Improving the genetic potential of okra (Abelmoschus esculentus L.) germplasm to tolerate salinity stress.

Okra (Abelmoschus esculentus L.) is the most consumed vegetable worldwide with the potential for diverse ecological adaptation. However, increasing salinization and changing climatic conditions are posing serious threats to the growth, yield, and quality of okra. Therefore, to mitigate increasing soil salinization and ensure sustainable okra production under rapidly changing climatic conditions, evaluation of new okra germplasm to develop salt tolerant cultivars is direly needed. The present study was designed to evaluate the genetic resources of okra genotypes for salt tolerance at growth and reproductive phases. Based on mophological and physio-biochemical responses of plants under stress condition, genotypes were divided into salt tolerant and succeptible groups. The experiment was comprised of 100 okra genotypes and each genotype was grown under control conditions and 6.5 dS m-1 NaCl concentration in a pot having 10 kg capacity. The experiment was conducted in a completely randomized design and each treatment was replicated three times. The results showed vast genetic variability among the evaluated okra germplasm traits like days to emergence, pod length, pod diameter, plant height, stem girth, and other yield-related parameters. Correlation analysis showed a highly significant positive association among the number of leaves at first flower and plant height at first flower.Likewise, pod weight also revealed a highly significant positive relationship for pod weight plant-1, pod length, and K+: Na+. Principal Component Analysis (PCA) revealed that out of 16 principal components (PCs), five components showed more than one eigenvalue and the first six PCs contributed 67.2% of the variation. Bi-plot analysis illustrated that genotypes 95, 111, 133, 99, and 128, under salt stress conditions, exhibited both high yield per plant and salt-tolerant behavior in other yield-related traits. On the basis of all studied traits, a salt susceptible group and a salt-tolerant group were formed. The salt tolerant group comprised of 97, 68, 95, 114, 64, 99, 111, 133, 128, and 109 genotypes, whereas, the salt susceptible group contained 137, 139, 130, 94, and 125 genotypes. Salt-tolerant okra genotypes were suggested to be used in further breeding programs aimed to develop salt tolerance in okra. These insights will empower precision breeding, underscore the importance of genetic diversity, and bear the potential to address the challenges of salt-affected soils while promoting broader agricultural resilience, economic prosperity, and food security.

Sublethal and transgenerational effects of synthetic insecticides on the biological parameters and functional response of Coccinella septempunctata (Coleoptera: Coccinellidae) under laboratory conditions.

Synthetic insecticides have been an inevitable part of plant protection throughout the world. Sublethal effects of these chemicals on beneficial insect species are one of the contemporary issues these days. Using the age-stage, two-sex life table model, this study evaluated the sublethal and transgenerational effects of six synthetic insecticides (imidacloprid, thiamethoxam, lambda-cyhalothrin, cypermethrin, chlorpyrifos and profenofos) commonly applied to winter vegetables, on the fitness and predation of the seven-spotted ladybeetle, Coccinella septempunctata, which is an efficient predator of aphids worldwide. According to results, all insecticides at their sublethal doses (LC30) significantly suppressed the emergence of adults, adult weight, fertility and fecundity of the parental generation compared to control treatment. The larval stage was prolonged and oviposition, fecundity and total longevity of the adult beetles were decreased in unexposed progeny whose parents were exposed to sublethal doses of all insecticides. Moreover, the biological parameters of adults, including the intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R 0) were significantly reduced when exposed to sublethal doses of insecticides. The predation rate of the F1 generation adults was also decreased after exposure to the sublethal doses of insecticides. However, chlorpyrifos, profenofos, lambda-cyhalothrin and cypermethrin exhibited more deleterious effects on the fitness and population parameters of beetles than imidacloprid and thiamethoxam.

Molecular bases of P450-mediated resistance to the neonicotinoid insecticide imidacloprid in the mosquito Ae. aegypti.

