Ethylene-responsive miRNAs in roots of Medicago truncatula identified by high-throughput sequencing at whole genome level

Ethylene is one of the classical plant hormones with a diverse function in plant growth and development. Root elongation is sensitive to ethylene such that treatments with ethylene and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) inhibit root growth. MicroRNA as one type of endogenous, non-coding small RNAs, plays an important role in regulation of plant growth, development and hormonal signaling by affecting expression of target genes. However, there has been no detailed study to evaluate the role of microRNAs in mediation of ethylene-dependent physiological processes in plants. Medicago truncatula is a model plant widely used for investigation of molecular biology in legume species. In this study, we constructed two small RNA libraries from roots of M. truncatula treated with and without ACC. High-throughput sequencing was employed to sequence the small RNA libraries, and more than 30 M raw reads were obtained. We annotated 301 known miRNAs and identified 3 new miRNAs in the two libraries. Treatment of M. truncatula with 10 μM ACC led to changes in expression of 8 miRNAs. The targets of the ethylene-responsive miRNAs were predicted by bioinformatic approach. The potential role of the ethylene-responsive miRNAs in the ethylene-induced inhibition of root elongation is discussed. These results are useful for functional characterization of miRNAs in mediation of ethylene-dependent physiological processes in general and root elongation in particular.

► The involvement of microRNAs in response of Medicago truncatula to ethylene was studied by high-throughput sequencing at whole genome level.
► Three hundred and one known miRNAs were annotated and 4 new miRNAs were identified in the two libraries constructed from M. truncatula roots treated with and without ethylene synthesis precursor ACC.
► Ten miRNAs were identified to be responsive to ACC, and the targets of the ethylene-responsive miRNAs were predicted by bioinformatic approach.
► The potential role of the ethylene-responsive miRNAs in ethylene-induced inhibition of root elongation was discussed.