Johnnie Van den Berg

Dominant inheritance of field-evolved resistance to Bt corn in Busseola fusca

Transgenic crops expressing Bacillus thuringiensis (Bt) toxins have been adopted worldwide, notably in developing countries. In spite of their success in controlling target pests while allowing a substantial reduction of insecticide use, the sustainable control of these pest populations is threatened by the evolution of resistance. The implementation of the “high dose/refuge” strategy for managing insect resistance in transgenic crops aims at delaying the evolution of resistance to Bt crops in pest populations by promoting survival of susceptible insects. However, a crucial condition for the “high dose/refuge” strategy to be efficient is that the inheritance of resistance should be functionally recessive. Busseola fusca developed high levels of resistance to the Bt toxin Cry 1Ab expressed in Bt corn in South Africa. To test whether the inheritance of B . fusca resistance to the Bt toxin could be considered recessive we performed controlled crosses with this pest and evaluated its survival on Bt and non-Bt corn. Results show that resistance of B . fusca to Bt corn is dominant, which refutes the hypothesis of recessive inheritance. Survival on Bt corn was not lower than on non-Bt corn for both resistant larvae and the F1 progeny from resistant × susceptible parents. Hence, resistance management strategies of B . fusca to Bt corn must address non-recessive resistance.

Ecology of the African Maize Stalk Borer, Busseola fusca (Lepidoptera: Noctuidae) with Special Reference to Insect-Plant Interactions

Busseola fusca (Lepidoptera: Noctuidae) is an important pest of maize and sorghum in sub-Saharan Africa. One century after its first description by Fuller in 1901, inaccurate information based on earlier reports are still propagated on its distribution (e.g., absent from the lower altitudes in East Africa) and host plant range (e.g., feeding on a large range of wild grass species). This review provides updated information on the biology, distribution and genetics of B. fusca with emphasis on insect-plant interactions. Related to this, new avenues of stem borer management are proposed.

Diversity and comparative phenology of Lepidoptera on Bt and non-Bt maize in South Africa

Abstract The use of Bt maize could directly or indirectly affect non-target organisms. Assessment of the impacts of Bt crops is hampered by the lack of even the most basic checklist of the species present in most systems. In South Africa the maize agroecosystem is of particular interest because of the large area planted with Bt maize (Event MON810) for the control of Busseola fusca (Fuller) (Lepidoptera: Noctuidae) and Chilo partellus (Swinhoe) (Lepidoptera: Crambidae) populations. The aims of this study were to determine the diversity of Lepidoptera species that feed on or are closely associated with maize, and to assess the possible effects of Bt maize on the incidence of Lepidoptera damage under field conditions. Field surveys were conducted at 24 sites during the 2004/2005 and 2005/2006 cropping seasons. Fifteen species of Lepidoptera were recorded on maize. Six of these species were recorded to feed on Bt maize. Limited information is available on host plants and biology of some of these species. The...Abstract The use of Bt maize could directly or indirectly affect non-target organisms. Assessment of the impacts of Bt crops is hampered by the lack of even the most basic checklist of the species present in most systems. In South Africa the maize agroecosystem is of particular interest because of the large area planted with Bt maize (Event MON810) for the control of Busseola fusca (Fuller) (Lepidoptera: Noctuidae) and Chilo partellus (Swinhoe) (Lepidoptera: Crambidae) populations. The aims of this study were to determine the diversity of Lepidoptera species that feed on or are closely associated with maize, and to assess the possible effects of Bt maize on the incidence of Lepidoptera damage under field conditions. Field surveys were conducted at 24 sites during the 2004/2005 and 2005/2006 cropping seasons. Fifteen species of Lepidoptera were recorded on maize. Six of these species were recorded to feed on Bt maize. Limited information is available on host plants and biology of some of these species. The incidences of Lepidoptera-infested plants and infestation levels were generally lower in Bt fields than in non-Bt fields possibly indicating that these species may be affected by Bt maize. Results showed that B. fusca larvae were able to survive on Bt maize at low population levels. On the basis of exposure and risk of resistance development, Helicoverpa armigera and Acantholeucania loreyi can be considered to be important non-target lepidopteran herbivores on maize in South Africa.

