Samuel Nibouche

Assessing the role of non-cotton refuges in delaying Helicoverpa armigera resistance to Bt cotton in West Africa

Non‐cotton host plants without Bacillus thuringiensis (Bt) toxins can provide refuges that delay resistance to Bt cotton in polyphagous insect pests. It has proven difficult, however, to determine the effective contribution of such refuges and their role in delaying resistance evolution. Here, we used biogeochemical markers to quantify movement of Helicoverpa armigera moths from non‐cotton hosts to cotton fields in three agricultural landscapes of the West African cotton belt (Cameroon) where Bt cotton was absent. We show that the contribution of non‐cotton hosts as a source of moths was spatially and temporally variable, but at least equivalent to a 7.5% sprayed refuge of non‐Bt cotton. Simulation models incorporating H. armigera biological parameters, however, indicate that planting non‐Bt cotton refuges may be needed to significantly delay resistance to cotton producing the toxins Cry1Ac and Cry2Ab. Specifically, when the concentration of one toxin (here Cry1Ac) declined seasonally, resistance to Bt cotton often occurred rapidly in simulations where refuges of non‐Bt cotton were rare and resistance to Cry2Ab was non‐recessive, because resistance was essentially driven by one toxin (here Cry2Ab). The use of biogeochemical markers to quantify insect movement can provide a valuable tool to evaluate the role of non‐cotton refuges in delaying the evolution of H. armigera resistance to Bt cotton.

Low genetic diversity in Melanaphis sacchari Aphid populations at the worldwide scale

Numerous studies have examined the genetic diversity and genetic structure of invading species, with contrasting results concerning the relative roles of genetic diversity and phenotypic plasticity in the success of introduced populations. Increasing evidence shows that asexual lineages of aphids are able to occupy a wide geographical and ecological range of habitats despite low genetic diversity. The anholocyclic aphid Melanaphis sacchari is a pest of sugarcane and sorghum which originated in the old world, was introduced into the Americas, and is now distributed worldwide. Our purpose was to assess the genetic diversity and structuring of populations of this species according to host and locality. We used 10 microsatellite markers to genotype 1333 individuals (57 samples, 42 localities, 15 countries) collected mainly on sugarcane or sorghum. Five multilocus lineages (MLL) were defined, grouping multilocus genotypes (MLG) differing by only a few mutations or scoring errors. Analysis of a 658 bp sequence of mitochondrial COI gene on 96 individuals revealed five haplotypes, with a mean divergence of only 0.19 %. The distribution of MLL appeared to be strongly influenced by geography but not by host plant. Each of the five MLL grouped individuals from (A) Africa, (B) Australia, (C) South America, the Caribbean and the Indian Ocean including East Africa, (D) USA, and (E) China. The MLL A and C, with a wide geographic distribution, matched the definition of superclone. Among aphids, M. sacchari has one of the lowest known rates of genetic diversity for such a wide geographical distribution.

First step towards integrated pest management on cotton in Burkina Faso

Abstract The ‘lutte etagee ciblee’ (LEC), a new spray programme, was tested in Burkina Faso. Each routine insecticide application at a fortnightly interval was followed by scouting and, if necessary, another application based on a threshold, 7 days later. The dosages of the active ingredients used were about half those in the previously recommended 14-day routine treatment programme. On-station trials revealed that the new programme provided the same level of pest control as the previously recommended routine programme. Similar results were obtained in smallholder field trials with yields greater than the fortnightly spraying programme used by the farmers. The amount of insecticide saved by this new programme ranged between 44% and 54%.

Beauveria brongniartii on white grubs attacking sugarcane in South Africa.