Resistance to chemical insecticides including pyrethroids, the main insecticide class used against mosquitoes, has re-kindled interest in the use of neonicotinoids. In this context, the present study aimed to characterize the molecular basis of neonicotinoid resistance in the mosquito Aedes aegypti. Resistance mechanisms were studied by combining transcriptomic and genomic data obtained from a laboratory strain selected at the larval stage after 30 generations of exposure to imidacloprid (Imida-R line). After thirty generations of selection, larvae of the Imida-R line showed an 8-fold increased resistance to imidacloprid and a significant cross-tolerance to the pyrethroids permethrin and deltamethrin. Cross-resistance to pyrethroids was only observed in adults when larvae were previously exposed to imidacloprid suggesting a low but inducible expression of resistance alleles at the adult stage. Resistance of the Imida-R line was associated with a slower larval development time in females. Multiple detoxification enzymes were over-transcribed in larvae in association with resistance including the P450s CYP6BB2, CYP9M9 and CYP6M11 previously associated with pyrethroid resistance. Some of them together with their redox partner NADPH P450 reductase were also affected by non-synonymous mutations associated with resistance. Combining genomic and transcriptomic data allowed identifying promoter variations associated with the up-regulation of CYP6BB2 in the resistant line. Overall, these data confirm the key role of P450s in neonicotinoid resistance in Ae. aegypti and their potential to confer cross-resistance to pyrethroids, raising concerns about the use of neonicotinoids for resistance management in this mosquito species.

Dechlorane Plus as an emerging environmental pollutant in Asia: a review.

Dechlorane Plus (DP) is an unregulated, highly chlorinated flame retardant. It has been manufactured from past 40 years but its presence in the environment was initially reported in 2006. Later, it has been found in various biotic and abiotic environmental matrices. However, little attention has been paid to monitor its presence in Asia. Many studies have reported the occurrence of DP in the environment of Asia, yet the data are scarce, and studies are limited to few regions. The objective of present review is to summarize the occurrence, distribution, and toxicity of this ubiquitous pollutant in various environmental matrices (biotic and abiotic). DP has also been reported in the areas with no emission sources, which proves its long-range transport. Moreover, urbanization and industrialization also affect the distribution of DP, i.e., high levels of DP have been found in urban areas relative to the rural. Tidal movement also incorporates in transport of DP across the aquatic system. Further, bioaccumulation trend of DP in various tissues is kidney > liver > muscle tissues, whereas, blood brain barrier resists its accumulation in brain tissues. Additionally, gender-based accumulation trends revealed high DP levels in females in comparison to males due to strong metabolism of males. Furthermore, methodological aspects and instrumental analysis used in previous studies have also been summarized here. However, data on biomagnification in aquatic ecosystem and bioaccumulation of DP in terrestrial food web are still scarce. Toxicity behavior of syn-DP and anti-DP is still unknown which might gain the interest for future studies.

Identification of Lysinibacillus sphaericus Binary toxin binding proteins in a malarial mosquito cell line by proteomics: A novel approach towards improving mosquito control.

Bacterial insecticidal proteins, such as the Bin toxin from Lysinibacillus sphaericus, could be used more extensively to control insecticide resistant mosquitoes. This study was aimed at identification of mosquito cell proteins binding Bin toxin. Results showed that purified toxin was toxic to Anopheles gambiae larvae and Ag55 cultured cells. Clathrin heavy chain (an endocytosis protein) and glycolytic enzymes such as pyruvate kinase, enolase and dihydrolipoamide dehydrogenase were identified as binders of Bin toxin. The viability of Ag55 cells in the presence of endocytosis inhibitor, pitstop2, was significantly decreased upon Bin treatment, while the inhibitor chlorpromazine did not affect Bin toxicity. Bin toxin treatment decreased ATP production and mitochondrial respiration in Ag55 cells, whereas non-mitochondrial oxygen consumption significantly increased after Bin toxin treatment. These findings are steps towards understanding how Bin toxin kills mosquitoes. SIGNIFICANCE: Mosquitoes are vectors of pathogens causing human diseases such as dengue fever, yellow fever, zika virus and malaria. An insecticidal toxin from Lysinibacillus sphaericus called Binary, or Bin, toxin could be used more extensively to control insecticide resistant mosquitoes. Bin toxin enter cells in susceptible mosquitoes and induces apoptosis or autophagy. In the current research, we used the malaria mosquito Anopheles gambiae Ag55 cell line as a model. A proteomic-based approach identified proteins that interact with Bin toxin. Interacting proteins include clathrin heavy chain (endocytosis protein) and glycolysis enzymes such as pyruvate kinase, enolase and dihydrolipoamide dehydrogenase. In Ag55 cell toxicity assays, an endocytosis inhibitor, pitstop2, increased Bin toxicity. Real time assays with a Seahorse™ flux analyzer showed that Bin significantly affects mitochondrial respiration, a result consistent with cell death via apoptosis or autophagy. These research findings add insights into how an unusual binary protein exploits cellular machinery to kill mosquitoes.

