Fluorochromy%20pou

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Laboratory of flow cytometry Pruhonice
J. Suda1,2 A. Krahulcová2 1Department of
Botany, Faculty of Science, Charles
University 2Institute of Botany, Academy of
Sciences of the Czech Republic
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E q u i p m e n t
Partec PA II HBO lamp (UV, DAPI
staining) air-cooled argon ion laser (488 nm, PI
staining) 2 parameters - FL1, FL2
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B a s i c d a t a
Startup September 2000 Total number of samples
10 517 Botanical Institute 5 509 (cca 52
) Department of Botany 5 008 (cca 48 )
Operating ca 940 days Out of order ca 200 days
(!) (13.9. 02 - 23.1. 03) (6.6. 03 - 14.8. 03)
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M a t e r i a l a n a l y z e d
Other taxa Agropyron, Agrostis, Allium,
Asplenium, Aster, Betula, Butomus, Carduus,
Centaurea, Dryopteris, Echium, Lythrum, Orchis,
Phragmites, Poa, Ranunculus, Tragopogon,
Vaccinium, Valerianella
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A p p l i c a t i o n s
Nuclear DNA amount analysis
Ploidy estimation Determination of closely
related taxa Determination of hybrid
plants Detection of rare cytotypes Sympatric
growth of various cytotypes Breeding mode
detection
Absolute nuclear DNA content Correlation with
environmental conditions and/or
evolutionary history Discrimination of plants
with the same chromosome number Genome
composition in allopolyploid taxa
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M e t h o d s
Nuclei isolation (chopping in hypotonic buffer)
Two step procedure Otto I II (Otto 1990)
(with centrifugation) Simplified Otto I II
(without centrifugation) LB01 (Doležel et al.
1989) Matzk (seeds) (Matzk et al.
2000) Tris-MgCl2 (Pfosser et al. 1995)
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G r a n t p r o j e c t s
2002-6 (GAAV) The influence of environmental
parameters on the genetic structure of young
agamic complex Hieracium subgen.
Pilosella (project leader F. Krahulec) 2002-4
(GACR) Evolution within hybridogenous population
of Tragopogon does Czech population
differ from those ones in US? (project leader
F. Krahulec) 2002 (FRVŠ) An employment of flow
cytometry and isozyme analyses in the study of
the genus Taraxacum (project leader L.
Záveský) 2003 (FRVŠ) Genome size in the genus
Reynoutria stable or variable? (project
leader J. Suda, researcher Z. Hoták) 2002
(FRVŠ) A biosystematic study of the polyploid
Campanula patula L. (project leader J.
Suda, researcher H. Látalová) 2001 (FRVŠ) An
employment of flow cytometry in the taxonomic
study of the genus Galeodolon (project
leader J. Suda, researcher R.
Rosenbaumová) 2002-4 (GAUK) Genome size a key
to the knowledge of relationships among central
European taxa from Hieracium subg.
Pilosella (project leader J.
Suda, researchers A. Krahulcová, T. Peckert, R.
Rosenbaumová) 2001-3 (GAUK) A biosystematic
study of the genus Empetrum (project leader J.
Suda)
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M a s t e r T h e s e s
2000 - 2 R. Rosenbaumová A taxonomic study of
the genus Galeobdolon Hudson
(supervisor J. Suda) 2000 - 2 S. Papoušková
Reproduction mode variability in the
facultatively apomictic hawkweed
Hieracium rubrum (supervisor A.
Krahulcová) 2001 - ? V. Koštálová Hybridization
between Hieracium bauhinii and H. pilosella in
mixed populations (supervisor F.
Krahulec) 2003 - ? T. Urfus Sources of
variation in Hieracium pilosella (supervisor
F. Krahulec) 2003 - ? T. Mandáková
Biosystematic and ecological study of various
cytotypes in Aster amellus (supervisor Z.
Münzbergová) 2003 - ? K. Mozolová Cytogeography
and population structure in Pimpinella
saxifraga (supervisor J. Suda) 2003 - ? L.
Grattová Morphological and phytocoenological
differentiation of the taxa in the
genus Empetrum (supervisor J. Suda) 2003 -
? R. Holubová Morphological and karyological
study of the Dryopetris dilatata alliance
(supervisor J. Suda) 2003 - ? P. Vít
Biosystematic study of the Czech endemic apomicts
from the genus Sorbus (supervisor J.
Suda)
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P h. D. T h e s e s
2000 - 3/4 J. Suda An employment of flow
cytometry in the biosystematics of vascular
plants (supervisor J.
Kirschner) 2001 - ? T. Peckert A taxonomic
study of Hieracium sect. Echinina in Central
Europe (supervisor J.
Chrtek jun.) 2001 - ? L. Záveský Apomixis vs.
sexuality different approaches to the
investigation in the genus Taraxacum Wigg
(supervisor J. Štepánek) 2001 - ? V. Mahelka
An effect of potential hybridization between
crops and weedy species a risk of genetic
material transfer (with Elytrigia spp. as a model
organism) 2002 - ? R. Rosenbaumová
Relationships between breeding mode and
population structure in Hieracium
subg. Pilosella (supervisor F. Krahulec)
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P r o s a n d C o n s
? high accuracy of measurements (when properly
adjusted)
? sub-optimal design of the instrument non-profes
sional Partec service absolute incompetence of
the Biotech (Czech representative)
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P e r s p e c t i v e s

• new instrument (Cy Flow with green laser)
• utilization of forward scatter
• image cytometry
• volumetric measurements (Coulter counter)

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