The potential global distribution of Chilo partellus, including consideration of irrigation and cropping patterns

Chilo partellus is a major crop pest in Asia and Africa, and has recently spread to the Mediterranean region. Knowledge of its potential distribution can inform biosecurity policies aimed at limiting its further spread and efforts to reduce its impact in areas that are already invaded. Three models of the potential distribution of this insect have been published, each with significant shortcomings. We re-parameterized an existing CLIMEX model to address some parameter inconsistencies and to improve the fit to the known distribution of C. partellus. The resulting model fits the known distribution better than previous models, highlights additional risks in equatorial regions and reduces modelled risks in wet and extremely dry regions. We bring new insights into the role of irrigation in the potential spread of this invasive insect and compare its potential distribution with the present known distribution of its hosts. We also distinguish regions that are suitable for supporting persistent populations from those that may be at risk from ephemeral populations during favourable seasons. We present one of the first demonstrations of a new capability in CLIMEX to automatically estimate parameter sensitivity and model uncertainty. Our CLIMEX model highlights the substantial invasion risk posed by C. partellus to cropping regions in the Americas, Australia, China, Europe, New Zealand and West Africa. Its broad host range and reported impacts suggest that it should be a pest of significant concern to biosecurity agencies in these presently uninvaded regions.

Integrative taxonomy of Acrapex stem borers (Lepidoptera : Noctuidae : Apameini): combining morphology and Poisson Tree Process analyses

Abstract. Ten morphologically similar species of Acrapex from eastern and south-eastern Africa belonging to the A. stygiata and A. albivena groups are reviewed. Six species are described as new: A. brunneella, A. mitiwa, A. mpika, A. salmona, A. sporobola and A. yakoba. The Poaceae host plants of eight species are recorded; four species, A. mitiwa. A. subalbissima, A. syscia and A. yakoba, were found developing exclusively on Imperata cylindrica (L.) Beauv., (Andropogoneae); two species, A. sporobola and A. salmona, on I. cylindrica and Sporobolus macranthelus Chiov. (Zoysieae); and A. albivena on I. cylindrica, Miscanthus capensis (Nees) Andersson (Andropogoneae) and Cymbopogon sp. (Andropogoneae). Acrapex stygiata larvae developed on M. capensis and Cymbopogon sp. The host plants of A. brunneella and A. mpika remain unknown. We also conducted molecular phylogenetics and molecular species delimitation analyses on a comprehensive sample of 49 specimens belonging to nine of the studied species. Molecular phylogenetics and molecular species delimitation analyses provided additional evidence of the validity of the six newly described species but also suggested a level of hidden biodiversity for one of them.

Responses of Parasitoids to Volatiles Induced by Chilo partellus Oviposition on Teosinte, a Wild Ancestor of Maize

Maize, a genetically diverse crop, is the domesticated descendent of its wild ancestor, teosinte. Recently, we have shown that certain maize landraces possess a valuable indirect defense trait not present in commercial hybrids. Plants of these landraces release herbivore-induced plant volatiles (HIPVs) that attract both egg [Trichogramma bournieri Pintureau & Babault (Hymenoptera: Trichogrammatidae)] and larval [Cotesia sesamiae Cameron (Hymenoptera: Braconidae)] parasitoids in response to stemborer egg deposition. In this study, we tested whether this trait also exists in the germplasm of wild Zea species. Headspace samples were collected from plants exposed to egg deposition by Chilo partellus Swinhoe (Lepidoptera: Crambidae) moths and unexposed control plants. Four-arm olfactometer bioassays with parasitic wasps, T. bournieri and C. sesamiae, indicated that both egg and larval parasitoids preferred HIPVs from plants with eggs in four of the five teosinte species sampled. Headspace samples from oviposited plants released higher amounts of EAG-active compounds such as (E)-4,8-dimethyl-1,3,7-nonatriene. In oviposition choice bioassays, plants without eggs were significantly preferred for subsequent oviposition by moths compared to plants with prior oviposition. These results suggest that this induced indirect defence trait is not limited to landraces but occurs in wild Zea species and appears to be an ancestral trait. Hence, these species possess a valuable trait that could be introgressed into domesticated maize lines to provide indirect defense mechanisms against stemborers.

Vetiver grass (Vetiveria zizanioides (L.) Nash) as trap plant for Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae) and Busseola fusca (Fuller) (Lepidoptera: Noctuidae)

Abstract The preference of lepidopterous stem borer moths to oviposit on certain wild host plants can be exploited in habitat management systems by using those hosts as trap crops. Vetiver grass (Vetiveria zizanioides (L.) Nash) was evaluated for its attractiveness and suitability to the pyralid Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae) and the noctuid Busseola fusca (Fuller) (Lepidoptera: Noctuidae). Two choice tests were conducted in the laboratory and in the greenhouse to determine oviposition choice of C. partellus for maize, Vetiver and rice (Oryza sativa L.), and of B. fusca for Vetiver and maize. C. partellus larval survival was evaluated in green house studies. Results indicated that C. partellus chose Vetiver grass over maize though larval survival on Vetiver was extremely low. B. fusca did not show any host preference.Abstract The preference of lepidopterous stem borer moths to oviposit on certain wild host plants can be exploited in habitat management systems by using those hosts as trap crops. Vetiver grass (Vetiveria zizanioides (L.) Nash) was evaluated for its attractiveness and suitability to the pyralid Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae) and the noctuid Busseola fusca (Fuller) (Lepidoptera: Noctuidae). Two choice tests were conducted in the laboratory and in the greenhouse to determine oviposition choice of C. partellus for maize, Vetiver and rice (Oryza sativa L.), and of B. fusca for Vetiver and maize. C. partellus larval survival was evaluated in green house studies. Results indicated that C. partellus chose Vetiver grass over maize though larval survival on Vetiver was extremely low. B. fusca did not show any host preference.

