Genetic mapping and candidate gene analysis for melon resistance to Phytophthora capsici.

Phytophthora blight is one of the most serious diseases affecting melon (Cucumis melo) production. Due to the lack of highly resistant germplasms, the progress on disease-resistant research is slow. To understand the genetics of melon resistance to Phytophthora capsici, an F2 population containing 498 individuals was developed by crossing susceptible line E31 to highly resistant line ZQK9. Genetic analysis indicated that the resistance in ZQK9 was controlled by a dominant gene, tentatively named MePhyto. Through bulked-segregant analysis (BSA-Seq) and chromosome walking techniques, the MePhyto gene was mapped to a 52.44 kb interval on chromosome 12. In this region, there were eight genes and their expression patterns were validated by qRT-PCR. Among them, one wall-associated receptor kinase (WAK) gene MELO3C002430 was significantly induced in ZQK9 after P. capsici inoculation, but not in E31. Based on the non-synonymous mutation site in MELO3C002430, a cleaved amplified polymorphic sequence (CAPS) marker, CAPS2430, was developed and this maker was co-segregated with MePhyto in both F2 population and a collection of 36 melon accessions. Thus MELO3C002430 was considered as the candidate gene and CAPS2430 was a promising marker for marker-assisted selection (MAS) in breeding. These results lay a foundation for revealing the resistance mechanism of melon to P. capsici.

Fitness costs in clothianidin-resistant population of the melon aphid, Aphis gossypii.

Clothianidin is a second-generation neonicotinoid insecticide, widely used against sap-sucking insect pest including melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae). This pest causes severe economic damage to Cucurbitaceae plants worldwide. In this study, we investigated clothianidin resistance development under continuous selection pressure. Moreover, the age-stage, two-sex life table approach was used to evaluate the impact of clothianidin resistance on the fitness of A. gossypii. A clothianidin resistant strain (CT-R) with a 23.17-fold resistance level was developed from a susceptible strain (CT-S) after continuous selection for 24 generations. Life table results showed a significant reduction in the relative fitness (0.847) of CT-R strain compared to the CT-S strain of A. gossypii. The developmental duration, oviposition days, total pre-oviposition period (TPOP), longevity, and fecundity of CT-R strain were found to be significantly lower when compared to CT-S strain. The demographic parameters, including the intrinsic rate of increase (r), finite rate of increase (λ), net reproductive rate (R0), and mean generation time (T) were also significantly decreased in CT-R strain compared to the CT-S strain. Both the reproductive and survival rates were affected by clothianidin resistance in CT-R strain compared with the CT-S strain of A. gossypii. Overall, our results demonstrate that in-depth knowledge about the trade-off at play between resistance degree and fitness cost might be useful to design resistance management strategies against A. gossypii.

Can Winged Aphid Abundance Be a Predictor of Cucurbit Aphid-Borne Yellows Virus Epidemics in Melon Crop?

Aphid-borne viruses are frequent yield-limiting pathogens in open field vegetable crops. In the absence of curative methods, virus control relies exclusively on measures limiting virus introduction and spread. The efficiency of control measures may greatly benefit from an accurate knowledge of epidemic drivers, in particular those linked with aphid vectors. Field experiments were conducted in southeastern France between 2010 and 2019 to investigate the relationship between the epidemics of cucurbit aphid-borne yellows virus (CABYV) and aphid vector abundance. Winged aphids visiting melon crops were sampled daily to assess the abundance of CABYV vectors (Aphis gossypii, Macrosiphum euphorbiae and Myzus persicae) and CABYV was monitored weekly by DAS-ELISA. Epidemic temporal progress curves were successfully described by logistic models. A systematic search for correlations was undertaken between virus variables including parameters µ (inflection point of the logistic curve) and γ (maximum incidence) and aphid variables computed by aggregating abundances on periods relative either to the planting date, or to the epidemic peak. The abundance of A. gossypii during the first two weeks after planting was found to be a good predictor of CABYV dynamics, suggesting that an early control of this aphid species could mitigate the onset and progress of CABYV epidemics in melon crops.

Chromosome-level assembly of the melon thrips genome yields insights into evolution of a sap-sucking lifestyle and pesticide resistance.

