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 Citations (Scopus)

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

Fingerprint

Bisbenzimidazole
Plant Development
Protoplasts
protoplasts
cell cycle
Cell Cycle
Mitosis
Polyploidy
DNA
mitosis
Diploidy
polyploidy
nuclear genome
cells
diploidy
Cell Size
synthesis
Cell Division
Tobacco
Coloring Agents

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science
  • Cell Biology

Cite this

Analysis of the initial stages of plant protoplast development using 33258 Hoechst : reactivation of the cell cycle. / Galbraith, David W.; Mauch, Teri J; Shields, Barbara A.

In: Physiologia Plantarum, Vol. 51, No. 4, 04.1981, p. 380-386.

Research output: Contribution to journalArticle

@article{ef88a1d4334e427d8b8acacc11a873db,
title = "Analysis of the initial stages of plant protoplast development using 33258 Hoechst: reactivation of the cell cycle",
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.",
author = "Galbraith, {David W.} and Mauch, {Teri J} and Shields, {Barbara A.}",
year = "1981",
month = "4",
doi = "10.1111/j.1399-3054.1981.tb05573.x",
language = "English (US)",
volume = "51",
pages = "380--386",
journal = "Physiologia Plantarum",
issn = "0031-9317",
publisher = "Wiley-Blackwell",
number = "4",

}

TY - JOUR

T1 - Analysis of the initial stages of plant protoplast development using 33258 Hoechst

T2 - reactivation of the cell cycle

AU - Galbraith, David W.

AU - Mauch, Teri J

AU - Shields, Barbara A.

PY - 1981/4

Y1 - 1981/4

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0000472740&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0000472740&partnerID=8YFLogxK

U2 - 10.1111/j.1399-3054.1981.tb05573.x

DO - 10.1111/j.1399-3054.1981.tb05573.x

M3 - Article

AN - SCOPUS:0000472740

VL - 51

SP - 380

EP - 386

JO - Physiologia Plantarum

JF - Physiologia Plantarum

SN - 0031-9317

IS - 4

ER -