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

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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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1 Biotechnologia

1 2004

1 Trejgell A.

1 Tretyn A.

1 Wisniewska J.

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