Thrips are tiny insects from the order Thysanoptera (Hexapoda: Condylognatha), including many sap-sucking pests that are causing increasing damage to crops worldwide. In contrast to their closest relatives of Hemiptera (Hexapoda: Condylognatha), including numerous sap-sucking species, there are few genomic resources available for thrips. In this study, we assembled the first thrips genome at the chromosomal level from the melon thrips, Thrips palmi, a notorious pest in agriculture, using PacBio long-read and Illumina short-read sequences. The assembled genome was 237.85 Mb in size, with 1,324 contigs and a contig N50 of 567 kb. All contigs were assembled into 16 linkage groups assisted by the Hi-C technique. In total, 16,333 protein-coding genes were predicted, of which 88.13% were functionally annotated. Among sap-sucking insects, polyphagous species (e.g., T. palmi and Bemisia tabaci) usually possess more detoxification genes than oligophagous species (e.g., Diaphorina citri). The polyphagous thrips genomes characterized so far have relatively more detoxification genes in the GST and CCE families than polyphagous aphids, but they have fewer UGTs. HSP genes, especially from the Hsp70s group, have expanded in thrips compared to other hemipterans. These differences point to different genetic mechanisms associated with detoxification and stress responses in these two groups of sap-sucking insects. The expansion of these gene families may contribute to the rapid development of pesticide resistance in thrips, as supported by a transcriptome comparison of resistant and sensitive populations of T. palmi. The high-quality genome developed here provides an invaluable resource for understanding the ecology, genetics, and evolution of thrips as well as their relatives more generally.

Elevated CO2 concentration affects the defense of tobacco and melon against lepidopteran larvae through the jasmonic acid signaling pathway.

The massive use of fossil fuels since the industrial revolution has led to a rapid increase in the concentration of carbon dioxide (CO2) in the atmosphere. What effects elevated CO2 concentrations (ECO2) have on the defense mechanisms plants employ against insects remains poorly understood. This study showed that ECO2 of 750 ± 20 mmol/mol, increased the photosynthetic rate and biomass gain of tobacco and melon plants. However, while mass gain of Spodoptera litura, a nocturnal moth in the Noctuidae family, was higher when feeding on tobacco plants under ECO2, mass gain of Diaphania indica was reduced when feeding on melon plant at ECO2 compared to ambient CO2. Plants have many mechanisms to defend themselves against insects. Jasmonic acid (JA) is a crucial element of plant defense against lepidopteran insects. Our study showed that JA levels increased in tobacco plants under ECO2 but decreased in melon plants. It is speculated that ECO2 changes plant resistance to insects mainly by affecting the JA signaling pathway. Nutrient analysis suggested defensive metabolites rather than changes in the total nitrogen or protein content of the plants led to the changes in plant defense levels under ECO2. In summary, ECO2 affects the interaction between plants and insects. The results may provide a theoretical basis for studying the changes in crop resistance to pests under ECO2 and predicting the impact of ECO2 on future agro-ecosystems.

Bacterial communities in the gut of wild and mass-reared Zeugodacus cucurbitae and Bactrocera dorsalis revealed by metagenomic sequencing.

