草地贪夜蛾雄性成虫和5龄幼虫的转录组比较分析

【目的】草地贪夜蛾Spodoptera frugiperda是一种新近入侵我国的重要害虫。本研究旨在对草地贪夜蛾雄性成虫和5龄幼虫两个不同发育阶段的转录组进行比较分析。【方法】利用高通量测序技术对草地贪夜蛾雄性成虫和5龄幼虫进行转录组测序和数据组装,并对转录组数据进行功能注释和比较分析。【结果】经de novo组装共获得209 002条转录本,平均长度为687.55 bp,N50为982 bp。共有46 198条(57.43%) unigene在至少一个数据库中获得功能注释,其中1 713条(2.13%) unigene在所有数据库中均能获得注释。在GO数据库中获得205 269条unigene的注释,主要包括68个功能分类;在KEGG数据库中共有3 408条unigene得到注释,涉及277个代谢通路。共鉴定到424个嗅觉相关的基因,并且在雄性成虫和5龄幼虫之间的表达存在差异。通过比较转录组分析,在雄性成虫中鉴定到9 162个上调和6 399个下调差异表达基因(DEGs);功能富集分析发现在上调DEGs中涉及信息素以及信号转导的代谢通路显著富集,而下调DEGs中涉及解毒相关的通路显著富集。【结论】这些转录组数据为探究草地贪夜蛾的生长发育、嗅觉相关功能基因以及候选分子靶标提供资源信息。     

De novo transcriptome analysis of Taxus chinensis var. mairei to identify significant pathways associated with the fruit color of this species

The goal of this study was to find significant pathways associated with fruit color in Taxus chinensis var. mairei. Two Taxus chinensis var. mairei with fruits of different colors were sequenced using high-throughput sequencing. Then, unigene assembly, unigene annotation, and classification of Clusters of Orthologous Groups of proteins (COGs) were performed. Gene ontology (GO) classification and pathway enrichment analysis for unigenes were also conducted. Protein coding sequences (CDS) were predicted. Subsequently, differentially expressed unigenes, functional enrichment pathways of differentially expressed unigenes, single nucleotide polymorphisms (SNPs), and simple sequence repeats (SSRs) were analyzed. The data of Taxus chinensis var. mairei were assembled into 45,112 unigenes with an average size of 1232 bp and an N50 of 1882 bp. The significantly enriched biological processes (BP) were metabolic processes, and cellular processes, followed by single-organism processes, and reproductive processes. Significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were metabolic pathways, pathways related to the biosynthesis of secondary metabolites, and RNA transport pathways. Transition had a higher number of SNP sites than transversion. Trinucleotide had the highest SSR motif numbers. Metabolic pathways and the biosynthesis of secondary metabolites may play significant roles in determining fruit color in Taxus chinensis var. mairei. In addition, the present dataset would provide a basis for future functional genomic research on Taxus chinensis var. mairei.     

沙葱萤叶甲卵滞育的转录组学分析

【目的】建立沙葱萤叶甲Galeruca daurica滞育卵转录组数据库,挖掘卵滞育相关的基因以及代谢和信号通路,在转录组水平探讨卵滞育的分子机制。【方法】采用Illumina NovaSeq6000高通量测序平台对沙葱萤叶甲滞育卵与解除滞育卵进行转录组测序,并进行生物信息学分析;利用DESeq软件分析沙葱萤叶甲滞育卵与解除滞育卵中的差异表达基因,对差异表达基因进行KEGG通路富集分析;利用qRT PCR技术对10个差异表达基因的表达模式进行验证。【结果】基于沙葱萤叶甲滞育卵与解除滞育卵转录组测序结果,共获得53 389个unigene,其中差异表达基因2 145个,24个差异表达基因与保幼激素信号及脂肪酸生物合成和降解相关。与解除滞育卵相比,滞育卵转录组中1 297个基因上调表达,富集于124条KEGG通路,其中核糖体通路显著富集;848个基因下调表达,富集于73条KEGG通路,其中MAPK信号通路和糖胺聚糖生物合成通路显著富集。qRT-PCR结果表明,随机选取的10个差异表达基因的表达趋势与RNA-Seq转录组测序结果完全一致。【结论】保幼激素,脂肪酸生物合成和降解,核糖体,MAPK信号及糖胺聚糖生物合成等通路可能在沙葱萤叶甲卵滞育调控中起着重要的作用。     

三峡库区消落带野生狗牙根对水淹适生性的转录组分析

为探究狗牙根对水淹环境的适生性机制,取长势一致的狗牙根分为未水淹对照组(A1)和30 cm沉水处理组(A2),A2水淹15 d后两组同时采样,提取样本总RNA作转录组测序。A1和A2测序数据经de novo组装分别获得128031和83363条高质量Unigene;A1和A2对比发现28844条差异表达基因(differentially expressed genes, DEGs),其中11858条DEGs上调表达,16986条DEGs下调表达;功能注释显示,这些DEGs主要集中在转录、翻译、碳水化合物代谢和环境适应等途径;信号通路富集显示,包括植物对病原体的防御、物质代谢、光合作用、形态发育和抗氧化等过程的103个信号通路与狗牙根对水淹生境的适生性有关;随机挑选4个DEGs作qRT-PCR分析,荧光定量结果与测序数据基本一致,说明基于转录组分析DEGs的表达情况较为可靠。该工作或可为进一步挖掘与狗牙根水淹适生性相关的调控基因和功能基因提供基础。     

三叶木通转录组分析及三萜皂苷生物合成途径关键酶基因的发掘

为了解三叶木通(Akebia trifoliata(Thunb.)Koidz.)的三萜皂苷合成途径及其关键酶,本研究对其花、叶、根、茎进行转录组测序,组装获得了57.25 Gb数据,含140 859个unigenes,序列平均长度为1350 bp。KEGG代谢通路富集结果显示,517个unigenes参与三萜皂苷合成相关的3条代谢途径,其中415个unigenes编码三萜皂苷生物合成途径的19个关键酶。对三萜皂苷生物合成过程中的关键酶角鲨烯环氧酶(SE)进行序列分析和同源建模,发现其具有保守的底物结合结构域。将三叶木通茎与花、叶、根的基因表达水平进行比较,发现茎与根相比较其上调基因数目最多,其中295个差异表达基因(DEGs)与三萜皂苷生物合成途径相关。