biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 44:339-345, 2001 | DOI: 10.1023/A:1012426306493

Assessment of Age-Related Polyploidy in Quercus Robur L. Somatic Embryos and Regenerated Plants Using DNA Flow Cytometry

M. Endemann1, K. Hristoforoglu1, T. Stauber2, E. Wilhelm1
1 Division of Life Sciences, Austrian Research Centers Seibersdorf, Seiberdorf, Austria
2 Institute for Forest Tree Breeding, Federal Research Centre for Forestry and Forest Products, Waldsieversdorf, Germany

Flow cytometric analysis with 4,6-diamidino-2-phenylindole (DAPI) staining was used to screen for chromosomal changes in Quercus robur during in vitro culture. The initiated cell lines (1992 until 1999) were maintained via secondary embryogenesis on P24 medium with 0.9 μM 6-benzylaminopurine (BAP) in regular subculture intervals of 6 weeks. Regenerated plants established in the greenhouse and in vitro plantlets derived from encapsulated somatic embryos were screened. The embryogenic cell lines were characterized as individual clones by isoenzyme analysis. Flow cytometric relative DNA content analysis of the first screening period revealed that somaclonal variation in form of tetraploidy occurred in two out of 26 tested somatic embryo clones (Alt and Jung). These two clones lost their ability to convert into plantlets. Intraspecific relative DNA content variation including technical variation was below 3 %. In the second screening period, however, 3 out of 37 clones (Alt, E4.31H9 and P3.27H) contained tetraploid cells leading to the assumption that the frequency of tetraploidy seems to be correlated with the duration of in vitro culture. No chromosomal differences were detected in regenerated plants. However, tetraploidy occurred in 8 % of the tested clones over a culture period of 7 years.

Keywords: ploidy level; oak; somaclonal variation; synthetic seeds
Subjects: age-related polyploidy in somatic embryos; auxins; cytokinins; DNA flow cytometry, chromosomal changes in in vitro culture; in vitro culture, chromosomal changes; in vitro culture, somaclonal variation, salt stress; oak; ploidy level; Quercus robur; synthetic seeds, ploidy level

Published: September 1, 2001  Show citation

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Endemann, M., Hristoforoglu, K., Stauber, T., & Wilhelm, E. (2001). Assessment of Age-Related Polyploidy in Quercus Robur L. Somatic Embryos and Regenerated Plants Using DNA Flow Cytometry. Biologia plantarum, 44(3), 339-345. doi: 10.1023/A:1012426306493
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