BACKGROUND: Insect pests belonging to genus Bactrocera sp. (Diptera: Tephritidae) pose major biotic stress on various fruits and vegetable crops around the world. Zeugodacus and Bactrocera sp. are associated with diverse bacterial communities which play an important role in the fitness of sterile insects. The wild populations of melon fly, Zeugodacus cucurbitae (Coquillett) and Oriental fruit fly, Bactrocera dorsalis (Hendel) were collected from pumpkin and mango fields, respectively. The laboratory populations of Z. cucurbitae and B. dorsalis were mass-reared on bottle gourd and sweet banana, respectively. Bacterial communities present in the gut of wild and mass-reared mature (~ 12 days old) and newly emerged (< 1 h after emergence) male and female adults of Z. cucurbitae and B. dorsalis were assessed. We used Illumina HiSeq next-generation sequencing of 16S rRNA gene to profile the gut bacterial communities of wild and mass-reared mature and newly emerged Z. cucurbitae and B. dorsalis adults. RESULTS: We found diverse bacterial composition in the gut of wild and mass-reared Z. cucurbitae (ZC) and B. dorsalis (BD) with varied relative abundance. Few taxonomic groups were common to both the species. The most dominant phyla in all samples of Z. cucurbitae and B. dorsalis adults were Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria. The phylum Proteobacteria occurred more in wild Z. cucurbitae (~ 87.72%) and B. dorsalis (~ 83.87%) as compared to mass-reared Z. cucurbitae (64.15%) and B. dorsalis (~ 80.96%). Higher relative abundance of Phylum Firmicutes was observed in mass-reared fruit fly than wild adults. Cyanobacteria/Chloroplast and Actinobacteria were also present with very low relative abundance in both wild as well as mass-reared melon fly and Oriental fruit fly. Enterobacteriaceae (61.21%) was dominant family in the gut of both wild and mass-reared adults. Providencia and Lactococcus were dominant genera with varied relative abundance in wild as well as in mass-reared mature and newly emerged fruit fly adults of both species. Some of the genera like Morganella and Serratia were only detected in mass-reared mature and newly emerged Z. cucurbitae and B. dorsalis adults. Principal Coordinate Analysis (PCoA) showed that fruit fly adult samples were grouped based on species and age of the adults while no grouping was observed on the basis of sex of the adult fruit fly. CONCLUSIONS: The gut bacterial communities associated with wild and mass-reared mature and newly emerged adults of Z. cucurbitae and B. dorsalis showed variation that depends on species and age of the insects. Understanding the gut microbiota of wild and mass-reared Z. cucurbitae and B. dorsalis using high throughput technology will help to illustrate microbial diversity and this information could be used to develop efficient mass-rearing protocols for successful implementation of sterile insect technique (SIT).

The effect of diet and radiation on the bacterial symbiome of the melon fly, Zeugodacus cucurbitae (Coquillett).

BACKGROUND: Symbiotic bacteria contribute to a multitude of important biological functions such as nutrition and reproduction and affect multiple physiological factors like fitness and longevity in their insect hosts. The melon fly, Zeugodacus cucurbitae (Coquillett), is an important agricultural pest that affects a variety of cultivated plants belonging mostly to the Cucurbitaceae family. It is considered invasive and widespread in many parts of the world. Several approaches are currently being considered for the management of its populations including the environmentally friendly and effective sterile insect technique (SIT), as a component of an integrated pest management (IPM) strategy. In the present study, we examined the effect of diet and radiation on the bacterial symbiome of Z. cucurbitae flies with the use of Next Generation Sequencing technologies. RESULTS: Melon flies were reared on two diets at the larval stage, an artificial bran-based diet and on sweet gourd, which affected significantly the development of the bacterial profiles. Significant differentiation was also observed based on gender. The effect of radiation was mostly diet dependent, with irradiated melon flies reared on the bran diet exhibiting a significant reduction in species diversity and richness compared to their non-irradiated controls. Changes in the bacterial symbiome of the irradiated melon flies included a drastic reduction in the number of sequences affiliated with members of Citrobacter, Raoultella, and Enterobacteriaceae. At the same time, an increase was observed for members of Enterobacter, Providencia and Morganella. Interestingly, the irradiated male melon flies reared on sweet gourd showed a clear differentiation compared to their non-irradiated controls, namely a significant reduction in species richness and minor differences in the relative abundance for members of Enterobacter and Providencia. CONCLUSIONS: The two diets in conjunction with the irradiation affected significantly the formation of the bacterial symbiome. Melon flies reared on the bran-based artificial diet displayed significant changes in the bacterial symbiome upon irradiation, in all aspects, including species richness, diversity and composition. When reared on sweet gourd, significant changes occurred to male samples due to radiation, only in terms of species richness.

Impact of low lethal concentrations of buprofezin on biological traits and expression profile of chitin synthase 1 gene (CHS1) in melon aphid, Aphis gossypii.

