acta societatis botanicorum poloniae - LV Zjazd Polskiego ...

55th Meeting of the Polish Botanical Society, Warsaw 2010
in vitro CULTUrE OF PLANTAGINACEAE SPECIES
Andrzejewska-Golec Emilia, Makowczyńska Joanna. Medical
University, Division of Biology and Pharmaceutical Botany,
Department of Biology and Pharmaceutical Biotechnology,
1 Muszyńskiego St., 90-151 Łódź, Poland, andrzejewska@
gmail.com
In vitro cultures of only some species among the known nearly
260 species of the Plantaginaceae family have been described.
Our study was concerned with the unknown species in Poland.
They are the East Asian medicinal plants: Plantago asiatica L.
and P. camtschatica Link, also the species: P. maritima L. and
P. coronopus L. which are very rare in Poland. Micropropagation
of these taxa was worked out. In the case of the P. asiatica
culture, the early stages embryogenesis and organogenesis, histochemical
structure, ultrastructure and nuclear DNA content
were also examined.
in vitro CULTUrES OF tussilAGo fArfArA L.
Buchwald Waldemar, Krajewska-Patan Anna, dreger Mariola,
Stanisławska Marzena, Górska-Paukszta Małgorzata,
Adamczak Artur, Mielcarek Sebastian, Gryszczyńska
Agnieszka. The Branch of Medicinal Plants of the Institute of
Natural Fibres and Medicinal Plants, 27 Libelta St., 61-707 Poznań,
Poland, wbuchwald@iripz.pl
Consumption of herbal medicinal products containing coltsfoot
(Tussilago farfara L.) is limited due to the presence of the
hepatotoxic pyrrolizidine alkaloids (recommendation of Commission
E). Coltsfoot products are recommended for upper respiratory
tract diseases. The way to eliminate the alkaloid content
problem seems to be by attainment of low alkaloid content
plants which originate from the natural populations. The presented
investigations are part of the work involving the selection
of naturally occurring Polish populations. Determination of
the genetic and phytochemical diversity of the natural populations,
in order to obtain breeding lines with low content of pyrrolizidine
alkaloids, must be done. In vitro cultures of coltsfoot
were established, after elaboration of the shoots multiplication
and rhizogenesis methods, from the collection of harvested cultures.
The chemical analysis (the method of instrumental analysis;
techniques HPLC with DAD detection) of in vitro cultured
shoots showed the content of senkirkine ranged from 1.008 to
8.59 mg/100g dry raw material depending on the initial population.
The in vitro established collection constitutes a base for
the obtaining of coltsfoot lines by biotechnological methods,
characterized by low content of alkaloids. This study was financed
by the Ministry of Science and Higher Education, grant
No. NN 405 306236.
rOOT CULTUrES OF PlAntAGo ovAtA FOrSSK.
Budzianowska Anna. Poznań University of Medical Sciences,
Department of Pharmaceutical Botany and Plant Biotechnology,
14 Św. Marii Magdaleny St., 61-213 Poznań, Poland, abudzian@
ump.edu.pl
Plantago ovata Forssk. (Ispagula, Isubgol) (Plantaginaceae) is
native to Asia and it is cultivated for seeds and seed husks (Plantaginis
ovatae semen, Plantaginis ovatae seminis tegumentum)
used mainly as anticonstipant remedies. Micropropagation of
that species was achieved through induction of axillary buds,
direct organogenesis and somatic embryogenesis [Fons et al.
2008]. In our previous studies we had already obtained the
shoot and callus cultures [Budzianowska and Budzianowski
2009]. When new media were tested for callus initiation and
indirect organogenesis, strong rhizogenesis was observed. This
was true, particularly on media containing NAA with BAP or
NAA with KIN. Chromatography analysis for phenolic compounds
of the extract from roots showed a presence of phenyle-
108
thanoids (phenethyl glycosides) similar to those found in callus
(verbascoside, plantamajoside). Then, the root cultures were established
in a liquid MS medium supplemented with NAA 0.25
mgl -1 and BAP 0.5 mgl -1 , in which root growth proved very fast
and significantly higher than that on the solid medium. The root
cultures of P.ovata may become an efficient source of phenylethanoids
characteristic for the genus Plantago L. Fons F., Gargadennec
A., Rapior S. (2008) Acta Biol. Gallica 155: 277– 300.
