Unit

UNIT 7.29 Nuclear DNA Content Analysis of Plant Seeds by Flow Cytometry

  1. Elwira Sliwinska

Published Online: 1 FEB 2006

DOI: 10.1002/0471142956.cy0729s35

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Sliwinska, E. 2006. Nuclear DNA Content Analysis of Plant Seeds by Flow Cytometry. Current Protocols in Cytometry. 35:7.29:7.29.1–7.29.13.

Author Information

  1. University of Technology and Agriculture, Bydgoszcz, Poland

Publication History

  1. Published Online: 1 FEB 2006
  2. Published Print: JAN 2006

Abstract

Procedures describing the utilization of seeds or their parts for flow cytometric determination of plant ploidy and endopolyploidy, genome size, and cell cycle activity are presented. The methods have been developed for a single-fluorescence-parameter flow cytometer, equipped with light sources for 488-nm and UV-light illumination. The procedures presented in this unit utilize the two most widely used fluorochromes for plant DNA content analysis, propidium iodide (PI) and 4′,6-diamidino-2-phenylindole (DAPI). These methods provide an alternative to estimation of DNA content based on the fluorescence of DNA in cell nuclei isolated from plant leaves. In some instances seeds are more suitable for analysis than leaves, e.g., when plant material must be transported for a long distances or stored for prolonged periods before flow cytometric analysis, or when leaves contain fluorochrome-staining inhibitors. In addition, flow cytometric determination of nuclear replication stages in seeds gives information about their physiological status (e.g., maturity, advancement of germination), which is valuable to seed producers and technologists.

Keywords:

  • flow cytometry;
  • ploidy;
  • genome size;
  • endoreplication;
  • cell cycle;
  • seed;
  • embryo;
  • endosperm