Buprofezin, a chitin synthesis inhibitor that can be used for the control of hemipteran pests, especially melon aphid, Aphis gossypii. The impact of low lethal concentrations of buprofezin on the biological parameters and expression profile of CHS1 gene were estimated for two successive generations of A. gossypii. The present result shows that the LC15 and LC30 of buprofezin significantly decreased the fecundity and longevity of both generations. Exposure of F0 individuals to both concentrations delay the developmental period in F1. Furthermore, the survival rate, intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0) were reduced significantly in progeny generation at both concentrations. However, the reduction in gross reproductive rate (GRR) was observed only at LC30. Although, the mean generation time (T) prolonged substantially at LC30. Additionally, expression of the CHS1 gene was significantly increased in F0 adults. Significant increase in the relative abundance of CHS1 mRNA transcript was also observed at the juvenile and adult stages of F1 generation following exposure to LC15 and LC30. Therefore, our results show that buprofezin could affect the biological traits by diminishing the chitin contents owing to the inhibition of chitin synthase activity in the succeeding generation of melon aphid.

Plant Growth Promotion and Biocontrol of Pythium ultimum by Saline Tolerant Trichoderma Isolates under Salinity Stress.

This present study evaluates three isolates of Trichoderma as plant growth promoting or biological control agents: Trichoderma aggressivum f. sp. europaeum, Trichoderma saturnisporum, and the marine isolate obtained from Posidonia oceanica, Trichoderma longibrachiatum. The purpose is to contribute to an overall reduction in pesticide residues in the fruit and the environment and to a decrease in chemical fertilizers, the excess of which aggravates one of the most serious abiotic stresses, salinity. The tolerance of the different isolates to increasing concentrations of sodium chloride was evaluated in vitro, as well as their antagonistic capacity against Pythium ultimum. The plant growth promoting capacity and effects of Trichoderma strains on the severity of P. ultimum on melon seedlings under saline conditions were also analysed. The results reveal that the three isolates of Trichoderma, regardless of their origin, alleviate the stress produced by salinity, resulting in larger plants with an air-dry weight percentage above 80% in saline stress conditions for T. longibrachiatum, or an increase in root-dry weight close to 50% when T. aggressivum f. sp. europaeum was applied. Likewise, the three isolates showed antagonistic activity against P. ultimum, reducing the incidence of the disease, with the highest response found for T. longibrachiatum. Biological control of P. ultimum by T. aggressivum f. sp. europaeum and T. saturnisporum is reported for the first time, reducing disease severity by 62.96% and 51.85%, respectively. This is the first description of T. aggressivum f. sp. europaeum as a biological control agent and growth promoter. The application of these isolates can be of enormous benefit to horticultural crops, in both seedbeds and greenhouses.

Sustainable alternatives to 1,3-dichloropropene for controlling root-knot nematodes and fungal pathogens in melon crops in Mediterranean soils: Efficacy and effects on soil quality.

The control of agricultural pests is key to maintain economically viable crops. Increasing environmental awareness, however, is leading to more restrictive European policies regulating the use of certain pesticides due to their impact on human health and the soil system. Given this context, we evaluated the efficacy of three alternatives to the soil fumigant 1,3-dichloropropene (1,3-D), which is currently banned in Europe: two non-fumigant nematicides [oxamyl (OX) and fenamiphos (FEN)] and the soil fumigant dimethyl disulfide (DMDS). We analysed the efficiency of these pesticides against root-knot nematodes and soil fungal pathogens (determined by qPCR) as well as the soil biological quality after treatments application (estimated by enzyme activities). Among treatments, 1,3-D and DMDS significantly reduced nematode populations. FEN was more effective in sandy soil, while OX had no effect in any soil. OX and FEN had no effect on fungal pathogens, whereas DMDS reduced the abundance of Rhizoctonia solani and Fusarium solani at the root level in clay-loam soil. Soil quality decreased after treatment application but then recovered throughout the experiment, indicating the possible dissipation of the pesticides. Our findings support DMDS as a potential sustainable alternative for controlling root-knot nematodes and fungal pathogens due to its effectiveness in both studied soils, although its negative impact on soil biological quality in sandier soils must be taken into account. Main finding of the work. DMDS is a reliable alternative to 1,3-D for controlling agricultural pest but its inhibitory effect on soil enzyme activities varied according to the soil characteristics.

Cucumis metuliferus reduces Meloidogyne incognita virulence against the Mi1.2 resistance gene in a tomato-melon rotation sequence.