Comparative efficacy of mitigation techniques for the detoxification of Prunus persica (L.) from selected pesticide residues.

Fruits are the valuable and important components of human diet. Among them, Prunus persica is a rich source of different minerals and dietary fibers. In Pakistan, the total annual production of P. persica is approximately 837,000 tons. In order to enhance agricultural yield and quality, the plant protection agents are employed during fruit production. Ultimately, this in turn leads to the incorporation of pesticide residues in fruits. In present study, an effort has been made for the determination of three selected pesticide residues, i.e., chlorpyrifos (CPF), difenoconazole (DFN), and carbendazim (CRB) in samples of P. persica collected from Swat territory. Samples were analyzed through high performance liquid chromatography (HPLC). Results revealed the occurrence of all three pesticides in studied samples; however, levels of CPF and DFN were found to be higher than MRLs. Moreover, the effects of different mitigation techniques revealed that highest reduction of CPF, DFN, and CRB (86%, 97%, 89%) residues was obtained by treatment with 10% CH3COOH followed by 10% NaCl (74%, 78%, 84%). The lowest reduction was obtained by treatment with 10% solution of NaOH (52%, 55%, 63%).

Seasonal variations of soil phosphorus and associated fertility indicators in wastewater-irrigated urban aridisol.

Use of wastewater is known to provide nutrients for crop plants, but its potential to improve phosphorus (P) availability in semi-arid regions is poorly understood. In this study, seasonal changes in soil P availability as well as associated phyiscochemical and biochemical indicators were investigated from the wastewater irrigated urban soils of Faisalabad, Pakistan. Soil sampling was carried out during summer and winter season from four wastewater irrigated sites of varied stream flow i.e. upstream wastewater (UWW), midstream wastewater (MWW), lowerstream wastewater (LWW) and downstream wastewater (DWW), and canal water irrigation (CWI) as a reference site. Across seasons, MWW site had significantly higher soil organic carbon (SOC), water extractable organic carbon (WEOC), microbial biomass carbon (MBC), microbial biomass phosphorus (MBP) as well as the availability of phosphorus i.e. NaHCO3-P and H2O-P compared to CWI site. In both sampling seasons, MWW site also recorded significantly higher soil enzyme activities compared to the rest of wastewater sites. Moreover, significantly higher total P and electrical conductivity (EC) of soil was noticed at DWW site across both summer and winter seasons. Biplot principle component analysis also indicated seasonally a stronger shift in soil total P and EC at DWW site. On the other hand, availability of P was closely related to soil active carbon pools at MWW site. However, buildup of soil salinity particularly at DWW site along with lower P availability and associated changes in other soil properties, call for careful assessment of wastewater use in these urban soils.

Determination of drought tolerance related traits in Helianthus argophyllus, Helianthus annuus, and their hybrids.

