acta societatis botanicorum poloniae - LV Zjazd Polskiego ...

55th Meeting of the Polish Botanical Society, Warsaw 2010
the highest frequency of transformation (61%) was achieved on
the shoot explants, 6 weeks after infection. Thus, 40 clones of
hairy roots were obtained. Each of the obtained clones of hairy
roots were able to produce verbascoside, isoverbascoside and
catalposide, but at different levels. The highest total amount of
these compounds (211.88 mg per flask) produced clone no 38
after 4 weeks of growth in liquid Woody Plant hormone-free
medium. Financial support 502-13-621.
HISTOLOGICAL ANd IMMUNOCYTOCHEMICAL
STUdIES ON SOMATIC EMBrYOGENESIS IN trifolium
niGrescens VIV.
Pilarska Maria1 , Knox John P. 2 , Konieczny robert1 , Kuta
Elżbieta1 . 1Jagiellonian University, Department of Plant Cytology
and Embryology, 52 Grodzka St., 31-044 Cracow, Poland,
m.pilarska@iphils.uj.edu.pl; 2University of Leeds, Centre for
Plant Sciences, Leeds LS2 9JT, United Kingdom
Somatic embryo formation from hypocotyl of zygotic embryos
in cotyledonary stage (CsE) of Trifolium nigrescens was studied
with the aid of light microscopy, scanning electron microscopy
(SEM) and the immunolabelling technique. Histological
slides indicated direct somatic embryogenesis (SE) on MS medium
with 0.5 mg/l NAA and 2 mg/l 2iP. Most proembryoids
developed into mature somatic embryos with typical bilateral
symmetry, easily distinguishable root and two cotyledons. Nonregenerative
callus accompanying SE was heterogenous tissue,
formed by loosely attached parenchymatous cells at the periphery
and more compact cells in its inner region. Extracellular
surface matrix network (ECMSN) visible in SEM as reticulated
fibrils and strands, covered the surface of callus and epidermis
of somatic embryos in torpedo- and cotyledonary stages. The
extracellular expression of specific pectin epitopes JIM5 and
JIM7 coincided with ECMSN localization suggesting the putative
pectic nature of the observed fibrills and strands. Development
of somatic embryos was accompanied by a gradual diversity
loss of AGPs epitopes in embryonic tissue.
THE ExPLOITATION OF SINGLE CELL ExPLANTS
IN STUdIES ON PLANT SPECIES rECALCITrANT in
vitro
Pindel Anna, Wiszniewska Alina, Piwowarczyk Barbara.
Agricultural University in Cracow, Department of Botany and
Plant Physiology, 54 29-Listopada St., 31-425 Cracow, Poland,
a.pindel@ogr.ur.krakow.pl
In studies aimed at explanation of limited morphogenetic response
in vitro, exhibited by some agriculturally important
plant species, single protoplasts and cells were used. These
were isolated from standard explants, exploited in previously
determined regeneration pathways. Examined species were:
ornamental asparagus ‘Sprengeri’, hyacinth ‘Anna Lisa’, cymbidium
hybrid, yellow lupin ‘Parys’ and grass pea ‘Derek’.
We investigated the influence of numerous factors, including
nurse cultures and conditioned medium application, on the
induction of morphogenetic potential of the species. Studies
on cellular events, like organization of tubulin cytoskeleton,
cell wall regeneration process, presence of arabinogalactan
proteins (AGP) in the cell wall structure, as well as cell division
process itself, were also conducted. The results revealed
strong sensitivity of examined plant material, especially on osmotic
stress and type of gelling agents. The recalcitrance was
manifested by cytoskeletal disorders, disturbances in cell wall
component distribution that resulted in unsettled cell morphology,
and retardation of divisions that were often of an atypical
character.
112
in vitro CHrOMOSOME dOUBLING IN TULIP
Podwyszyńska Małgorzata. Research Institute of Pomology
and Floriculture Department of Physiology and Morphogenesis
of Ornamental Plants, 18 Pomologiczna St., 96-100 Skierniewice,
Poland, mpodwysz@insad.pl
Polyploids, especially triploids and tetraploids are widely used
in the breeding programs, due to their desirable traits such as
vigorous growth, large leaves, flowers, tubers, roots or fruits.
There is considerable interest in obtaining polyploidization
of tulip. The majority of tulip cultivars were derived from
Tulipa gesneriana with a diploid chromosome number of
2n = 2x = 24. Within the tulip polyploids, in horticulture production,
the most important are the triploids. The creation of
the new cultivars; tetraploids, especially allotetraploids as fertile
plants are more prized by breeders than triploids which are
usually sterile. Tetraploids themselves can form the new cultivars
but, first of all, they can be utilized for further breeding in
order to obtain triploids by crossing tetraploids with diploids.
Polish diploid cultivars of tulip were use for the study. The initial
explants (flower stem fragments) and the multiplied cyclic
adventitious shoot cultures were subjected to polyploidization.
The plant material was treated with antimitotic agents such
as colchicine, oryzalin, amiprophos methyl and trifluralin.
Treatment used incubation of plant material for several days
on the regeneration media containing the above mentioned
chemicals. The ploidy level evaluation, using flow cytometry,
revealed that a significantly higher number of tetraploids was
obtained when the cultures of adventitious shoots were used
for antimitotic treatment. Each of the antimitotic agents induced
tetraploid production.
SYNTHETIC SEEdS OF sAlviA PrzewAlskii MAxIM.
Skała Ewa, Wysokińska Halina. Medical University of
Łódź, Department of Biology and Pharmaceutical Botany,
1 Muszyńskiego St., 90-151 Łódź, Poland, ewa.skala@umed.
lodz.pl
Salvia przewalskii is endemic species in north-western China.
The roots of this species have been used for the long time in Far
East medicine, because of the presence of some abietane diterpenoids
called tanshinones. The aim of this work was to produce
synthetic seeds of S. przewalskii and study their germination
and their conversion into plants. The shoot tips or axillary
shoot buds from in vitro cultured plants were used as explants.
The explants were encapsulated in 3% sodium alginate solution
with or without sucrose (1.5 or 3%) and/or gibberellic acid (0.5
or 1.0 mg/l). The obtained (as above described) artificial seeds
were cultured on full-strength Murashige and Skoog (MS) agar
medium directly after encapsulation or after storage in the refrigerator
at 4– 6ºC for 4 and 8 weeks. It was found that synthetic
seeds showed a high capacity to form shoots (97%) even after
8 weeks of storage. The alginate matrix was supplemented with
3% sucrose and 0.5 mg/l gibberellic acid. Shoots were rooted
successfully ex vitro, during acclimatization in pots, in greenhouse.
After 10 weeks, 90% of the plants survived the transfer
to pots.
ATTEMPT AT STIMULATING THE PHENOLIC ACId
ACCUMULATION IN in vitro CULTUrES OF ex-
Acum Affine BALF. F.
Skrzypczak-Pietraszek Ewa, Sokołowska Monika. Jagiellonian
University, Collegium Medicum, Department of Pharmceutical
Botany, 9 Medyczna St., 30-688 Cracow, Poland, ewa.
skrzypczakpietraszek@gmail.com
The aim of this study was an attempt at increasing the phenolic
acid accumulation in agitated shoot cultures of Exacum affine
Balf. f., after the addition of L-phenylalanine as the precur-