BACKGROUND: Susceptible tomato cv. Durinta, ungrafted or grafted onto cv. Aligator resistant rootstock, both followed by the susceptible melon cv. Paloma, ungrafted or grafted onto Cucumis metuliferus BGV11135, and in the reverse order, were cultivated from 2015 to 2017 in the same plots in a plastic greenhouse, infested or not with Meloidogyne incognita. For each crop, soil nematode densities, galling index, number of eggs per plant and crop yield were determined. Virulence selection was evaluated in pot experiments. RESULTS: In the tomato-melon rotation, nematode densities increased progressively for the grafted tomato, being higher than for ungrafted plants at the end of the study; this was not the case in the melon-tomato rotation. Grafted crops yielded more than ungrafted crops in the infested plots. Virulence against the Mi1.2 gene was detected, but not against C. metuliferus. Reproduction of M. incognita on the resistant tomato was ∼ 120% that on the susceptible cultivar after the first grafted tomato crop, but this decreased to just 25% at the end of the experiment. CONCLUSION: Alternating different resistant plant species suppresses nematode population growth rate and yield losses. Although this strategy does not prevent virulence selection, the level was reduced. © 2018 Society of Chemical Industry.

Impact of the secondary plant metabolite Cucurbitacin B on the demographical traits of the melon aphid, Aphis gossypii.

Cucurbitacin B is a natural triterpene present in plants of Cucurbitaceae family, which are among the host plants for melon aphid, Aphis gossypii. In present study we characterized the effects of two cucurbitacin B concentrations on the biological parameters of adults (F0) and of juveniles and adults of their progeny (F1). The results showed that cucurbitacin B at 25 ppm significantly reduced the adult longevity and fecundity of both F0 and F1 generation. Exposure of F0 generation to 25 ppm though reduced the demographic traits of F1 including the intrinsic rate of increase r (day-1), generation time T (day), finite rate of increase λ (day-1), however, only net reproductive rate R0 (offspring/individual) decreased significantly. While 100 ppm reduced not only the longevity and fecundity of F0 generation but also the longevity of F1 generation. Fecundity of F1 was not affected by 100 ppm of cucurbitacin B, however, R0 (offspring/individual) and T (day) of F1 generation were lower than the control population. These results support the hypothesis that high contents of cucurbitacin B caused negative impact on melon aphid and could be used as a lead for classical selection of resistant varieties of plants that are main hosts for the melon aphid.

Presence of Native Prey Does Not Divert Predation on Exotic Pests by Harmonia axyridis in Its Indigenous Range.

In China, two invasive pests, Bemisia tabaci MEAM1 (Gennadius) and Frankliniella occidentalis (Pergande), often co-occur with the native pest, Aphis gossypii (Glover), on plants of Malvaceae and Cucurbitaceae. All three are preyed on by the native ladybird, Harmonia axyridis (Pallas); however, the native predator might be expected to prefer native prey to the exotic ones due to a shared evolutionary past. In order to clarify whether the presence of native prey affected the consumption of these two invasive species by the native predator, field-cage experiments were conducted. A duplex qPCR was used to simultaneously detect both non-native pests within the gut of the predator. H. axyridis readily accepted both invasive prey species, but preferred B. tabaci. With all three prey species available, H. axyridis consumption of B. tabaci was 39.3±2.2% greater than consumption of F. occidentalis. The presence of A. gossypii reduced (by 59.9% on B. tabaci, and by 60.6% on F. occidentalis), but did not stop predation on the two exotic prey when all three were present. The consumption of B. tabaci was similar whether it was alone or together with A. gossypii. However, the presence of aphids reduced predation on the invasive thrips. Thus, some invasive prey may be incorporated into the prey range of a native generalist predator even in the presence of preferred native prey.

Using Next-Generation Sequencing to Develop Molecular Diagnostics for Pseudoperonospora cubensis, the Cucurbit Downy Mildew Pathogen.

