biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 60:427-434, 2016 | DOI: 10.1007/s10535-016-0617-3

Characterization and expression analysis of conserved miRNAs and their targets in Pinus densata

Z.-B. Qiu1,*, M.-M. Yuan1, B.-Z. Hai1, L. Wang1, L. Zhang1
1 College of Life Science, Henan Normal University, Xinxiang, P.R. China

MicroRNAs (miRNAs) are small non-coding RNAs that play crucial regulatory roles in diverse developmental processes via cleavage or translational inhibition of their target mRNAs. Although a growing number of miRNAs and their targets have been predicted and discovered via experimentation in many plants, little is known about conserved miRNAs and their target genes in Pinus densata. In the present study, the conserved miRNAs, miR171 and miR482, from Pinus densata were characterized. Analysis of miR171 and miR482 reveal that these miRNAs were highly conserved in other plant species. In addition, the precursors of miR171 and miR482 were validated by real time-PCR and sequencing. Using real-time quantitative PCR, miR171 and miR482 as well as their corresponding targets were found to be differentially expressed in needles, stems, and roots of Pinus densata. Furthermore two target genes, one GRAS family transcription factor protein gene and one nucleotide-binding site leucine-rich repeat (NBS-LRR) resistance protein gene, were experimentally verified to be the targets of pde-miR171 and pde-miR482, respectively, using RNA ligase-mediated 5'-rapid amplification of cDNA ends (RLM-RACE).

Keywords: resistance protein; RLM-RACE; real time-qPCR; transcription factor
Subjects: expression analysis; miRNA; target genes; RLM-RACE; transcription factor; phylogenetic tree

Received: June 14, 2015; Revised: October 16, 2015; Accepted: November 3, 2015; Published: September 1, 2016  Show citation

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Qiu, Z.-B., Yuan, M.-M., Hai, B.-Z., Wang, L., & Zhang, L. (2016). Characterization and expression analysis of conserved miRNAs and their targets in Pinus densata. Biologia plantarum, 60(3), 427-434. doi: 10.1007/s10535-016-0617-3
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