Abstract
Globally, Aedes aegypti is one of the most dangerous mosquitoes that plays a crucial role as a vector for human diseases, such as yellow fever, dengue, and chikungunya. To identify (1) transcriptomic basis of midgut (2) key genes that are involved in the toxicity process by a comparative transcriptomic analysis between the control and Bacillus thuringiensis (Bt) toxin (LLP29 proteins)-treated groups. Next-generation sequencing technology was used to sequence the midgut transcriptome of A. aegypti. A total of 17130 unigenes, including 574 new unigenes, were identified containing 16358 (95.49%) unigenes that were functionally annotated. According to differentially expressed gene (DEG) analysis, 557 DEGs were annotated, including 226 upregulated and 231 downregulated unigenes in the Bt toxin-treated group. A total of 442 DEGs were functionally annotated; among these, 33 were specific to multidrug resistance, 6 were immune-system-related (Lectin, Defensin, Lysozyme), 28 were related to putative proteases, 7 were lipase-related, 8 were related to phosphatases, and 30 were related to other transporters. In addition, the relative expression of 28 DEGs was further confirmed through quantitative real time polymerase chain reaction. The results provide a transcriptomic basis for the identification and functional authentication of DEGs in A. aegypti.
Highlights
Bacillus thuringiensis (Bt) plays an important role in pest control management and in public health and is widely used[15,16,17]
In order to further understand the transcriptomic basis of midgut and its main functional genes after B. thuringiensis subsp. israelensis (Bti) LLP29 infection, transcriptomic analysis of A. aegypti in response to LLP29 was carried out in the present research, other than only single Cry toxin, such as Cry11Aa30
These unigenes were compared by Basic Local Alignment Search Tool (BLAST) with those in Swiss-Prot[31], GO32, NR33, COG34, and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases[35]
Summary
Bacillus thuringiensis (Bt) plays an important role in pest control management and in public health and is widely used[15,16,17]. Israelensis (Bti) produces Cry toxins that have been used for mosquito control. These toxins are highly toxic to mosquitoes, such as Aedes, Anopheles, Culex, Mansonia, and Simulium larvae[18]. It is important to understand the interaction of Cry toxins with other midgut proteins. As toxin binding is partially dependent on receptor glycosylation[25], proteins that interact with carbohydrate-recognition domains (CRD) might affect toxicity. Next-generation sequencing technology was used to sequence the transcriptome of A. aegypti exposed to LLP29 protein, and successfully built a transcript database; In addition, putative insecticide resistance transcript, immune-system related transcript, stressed-related transcript, detoxification-related molecules, and selected DEGs were confirmed through quantitative real time polymerase chain reaction (qPCR). The present study provides valuable information that can be used to develop new genetics-based strategies and novel molecular tools to control A. aegypti
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