Evaluation of Reference Genes and Expression Level of Genes Potentially Involved in the Mode of Action of Cry1Ac and Cry1F in a Susceptible Reference Strain of Chrysodeixis includens.

Abstract

Simple SummarySoybean looper (a moth species) is a major pest of soybean plants in the American continent and its larvae need to be kept under economic damage thresholds to guarantee sustainable yields. Soybean looper control relies mostly on the use of insecticides and genetically modified crops expressing Bacillus thuringiensis (Bt) insecticidal proteins. Due to the high selection pressure exerted by these control measures, resistance has developed to different insecticides and Bt proteins. Here, we tested the basal sensitivity of a soybean looper laboratory reference strain against two insecticidal proteins and determined the level of expression of potential receptors of these proteins across all (six) larval stages. Furthermore, we identified stable reference genes across all larval stages to normalize gene expression data obtained by quantitative polymerase chain reaction (qPCR). The results presented in this communication are useful to support future studies on insecticide and insecticidal protein resistance in soybean looper.Soybean looper (SBL), Chrysodeixis includens (Walker), is one of the major lepidopteran pests of soybean in the American continent. SBL control relies mostly on the use of insecticides and genetically modified crops expressing Bacillus thuringiensis (Bt) insecticidal Cry proteins. Due to the high selection pressure exerted by these control measures, resistance has developed to different insecticides and Bt proteins. Nevertheless, studies on the mechanistic background are still scarce. Here, the susceptibility of the laboratory SBL-Benzon strain to the Bt proteins Cry1Ac and Cry1F was determined in diet overlay assays and revealed a greater activity of Cry1Ac than Cry1F, thus confirming results obtained for other sensitive SBL strains. A reference gene study across larval stages with four candidate genes revealed that RPL10 and EF1 were the most stable genes for normalization of gene expression data obtained by RT-qPCR. Finally, the basal expression levels of eight potential Bt protein receptor genes in six larval instars were analyzed, including ATP-binding cassette (ABC) transporters, alkaline phosphatase, aminopeptidases, and cadherin. The results presented here provide fundamental knowledge to support future SBL resistance studies.