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Background  Switchgrass ( Panicum virgatum  L.), a C 4  perennial grass, has been recognized as one of the most potentially important lignocellulose biofuel crops. MicroRNA319 (miR319) plays a key role in plant development, abiotic resistance, and cell wall biosynthesis by repressing expression of its target  TCP  genes. We hypothesized miR319– TCP  pathway could play important roles in switchgrass feedstock characteristics for biofuel production, and produced switchgrass transgenic plants overexpressing miR319 (by ectopic expressing  Osa - MIR319b  gene), blocking miR319 (by overexpressing a target mimicry of miR319/ MIM319 ) and repression of miR319 target gene  PvPCF5 . Plant phenotype, biomass yield, and feedstock quality of transgenic plants were analyzed.    Results  Overexpression of miR319 in switchgrass promoted leaf elongation and expansion of transgenic plants, increased plant height, stem diameter, and resulted in a significant increase in plant biomass yield. Transgenic plants overexpressing of miR319 reduced lignin content, showed significantly higher enzymatic hydrolysis efficiency compared to the wild type plant. However, opposite results were observed in the  MIM319  plants. Furthermore, suppression of miR319 target gene  PvPCF5  activity also reduced lignin content, increased lignin monomer S/G ratio and the proportion of β- O -4 linkages, while significantly improving the sugar production per plant. Quantitative real-time (qRT-PCR) analysis indicated that expression of  PvMYB58/63B  and  PvHCT  with predicted TCP binding sites in their promoter regions was negatively regulated by miR319– PvPCF5  module.    Conclusions  MiR319– PvPCF5  module plays positive roles in regulating biomass yield and quality of switchgrass. It can be utilized as a candidate molecular tool in regulating biomass yield and feedstock quality. The finding could also be transferred to other grasses for forage quality improvement through genetic manipulation.

Heteroexpression of Osa-miR319b improved switchgrass biomass yield and feedstock quality by repression of PvPCF5