Detection of transgenes in local maize varieties of small-scale farmers in Eastern Cape, South Africa

Small-scale subsistence farmers in South Africa have been introduced to genetically modified (GM) crops for more than a decade. Little is known about i) the extent of transgene introgression into locally recycled seed, ii) what short and long-term ecological and socioeconomic impacts such mixing of seeds might have, iii) how the farmers perceive GM crops, and iv) to what degree approval conditions are followed and controlled. This study conducted in the Eastern Cape, South Africa, aims primarily at addressing the first of these issues. We analysed for transgenes in 796 individual maize plants (leaves) and 20 seed batches collected in a village where GM insect resistant maize was previously promoted and grown as part of an governmental agricultural development program over a seven year period (2001–2008). Additionally, we surveyed the varieties of maize grown and the farmers’ practices of recycling and sharing of seed in the same community (26 farmers were interviewed). Recycling and sharing of seeds were common in the community and may contribute to spread and persistence of transgenes in maize on a local or regional level. By analysing DNA we found that the commonly used transgene promoter p35s occurred in one of the 796 leaf samples (0.0013%) and in five of the 20 seed samples (25%). Three of the 20 seed samples (15%) included herbicide tolerant maize (NK603) intentionally grown by the farmers from seed bought from local seed retailers or acquired through a currently running agricultural development program. The two remaining positive seed samples (10%) included genes for insect resistance (from MON810). In both cases the farmers were unaware of the transgenes present. In conclusion, we demonstrate that transgenes are mixed into seed storages of small-scale farming communities where recycling and sharing of seeds are common, i.e. spread beyond the control of the formal seed system.

Impact of violated high-dose refuge assumptions on evolution of Bt resistance.

Transgenic crops expressing Bacillus thuringiensis (Bt) toxins have been widely and successfully deployed for the control of target pests, while allowing a substantial reduction in insecticide use. The evolution of resistance (a heritable decrease in susceptibility to Bt toxins) can pose a threat to sustained control of target pests, but a high‐dose refuge (HDR) management strategy has been key to delaying countervailing evolution of Bt resistance. The HDR strategy relies on the mating frequency between susceptible and resistant individuals, so either partial dominance of resistant alleles or nonrandom mating in the pest population itself could elevate the pace of resistance evolution. Using classic Wright‐Fisher genetic models, we investigated the impact of deviations from standard refuge model assumptions on resistance evolution in the pest populations. We show that when Bt selection is strong, even deviations from random mating and/or strictly recessive resistance that are below the threshold of detection can yield dramatic increases in the pace of resistance evolution. Resistance evolution is hastened whenever the order of magnitude of model violations exceeds the initial frequency of resistant alleles. We also show that the existence of a fitness cost for resistant individuals on the refuge crop cannot easily overcome the effect of violated HDR assumptions. We propose a parametrically explicit framework that enables both comparison of various field situations and model inference. Using this model, we propose novel empiric estimators of the pace of resistance evolution (and time to loss of control), whose simple calculation relies on the observed change in resistance allele frequency.

Gut microbiota of Busseola fusca (Lepidoptera: Noctuidae)

Busseola fusca (Fuller) (Lepidoptera: Noctuidae) is a stemborer pest that attacks maize (Zea mays) throughout sub-Saharan Africa. Genetically modified maize has been shown to be effective against B. fusca. However, resistance of B. fusca against Bt-maize has developed and spread throughout South Africa. Previous studies suggested that gut microbiota contribute to mortality across a range of Lepidoptera. To fully assess the role of microbiota within the gut, it is essential to understand the microbiota harboured by natural B. fusca populations. This study aimed to identify the gut-associated bacteria by 16S rRNA gene sequencing. A total of 78 bacterial strains were characterised from the midgut of B. fusca larvae that were collected from 30 sites across the maize producing region of South Africa. Molecular phylogenetic analyses revealed bacteria affiliated to Proteobacteria, Actinobacteria, and Firmicutes. Taxonomic distribution placed these isolates into 15 different genera representing 20 species. The majority of bacteria identified belong to the genera Bacillus, Enterococcus, and Klebsiella. The B. fusca gut represents an intriguing and unexplored niche for analysing microbial ecology. The study could provide opportunities for developing new targets for pest management and contribute to understanding the phenomenon of resistance evolution of this species.Graphical Abstract