New Insights on Coffea miRNAs: Features and Evolutionary Conservation

被引:0
|
作者
S. S. Chaves
C. N. Fernandes-Brum
G. F. F. Silva
B. C. Ferrara-Barbosa
L. V. Paiva
F. T. S. Nogueira
T. C. S. Cardoso
L. R. Amaral
M. de Souza Gomes
A. Chalfun-Junior
机构
[1] Federal University of Lavras (UFLA),Plant Molecular Physiology Laboratory, Biology Department
[2] Agriculture College “Luiz de Queiroz” (ESALQ)/USP,Agricultural Biotechnology Center
[3] Federal University of Lavras (UFLA),Central Laboratory of Molecular Biology (LCBM), Chemistry Department
[4] Federal University of Uberlandia,Laboratory of Bioinformatics and Molecular Analysis—INGEB/FACOM
[5] Campus Patos de Minas,undefined
来源
Applied Biochemistry and Biotechnology | 2015年 / 177卷
关键词
Coffee; MicroRNAs; Evolutionary conservation; Stem-loop RT-PCR; 5′ RACE; PsRNATarget;
D O I
暂无
中图分类号
学科分类号
摘要
Small RNAs influence the gene expression at the post-transcriptional level by guiding messenger RNA (mRNA) cleavage, translational repression, and chromatin modifications. In addition to model plants, the microRNAs (miRNAs) have been identified in different crop species. In this work, we developed a specific pipeline to search for coffee miRNA homologs on expressed sequence tags (ESTs) and genome survey sequences (GSS) databases. As a result, 36 microRNAs were identified and a total of 616 and 362 potential targets for Coffea arabica and Coffea canephora, respectively. The evolutionary analyses of these molecules were performed by comparing the primary and secondary structures of precursors and mature miRNAs with their orthologs. Moreover, using a stem-loop RT-PCR assay, we evaluated the accumulation of mature miRNAs in genomes with different ploidy levels, detecting an increase in the miRNAs accumulation according to the ploidy raising. Finally, a 5′ RACE (Rapid Amplification of cDNA Ends) assay was performed to verify the regulation of auxin responsive factor 8 (ARF8) by MIR167 in coffee plants. The great variety of target genes indicates the functional plasticity of these molecules and reinforces the importance of understanding the RNAi-dependent regulatory mechanisms. Our results expand the study of miRNAs and their target genes in this crop, providing new challenges to understand the biology of these species.
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页码:879 / 908
页数:29
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