洋葱离体雌核诱导与单倍体愈伤再生体系建立的研究

Study on Establishment of in Vitro Gynogenetic Induction and Haploid Callus Regeneration System of Onion

以不同洋葱品种的离体花蕾为外植体,开展了雌核胚状体诱导及单倍体愈伤再生的培养试验。试验结果表明:洋葱的离体花蕾适宜采用0.1%HgCl_(2)溶液消毒;在B5培养基中,2.0 mg/L 2,4-D+2.0 mg/L 6-BA的激素配比最适于洋葱离体花蕾雌核胚状体的诱导,出胚率最高达4.6%;在MS培养基中,1.5 mg/L 2,4-D+2.5 mg/L 6-BA的激素配比最适于洋葱雌核胚状体的愈伤诱导,诱导率接近100%;在不含任何激素的MS培养基中,再生芽的诱导成功率最高,达75%左右;在只含0.5 mg/L 2,4-D的MS培养基中,再生芽均能诱导出根系。经流式细胞仪进行倍性鉴定,洋葱离体雌核单倍体的诱导成功率达97%,洋葱单倍体植株经炼苗、定植后的大田存活率在90%以上。

The onion flower buds in vitro were used to carry out the gynogenetic embryoid induction and haploid callus regeneration culture experiment.The results showed that the onion flower buds in vitro should be disinfected with 0.1%HgCl_(2)solution.The concentrations of 2.0 mg/L 2,4-D and 2.0 mg/L 6-BA in B5 medium were suitable for the induction and development of gynogenic embryoids of flower buds in vitro,and the embryo emergence rate was 4.6%.The success rate of gynogenic embryoid callus induction was close to 100%in MS medium with 1.5 mg/L 2,4-D and 2.5 mg/L 6-BA hormone concentrations.The highest success rate of bud induction was about 75%in MS medium without hormone,and the roots were successfully induced by regenerated buds at 0.5 mg/L 2,4-D and 0 mg/L 6-BA concentrations.The success rate of haploid induction in vitro gynogenesis of onion was 97%,and the survival rate of onion haploid plants was more than 90%after acclimatization and transplanting.