Drought is a major constraint for sunflower (Helianthus annuus) production worldwide. Drought tolerance traits have been identified in the related wild species Helianthus argophyllus. This study was initiated to develop sunflower drought-tolerant genotypes by crossing cultivated sunflower with this species and analyze drought tolerance traits in the H. annuus and H. argophyllus populations, H. annuus intraspecific hybrids, and H. annuus × H. argophyllus interspecific hybrids along with the commercial hybrid Hysun-33 under three stress regimes: exogenous application of ABA, both by foliar spray and irrigation, and 5% PEG-induced osmotic stress. H. argophyllus populations had a significantly lower leaf area and higher water-use efficiency and leaf cuticular wax content under all treatments, and maintained a higher net photosynthetic rate and stomatal conductance under osmotic stress. Small leaf area and high cuticular waxes content of the wild species were, however, not inherited in interspecific hybrids which suggested for selection in F2 for these traits. Therefore, transgressive plants were selected in the F2 population to establish F3 plant progenies with silver-leafed canopy of H. argophyllus which showed higher achene yield under stress condition. These results are discussed with a view to using H. argophyllus to improve drought tolerance in cultivated sunflower.

Characterization of N2O emission and associated bacterial communities from the gut of wood-feeding termite Nasutitermes voeltzkowi.

Xylophagous termites rely on nitrogen deficient foodstuff with a low C/N ratio. Most research work has focused on nitrogen fixation in termites highlighting important inflow and assimilation of atmospheric nitrogen into their bodies fundamentally geared up by their intestinal microbial symbionts. Most of termite body nitrogen is of atmospheric origin, and microbially aided nitrification is the principal source of this nitrogen acquisition, but contrarily, the information regarding potent denitrification process is very scarce and poorly known, although the termite gut is considered to carry all favorable criteria necessary for microbial denitrification. Therefore, in this study, it is hypothesized that whether nitrification and denitrification processes coexist in intestinal milieu of xylophagous termites or not, and if yes, then is there any link between the denitrification product, i.e., N2O and nitrogen content of the food substrate, and moreover where these bacterial communities are found along the length of termite gut. To answer these questions, we measured in vivo N2O emission by Nasutitermes voeltzkowi (Nasutitermitinae) maintained on different substrates with varying C/N ratio, and also, molecular techniques were applied to study the diversity (DGGE) and density (qPCR) of bacterial communities in anterior and posterior gut portions. Rersults revealed that xylophagous termites emit feeble amount of N2O and molecular studies confirmed this finding by illustrating the presence of an ample density of N2O-reductase (nosZ) gene in the intestinal tract of these termites. Furthermore, intestinal bacterial communities of these termites were found more dense and diverse in posterior than anterior portion of the gut.

Chemical and biological insecticides select distinct gene expression patterns in Aedes aegypti mosquito.

Worldwide evolution of mosquito resistance to chemical insecticides represents a major challenge for public health, and the future of vector control largely relies on the development of biological insecticides that can be used in combination with chemicals (integrated management), with the expectation that populations already resistant to chemicals will not become readily resistant to biological insecticides. However, little is known about the metabolic pathways affected by selection with chemical or biological insecticides. Here we show that Aedes aegypti, a laboratory mosquito strain selected with a biological insecticide (Bacillus thuringiensis israelensis, Bti) evolved increased transcription of many genes coding for endopeptidases while most genes coding for detoxification enzymes were under-expressed. By contrast, in strains selected with chemicals, genes encoding detoxification enzymes were mostly over-expressed. In all the resistant strains, genes involved in immune response were under-transcribed, suggesting that basal immunity might be a general adjustment variable to compensate metabolic costs caused by insecticide selection. Bioassays generally showed no evidence for an increased susceptibility of selected strains towards the other insecticide type, and all chemical-resistant strains were as susceptible to Bti as the unselected parent strain, which is a good premise for sustainable integrated management of mosquito populations resistant to chemicals.

Comparative analysis of response to selection with three insecticides in the dengue mosquito Aedes aegypti using mRNA sequencing.

