Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 99, Issue 3, pp 353–357 | Cite as

Induction of tetraploid poplar and black locust plants using colchicine: chloroplast number as an early marker for selecting polyploids in vitro

  • D. Ewald
  • K. Ulrich
  • G. Naujoks
  • M.-B. Schröder
Research Note


Tetraploid plants were produced by inducing chromosome doubling using colchicine in in vitro shoot tips of poplar and black locust clones. Many of the plants treated with colchicine showed modified morphological characteristics like stunted growth, thicker leaves and modified leaf morphology. The counting of chloroplast number in the epidermal guard cells of stomata was used for the rapid screening of tetraploids. The differences in mean chloroplast numbers between diploid and tetraploid plants were highly significant. For all plants tested, the tetraploid genotype had almost double the number of chloroplasts per guard cell compared to the diploid origin. Some plants were further analysed by flow cytometry to verify their ploidy status that was determined by chloroplast numbers. The results of this study demonstrated for the first time that chloroplast counting in poplar and black locust could be an effective and reliable method for pre-screening large numbers of plants for their ploidy level. The protocol might be applicable in a wide scope of breeding programs.


Chloroplast number Colchicine In vitro Flow cytometry Poplar breeding Tetraploid 







This project was partially funded by the Agency for Renewable Resources e.V. (FNR) within the framework of the project “Breeding of fast growing trees for production of renewable resources in short rotation periods (FastWOOD). The authors are grateful to Mr. Matthias Steinberg, Partec GmbH, Münster, Germany for a part of the flow cytometry analyses. We also thank Ms Sarah Görß for counting chloroplasts.


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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • D. Ewald
    • 1
  • K. Ulrich
    • 1
  • G. Naujoks
    • 1
  • M.-B. Schröder
    • 2
  1. 1.Johann Heinrich von Thünen-Institute (vTI), Federal Research Institute for Rural Areas, Forestry and FisheriesInstitute of Forest GeneticsWaldsieversdorfGermany
  2. 2.The State Research Centre Geisenheim and Campus GeisenheimWiesbaden University of Applied SciencesGeisenheimGermany

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