Beauveria brongniartii (Saccardo) Petch fungal infections were observed on the melolonthid Hypopholis sommeri Burmeister (Coleoptera: Scarabaeidae) at two sites (Harden Heights and Canema) in the sugarcane producing area of the northern KwaZulu-Natal Midlands of South Africa. To initially identify the disease-causing fungus, 17 different fluorescently-labelled microsatellite PCR primers were used to target 78 isolates of Beauveria spp. DNA. Microsatellite data resolved two distinct clusters of Beauveria isolates which represented the Beauveria bassiana s.s. (Balsamo) Vuillemin (17 isolates) and B. brongniartii (60 isolates) species groups. These groupings were supported by two gene regions, the nuclear ribosomal Internal Transcribed Spacer (ITS) and the nuclear Bloc gene of which 23 exemplar Beauveria isolates were represented and sequenced. When microsatellite data were analysed, 26 haplotypes among 58 isolates of B. brongniartii were distinguished. Relatively low levels of genetic diversity were detected in B. brongniartii and isolates were shown to be closely related. No genetic differentiation was observed between the Harden Heights and Canema populations; they thus may be considered one, structured and fragmented population over a distance of 5.5 km. Historically high levels of gene flow from swarming H. sommeri beetles is the proposed mechanism for this observed lack of genetic differentiation between populations. Microsatellite analyses also showed that B. brongniartii conidia were being cycled from arboreal forest to subterranean sugarcane habitats and vice versa in the environment by H. sommeri life stages. This is the first record of this species of fungus infecting H. sommeri larvae and adults in South Africa.

Resistance to Melanaphis sacchari in the sugarcane cultivar R 365

This study focuses on the resistance of sugarcane, Saccharum spec. (Poaceae), to the sugarcane aphid, Melanaphis sacchari (Zehntner) (Hemiptera: Aphididae), which vectors Sugarcane yellow leaf virus (SCYLV). Resistance was characterized in cultivar R 365, using a 3‐year field trial and laboratory experiments on potted plantlets and excised leaves. R 365 reduced aphid populations in the field by antixenosis and antibiosis. Using the electrical penetration graph technique, we detected delayed aphid salivation in phloem and inhibition of passive phloem sap uptake in R 365. The resistance factors also proved to be effective against the corn leaf aphid, Rhopalosiphum maidis (Fitch) (Hemiptera: Aphididae), another vector of SCYLV.

Description and phylogenetic placement of Beauveria hoplocheli sp. nov. used in the biological control of the sugarcane white grub, Hoplochelus marginalis, on Reunion Island

On Reunion Island successful biological control of the sugarcane white grub Hoplochelus marginalis Fairmaire (Coleoptera: Melolonthidae) has been conducted for decades with strains from the entomopathogenic fungal genus Beauveria (Ascomycota: Hypocreales). A study based on morphological characters combined with a multisequence phylogenetic analysis of genes that encode the translation elongation factor 1-alpha (TEF1), RNA polymerase II largest subunit (RPB1), RNA polymerase II second largest subunit (RPB2) and the Bloc nuc intergenic region was carried out on Beauveria strains isolated on Reunion and Madagascar from H. marginalis. This study revealed that these strains, previously identified as Beauveria brongniartii, did not match that species and are closely related to but still distinct from B. malawiensis strains. Therefore we describe the Reunion Island fungus as the new species B. hoplocheli.

Modeling Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) damages on cotton.

Abstract We studied and modeled damage caused by Helicoverpa armigera larvae on cotton with the aim of developing a coupled crop pest model. Two damage components were studied: the voracity (quantity of fresh matter and number of organs consumed) and feeding preferences (type of organ infested). The laboratory no-choice study of voracity on excised squares and bolls revealed that an H. armigera larva consumes 2,856 mg of fresh matter throughout its larval life, with the sixth instar consuming 86% of this quantity. This consumption rate corresponded to 23.6 squares, or 7.8 bolls. We developed equations to predict the quantity of fresh matter uptake from an individual plant organ, according to the organ mass and the larval instar. The field study of feeding preference confirmed previous findings that larvae prefer squares to bolls, with this preference decreasing as the larval age increases. However, no significant relationship was noted between the age of larvae and the size of infested organs within each organ class (square or boll). We developed a logistic model to predict the probability of a larva infesting a boll rather than a square. According to this model, the relative organ availability in the field and the larval instar were found to be significant factors.

