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生物燃料作物甘蔗中 microRNAs 的鉴定和表达分析。

Identification and expression analysis of microRNAs and targets in the biofuel crop sugarcane.

机构信息

Centro de Biologia Molecular e Engenharia Genetica (CBMEG), Universidade Estadual de Campinas, Campinas, SP, Brazil.

出版信息

BMC Plant Biol. 2010 Nov 24;10:260. doi: 10.1186/1471-2229-10-260.

Abstract

BACKGROUND

MicroRNAs (miRNAs) are small regulatory RNAs, some of which are conserved in diverse plant genomes. Therefore, computational identification and further experimental validation of miRNAs from non-model organisms is both feasible and instrumental for addressing miRNA-based gene regulation and evolution. Sugarcane (Saccharum spp.) is an important biofuel crop with publicly available expressed sequence tag and genomic survey sequence databases, but little is known about miRNAs and their targets in this highly polyploid species.

RESULTS

In this study, we have computationally identified 19 distinct sugarcane miRNA precursors, of which several are highly similar with their sorghum homologs at both nucleotide and secondary structure levels. The accumulation pattern of mature miRNAs varies in organs/tissues from the commercial sugarcane hybrid as well as in its corresponding founder species S. officinarum and S. spontaneum. Using sugarcane MIR827 as a query, we found a novel MIR827 precursor in the sorghum genome. Based on our computational tool, a total of 46 potential targets were identified for the 19 sugarcane miRNAs. Several targets for highly conserved miRNAs are transcription factors that play important roles in plant development. Conversely, target genes of lineage-specific miRNAs seem to play roles in diverse physiological processes, such as SsCBP1. SsCBP1 was experimentally confirmed to be a target for the monocot-specific miR528. Our findings support the notion that the regulation of SsCBP1 by miR528 is shared at least within graminaceous monocots, and this miRNA-based post-transcriptional regulation evolved exclusively within the monocots lineage after the divergence from eudicots.

CONCLUSIONS

Using publicly available nucleotide databases, 19 sugarcane miRNA precursors and one new sorghum miRNA precursor were identified and classified into 14 families. Comparative analyses between sugarcane and sorghum suggest that these two species retain homologous miRNAs and targets in their genomes. Such conservation may help to clarify specific aspects of miRNA regulation and evolution in the polyploid sugarcane. Finally, our dataset provides a framework for future studies on sugarcane RNAi-dependent regulatory mechanisms.

摘要

背景

微小 RNA(miRNA)是一类小的调控 RNA,其中一些在不同植物基因组中保守。因此,从非模式生物中计算鉴定和进一步实验验证 miRNA 是可行的,并且对于解决 miRNA 为基础的基因调控和进化问题具有重要意义。甘蔗(Saccharum spp.)是一种重要的生物燃料作物,具有公开的表达序列标签和基因组调查序列数据库,但对于这个高度多倍体物种中的 miRNA 及其靶标知之甚少。

结果

在这项研究中,我们通过计算鉴定了 19 个不同的甘蔗 miRNA 前体,其中一些在核苷酸和二级结构水平上与高粱同源物高度相似。成熟 miRNA 在商业甘蔗杂种及其相应的祖种甜高粱和甘蔗中的器官/组织中的积累模式不同。以甘蔗 MIR827 为查询,我们在高粱基因组中发现了一个新的 MIR827 前体。基于我们的计算工具,共鉴定了 19 个甘蔗 miRNA 的 46 个潜在靶标。一些高度保守 miRNA 的靶标是在植物发育中起重要作用的转录因子。相反,谱系特异性 miRNA 的靶基因似乎在多种生理过程中发挥作用,例如 SsCBP1。实验证实 SsCBP1 是单子叶特异性 miR528 的靶标。我们的研究结果支持这样一种观点,即 miR528 对 SsCBP1 的调控至少在禾本科单子叶植物中是共享的,并且这种 miRNA 为基础的转录后调控是在单子叶植物从真双子叶植物分化后特有的。

结论

利用公开的核苷酸数据库,鉴定了 19 个甘蔗 miRNA 前体和一个新的高粱 miRNA 前体,并将其分为 14 个家族。甘蔗和高粱之间的比较分析表明,这两个物种在其基因组中保留了同源 miRNA 和靶标。这种保守性可能有助于阐明多倍体甘蔗中 miRNA 调控和进化的特定方面。最后,我们的数据集为未来甘蔗 RNAi 依赖性调控机制的研究提供了框架。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6bdc/3017846/9b4f377e8161/1471-2229-10-260-1.jpg

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