Advances in next-generation sequencing (NGS) allow for rapid development of genomics resources needed to generate molecular diagnostics assays for infectious agents. NGS approaches are particularly helpful for organisms that cannot be cultured, such as the downy mildew pathogens, a group of biotrophic obligate oomycetes that infect crops of economic importance. Unlike most downy mildew pathogens that are highly host-specific, Pseudoperonospora cubensis causes disease on a broad range of crops belonging to the family Cucurbitaceae. In this study, we identified candidate diagnostic markers for P. cubensis by comparing NGS data from a diverse panel of P. cubensis and P. humuli isolates, two very closely related oomycete species. P. cubensis isolates from diverse hosts and geographical regions in the United States were selected for sequencing to ensure that candidates were conserved in P. cubensis isolates infecting different cucurbit hosts. Genomic regions unique to and conserved in P. cubensis isolates were identified through bioinformatics. These candidate regions were then validated using PCR against a larger collection of isolates from P. cubensis, P. humuli, and other oomycetes. Overall seven diagnostic markers were found to be specific to P. cubensis. These markers could be used for pathogen diagnostics on infected tissue, or adapted for monitoring airborne inoculum with real-time PCR and spore traps.

NBS-LRR-mediated resistance triggered by aphids: viruses do not adapt; aphids adapt via different mechanisms.

BACKGROUND: Aphids are serious pest on crops. By probing with their stylets, they interact with the plant, they vector viruses and when they reach the phloem they start a continuous ingestion. Many plant resistances to aphids have been identified, several have been deployed. However, some resistances breaking down have been observed. In the melon, a gene that confers resistance to aphids has been deployed in some melon-producing areas, and aphid colony development on Vat-carrying plants has been observed in certain agrosystems. The Vat gene is a NBS-LRR gene that confers resistance to the aphid species Aphis gossypii and exhibits the unusual characteristic of also conferring resistance to non-persistently transmitted viruses when they are inoculated by the aphid. Thus, we characterized patterns of resistance to aphid and virus using the aphid diversity and we investigated the mechanisms by which aphids and viruses may adapt to the Vat gene. RESULTS: Using a Vat-transgenic line built in a susceptible background, we described the Vat- spectrum of resistance to aphids, and resistance to viruses triggered by aphids using a set of six A. gossypii biotypes. Discrepancies between both resistance phenotypes revealed that aphid adaptation to Vat-mediated resistance does not occur only via avirulence factor alterations but also via adaptation to elicited defenses. In experiments conducted with three virus species serially inoculated by aphids from and to Vat plants, the viruses did not evolve to circumvent Vat-mediated resistance. We confirmed discrepancies between both resistance phenotypes by testing each aphid biotype with a set of thirteen melon accessions chosen to reflect the natural diversity of the melon. Inheritance studies revealed that patterns of resistance to virus triggered by aphids are controlled by different alleles at the Vat locus and at least another locus located at a short genetic distance. Therefore, resistance to viruses triggered by aphids is controlled by a gene cluster. CONCLUSIONS: Under the Flor model, changes in the avirulence gene determine the ability of the pathogen to overcome the resistance conferred by a plant gene. The Vat gene belongs to a resistance gene family that fits this pest/pathogen-plant interaction, and we revealed an additional mechanism of aphid adaptation that potentially exists in other interactions between plants and pests or pathogens.

Identification and preliminary characterization of chemosensory perception-associated proteins in the melon fly Bactrocera cucurbitae using RNA-seq.

An investigation into proteins involved in chemosensory perception in the melon fly, Bactrocera cucurbitae (Diptera: Tephritidae) is described here using a newly generated transcriptome dataset. The melon fly is a major agricultural pest, widely distributed in the Asia-Pacific region and some parts of Africa. For this study, a transcriptome dataset was generated using RNA extracted from 4-day-old adult specimens of the melon fly. The dataset was assembled and annotated via Gene Ontology (GO) analysis. Based on this and similarity searches to data from other species, a number of protein sequences putatively involved in chemosensory reception were identified and characterized in the melon fly. This included the highly conserved "Orco" along with a number of other less conserved odorant binding protein sequences. In addition, several sequences representing putative ionotropic and gustatory receptors were also identified. This study provides a foundation for future functional studies of chemosensory proteins in the melon fly and for making more detailed comparisons to other species. In the long term, this will ultimately help in the development of improved tools for pest management.

Novel toxicity of Bacillus thuringiensis strains against the melon fruit fly, Bactrocera cucurbitae (Diptera: Tephritidae).