Host plant specialization in the sugarcane Aphid Melanaphis sacchari

Most aphids are highly specialized on one or two related plant species and generalist species often include sympatric populations adapted to different host plants. Our aim was to test the hypothesis of the existence of host specialized lineages of the aphid Melanaphis sacchari in Reunion Island. To this end, we investigated the genetic diversity of the aphid and its association with host plants by analyzing the effect of wild sorghum Sorghum bicolor subsp. verticilliflorum or sugarcane as host plants on the genetic structuring of populations and by performing laboratory host transfer experiments to detect trade-offs in host use. Genotyping of 31 samples with 10 microsatellite loci enabled identification of 13 multilocus genotypes (MLG). Three of these, Ms11, Ms16 and Ms15, were the most frequent ones. The genetic structure of the populations was linked to the host plants. Ms11 and Ms16 were significantly more frequently observed on sugarcane, while Ms15 was almost exclusively collected in colonies on wild sorghum. Laboratory transfer experiments demonstrated the existence of fitness trade-offs. An Ms11 isofemale lineage performed better on sugarcane than on sorghum, whereas an Ms15 lineage developed very poorly on sugarcane, and two Ms16 lineages showed no significant difference in performances between both hosts. Both field and laboratory results support the existence of host plant specialization in M. sacchari in Reunion Island, despite low genetic differentiation. This study illustrates the ability of asexual aphid lineages to rapidly undergo adaptive changes including shifting from one host plant to another.

Within-plant distribution of cotton boll-infesting lepidoptera: Application to Sampling

Abstract Sampling of Lepidoptera-infesting cotton bolls [Helicoverpa armigera (Hübner), Diparopsis watersi (Rothschild), Spodoptera littoralis (Boisduval), and Earias spp.] in Cameroon is based on examination of whole plants. To reduce the time taken by sampling procedures, we studied the suitability of a subsampling plan based on examination of plant terminals. The development of such a subsampling plan requires that the proportion of larvae located on plant terminals should remain constant. However, our study of the within-plant distribution of larvae revealed that their proportion on the five upper fruiting branches was too variable to allow the development of a sampling plan. Examination of the 10 upper branches led to less variable results, but the proportion was still significantly influenced by the pest species and time after crop emergence. We designed a conservative sampling plan in which the proportion was constant and equal to the lower bound of the 90% confidence interval for its predicted minimum (0.594). With this underestimation, the probability distribution of the number of larvae on the ten upper branches of a n plant sample was a compound of a negative binomial and a binomial distribution. These results enabled the design of a sampling plan that reduces sampling time by up to 60%, but the cost of this reduction is an increase in the risk in deciding on intervention when the mean infestation is lower than the critical density.

Diapause incidence and duration in the pest mango blossom gall midge, Procontarinia mangiferae (Felt), on Reunion Island.

The mango blossom gall midge, Procontarinia mangiferae, is a multivoltine species that induces galls in inflorescences and leaves of the mango tree, Mangifera indica. In subtropical Reunion Island, populations of P. mangiferae are observed all-year round, but the pattern and the role of dormancy in their life cycle have never been documented. We performed field and laboratory experiments using more than 15,000 larvae. We demonstrated that a larval diapause may affect a part of the midge population, regardless of the season. The total duration of the diapause varied from 6 weeks to more than 1 year. One year of field monitoring showed that the highest incidence of diapause was observed in larvae collected during the summer from mango leaves, where it affected approximately one-third of the individuals. This facultative diapause allows the permanent presence of P. mangiferae in the orchards. By recording diapause duration during 22 weeks under controlled conditions, we showed that high temperatures (26 °C) increased diapause duration and extended the range of the dates of diapause emergence, whereas cool temperatures (20 °C) shortened diapause duration and shortened the range of the dates of emergence from diapause. A temperature decrease from 26 to 20 °C triggered the emergence of diapausing individuals. These mechanisms ensure the synchronization of the emergence of diapausing individuals with the appearance of mango inflorescences, which is also induced by cool winter temperatures.