Molecular Manipulation of MicroRNA397 Abundance Influences the Development and Salt Stress Response of Arabidopsis thaliana

Int J Mol Sci. 2020 Oct 23;21(21):7879. doi: 10.3390/ijms21217879.

Abstract

Arabidopsis thaliana (Arabidopsis) has been used extensively as a heterologous system for molecular manipulation to genetically characterize both dicotyledonous and monocotyledonous plant species. Here, we report on Arabidopsis transformant lines molecularly manipulated to over-accumulate the small regulatory RNA microRNA397 (miR397) from the emerging C4 monocotyledonous grass model species Setaria viridis (S. viridis). The generated transformant lines, termed SvMIR397 plants, displayed a range of developmental phenotypes that ranged from a mild, wild-type-like phenotype, to a severe, full dwarfism phenotype. Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR)-based profiling of the SvMIR397 transformant population revealed a strong correlation between the degree of miR397 over-accumulation, repressed LACCASE (LAC) target gene expression, reduced lignin content, and the severity of the developmental phenotype displayed by SvMIR397 transformants. Further, exposure of SvMIR397 transformants to a 7-day regime of salt stress revealed the SvMIR397 transformant lines to be more sensitive to the imposed stress than were wild-type Arabidopsis plants. Taken together, the findings reported here via the use of Arabidopsis as a heterologous system show that the S. viridis miR397 small regulatory RNA is able to repress the expression of three Arabidopsis LAC genes which led to reduced lignin content and increased salt stress sensitivity.

Keywords: Arabidopsis thaliana; LACCASE (LAC); RT-qPCR; Setaria viridis; gene expression regulation; lignin; microRNA397 (miR397); salt stress.

MeSH terms

  • Arabidopsis / genetics*
  • Arabidopsis / growth & development
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics
  • Gene Expression Regulation, Plant
  • Laccase / genetics*
  • Lignin / metabolism
  • MicroRNAs / genetics*
  • Phenotype
  • Plants, Genetically Modified / growth & development
  • Plants, Genetically Modified / metabolism
  • RNA, Plant / genetics
  • Salt Stress
  • Setaria Plant / genetics*

Substances

  • Arabidopsis Proteins
  • MicroRNAs
  • RNA, Plant
  • Lignin
  • Laccase