Efficient production of doubled haploid plants by immediate colchicine treatment of isolated microspores in winter Brassica napus |
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Authors: | WJ Zhou GX Tang P Hagberg |
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Institution: | (1) College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310029, China;(2) Nilsson-Ehle Laboratory, Svalof Weibull AB, 26881 Svalov, Sweden |
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Abstract: | The effect of colchicine on embryogenesis induction and chromosomedoubling during microspore culture was evaluated in two F1 hybridsofwinter oilseed rape (Brassica napus L.). Colchicinetreatment (50 and 500 mg/L) of isolated microspores during thefirst 15 h in culture stimulated embryogenesis and produced large amounts ofhealthy-looking embryos. These normal embryos germinated well at 24°C after being transferred to solid regeneration medium and aninitial period of low temperature (2 °C) for 10 days, andcoulddirectly and rapidly regenerate vigorous plants. A high doubling efficiency of84–88% was obtained from 500 mg/L colchicine treatment for15h with low frequency of polyploid and chimeric plants. Acolchicinetreatment duration of 6 h was less effective on embryogenesis anddoubling efficiency. The present experiment also showed that changing of induction medium 15h after microspore isolation produced higher spontaneous doublingefficiency, as compared with medium change 6 h after isolation. |
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Keywords: | Brassica napus L Chromosome doubling Colchicine Doubled haploid Embryogenesis Microspore culture |
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