Abstract
Due to its high biomass yield, low environmental impact, and widespread adaptability to poor soils and harsh conditions, switchgrass (Panicum virgatum L.), a warm-region perennial herbaceous plant, has attracted much attention in recent years. However, little is known about microRNAs (miRNAs) and their functions in this bioenergy grass. Here, we identified and characterized a miRNA gene, Pvi-MIR319a, encoding microRNA319a in switchgrass. Transgenic rice lines generated by overexpressing the Pvi-MIR319a precursor gene exhibited broader leaves and delayed flowering compared with the control. Gene expression analysis indicated at least four putative target genes were downregulated. Additionally, we cloned a putative target gene (PvPCF5) of Pvi-MIR319a from switchgrass. PvPCF5, a TCP transcription factor, is a nuclear-localized protein with transactivation activity and control the development of leaf. Our results suggest that Pvi-MIR319a and its target genes may be used as potential genetic regulators for future switchgrass genetic improvement.
Highlights
Switchgrass (Panicum virgatum L.), which is highly adaptable and resistant to stress, can survive in many climatic conditions, including in a variety of soil conditions (Vogel et al, 2010)
The specific primers used in this study were listed in stem-loop sequences of the rice miR319 precursor genes as a reference, we found an EST (ID: FL985594) with a length of 748 bp from switchgrass (P. virgatum) by BLAST search7 (Supplementary Figure S1)
High levels of miR319 or downregulation of its target genes leads to a broader leaf phenotype in arabidopsis (Palatnik et al, 2003), and similar results were found in snapdragons, tomatoes, rice, and other plants (Ori et al, 2007; Yang et al, 2013)
Summary
Switchgrass (Panicum virgatum L.), which is highly adaptable and resistant to stress, can survive in many climatic conditions, including in a variety of soil conditions (Vogel et al, 2010). We isolated and identified a putative target gene (PvPCF5) of PviMIR319a in switchgrass; this is a nuclear localization protein with transcriptional activation activity that is negatively regulated by Pvi-MIR319a. Cloning of the Pvi-MIR319a Gene, Construction of Plant Expression Vectors, and Rice Transformation
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