穿心莲海藻糖-6-磷酸合酶1(ApTPS1)生物信息学与表达分析

Bioinformatics and Expression Analysis of Trehalose-6-phosphate Synthase 1(ApTPS1)in Andrographis paniculata

以穿心莲(Andrographis paniculata)为材料,基于全长转录组数据,筛选和克隆穿心莲海藻糖-6-磷酸合酶1基因(命名为ApTPS1)。生物信息学分析显示,ApTPS1基因的开放阅读框长度为2787 bp,编码928个氨基酸,分子质量为105178.82 u,理论等电点为6.67,为亲水蛋白,无信号肽。多重序列比对显示,ApTPS1较为保守,系统进化树分析表明该蛋白与凤梨亲缘关系最近,与拟南芥和烟草亲缘关系相对较远。利用实时荧光定量PCR分析穿心莲不同生态型、不同器官以及不同光周期条件下ApTPS1基因的表达特性,结果表明,早花生态型穿心莲的ApTPS1表达水平高于晚花生态型,且这种关系不受光周期的影响;ApTPS1在穿心莲的不同器官中的表达水平依次为花>果实>花蕾≈新叶≈老叶。以上结果说明,ApTPS1可能与穿心莲开花时间及生殖器官发育有密切关系。ApTPS1基因的克隆及差异表达分析为穿心莲开花期调控及优良品种的选育提供了理论基础。

Trehalose-6-phosphate synthase 1 gene(named ApTPS1)from Andrographis paniculata was screened and cloned based on the full-length transcriptome data.The bioinformatics analysis showed that the open reading frame(ORF)length of ApTPS1 gene was 2787 bp,encoding 928 amino acids,the molecular mass was 105178.82 u,and the theoretical isoelectric point PI was 6.67,which is a hydrophilic protein without signal peptide.Multiple sequence alignment showed that ApTPS1 was relatively conservative.Phylogenetic tree analysis showed that the protein was closest to pineapple but relatively far from model plants Arabidopsis thaliana and tobacco.The expression characteristics of ApTPS1 gene in different ecotypes,plant organs and under different photoperiod induced conditions of A.paniculata were analyzed by real-time fluorescence quantitative PCR(q-PCR).The results showed that the expression level of ApTPS1 gene in early flowering ecotypes of A.paniculata was higher than that in late flowering ecotypes,its relationship was not affected by photoperiod.The expression levels of ApTPS1 in different organs of A.paniculata were flower>fruit>flower bud≈new leaf≈old leaf.The results suggested that ApTPS1 is closely related to the flowering time of A.paniculata.The cloning and differential expression analysis of ApTPS1 gene provides a theoretical basis for the flowering regulation and breeding of A.paniculata.