Maize histone H2B-mCherry: a new fluorescent chromatin marker for somatic and meiotic chromosome research

DNA Cell Biol. 2012 Jun;31(6):925-38. doi: 10.1089/dna.2011.1514. Epub 2012 Jun 4.

Abstract

Cytological studies of fluorescent proteins are rapidly yielding insights into chromatin structure and dynamics. Here we describe the production and cytological characterization of new transgenic maize lines expressing a fluorescent histone fusion protein, H2B-mCherry. The transgene is expressed under the control of the maize ubiquitin1 promoter, including its first exon and intron. Polymerase chain reaction-based genotyping and root-tip microscopy showed that most of the lines carrying the transgene also expressed it, producing bright uniform staining of nuclei. Further, plants showing expression in root tips at the seedling stage also showed expression during meiosis, late in the life cycle. Detailed high-resolution three-dimensional imaging of cells and nuclei from various somatic and meiotic cell types showed that H2B-mCherry produced remarkably clear images of chromatin and chromosome fiber morphology, as seen in somatic, male meiotic prophase, and early microgametophyte cells. H2B-mCherry also yielded distinct nucleolus staining and was shown to be compatible with fluorescence in situ hybridization. We found several instances where H2B-mCherry was superior to DAPI as a generalized chromatin stain. Our study establishes these histone H2B-mCherry lines as new biological reagents for visualizing chromatin structure, chromosome morphology, and nuclear dynamics in fixed and living cells in a model plant genetic system.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Cell Nucleus / metabolism
  • Chromatin / genetics
  • Chromatin / metabolism*
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / metabolism
  • Germ Cells, Plant / cytology
  • Germ Cells, Plant / metabolism
  • Histones / chemistry
  • Histones / genetics*
  • Histones / metabolism*
  • In Situ Hybridization, Fluorescence
  • Luminescent Proteins / analysis
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Meiosis*
  • Metaphase
  • Molecular Sequence Data
  • Plants, Genetically Modified
  • Protein Transport
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Zea mays / cytology*
  • Zea mays / genetics*
  • Zea mays / metabolism

Substances

  • Chromatin
  • Fluorescent Dyes
  • Histones
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • red fluorescent protein