Functional Analysis of Three miRNAs in <i>Agropyron mongolicum</i> Keng under Drought Stress

• Main Authors: Xuting Zhang, Bobo Fan, Zhuo Yu, Lizhen Nie, Yan Zhao, Xiaoxia Yu, Fengcheng Sun, Xuefeng Lei, Yanhong Ma
• Format: Article
• Language: English
• Published: MDPI AG 2019-10-01
• Series: Agronomy
• Subjects: <i>agropyron mongolicum</i> keng; drought-responsive microrna; next-generation sequencing; target gene prediction; functional analysis
• Online Access: https://www.mdpi.com/2073-4395/9/10/661

<i>Agropyron mongolicum</i> Keng, a perennial diploid grass with high drought tolerance, belongs to the genus <i>Agropyron</i>, tribe <i>Triticeae</i>. It has made tremendous contributions toward reseeding natural pasture and seeding artificial grassland in China, especially in the arid and semi-arid area of northern China. As a wild relative of wheat, <i>A. mongolicum</i> is also a valuable resource for the genetic improvement of wheat crops. MicroRNAs are small non-coding RNA molecules ubiquitous in plants, which have been involved in responses to a wide variety of stresses including drought, salinity, chilling temperature. To date, little research has been done on drought-responsive miRNAs in <i>A. mongolicum</i>. In this study, two miRNA libraries of <i>A. mongolicum</i> under drought and normal conditions were constructed, and drought-responsive miRNAs were screened via Solexa high throughput sequencing and bioinformatic analysis. A total of 114 new miRNAs were identified in <i>A. mongolicum</i> including 53 conservative and 61 unconservative miRNAs, and 1393 target genes of 98 miRNAs were predicted. Seventeen miRNAs were found to be differentially expressed under drought stress, seven (amo-miR21, amo-miR62, amo-miR82, amo-miR5, amo-miR77, amo-miR44 and amo-miR17) of which were predicted to target on genes involved in drought tolerance. QRT-PCR analysis confirmed the expression changes of the seven drought related miRNAs in <i>A. mongolicum</i>. We then transformed the seven miRNAs into <i>Arabidopsis thaliana</i> plants, and three of them (amo-miR21, amo-miR5 and amo-miR62) were genetically stable. The three miRNAs demonstrated the same expression pattern in <i>A. thaliana</i> as that in <i>A. mongolicum</i> under drought stress. Findings from this study will better our understanding of the molecular mechanism of miRNAs in drought tolerance and promote molecular breeding of forage grass with improved adaption to drought.