BACKGROUND: Mosquito control programmes using chemical insecticides are increasingly threatened by the development of resistance. Such resistance can be the consequence of changes in proteins targeted by insecticides (target site mediated resistance), increased insecticide biodegradation (metabolic resistance), altered transport, sequestration or other mechanisms. As opposed to target site resistance, other mechanisms are far from being fully understood. Indeed, insecticide selection often affects a large number of genes and various biological processes can hypothetically confer resistance. In this context, the aim of the present study was to use RNA sequencing (RNA-seq) for comparing transcription level and polymorphism variations associated with adaptation to chemical insecticides in the mosquito Aedes aegypti. Biological materials consisted of a parental susceptible strain together with three child strains selected across multiple generations with three insecticides from different classes: the pyrethroid permethrin, the neonicotinoid imidacloprid and the carbamate propoxur. RESULTS: After ten generations, insecticide-selected strains showed elevated resistance levels to the insecticides used for selection. RNA-seq data allowed detecting over 13,000 transcripts, of which 413 were differentially transcribed in insecticide-selected strains as compared to the susceptible strain. Among them, a significant enrichment of transcripts encoding cuticle proteins, transporters and enzymes was observed. Polymorphism analysis revealed over 2500 SNPs showing > 50% allele frequency variations in insecticide-selected strains as compared to the susceptible strain, affecting over 1000 transcripts. Comparing gene transcription and polymorphism patterns revealed marked differences among strains. While imidacloprid selection was linked to the over transcription of many genes, permethrin selection was rather linked to polymorphism variations. Focusing on detoxification enzymes revealed that permethrin selection strongly affected the polymorphism of several transcripts encoding cytochrome P450 monooxygenases likely involved in insecticide biodegradation. CONCLUSIONS: The present study confirmed the power of RNA-seq for identifying concomitantly quantitative and qualitative transcriptome changes associated with insecticide resistance in mosquitoes. Our results suggest that transcriptome modifications can be selected rapidly by insecticides and affect multiple biological functions. Previously neglected by molecular screenings, polymorphism variations of detoxification enzymes may play an important role in the adaptive response of mosquitoes to insecticides.

The central role of mosquito cytochrome P450 CYP6Zs in insecticide detoxification revealed by functional expression and structural modelling.

The resistance of mosquitoes to chemical insecticides is threatening vector control programmes worldwide. Cytochrome P450 monooxygenases (CYPs) are known to play a major role in insecticide resistance, allowing resistant insects to metabolize insecticides at a higher rate. Among them, members of the mosquito CYP6Z subfamily, like Aedes aegypti CYP6Z8 and its Anopheles gambiae orthologue CYP6Z2, have been frequently associated with pyrethroid resistance. However, their role in the pyrethroid degradation pathway remains unclear. In the present study, we created a genetically modified yeast strain overexpressing Ae. aegypti cytochrome P450 reductase and CYP6Z8, thereby producing the first mosquito P450-CPR (NADPH-cytochrome P450-reductase) complex in a yeast recombinant system. The results of the present study show that: (i) CYP6Z8 metabolizes PBAlc (3-phenoxybenzoic alcohol) and PBAld (3-phenoxybenzaldehyde), common pyrethroid metabolites produced by carboxylesterases, producing PBA (3-phenoxybenzoic acid); (ii) CYP6Z8 transcription is induced by PBAlc, PBAld and PBA; (iii) An. gambiae CYP6Z2 metabolizes PBAlc and PBAld in the same way; (iv) PBA is the major metabolite produced in vivo and is excreted without further modification; and (v) in silico modelling of substrate-enzyme interactions supports a similar role of other mosquito CYP6Zs in pyrethroid degradation. By playing a pivotal role in the degradation of pyrethroid insecticides, mosquito CYP6Zs thus represent good targets for mosquito-resistance management strategies.

Molecular mechanisms associated with increased tolerance to the neonicotinoid insecticide imidacloprid in the dengue vector Aedes aegypti.

