بهينه سازي باززايي گياه در كشت ميكروسپورهاي ايزوله كلزا

Optimization of plant regeneration in isolated microspores culture of Rapeseed (Brassica napus L.cv. PF704)

In this research, we designed three independent experiments in order to optimize conditions for regeneration of Rapeseed haploid embryos. Donor plants were grown in a growth chamber until their blooming at 15/10°C (day/night) with 16/8 (lightness/darkness) photoperiod. Microspores were isolated from floral buds and were cultured until cotyledon stage in NLN-13 liquid medium. Then these embryos were transferred to B5 regeneration medium. In the first experiment, we evaluated effect of 4°C chilling with different times (0, 7, 10 and 13 days) on the rate of normal regeneration. The result showed that 7 days with 61% frequency of regeneration had the best effect. In the second experiment, we studied effect of different concentrations of active charcoal (0, 0/125, 0/25, 0/5, 1 and 2 gl"1) in regeneration medium. The result showed that 0/125gl_1 with 40% frequency of regeneration had the best effect. In the late experiment, the effect of agar and Watman paper in culture medium (solid and liquid media) on the rate of normal regeneration was evaluated and we found that control (B5 medium with 0/8% agar) was the best one.