race, parasitic wasps frequently display high levels of specialization on very few host species. For instance, in braconid wasps very few generalist species have been described. However, within this family, Cotesia sesamiae is a generalist species that is widespread in sub-Saharan Africa and develops on several lepidopteran hosts. In this study, we tested the hypothesis that C. sesamiae may be a cryptic specialist when examined at the intraspecific level. We sequenced exon 2 of CrV1, a gene of the symbiotic polyDNAvirus that is integrated into the wasp genome and is associated with host immune suppression. We found that CrV1 genotype was more closely associated with the host in which the parasitoid developed than any abiotic environmental factor tested. We also tested a correlation between CrV1 genotype and an infection with Wolbachia bacteria, which are known for their ability to induce reproductive isolation. The Wolbachia bacteria infection polymorphism was also found as a major factor explaining the genetic structure of CrV1, and, in addition, the best model explaining CrV1 genetic structure involved an interaction between Wolbachia infection and host species. We suggest that Wolbachia could act as an agent capable of maintaining advantageous alleles for host specialization in different populations of C. sesamiae. This mechanism could be applicable to other insect models because of the high prevalence of Wolbachia in insects.

Factors affecting stem borer parasitoid species diversity and parasitism in cultivated and natural habitats.

The effects of biotic and abiotic factors on stem borer parasitoid diversity, abundance, and parasitism were studied in cultivated and natural habitats in four agroecological zones in Kenya. Comparing habitat types, we found partial support for the "natural enemy" hypothesis, whereby, across all localities, parasitoid diversity was higher in more diverse host plant communities in natural habitats, whereas parasitoid abundance was higher in cultivated habitats. For both habitats, parasitoid richness was mainly influenced by stem borer density and/or its interaction with stem borer richness, whereas parasitoid abundance was mainly affected by stem borer abundance. Parasitoid richness was higher in localities (with bimodal rainfall distribution) with increased spatial and temporal availability of host plants that harbored the borers. Across seasons, parasitoid richness was lower in both cultivated and natural habitats in the driest locality, Mtito Andei. Overall, parasitoid diversity was low in Suam and Mtito Andei, where maize cultivation was practiced on a commercial scale and intense grazing activities persist across seasons, respectively. Across localities, habitats, and seasons, stem borer parasitism was positively correlated with parasitoid richness and abundance. Furthermore, the interaction of rainfall and altitude influenced the presence and absence of parasitoids, and consequently, stem borer parasitism. Parasitism was positively and negatively correlated with temperature in cultivated and natural habitats, respectively. Overall, natural habitats seem to serve as important refugia for sustaining parasitoid diversity, which in turn can affect stem borer parasitism in the cereal cropping system.

Evolution of a polydnavirus gene in relation to parasitoid-host species immune resistance.

CrV1, a polydisperse DNA virus (polydnavirus or PDV) gene contributes to the suppression of host immunity in Cotesia genus parasitoids. Its molecular evolution was analyzed in relation to levels of resistance in the sympatric host species. Natural selection for nonsynonymous substitutions (positive Darwinian selection) was observed at specific amino acid sites among CrV1 variants; particularly, between parasitoid strains immune suppressive and nonimmune suppressive to the main resistant stem borer host, Busseola fusca. In Cotesia sesamiae, geographic distribution of CrV1 alleles in Kenya was correlated to the relative abundance of B. fusca. These results suggest that PDV genes evolve through natural selection and are genetically linked to factors of suppression of local host resistance. We discuss the forces driving the evolution of CrV1 and its use as a marker to understand parasitoid adaptation to host resistance in biological control.