Mosquitoes are vectors of several major human diseases and their control is mainly based on the use of chemical insecticides. Resistance of mosquitoes to organochlorines, organophosphates, carbamates and pyrethroids led to a regain of interest for the use of neonicotinoid insecticides in vector control. The present study investigated the molecular basis of neonicotinoid resistance in the mosquito Aedes aegypti. A strain susceptible to insecticides was selected at the larval stage with imidacloprid. After eight generations of selection, larvae of the selected strain (Imida-R) showed a 5.4-fold increased tolerance to imidacloprid while adult tolerance level remained low. Imida-R larvae showed significant cross-tolerance to other neonicotinoids but not to pyrethroids, organophosphates and carbamates. Transcriptome profiling identified 344 and 108 genes differentially transcribed in larvae and adults of the Imida-R strain compared to the parental strain. Most of these genes encode detoxification enzymes, cuticle proteins, hexamerins as well as other proteins involved in cell metabolism. Among detoxification enzymes, cytochrome P450 monooxygenases (CYPs) and glucosyl/glucuronosyl transferases (UDPGTs) were over-represented. Bioassays with enzyme inhibitors and biochemical assays confirmed the contribution of P450s with an increased capacity of the Imida-R microsomes to metabolize imidacloprid in presence of NADPH. Comparison of substrate recognition sites and imidacloprid docking models of six CYP6s over-transcribed in the Imida-R strain together with Bemisia tabaci CYP6CM1vQ and Drosophila melanogaster CYP6G1, both able to metabolize imidacloprid, suggested that CYP6BB2 and CYP6N12 are good candidates for imidacloprid metabolism in Ae. aegypti. The present study revealed that imidacloprid tolerance in mosquitoes can arise after few generations of selection at the larval stage but does not lead to a significant tolerance of adults. As in other insects, P450-mediated insecticide metabolism appears to play a major role in imidacloprid tolerance in mosquitoes.

Do pollutants affect insecticide-driven gene selection in mosquitoes? Experimental evidence from transcriptomics.

The control of mosquitoes transmitting infectious diseases relies mainly on the use of chemical insecticides. However, the emergence of insecticide resistance threatens mosquito control programs. Until now, most research efforts have been focused on elucidating resistance mechanisms caused by insecticide treatments. Less attention has been paid to the impact of the mosquito chemical environment on insecticide-driven selection mechanisms. Here the mosquito Aedes aegypti was used as a model species to conduct laboratory experiments combining the exposure of mosquito larvae to a sub-lethal concentration of xenobiotics and their selection with the insecticide permethrin. After 10 generations, bioassays and a transcriptome profiling with a 15 k microarray were performed comparatively on all strains. The three selected strains showed a small but significant increase of permethrin resistance compared to the susceptible parental strain. Microarray analysis revealed that the transcription of many genes was altered by insecticide selection. Exposing larvae to sub-lethal concentrations of the pollutant fluoranthene or the insecticide permethrin prior to selection at each generation affected the selection of several genes, including those involved in detoxification, transport and cell metabolism. Genes potentially involved in permethrin resistance and cross-responses between xenobiotics and insecticide were identified. The present study investigated for the first time the impact of the presence of pollutants in mosquito environment on insecticide-driven selection mechanisms. Our results revealed that mosquitoes exposed to xenobiotics show a different adaptive response to insecticide selection pressure. This suggests that insect chemical environment can shape the long-term selection of metabolic mechanisms leading to insecticide resistance.

Insecticide resistance in the dengue vector Aedes aegypti from Martinique: distribution, mechanisms and relations with environmental factors.

Dengue is an important mosquito borne viral disease in Martinique Island (French West Indies). The viruses responsible for dengue are transmitted by Aedes aegypti, an indoor day-biting mosquito. The most effective proven method for disease prevention has been by vector control by various chemical or biological means. Unfortunately insecticide resistance has already been observed on the Island and recently showed to significantly reduce the efficacy of vector control interventions. In this study, we investigated the distribution of resistance and the underlying mechanisms in nine Ae. aegypti populations. Statistical multifactorial approach was used to investigate the correlations between insecticide resistance levels, associated mechanisms and environmental factors characterizing the mosquito populations. Bioassays revealed high levels of resistance to temephos and deltamethrin and susceptibility to Bti in the 9 populations tested. Biochemical assays showed elevated detoxification enzyme activities of monooxygenases, carboxylesterases and glutathione S-tranferases in most of the populations. Molecular screening for common insecticide target-site mutations, revealed the presence of the "knock-down resistance" V1016I Kdr mutation at high frequency (>87%). Real time quantitative RT-PCR showed the potential involvement of several candidate detoxification genes in insecticide resistance. Principal Component Analysis (PCA) performed with variables characterizing Ae. aegypti from Martinique permitted to underline potential links existing between resistance distribution and other variables such as agriculture practices, vector control interventions and urbanization. Insecticide resistance is widespread but not homogeneously distributed across Martinique. The influence of environmental and operational factors on the evolution of the resistance and mechanisms are discussed.

