摘要
以油用牡丹‘凤丹白’为试材,基于已构建的雌蕊转录组数据库,筛选出与双受精相关的bHLH基因同源性高的Unigene序列,命名为PobHLH。其开放阅读框(ORF) 1 338 bp,推测编码一个含有445个氨基酸的蛋白。序列分析表明, PobHLH编码的氨基酸序列中包含已确定的保守基序,包括HLH结构域、APB基序和核定位信号NLS。氨基酸序列比对发现PobHLH与拟南芥bHLH家族VII(a+b)亚组的AtUNE10相似度最高,其次是同亚组的AtSPT和AtALC,可能参与双受精中雌配子体的发育。荧光定量PCR分析表明PobHLH在授粉后24 h表达增量最高,在授粉后48 h表达丰度最大,与‘凤丹白’双受精时间进程相吻合,暗示PobHLH可能在双受精过程中起关键作用。
An unigene sequence shared high homology with bHLH putatively involved in plant double fertilization were obtained from previous-constructed oil tree peony(Paeonia ostii ‘Fengdanbai’) gynoecium transcriptome database and named as PobHLH. PobHLH contains a 1 338 bp-long ORF, putatively encoding 445 amino acid residues with typical bHLH domains. The putative amino acid sequence encoded by PobHLH contained conserved motifs, including HLH domain, APB motifs and NLS. By amino acid sequence comparison, it showed that PobHLH shared the highest similarity with that of AtUNE10 belonging to bHLH subgroup of VII(a+b) in Arabidopsis thaliana, followed by AtSPT and AtALC, which were confirmed to regulate the development of female gametophyte during double fertilization. qRT-PCR analysis indicated that PobHLH showed the highest increment appeared after fertilization for 24 hours, while, the highest abundance was detected after fertilization for 48 hours, which was consistent with process of the double fertilization, which implied its putative function in double fertilization.
引文
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