Biologia Plantarum

, Volume 51, Issue 1, pp 7–14 | Cite as

Micropropagation of Juniperus phoenicea from adult plant explants and analysis of ploidy stability using flow cytometry

  • J. Loureiro
  • A. Capelo
  • G. Brito
  • E. Rodriguez
  • S. Silva
  • G. Pinto
  • C. Santos
Original Papers

Abstract

We report here the successful micropropagation of adult Juniperus phoenicea L. with respective ploidy stability studies. Microcuttings with axillary buds were grown on five media supplemented with different growth regulator combinations. Best elongation rates were achieved on Driver and Kuniyuki (DKW) medium supplemented with kinetin alone or with naphthaleneacetic acid (NAA), while Rugini olive (OM) medium stimulated the development of new branches. Shoots growing on Murashige and Skoog (MS) medium browned and showed necrotic zones. Shoots of second to fourth subcultures usually had higher elongation rates than those of the first culture. For rooting assays, half strength DKW and OM media, different concentrations of growth regulators, auxin continuous exposure vs. dipping and the type of solid matrix were assessed. During rooting assays, two morphotypes were observed with one type having well developed internodes and the other showing hyperhydratation and no internode development. High rooting rates (40 %) were only obtained in the first morphotype shoots exposed for 5 min to 2.4 µM IBA and then transferred to OM medium without growth regulators. Plants were acclimatized in pots containing a mixture of peat and Perlite (3:2) in greenhouse with progressive reduction of relative humidity. A flow cytometric screening for major ploidy changes revealed no differences among the morphotypes and between them and the mother plant. Also the nuclear DNA content of this species was estimated for the first time using flow cytometry (2C = 24.71 pg).

Additional key words

in vitro culture nuclear DNA content plant acclimatization rooting studies 

Abbreviations

2,4-D

2,4-dichlorophenoxyacetic acid

BAP

benzylaminopurine

DKW

Driver and Kuniyuki medium

FCM

flow cytometry

IAA

indole-3-acetic acid

IBA

indole-3-butyric acid

KIN

kinetin

MS

Murashige and Skoog medium

NAA

naphthaleneacetic acid

OM

Rugini olive medium

PI

propidium iodide

SH

Schenk and Hildebrandt medium

TRIS

3-(hydroxymethyl)-aminomethane

WPM

McCown Woody plant medium

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Copyright information

© Institute of Experimental Botany, ASCR 2007

Authors and Affiliations

  • J. Loureiro
    • 1
  • A. Capelo
    • 1
  • G. Brito
    • 1
  • E. Rodriguez
    • 1
  • S. Silva
    • 1
  • G. Pinto
    • 1
  • C. Santos
    • 1
  1. 1.Laboratory of Biotechnology and Cytomics, Department of BiologyUniversity of AveiroAveiroPortugal

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