Bright Fluorescent Vacuolar Marker Lines Allow Vacuolar Tracing Across Multiple Tissues and Stress Conditions in Rice

Int J Mol Sci. 2020 Jun 12;21(12):4203. doi: 10.3390/ijms21124203.

Abstract

The vacuole is indispensable for cells to maintain their water potential and to respond to environmental changes. Nevertheless, investigations of vacuole morphology and its functions have been limited to Arabidopsis thaliana with few studies in the model crop rice (Oryza sativa). Here, we report the establishment of bright rice vacuole fluorescent reporter systems using OsTIP1;1, a tonoplast water channel protein, fused to either an enhanced green fluorescent protein or an mCherry red fluorescent protein. We used the corresponding transgenic rice lines to trace the vacuole morphology in roots, leaves, anthers, and pollen grains. Notably, we observed dynamic changes in vacuole morphologies in pollen and root epidermis that corresponded to their developmental states as well as vacuole shape alterations in response to abiotic stresses. Our results indicate that the application of our vacuole markers may aid in understanding rice vacuole function and structure across different tissues and environmental conditions in rice.

Keywords: Oryza sativa; fluorescent marker; tonoplast intrinsic protein; vacuole.

MeSH terms

  • Acyltransferases / genetics*
  • Acyltransferases / metabolism
  • Flowers / genetics
  • Flowers / growth & development
  • Flowers / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal
  • Oryza / genetics
  • Oryza / growth & development*
  • Oryza / metabolism
  • Plant Leaves / growth & development
  • Plant Leaves / metabolism
  • Plant Roots / growth & development
  • Plant Roots / metabolism
  • Plants, Genetically Modified / growth & development
  • Plants, Genetically Modified / metabolism
  • Pollen / genetics
  • Pollen / growth & development
  • Pollen / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Red Fluorescent Protein
  • Stress, Physiological
  • Vacuoles / metabolism
  • Vacuoles / ultrastructure*

Substances

  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Acyltransferases