小黑瓢虫雌成虫滞育基因的转录组测序分析

Using transcriptome sequencing to analyze the diapause-related genes of female adults of whitefly predatory lady beetle Delphastus catalinae

为明确小黑瓢虫Delphastus catalinae滞育的分子机制,使用Illumina HiSeq2500高通量测序平台分析小黑瓢虫雌成虫非滞育期、滞育期和滞育解除期的相对转录水平,从转录组测序数据中选取碱性磷酸酶(alkaline phosphatase,ALP)基因、过氧化氢酶(catalase,CAT)基因、乳酸脱氢酶(lactate dehydrogenase,LDH)基因、过氧化物酶(peroxidase,POD)基因、超氧化物歧化酶(superoxide dismutase,SOD)基因、海藻糖酶(trehalase,TRE)基因、促葡萄糖转运1蛋白亚基基因Ref和SCoAL琥珀酰辅酶连接酶基因GDP-forming,采用实时荧光定量PCR(quantitative real-time PCR,qRTPCR)技术检测上述基因在小黑瓢虫雌成虫3个不同时期的表达特征。结果显示,从9个样品中共筛选出67.86 Gb的clean data;936447条contig被组装成52255条unigene,所有的unigene都通过BLAST对Nr数据库进行了注释,有23539条unigene被匹配。通过对unigene进行COG、GO和KEGG分析,发现在非滞育组和滞育组之间以及滞育解除组和滞育组之间分别有3690条和4662条差异表达基因,其中571条差异表达基因在滞育期特异性表达,涉及116条KEGG富集通路。qRT-PCR验证了转录组测序的准确性,发现LDH基因在滞育期的表达量显著下降,而其余7个基因的表达量均上升。

To clarify the molecular mechanism of whitefly predatory lady beetle Delphastus catalinae during diapause,Illumina HiSeq2500 high-throughput sequencing platform was used to reveal the relative transcriptional levels of non-diapause,diapause and termination of diapause(DT)of the female adults.The genes alkaline phosphatase(ALP),catalase(CAT),lactate dehydrogenase(LDH),peroxidase(POD),superoxide dismutase(SOD),trehalase(TRE),Ref,and GDP-forming were selected from the transcriptome database.Quantitative real-time PCR(qRT-PCR)was used to determine the expression patterns of these genes in three different stages.The results showed that a total of 67.86 Gb clean reads were filtered from nine samples.Additionally,936447 contigs were assembled into 52255 unigenes.All of the unigenes were annotated through BLAST alignment against the Nr database,and 23539 unigenes were matched.Further analysis of unigenes using the COG,GO and KEGG databases was performed.Through pairwise comparisons of the non-diapause,diapause,and diapause-termination groups,3690 and 4662 differentially expressed genes(DEGs)were identified between non-diapause and dia pause,and between diapause-termination and diapause groups,respectively.Moreover,571 DEGs were specifically expressed during the diapause period,involving 116 KEGG enrichment pathways.In addition,qRT-PCR verified the accuracy of the transcriptome.It was found that the expression of LDH decreased significantly during the diapause stage,while the expression of the other seven genes increased.