Analysis of the initial stages of plant protoplast development using 33258 Hoechst: reactivation of the cell cycle

David W. Galbraith, Teri J. Mauch, Barbara A. Shields

Research output: Contribution to journalArticle

54 Scopus citations

Abstract

A microfluorimetric procedure, employing the fluorescent stain 33258 Hoechst, has been developed for the investigation of the process of DNA synthesis during the initial stages of culture of tobacco (N. tabacum cv. Xanthi) leaf protoplasts. In this system, the freshly‐isolated protoplasts exhibited a unimodal distribution of nuclear DNA content characteristic of the diploid state. The almost immediate onset of DNA synthesis during culture resulted in a doubling of nuclear DNA levels prior to the first mitoses. Although the majority of the protoplasts subsequently entered into synchronous mitosis and cell division, a proportion of the remainder developed into large polyploid cells. Upon further culture, the polyploid cells became subdivided into clusters of small diploid cells. Measurement of total cell protein and cell volumes during culture indicated that a relationship existed between these parameters and the initiation of mitosis. The significance of these observations is discussed.

Original languageEnglish (US)
Pages (from-to)380-386
Number of pages7
JournalPhysiologia Plantarum
Volume51
Issue number4
DOIs
StatePublished - Apr 1981

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science
  • Cell Biology

Fingerprint Dive into the research topics of 'Analysis of the initial stages of plant protoplast development using 33258 Hoechst: reactivation of the cell cycle'. Together they form a unique fingerprint.

  • Cite this