In Vitroand cellular responses of jack pine embryos to three imbibition parameters

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Abstract

Eighteen imbibition treatments with differing parameters of light conditions, temperature and duration were applied to jack pine seeds. After imbibition, embryos were excised and cultured in a liquid medium for 4 wk under continous agitation. At the end of the culture period, the embryos were classified according to five categories: nodule-forming, callus-forming, nodule plus callus-forming, white (non-responsive), and necrotic. Our results showed variation in the in vitro responsiveness of the embryos for the three parameters of imbibition and interactions among them. Three statistical models were tested to analyze the data on nodule formation, and two of them showed that the interaction between time and temperature, and between time and the combined variable temperature/light, were significant, while the light and duration variables had a significant effect only as single factors. The different morphogenic responses observed might indicate specific metabolic requirements of the embryos for the different developmental pathways. Afterwards, nine imbibition treatments were selected to evaluate the effects of the three parameters on the cell cycle and absolute DNA amount per nucleus. Following imbibition the number of cells in the G0-G1 phase decreased compared to the cells in dry seed, while the number of cells increased in the G2 phase after all the treatments except one. The percentage of cells in the different cell cycle phases varied significantly among the treatments. DNA amount fluctuated from 35.5 to 40.37 pg per nucleus. Compared to dry seeds with 40.19 pg DNA per nucleus, embryos from four imbibition treatments showed a pronounced decrease in the DNA by 5.9-11.8%. This might indicate underreplication of DNA sequences and reflect DNA plasticity with regards to imbibition and environmental factors.

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