Effect of EMS on Microspore Embryogenesis Induction and Haploid Plant Regeneration in Rapeseed (Brassica napus L.)

Document Type : Research Paper

Authors

1 M. Sc. Student, Kermanshah University, Kermanshah, Iran.

2 Assistant Professor, Department of Tissue Culture and Gene Transformation, Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran;

Abstract

In this study, an experiment was designed for optimization of mutation application on microspore embryogenesis of canola, regeneration of embryos and producing haploid plants, from Hayola 401 and Hayola 402 hybrids using EMS mutagen treatments. EMS stocks were prepared for 0.1, 0.2 and 0.3 percent using NLN13 culture medium and then microspores were cultured in NLN13 medium. Then, they were stressed by heat shock (30°C for 14 days). The results showed that in both hybrids by 0.1 % mutagen treatment for 90 minutes, the highest numbers of embryos achieved. So in the 90 minutes durations all of three applied concentrations from EMS and the control had significant differences and by increasing the concentration along this duration, fewer embryos were achieved. Afterwards, embryos were transferred to B5 medium containing gibberellic acid and incubated at 25°C and after full growth they were transferred to B5 medium without gibberellic acid. The results in both hybrids showed that the highest numbers of regenerates were produced when EMS (0.3% for 90 min) was applied to isolated microspores. 

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