Imaging Mitosis in the Moss Physcomitrella patens

Methods Mol Biol. 2016:1413:263-82. doi: 10.1007/978-1-4939-3542-0_17.

Abstract

At first glance, mitosis in plants looks quite different from that in animals. In fact, terrestrial plants have lost the centrosome during evolution, and the mitotic spindle is assembled independently of a strong microtubule organizing center. The phragmoplast is a plant-specific mitotic apparatus formed after anaphase, which expands centrifugally towards the cell cortex. However, the extent to which plant mitosis differs from that of animals at the level of the protein repertoire is uncertain, largely because of the difficulty in the identification and in vivo characterization of mitotic genes of plants. Here, we discuss protocols for mitosis imaging that can be combined with endogenous green fluorescent protein (GFP) tagging or conditional RNA interference (RNAi) in the moss Physcomitrella patens, which is an emergent model plant for cell and developmental biology. This system has potential for use in the high-throughput study of mitosis and other intracellular processes, as is being done with various animal cell lines.

Keywords: Caulonemal apical cell; GFP tagging; Kinesin; Mitosis; Physcomitrella patens.

MeSH terms

  • Bryopsida / cytology*
  • Bryopsida / genetics*
  • Gene Expression
  • Gene Order
  • Gene Targeting
  • Genes, Reporter
  • Genetic Markers
  • Kinesins / genetics
  • Kinesins / metabolism
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence* / methods
  • Mitosis* / genetics
  • Molecular Imaging*
  • Plants, Genetically Modified
  • RNA Interference
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Genetic Markers
  • Recombinant Fusion Proteins
  • Kinesins