Transcriptome response to pollutants and insecticides in the dengue vector Aedes aegypti using next-generation sequencing technology.

BACKGROUND: The control of mosquitoes transmitting infectious diseases relies mainly on the use of chemical insecticides. However, mosquito control programs are now threatened by the emergence of insecticide resistance. Hitherto, most research efforts have been focused on elucidating the molecular basis of inherited resistance. Less attention has been paid to the short-term response of mosquitoes to insecticides and pollutants which could have a significant impact on insecticide efficacy. Here, a combination of LongSAGE and Solexa sequencing was used to perform a deep transcriptome analysis of larvae of the dengue vector Aedes aegypti exposed for 48 h to sub-lethal doses of three chemical insecticides and three anthropogenic pollutants. RESULTS: Thirty millions 20 bp cDNA tags were sequenced, mapped to the mosquito genome and clustered, representing 6850 known genes and 4868 additional clusters not located within predicted genes. Mosquitoes exposed to insecticides or anthropogenic pollutants showed considerable modifications of their transcriptome. Genes encoding cuticular proteins, transporters, and enzymes involved in the mitochondrial respiratory chain and detoxification processes were particularly affected. Genes and molecular mechanisms potentially involved in xenobiotic response and insecticide tolerance were identified. CONCLUSIONS: The method used in the present study appears as a powerful approach for investigating fine transcriptome variations in genome-sequenced organisms and can provide useful informations for the detection of novel transcripts. At the biological level, despite low concentrations and no apparent phenotypic effects, the significant impact of these xenobiotics on mosquito transcriptomes raise important questions about the 'hidden impact' of anthropogenic pollutants on ecosystems and consequences on vector control.

Impact of glyphosate and benzo[a]pyrene on the tolerance of mosquito larvae to chemical insecticides. Role of detoxification genes in response to xenobiotics.

The effect of exposure of Aedes aegypti larvae for 72h to sub-lethal concentrations of the herbicide glyphosate and the polycyclic aromatic hydrocarbon benzo[a]pyrene on their subsequent tolerance to the chemical insecticides imidacloprid, permethrin and propoxur, detoxification enzyme activities and transcription of detoxification genes was investigated. Bioassays revealed a significant increase in larval tolerance to imidacloprid and permethrin following exposure to benzo[a]pyrene and glyphosate. Larval tolerance to propoxur increased moderately after exposure to benzo[a]pyrene while a minor increased tolerance was observed after exposure to glyphosate. Cytochrome P450 monooxygenases activities were strongly induced in larvae exposed to benzo[a]pyrene and moderately induced in larvae exposed to imidacloprid and glyphosate. Larval glutathione S-transferases activities were strongly induced after exposure to propoxur and moderately induced after exposure to benzo[a]pyrene and glyphosate. Larval esterase activities were considerably induced after exposure to propoxur but only slightly induced by other xenobiotics. Microarray screening of 290 detoxification genes following exposure to each xenobiotic with the DNA microarray Aedes Detox Chip identified multiple detoxification and red/ox genes induced by xenobiotics and insecticides. Further transcription studies using real-time quantitative RT-PCR confirmed the induction of multiple P450 genes, 1 carboxy/cholinelesterase gene and 2 red/ox genes by insecticides and xenobiotics. Overall, this study reveals the potential of benzo[a]pyrene and glyphosate to affect the tolerance of mosquito larvae to chemical insecticides, possibly through the cross-induction of particular genes encoding detoxification enzymes.