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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

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.

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

88%

Trejgell A., Wisniewska J., Tretyn A.

Biotechnologia

2004

issue 2

219-224

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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2 Biotechnologia

1 2004

1 2001

2 Trejgell A.

2 Tretyn A.

1 Wisniewska J.

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

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