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利用高通量测序技术鉴定和表征亚洲棉()中的miRNA转录组

Identification and Characterization of miRNA Transcriptome in Asiatic Cotton () Using High Throughput Sequencing.

作者信息

Farooq Muhammad, Mansoor Shahid, Guo Hui, Amin Imran, Chee Peng W, Azim M Kamran, Paterson Andrew H

机构信息

Molecular Virology and Gene Silencing Laboratory, Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic EngineeringFaisalabad, Pakistan.

Plant Genome Mapping Laboratory, University of Georgia, AthensGA, United States.

出版信息

Front Plant Sci. 2017 Jun 15;8:969. doi: 10.3389/fpls.2017.00969. eCollection 2017.

Abstract

MicroRNAs (miRNAs) are small 20-24nt molecules that have been well studied over the past decade due to their important regulatory roles in different cellular processes. The mature sequences are more conserved across vast phylogenetic scales than their precursors and some are conserved within entire kingdoms, hence, their loci and function can be predicted by homology searches. Different studies have been performed to elucidate miRNAs using prediction methods but due to complex regulatory mechanisms or false positive predictions, not all of them express in reality and sometimes computationally predicted mature transcripts differ from the actual expressed ones. With the availability of a complete genome sequence of , it is important to annotate the genome for both coding and non-coding regions using high confidence transcript evidence, for this cotton species that is highly resistant to various biotic and abiotic stresses. Here we have analyzed the small RNA transcriptome of leaves and provided genome annotation of miRNAs with evidence from miRNA/miRNA transcripts. A total of 446 miRNAs clustered into 224 miRNA families were found, among which 48 families are conserved in other plants and 176 are novel. Four short RNA libraries were used to shortlist best predictions based on high reads per million. The size, origin, copy numbers and transcript depth of all miRNAs along with their isoforms and targets has been reported. The highest gene copy number was observed for gar-miR7504 followed by gar-miR166, gar-miR8771, gar-miR156, and gar-miR7484. Altogether, 1274 target genes were found in that are enriched for 216 KEGG pathways. The resultant genomic annotations are provided in UCSC, BED format.

摘要

微小RNA(miRNA)是一类长度为20 - 24个核苷酸的小分子,在过去十年中因其在不同细胞过程中的重要调控作用而得到了充分研究。其成熟序列在广泛的系统发育尺度上比前体更保守,有些甚至在整个界内都保守,因此,可以通过同源性搜索预测它们的位点和功能。人们已经开展了不同的研究,使用预测方法来阐明miRNA,但由于调控机制复杂或存在假阳性预测,并非所有预测的miRNA在实际中都表达,而且有时通过计算预测的成熟转录本与实际表达的转录本不同。对于这种对各种生物和非生物胁迫具有高度抗性的棉花物种,随着其完整基因组序列的可得,利用高可信度转录本证据对编码区和非编码区进行基因组注释非常重要。在这里,我们分析了棉花叶片的小RNA转录组,并根据miRNA/miRNA转录本的证据对miRNA进行了基因组注释。共发现446个miRNA聚集成224个miRNA家族,其中48个家族在其他植物中保守,176个是新的。使用了四个短RNA文库,根据每百万高读数来筛选最佳预测结果。报告了所有miRNA及其异构体和靶标的大小、来源、拷贝数和转录本深度。观察到gar-miR7504的基因拷贝数最高,其次是gar-miR166、gar-miR8771、gar-miR156和gar-miR7484。总共在棉花中发现了1274个靶基因,这些基因富集在216条KEGG通路中。最终的基因组注释以UCSC的BED格式提供。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1c2/5471329/f4736f6bc31f/fpls-08-00969-g001.jpg

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