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MicroRNAs and their putative targets in Brassica napus seed maturation

机译:甘蓝型油菜种子成熟中的微小RNA及其推定靶标

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Background MicroRNAs (miRNAs) are 20–21 nucleotide RNA molecules that suppress the transcription of target genes and may also inhibit translation. Despite the thousands of miRNAs identified and validated in numerous plant species, only small numbers have been identified from the oilseed crop plant Brassica napus (canola) – especially in seeds. Results Using next-generation sequencing technologies, we performed a comprehensive analysis of miRNAs during seed maturation at 9 time points from 10?days after flowering (DAF) to 50 DAF using whole seeds and included separate analyses of radicle, hypocotyl, cotyledon, embryo, endosperm and seed coat tissues at 4 selected time points. We identified more than 500 conserved miRNA or variant unique sequences with >300 sequence reads and also found 10 novel miRNAs. Only 27 of the conserved miRNA sequences had been previously identified in B. napus (miRBase Release 18). More than 180 MIRNA loci were identified/annotated using the B. rapa genome as a surrogate for the B.napus A genome. Numerous miRNAs were expressed in a stage- or tissue-specific manner suggesting that they have specific functions related to the fine tuning of transcript abundance during seed development. miRNA targets in B. napus were predicted and their expression patterns profiled using microarray analyses. Global correlation analysis of the expression patterns of miRNAs and their targets revealed complex miRNA-target gene regulatory networks during seed development. The miR156 family was the most abundant and the majority of the family members were primarily expressed in the embryo. Conclusions Large numbers of miRNAs with diverse expression patterns, multiple-targeting and co-targeting of many miRNAs, and complex relationships between expression of miRNAs and targets were identified in this study. Several key miRNA-target expression patterns were identified and new roles of miRNAs in regulating seed development are suggested. miR156, miR159, miR172, miR167, miR158 and miR166 are the major contributors to the network controlling seed development and maturation through their pivotal roles in plant development. miR156 may regulate the developmental transition to germination.
机译:背景MicroRNA(miRNA)是20-21个核苷酸的RNA分子,可抑制目标基因的转录,也可能抑制翻译。尽管已在众多植物物种中鉴定并验证了成千上万个miRNA,但从油料作物甘蓝型油菜(canola)中仅鉴定出少量-特别是在种子中。结果使用下一代测序技术,我们使用完整种子对开花后10 d(DAF)至50 DAF的9个时间点的种子成熟过程中的miRNA进行了全面分析,包括对胚根,下胚轴,子叶,胚, 4个选定的时间点的胚乳和种皮组织。我们鉴定了500多个保守的miRNA或具有> 300个序列读数的变体独特序列,还发现了10个新颖的miRNA。先前仅在甘蓝型油菜中鉴定了27个保守的miRNA序列(miRBase版本18)。使用油菜双歧杆菌基因组作为油菜双歧杆菌A基因组的替代物,鉴定/注释了180多个MIRNA基因座。许多miRNA以阶段或组织特异性方式表达,表明它们具有与种子发育过程中转录物丰度的微调有关的特定功能。预测了甘蓝型油菜中的miRNA靶标,并使用微阵列分析对其表达模式进行了分析。 miRNA及其靶标表达模式的全球相关性分析揭示了种子发育过程中复杂的miRNA-靶标基因调控网络。 miR156家族是最丰富的家族,大多数家族成员主要在胚胎中表达。结论在本研究中鉴定了大量的miRNA,这些miRNA具有不同的表达模式,许多miRNA的多靶点和共靶点以及miRNA的表达与靶标之间的复杂关系。确定了几种关键的miRNA靶标表达模式,并提出了miRNA在调节种子发育中的新作用。 miR156,miR159,miR172,miR167,miR158和miR166是通过其在植物发育中的关键作用来控制种子发育和成熟的网络的主要贡献者。 miR156可能调节发芽的发育过渡。

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