The Effect of Chilling on Regenera of Microspore Derived Embryos of Brassica napus L.
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Abstract
The effect of chilling on microspore-derived embryos regeneration was studied in Brassica napus L. For this aim, F1 seeds of PF7045/91×Quantum cross was planted in growth chamber under conditions of 19°C and 16/8-h photoperiod (200 µmol m-2 s-1). Small buds with 3-3.5 mm length, containing microspores at very late uninucleate stage were collected from racemes and cultured in modified NLN-13 medium in the dark at 32.5°C for 3 days. Cultures were then transferred to dark condition at 25°C and shaken at 70 rpm. Cell divisions were observed using an invert microscope 3 days after culture. After 14 and 20 days, tiny globular and heart-shaped embryos were visible with naked eyes, respectively. Finally after 28 days, cotyledonary embryos with big cotyledons were produced. Mature embryos were cultured on B5 medium supplemented with 0.1 g/lit GA3 and pretreated at 4°C for 3, 6, 9 and 12 days in the dark. The embryos regenerated at 25°C were as control. Then, pretreated cultures were transferred to the light (16-h light/ 8-h dark) at 25°C conditions. The experiment was carried out in a complete random design with 4 replication. There was no significant difference between the means of regeneration frequencies at 4°C for 3, 6, 9 days and the control. Chilling for 12 days showed significant difference at 1% probability level compared to the control as the frequency of regeneration decreased.
Keywords: microspore, embryogenesis, chilling, regeneration, Brassica napus
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References
[2] Chuong P.V. and W.D. Beversdorf. High frequency embrogenesis through isolated microspore culture in Brassica napus L. and B. Carinata Brawn. Plant Science. 39, 1985, 219-226.
[3] Keller, W.A., Arnison, P.G. and B.J. Cordy. Haploids from gametophytic cells-recent developments and future prospects. In: Plant Tissue and Cell Culture Alan R. Liss. New York 1987, 233-241.
[4] Pechan, P. M. and W. A. Keller. Induction of microspore embryogenesis in Brassica napus L. by gamma irradiation and athanol strees. In vitro Cell Development Biology. 25, 1989, 1073-1074.
[5] Kott, L.S., and W.D. Beversdorf. Enhanced plant regeneration from microspore-derived embryos of Brassica napus by chilling, partial desiccation and age selection. Pl. Cell Tissue Organ Cult. 23, 1990, 187-192.
[6] Chuong, P. V., Deslauriers, C., Kott, L. S. and W. D. Beversdorf. Effects of donor plant genotype and bud sampling on microspore culture of Brassica napus. Canadian Journal of Botany, 66, 1988, 1653-1657.
[7] Zhang, F. L. and Y. Takahata. Inheritance of microspore embryogenic ability in Brassica napus. Theor Appl Genet. 103, 2001, 254-258.