Part epidemic tissues of the fifth-instar larval mutants of four genes (white, brown, ok, lightoid) or the thorax and abdomen of adults (white, scarlet) and their corresponding wild types were dissected in phosphate buffer saline and then used to extract genomic DNA using TreliefTMAnimal Genomic DNA Kit (TsingKe, China) following the manufacturer’s protocols. The tissues of each individual were as a biological sample. At least three replicates were carried out for mutants of each gene. Except some adult mutants of white gene, part tissues of the same individual for the mutants and wild-types are also used for RNA extraction as described in the following part. Subsequently, primers flanking the target sites for each gene (Additional file 1: Table S4) were designed, and the PCR reaction were carried out using the 20 μl volumes, according to TransDirect PCR SuperMix (Trans, China). PCR products were TA-cloned into PMD19 vectors (Takara, Japan) and 10 clones were randomly picked up and sequenced for each individual. Sequence data were analyzed using SeqMan software (DNASTAR7.0) to determine the exact mutation type.

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