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1

Ability to make flowers of Pharbitis nil plants regenerated from flower buds and immature zygotic embryos

100%
Trejgell A., Tretyn A.
Biotechnologia
|
2001
|
issue 2
118-121
EN
Flower buds and immature embryos of P. nil Chois., which were grown in vivo , were material for the study. Flower bud (2-3 mm size) were treated with osmotic/trophic shock (12% sucrose) for 24h. After this time these explants were exposed on regeneration medium (MS supplemented with BAP in concentration 5 mg.dm-3 and NAA in concentration 0,1 mg.dm-3). After 4-6 weeks organogenesis of shoots was observed. Plantlets were isolated and transferred on MS medium with addition of GA3 [0,5 mg.dm-3] and NAA [0,1 mg.dm-3]. The plantlets developed into whole plants. Those plants were able to produce flower without photoperiodic induction, because these shoots regenerated from inducted tissue and ?remembered? this information. Immature embryos were isolated from previously sterilised fruit and afterwards transferred to MS without plant hormones. The embryos were cut across their axis. After 4-6 weeks of cultivation somatic embryos were formed in the injury place (hypocotyl region). These embryos were isolated and transferred on the same medium, which was used in shoots regenerated from flower buds. Embryos were converted into complete plants, but they weren?t able to flower without photoperiodic induction. However even the smallest embryos, which were used to our study (1 mm long) were able to flower, after a single induction cycle (16 hour of darkness), when the induction was given directly after isolation of embryos.
2

The effect of cytokinins on morphogenetic response of Polemonium coeruleum seedlings explants

100%
Trejgell A., Tretyn A.
Biotechnologia
|
2010
|
issue 2
125-131
EN
The aim of the presented research was to examine the morphogenetic response of Polemonium coeruleum explants. The donor material were 10-day-old seedlings. Surface sterilized seeds were germinated on MS medium supplemented with GA3 (1 mg?dm-3). Seedling explants (shoot tips, fragments of cotyledons, hipocotyls and roots) were isolated and transferred onto solidified MS medium supplemented with different types of cytokinins (BA, KN, ZEA, 2iP) at concentrations 1.0, 3.0 and 5.0 mg?dm-3 in combination with NAA (0.1 mg?dm-3). All explant types were characterized by callus proliferation. It was observed that calli developed on the entire surface of hipocotyl and root fragments. On the other hand, shoot tips and cotyledonary petioles formed callus tissue at the cut ends, and petioles only at abaxial ends. The growth of calli on all explant types was strongly stimulated by ZEA. Among the explants tested, only shoot tips exhibited shoot organogenesis. The highest frequency of shoot organogenesis was observed when the explants were cultured on a medium supplemented with 5.0 mg?dm-3 BA (100%) or 5.0 mg?dm-3 ZEA (97%). The highest shoot number per explant was obtained in the presence of 5.0 mg?dm-3 ZEA (8.4 on average). The presence of BA or ZEA in the proliferation medium inhibited rhizogenesis and the elongation growth of shoots. However, root organogenesis was supported by KN added into the medium.
3

Micropropagation in in vitro culture efficiency of selected protected species Asteraceae

100%
Trejgell A., Tretyn A.
Biotechnologia
|
2010
|
issue 3
202-209
EN
The aim of the present research was the assessment of efficiency of micropropagation system for selected species that belong to Asteraceae family, as well as analysis of morphological traits and plantlets ability to flower. The experimental material were shoot tips isolated from 7-day-old seedlings of Leontopodium alpinum, Senecio macrophyllus, Carlina acaulis, and Cirsium pannonicum. The shoot tips were transferred on the medium supplemented with BA (1 mg.dm-3), and NAA (0,1 mg.dm-3). The obtained shoots were transferred onto fresh medium with the same combination of growth regulator (3 subcultures). The shoots with 4 or more leaves were rooted on the half-strength MS medium without growth regulators for 4 weeks. The plantlets were acclimatized to ex vitro conditions and planted to the field. The analysis of flowering ability, leaves and flower morphology, and survival level were the objectives of the study. The plantlets were acclimatized in a greenhouse and planted to the field condition. From 10 seeds of initial material one can obtain 24 278 plants of Leontopodium alpinum, 1507 of Carlina acaulis, 1261 of Senecio macrophyllus, and 37 of Cirsium pannonicum taking into consideration the percentage of seed germination, proliferation rate in three subcultures, frequency of microshoots rooting and survival rate during acclimatization. The regenerated plants demonstrated traits of donor plants and were able to flower and produce viable seeds.
4

Cell suspension obtained from callus or directly from cotyledons of Pharbitis nil Chois

81%
Trejgell A., Wisniewska J., Tretyn A.
Biotechnologia
|
2004
|
issue 2
219-224
EN
An attempt has been made to obtain cell suspension from callus as well as directly from cotyledons P. nil. Cotyledons of 7-days old sterile seedlings and flower buds excised from 3-week old plants were the material for the induction of callus. The explants were laid out on MS medium supplemented with various combination of plant hormones: BAP (5 mg/l) and NAA (1 mg/l) and supplemented with 3% sucrose or 2% glucose and 1% sucrose, BAP (5 mg/l) and IAA (0,5 mg/l), BAP (0,5 mg/l) and Picloram (1 mg/l), BAP (5 mg/l) and Picloram (0,5 mg/l), 2,4-D (0,125 mg/l).The cultures were grown in continuous white light or in darkness. The callus obtained from cotyledons cultivated in darkness and callus from flower buds cultivated in light on MS medium with BAP (5 mg/l), NAA (1 mg/l), 2% glucose and 1% sucrose were proved useful for obtaining cell suspension. Moreover, an attempt was made to obtain cell suspension directly from cotyledons. Cotyledons were cut into small fragments or were subjected to enzymatic digestion. The cell suspensions were cultivated on MS medium with the addition of BAP (5 mg/l) and IAA (0,5 mg/l) on a shaker at 140 rpm. The increase of cell number was determined by counting the cells every 5 days. In the subsequent subcultures, a decrease of the number of cell divisions was observed.
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