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Effect of chemicals known as inducers or inhibitors of programmed cell death on the androgenesis in isolated microspore cultures of tobacco (Nicotiana tabacum L.) - preliminary experiments

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Zur I., Touraev A., Wedzony M.

Zeszyty Problemowe Postępów Nauk Rolniczych

2006

tom 509

Developmental pathway of isolated microspores in in vitro cultures of tobacco (Nicotiana tabacum L.), treated with chemical agents known as inducers or inhibitors of various forms of PCD was under the study. Microspores at unicellular stage of development were isolated and cultured according to the method described by Touraev and Heberle-Bors (1999). Chemical agents known as an inducer (tunikamycin, Tun) and inhibitor (leupeptin, Leu) of apoptosis, and inducer of autophagy (rapamycin, Rap) were applied to cultures continuing gamethopytic or inducing sporophytic developmental pathway and its effect on microspore development was observed. All chemicals were tested at several concentrations and several lengths of the treatment period, which were selected on the basis of literature data. Microspores treated with Rap continued its gametophytic development: majority of microspores divided asymmetrically and no significant influence of Rap could be detected. The effect of Tun was strongly depended on the duration of the Tun treatment. A short Tun treatment (1-5 h) increased slightly the number of symmetrically dividing microspores. A prolonged period of Tun treatment (12 h) resulted in a significant increase in the number of symmetrically dividing microspores (from 3% to 33%). Further prolongation of Tun treatment (24 h) had lethal influence: the number of symmetrically dividing microspores increased from 1 to 14% but the viability of cultures decreased from 57% to 20%. However, in any case further embryogenic development of dividing structures was observed - a subsequent culture resulted in its progressive degeneration and death. In cultures induced towards sporophytic development a clear evidence of Leu influence on the microspore development was found. The effect of Leu application was strongly correlated with the concentration and the time of treatment. In Leu treated cultures a significant decrease in the number of embryogenic microspores and stimulation of starch accumulation was noted. Also in this case an accumulative effect of DMSO could be observed. Longer than 2 day DMSO treatment had lethal influence and increased the number of degenerated microspores by about 18-32%. This effect was strengthened when higher concentrations of Leu were applied (30 versus 10 µmol·dm⁻³). The received results indicates that the application of autophagy inhibiting agent was essential for surviving the sucrose/nitrogen starvation period - the trigger of androgenesis in isolated microspore cultures of tobacco, and that inducer of apoptosis can inhibit gametophytic pathway but was not sufficient for a successful induction of sporophytic microspore development. These experiments are the introduction to further, more detailed examination.

Ograniczanie wyników

1 Zeszyty Problemowe Postępów Nauk Rolniczych

1 Touraev A.

1 Wedzony M.

1 Zur I.

1 2006

Wyniki wyszukiwania

Effect of chemicals known as inducers or inhibitors of programmed cell death on the androgenesis in isolated microspore cultures of tobacco (Nicotiana tabacum L.) - preliminary experiments