Abstract

BackgroundRadish (Raphanus sativus L.) is an economically important root vegetable crop, and the taproot-thickening process is the most critical period for the final productivity and quality formation. MicroRNAs (miRNAs) are a family of non-coding small RNAs that play an important regulatory function in plant growth and development. However, the characterization of miRNAs and their roles in regulating radish taproot growth and thickening remain largely unexplored. A Solexa high-throughput sequencing technology was used to identify key miRNAs involved in taproot thickening in radish.ResultsThree small RNA libraries from ‘NAU-YH’ taproot collected at pre-cortex splitting stage, cortex splitting stage and expanding stage were constructed. In all, 175 known and 107 potential novel miRNAs were discovered, from which 85 known and 13 novel miRNAs were found to be significantly differentially expressed during taproot thickening. Furthermore, totally 191 target genes were identified for the differentially expressed miRNAs. These target genes were annotated as transcription factors and other functional proteins, which were involved in various biological functions including plant growth and development, metabolism, cell organization and biogenesis, signal sensing and transduction, and plant defense response. RT-qPCR analysis validated miRNA expression patterns for five miRNAs and their corresponding target genes.ConclusionsThe small RNA populations of radish taproot at different thickening stages were firstly identified by Solexa sequencing. Totally 98 differentially expressed miRNAs identified from three taproot libraries might play important regulatory roles in taproot thickening. Their targets encoding transcription factors and other functional proteins including NF-YA2, ILR1, bHLH74, XTH16, CEL41 and EXPA9 were involved in radish taproot thickening. These results could provide new insights into the regulatory roles of miRNAs during the taproot thickening and facilitate genetic improvement of taproot in radish.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0427-3) contains supplementary material, which is available to authorized users.

Highlights

  • Radish (Raphanus sativus L.) is an economically important root vegetable crop, and the taprootthickening process is the most critical period for the final productivity and quality formation

  • GSS and EST sequence records in NCBI database to perfect the radish reference sequences for isolating miRNAs associated with radish taproot thickening and development

  • To identify miRNAs involved in radish taproot thickening and development, three small RNA libraries, stage1 (10 Day after sowing (DAS)), stage2 (20 DAS) and stage3 (40 DAS), were constructed, and sequenced by the Illumina Solexa system

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Summary

Introduction

Radish (Raphanus sativus L.) is an economically important root vegetable crop, and the taprootthickening process is the most critical period for the final productivity and quality formation. The characterization of miRNAs and their roles in regulating radish taproot growth and thickening remain largely unexplored. The taproot thickening phase is a critical period of root development. The taproot of radish is a storage root, the knowledge about gene regulation and the molecular mechanism is little known in storage root development, including radish. Radish genome sequencing and the radish root transcriptomics studies have facilitated the investigation of the molecular mechanisms in radish taproot development [11,12]. The key gene isolation and molecular mechanism underlying radish taproot thickening remain elusive

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