关键词: 菜粉蝶, 触角转录组, 高通量测序, 嗅觉相关基因, qRT-PCR

Abstract:  【Aim】To establish the antennal transcriptome database of the Pieris rapae adult, so as to deeply mine the gene data of P. rapae by using bioinformatics and molecular biology methods.【Methods】High-throughput sequencing platform Illumina NovaSeq 6000 was used to perform antennal transcriptome sequencing, sequence assembly, functional annotation and differentially expressed gene analysis of P. rapae adults. qRT-PCR was used to verify nine differentially expressed olfaction-related genes including PrapOR1, PrapOR2, PrapOR5, PrapOBP1, PrapOBP4, PrapOBP5, PrapSNMP1, PrapSNMP2 and PrapSNMP3.【Results】By sequencing of the antennal transcriptome of adults of P. rapae, a total of 17.65 GB sequencing data (NCBI registration number: PRJNA869896) were obtained. A total of 116 317 transcripts were obtained through filtering and sequence splicing. Then, 43 390 unigenes were obtained by Corset hierarchical clustering. BUSCO evaluation showed that the stitching quality had good integrity and high accuracy. The databases with the largest number of unigene annotations in a descending order were NT, NR, Pfam, GO, Swiss-Prot, KEGG and KOG/COG. Furthermore, 176 olfaction-related genes were screened by gene functional annotation analysis, among them 19 genes were differentially expressed, including 15 genes highly expressed in female adult antennae and four genes highly expressed in male adult antennae. qRT-PCR verification results showed that PrapOR1 and PrapOR2 were highly expressed in male adult antennae, PrapOR5, PrapOBP1, PrapOBP4, PrapOBP5, PrapSNMP1, PrapSNMP2 and PrapSNMP3 were highly expressed in female adult antennae, indicating the consistency with the transcriptome sequencing results.【Conclusion】In this study, a transcriptomic database of adult antennae of P. rapae was established, the olfaction-related genes were screened and the differential expression of olfaction-related genes was analyzed. The results provide a theoretical basis for further studies on the gene function and olfactory molecular mechanism of P. rapae.

Key words: Pieris rapae, antennal transcriptome, high-throughput sequencing, olfaction-related genes, qRT-PCR