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高通量测序小 RNA 与日本甜樱桃花柱发育相关差异表达 microRNAs 的分析。

High-throughput sequencing of small RNAs and analysis of differentially expressed microRNAs associated with pistil development in Japanese apricot.

机构信息

College of Horticulture, Nanjing Agricultural University, Nanjing, P R China.

出版信息

BMC Genomics. 2012 Aug 3;13:371. doi: 10.1186/1471-2164-13-371.

Abstract

BACKGROUND

MicroRNAs (miRNAs) are a class of endogenous, small, non-coding RNAs that regulate gene expression by mediating gene silencing at transcriptional and post-transcriptional levels in high plants. However, the diversity of miRNAs and their roles in floral development in Japanese apricot (Prunus mume Sieb. et Zucc) remains largely unexplored. Imperfect flowers with pistil abortion seriously decrease production yields. To understand the role of miRNAs in pistil development, pistil development-related miRNAs were identified by Solexa sequencing in Japanese apricot.

RESULTS

Solexa sequencing was used to identify and quantitatively profile small RNAs from perfect and imperfect flower buds of Japanese apricot. A total of 22,561,972 and 24,952,690 reads were sequenced from two small RNA libraries constructed from perfect and imperfect flower buds, respectively. Sixty-one known miRNAs, belonging to 24 families, were identified. Comparative profiling revealed that seven known miRNAs exhibited significant differential expression between perfect and imperfect flower buds. A total of 61 potentially novel miRNAs/new members of known miRNA families were also identified by the presence of mature miRNAs and corresponding miRNA*s in the sRNA libraries. Comparative analysis showed that six potentially novel miRNAs were differentially expressed between perfect and imperfect flower buds. Target predictions of the 13 differentially expressed miRNAs resulted in 212 target genes. Gene ontology (GO) annotation revealed that high-ranking miRNA target genes are those implicated in the developmental process, the regulation of transcription and response to stress.

CONCLUSIONS

This study represents the first comparative identification of miRNAomes between perfect and imperfect Japanese apricot flowers. Seven known miRNAs and six potentially novel miRNAs associated with pistil development were identified, using high-throughput sequencing of small RNAs. The findings, both computationally and experimentally, provide valuable information for further functional characterisation of miRNAs associated with pistil development in plants.

摘要

背景

microRNAs(miRNAs)是一类内源性的、小的、非编码 RNA,通过在高等植物中转录和转录后水平介导基因沉默来调节基因表达。然而,miRNAs 的多样性及其在日本杏(Prunus mume Sieb. et Zucc)花发育中的作用在很大程度上仍未得到探索。雌蕊败育的不完全花严重降低了产量。为了了解 miRNAs 在雌蕊发育中的作用,通过 Solexa 测序鉴定了日本杏雌蕊发育相关的 miRNAs。

结果

Solexa 测序用于鉴定和定量分析日本杏完美和不完全花蕾中的小 RNA。从两个由完美和不完全花蕾构建的小 RNA 文库中分别测序了 22,561,972 和 24,952,690 个reads。鉴定出 61 个已知 miRNA,属于 24 个家族。比较分析显示,在完美和不完全花蕾之间有 7 个已知 miRNA 表现出显著的差异表达。总共鉴定出 61 个潜在的新 miRNA/已知 miRNA 家族的新成员,这些新 miRNA/新成员在 sRNA 文库中存在成熟 miRNA 和相应的 miRNA*。比较分析显示,在完美和不完全花蕾之间有 6 个潜在的新 miRNA 表现出差异表达。13 个差异表达 miRNA 的靶基因预测导致 212 个靶基因。GO 注释显示,高排名 miRNA 靶基因与发育过程、转录调控和应激反应有关。

结论

本研究代表了首次对完美和不完全日本杏花之间的 miRNAome 进行比较鉴定。通过对小 RNA 的高通量测序,鉴定出与雌蕊发育相关的 7 个已知 miRNA 和 6 个潜在的新 miRNA。这些发现从计算和实验两方面为进一步研究与植物雌蕊发育相关的 miRNAs 的功能特征提供了有价值的信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/80ab/3464595/14b2279acd59/1471-2164-13-371-1.jpg

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