Bactrocera cucurbitae (melon fruit fly) is one of the most detrimental vegetable-damaging pests in Bangladesh. The toxicity of Bacillus thuringiensis (Bt) has been reported against a few genera of Bactrocera in addition to numerous other insect species. Bt strains, harbouring cry1A-type genes were, therefore, assayed in vivo against the 3(rd) instar larvae of B. cucurbitae in this study. The biotype-based prevalence of cry1 and cry1A genes was calculated to be 30.8% and 11.16%, respectively, of the test strains (n=224) while their prevalence was greatest in biotype kurstaki. Though three indigenous Bt strains from biotype kurstaki with close genetic relationship exhibited higher toxicity, maximum mortalities were recorded for Btk HD-73 (96%) and the indigenous Bt JSc1 (93%). LC50 and LC99 values were determined to be 6.81 and 8.32 for Bt JSc1, 7.30 and 7.92 for Bt SSc2, and 6.99 and 7.67 for Btk HD-73, respectively. The cause of toxicity and its variation among the strains was found to be correlated with the synergistic toxic effects of cry1, cry2, cry3 and cry9 gene products, i.e. relevant Cry proteins. The novel toxicity of the B. thuringiensis strains against B. cucurbitae revealed in the present study thus will help in developing efficient and eco-friendly control measures such as Bt biopesticides and transgenic Bt cucurbits.

Implementing a spinosad-based local bait station to control Bactrocera cucurbitae (Diptera: Tephritidae) in high rainfall areas of Reunion Island.

Three species of fruit flies cause serious damage to cucurbit crops on Reunion Island: Bactrocera cucurbitae (Diptera: Tephritidae) (Coquillett 1899), Dacus ciliatus (Loew 1901), and Dacus demmerezi (Bezzi 1917). To control them, a program of agroecological management of cucurbit flies has been implemented based on the application of Synéis-appât, especially spot sprays on corn borders. However, the high rainfall on Reunion Island limits the long-term efficiency of the bait; in addition, this method cannot be used for large chayote trellises, because corn borders cannot be planted around them. The aim of this study was to design a bait station adapted to prevailing conditions on Reunion Island. An 'umbrella trap' tested in Taiwan was used as a reference to compare its efficacy with our local bait station. Experiments were conducted in field cages on B. cucurbitae to test different characteristics of bait stations and to construct one using local materials. Results were validated in the field. The attractiveness of the bait station was related mainly to the color of the external surface, yellow being the most attractive color. The efficacy of the bait station with respect to fly mortality was found to be linked to the accessibility of the bait, and direct application of Synéis-appât on the bait station was found to be the most efficient. In the field, B. cucurbitae were more attracted to the local bait station than to the umbrella trap, while the two other fly species displayed equal attraction to both trap types. Our local bait station is a useful alternative to spot sprays of Synéis-appât and is now included in a local pest management program and is well accepted by farmers.

Patterns of species richness and diversity of insects associated with cucurbit fruits in the southern part of Cameroon.

Patterns of species diversity and community structure of insects associated with fruits of domesticated cucurbits were investigated from January 2009 to 2011 in three localities from two agroecological zones in the southern part of Cameroon. Rarefaction curves combined with nonparametric estimators of species richness were used to extrapolate species richness beyond our own data. Sampling efforts of over 92% were reached in each of the three study localities. Data collected revealed a total of 66 insect morphospecies belonging to 37 families and five orders, identified from a set of 57,510 insects. The orders Diptera (especially Tephritidae and Lonchaeidae) and Hymenoptera (mainly Braconidae and Eulophidae) were the most important, in terms of both abundance and species richness on the one hand, and effects on agronomic performance on the other. Values for both the species diversity (Shannon and Simpson) and the species richness indices (Margalef and Berger-Parker) calculated showed that the insect communities were species-rich but dominated, all to a similar extent, by five main species (including four fruit fly species and one parasitoid). Species abundance distributions in these communities ranged from the Zipf-Mandelbrot to Mandelbrot models. The communities are structured as tritrophic networks, including cucurbit fruits, fruit-feeding species (fruit flies) and carnivorous species (parasitoids). Within the guild of the parasitoids, about 30% of species, despite their low abundance, may potentially be of use in biological control of important pests. Our field data contribute in important ways to basic knowledge of biodiversity patterns in agrosystems and constitute baseline data for the planned implementation of biological control in Integrated Pest Management.