Budzianowska A., Budzianowski J. (2009) Acta. Biolog. Cracov.
ser. Bot. 51 suppl. 1: 34.
PHENOLIC COMPOUNdS OF CALLUS OF PlAntAGo
ovAtA FOrSSK.
Budzianowska Anna 1 , Budzianowski Jaromir. Poznań University
of Medical Sciences, Department of Pharmaceutical
Botany and Plant Biotechnology, 14 Św. Marii Magdaleny St.,
61-213 Poznań, Poland, 1 abudzian@ump.edu.pl
Plantago ovata – Ispaghula (Plantaginaceae) is native to Asia
and is cultivated for its seeds and seed husks. It is used mainly
for constipation because the seeds and seed husks have a high
content of mucilage. Phytochemical analyses showed the presence
of iridoids and phenylethanoids (verbascoside, plantamajoside)
in plants from cultivation and phenylethanoids (verbascoside,
forsythoside B) and flavonoids in seeds. In our previous
studies shoot and callus cultures of that species were obtained
[Budzianowska, Budzianowski 2009]. The subject of the current
studies were callus cultures from leaves, roots and hypocotyls
of seedlings on MS medium with lowered content of NH-
4NO 3 and supplemented with 2,4-D 1 mgl-1 and KIN 0.5 mgl-1.
In passages X, XI and XII, the growth indices were measured
(about 300%). Methanol extracts prepared from callus lines derived
from different organs, were analyzed for phenolic compounds
by TLC and 2D-TLC chromatography. Only phenylethanoids
were found – verbascoside and plantamajoside being
the dominant compounds, similarly as earlier observed in the
case of callus of P.lanceolata [Budzianowska et al. 2004]. Suspension
cultures obtained from callus were also performed. No
excrection of phenolic compounds to the medium was observed.
Budzianowska A., Budzianowski J. (2009) Acta. Biolog. Cracov.
ser. Bot. 51 suppl. 1: 34. Budzianowska A., Skrzypczak L.,
Budzianowski J. (2004) Planta Medica 70: 834– 840.
APPLICATION OF in vitro CULTUrE TECHNIqUES
IN OBTAINING frAGAriA SP. CLONES WITH IN-
CrEASEd TOLErANCE TO SALT STrESS
dziadczyk Ewa, Burian Maria, Bednara Józef. Maria-
Curie-Skłodowska University, Department of Plant Anatomy
and Cytology, 19 Akademicka St., 20-033 Lublin, Poland,
ewadziadczyk@interia.pl
In vitro selection methods provide the possibility of obtaining
lines with increased tolerance to different kinds of abiotic stresses,
especially drought, salinity or cold. Strawberry (Fragaria x
ananassa Duch.) is one of the crop species with low tolerance
to soil salinity. Salt stress reduces both, vegetative growth of
plants and fruit yields. Our aim was to use in vitro culture to
obtain strawberry clones with increased salinity tolerance. Selective
pressure was applied during seed germination and also
during vegetative growth of young seedlings. We tested seeds
collected from the self pollinated plants of cultivar ‘Elkat’ and
seeds collected after open pollination of cvs. ‘Senga Sengana’,
‘Selva’ and clone SSSR. The selection procedure was as follows:
1) Seeds were sterilized in a 4% (w/v) sodium hypochloride solution,
and placed on modified Murashige and Skoog (1962) vernalized
for 8 weeks at 5 ° C in the dark. 2) After vernalization,
they were transferred onto selective medium supplemented with
175mM of NaCl and maintained in a growth chamber at 23 ° C
and 16h/day photoperiod. Germinated seeds were counted in