Morphometric and molecular discrimination of the sugarcane aphid, Melanaphis sacchari, (Zehntner, 1897) and the sorghum aphid Melanaphis sorghi (Theobald, 1904).

Melanaphis sacchari (Zehntner, 1897) and Melanaphis sorghi (Theobald, 1904) are major worldwide crop pests causing direct feeding damage on sorghum and transmitting viruses to sugarcane. It is common in the scientific literature to consider these two species as synonyms, referred to as the 'sugarcane aphid', although no formal study has validated this synonymy. In this study, based on the comparison of samples collected from their whole distribution area, we use both morphometric and molecular data to better characterize the discrimination between M. sacchari and M. sorghi. An unsupervised multivariate analysis of morphometric data clearly confirmed the separation of the two species. The best discriminating characters separating these species were length of the antenna processus terminalis relative to length of hind tibia, siphunculus or cauda. However, those criteria sometimes do not allow an unambiguous identification. Bayesian clustering based on microsatellite data delimited two clusters, which corresponded to the morphological species separation. The DNA sequencing of three nuclear and three mitochondrial regions revealed slight divergence between species. In particular, the COI barcode region proved to be uninformative for species separation because one haplotype is shared by both species. In contrast, one SNP located on the nuclear EF1-α gene was diagnostic for species separation. Based on morphological and molecular evidence, the invasive genotype damaging to sorghum in the US, Mexico and the Caribbean since 2013 is found to be M. sorghi.

Few Sensory Cues Differentiate Host and Dead-End Trap Plant for the Sugarcane Spotted Borer Chilo sacchariphagus (Lepidoptera: Crambidae).

The use of Erianthus arundinaceus as a trap plant in association with sugarcane reduces populations of the spotted sugarcane stalk borer Chilo sacchariphagus (Bojer) (Lepidoptera: Crambidae). This grass acts as a dead-end trap crop because it is the preferred plant for oviposition relative to sugarcane, and it precludes larval development. We explored the chemical mechanisms involved in host choice by C. sacchariphagus. We showed that the insect's antennal receptors are particularly sensitive to the shared compounds found in the volatile emissions produced by sugarcane and E. arundinaceus. In accordance with their phylogenetic proximity, the two plant species share many physicochemical properties, which suggests that C. sacchariphagus has few sensory cues to differentiate between the two. The terpene (E)-ß-ocimene is constitutively emitted by E. arundinaceus, but not by sugarcane. It elicits an electroantennographic response and behavioral responses from female C. sacchariphagus in Y-tube bioassays. Our study suggests that the sensory confusion between host plants, combined with a marginal sensory difference orienting the choice of an egg-laying site, constitutes a mechanism that is relevant to trap cropping. Systems based on this type of mechanism could provide long-term protection for crops vulnerable to insect pests.

Variation in physiological host range in three strains of two species of the entomopathogenic fungus Beauveria.

Knowledge of the host range of a biocontrol agent (BCA) is fundamental. Host range determines the BCA's economic potential, as well as the possible risk for non-target organisms. Entomopathogenic fungal strains belonging to the genus Beauveria are widely used as BCA, but our knowledge of their physiological host range is only partial. The aim of this study was to improve our understanding of the physiological host range of three Beauveria strains belonging to two species, B. hoplocheli and B. bassiana. We performed laboratory mortality bioassays to assess their pathogenicity and virulence against nine insect pests, belonging to three orders: Lepidoptera, Coleoptera and Diptera. Mortality rate, mean survival time and mycosis rate were used to estimate virulence. Pathogenicity was assessed as the capacity to cause a disease and induce mortality. Virulence was assessed as the severity of the disease based on mortality rate, mean survival time and mycosis rate. The results of this study revealed significant differences in the physiological host range of the three Beauveria strains tested. The three strains were pathogenic to all Diptera and Lepidoptera species tested. In the case of the Coleoptera, only the B. hoplocheli strain was pathogenic to the white grub Hoplochelus marginalis and only the B. bassiana strains were pathogenic to Alphitobius diaperinus. The B. hoplocheli strain was less virulent on Lepidoptera and Diptera than the two B. bassiana strains. The latter both exhibited very similar virulence patterns. The fact that B. hoplocheli and B. bassiana strains have different host ranges means that they can be used as BCA to target different pests. Impacts on non-target insects across multiple orders cannot be ruled out in the absence of ecological host range studies.

Invasion of sorghum in the Americas by a new sugarcane aphid (Melanaphis sacchari) superclone.

In the United States (US), the sugarcane aphid (SCA) Melanaphis sacchari (Zehnter) (Hemiptera: Aphididae) was introduced in the 1970s, however at that time it was only considered a pest of sugarcane. In 2013, a massive outbreak of M. sacchari occured on sorghum, resulting in significant economic damage to sorghum grown in North America including the US, Mexico, and Puerto Rico. The aim of the present study was to determine if the SCA pest emergence in American sorghum resulted from the introduction of new genotypes. To this end we used microsatellite markers and COI sequencing to compare the genetic diversity of SCA populations collected in the Americas after the 2013 SCA outbreak on sorghum (during 2013-2017) to older samples collected before the pest outbreak (during 2007-2009). Our results show that the SCA outbreak in the Americas and the Caribbean observed since 2013 belong to populations exhibiting low genetic diversity and consisting of a dominant clonal lineage, MLL-F, which colonizes Sorghum spp. and sugarcane. The comparison of MLL-F specimens collected post-2013 with specimens collected in Louisiana in 2007 revealed that both populations are genetically distinct, according to COI sequencing and microsatellite data analyses. Our result suggest that MLL-F is a new invasive genotype introduced into the Americas that has spread rapidly across sorghum growing regions in the US, Mexico, Honduras and the Caribbean. The origin of this introduction is either Africa or Asia, with Asia being the most probable source.

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.

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.

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.

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.

Variation in Infection Capacity and in Virulence Exists Between Genotypes of Sugarcane yellow leaf virus.

Two experiments, one in Guadeloupe and one in Réunion Island, were performed to transmit different genotypes of Sugarcane yellow leaf virus (SCYLV) to eight sugarcane cultivars differing in resistance to infection by the virus and to yellow leaf. Transmission was attempted from SCYLV-infected sugarcane plants or leaves to healthy tissue-cultured plantlets grown in vitro and with the aphid vector Melanaphis sacchari. After inoculation and elimination of insects with an insecticide, plantlets were transferred to Montpellier, France and grown in a greenhouse. Plants were tested for presence of SCYLV by tissue-blot immunoassay and reverse-transcription polymerase chain reaction after 5 to 6 months of growth. SCYLV genotypes BRA-PER, CUB, and REU were detected in 47, 62, and 39% of plants inoculated with these genotypes in Guadeloupe, respectively. SCYLV genotypes BRA-PER and REU and a mixed infection of genotypes BRA-PER and REU were detected in 56, 33, and 42% of plants inoculated with these genotypes in Réunion Island, respectively. Genotypes BRA-PER and CUB could be transmitted to all eight sugarcane cultivars, but genotype REU could never be transmitted to resistant sugarcane cvs. H78-4153 and H78-3567. SCYLV genotype REU was transmitted successfully to sugarcane cv. R570 in Guadeloupe, but not in Réunion Island. Genotypes BRA-PER and CUB induced yellow leaf symptoms in susceptible or highly susceptible sugarcane cultivars, whereas genotype REU induced very few symptoms. SCYLV was not found in several symptomatic plants, suggesting an association of disease with undetectable populations of the virus or a nonviral cause. This is the first report of variation in infection capacity and in virulence of SCYLV.