Para Sodium Channel Expression Profiling of the pink bollworm Pectinophora Gossypiella (Saunders) (Lepidoptera : Gelichidae) Exposed to Some Insecticide Using Real-Time PCR
Keywords:
Malathion, Chlorpyrifos, Cypermethrin, Deltamethrin, PARA Voltage-Gated Sodium Channel, Real-Time PCR, Reference Gene, Stability Level, Insecticide Treatments, Pectinophora GossypiellaAbstract
Pyrethroid insecticides are widely used to control many agricultural pests, its neurotoxic mode of action works by modifying the normal function of voltage-gated sodium channels dynamic leading to insect paralysis and death. The Pink Bollworm Pectinophora gossypiella (Saunders) is a serious pest in attacking cotton in Egyptian fields. Insecticide control facing problems because of its biological features of hiding inside bolls and being protected from insecticide applications. Thus, a large number of bolls infested with larvae collected from fields of Sharkia and Benysuef governorate, and exposed to insecticide of pyrethroid and organophosphorus sub-lethal concentrations equal LC50 defined from laboratory bioassay toxicity features. Thus, the PARA voltage-gated sodium channel domain ll relative expression was investigated using the molecular marker Real-time PCR procedures via the steps of generating data was RNA isolation and characterization, cDNA synthesis, then generating normalization factors, and Ct data attained and analyzed using 2-ΔΔCT method to quantify the gene expression quantitatively of insecticide treated larvae tissue was extracted. Expression profiling, stability, ranking of reference gene, and validation of the optimum reference genes were calculated using the three optimal gene finders. Results showed that chlorpyrifos and malathion treated genes having lower mean weights and were considered transcriptionally stable and considered ideal reference genes. statistical analysis results were Hierarchical clustering, Heat map, and principle component analysis based on various clustering systems and algorithms as distance measures were completed and gene copy number was about 10 copies. Data showed the most expressed ratio of samples was chlorpyrifos followed by malathion compared with the control, cypermethrin, and deltamethrin. The most effective insecticide was cypermethrin followed by malathion and the least the other two insecticides according to CT values.
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