摘要
【目的】建立菜粉蝶Pieris rapae成虫触角转录组数据库,利用生物信息学和分子生物学手段深入挖掘菜粉蝶基因数据信息。【方法】采用高通量测序平台(Illumina NovaSeq 6000)对菜粉蝶成虫触角进行转录组测序、序列组装、功能注释及差异表达基因分析,对PrapOR1,PrapOR2,PrapOR5,PrapOBP1,PrapOBP4,PrapOBP5,PrapSNMP1,PrapSNMP2和PrapSNMP39个差异表达的嗅觉相关基因进行qRT-PCR验证。【结果】菜粉蝶成虫触角转录组共获得17.65 GB测序数据(NCBI登录号:PRJNA869896),经过滤和序列拼接共得到116317条转录本,随后进行Corset层次聚类获得43390条unigene,经BUSCO评估,拼接质量完整度好,准确性高。注释到的unigene数目从大到小排列的数据库依次为NT,NR,Pfam,GO,Swiss-Prot,KEGG和KOG/COG;进一步进行基因功能注释分析,筛选得到176个嗅觉相关基因,其中19个基因差异表达,包括在雌成虫触角中高表达的15个基因和在雄成虫触角中高表达的4个基因。qRT-PCR验证结果表明,PrapOR1和PrapOR2在雄成虫触角中高表达,PrapOR5,PrapOBP1,PrapOBP4,PrapOBP5,PrapSNMP1,PrapSNMP2和PrapSNMP3在雌成虫触角中高表达,与转录组测序结果相符。【结论】本研究建立了菜粉蝶成虫触角转录组数据库,筛选得到了嗅觉相关基因,并对嗅觉相关基因进行了差异表达分析,研究结果为进一步研究菜粉蝶的基因功能及嗅觉分子机制奠定了理论基础。
【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 116317 transcripts were obtained through filtering and sequence splicing.Then,43390 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.
作者
薛增昇
闫喜中
赵劲宇
宋程飞
刘丹
王韩
郝赤
马力
XUE Zeng-Sheng;YAN Xi-Zhong;ZHAO Jing-Yu;SONG Cheng-Fei;LIU Dan;WANG Han;HAO Chi;MA Li(College of Plant protection,Shanxi Agricultural University,Taigu 030801,China)
出处
《昆虫学报》
CAS
CSCD
北大核心
2023年第5期642-652,共11页
Acta Entomologica Sinica
基金
科技部高端外国专家引进计划(G2022004006L)
山西省自然科学基金项目(20210302123387,202103021223125)
山西省研究生教育创新项目(2021Y330)
山西农业大学“特”“优”农业高质量发展科技支撑工程(TYGC-09)。
