烟草新型体细胞胚结构类蛙卵体的诱导发生

The Induction System of a Novel Somatic Embryogenesis Structure,FELB,in Tobacco

通过对烟草外植体、植物生长调节剂类别与浓度的筛选,构建其类蛙卵体(FELB)的诱导发生体系,以期为烟草干细胞生物反应器的搭建奠定基础。选取烟草根、茎、叶为外植体,分别置于不同质量浓度的NAA和2,4-D的MS培养基上,进行特殊胚性愈伤与FELB结构的诱导。再将FELB置于不同质量浓度BAP的MS培养基中进行胚体萌发再生诱导。结果表明:黑暗条件下,不同烟草外植体的FELB诱导效率为叶>茎>根。其中,10 mg/L 2,4-D对烟草FELB的诱导效果最佳,5 mg/L与15 mg/L 2,4-D效果次之;而黑暗条件下的各种质量浓度NAA以及光照条件下的所有NAA和2,4-D的处理组均未能诱导出FELB结构。2.0 mg/L BAP对FELB的催萌效果最佳。通过对FELB不同发育时期的形态学和组织细胞学分析发现,其发生方式属于内起源。

The induction system of FELB(Frog egg-like body)was established through screening the explants,the varieties and concentrations of plant growth regulators(PGRs)in tobacco which will lay the foundation for the building of tobacco stem cell bioreactor.The root,stem and leaf of tobacco were used as explant materials,and were placed on MS basic medium supplemented with different concentrations of NAA and 2,4-D,respectively in order to induce special embryonic callus and FELB.Then the FELB structure was placed on MS media containing different concentrations of BAP for regeneration into plantlets culture.The results showed that the explants were induced into FELB with the efficiency of leaf>stem>root in the dark condition,and 10 mg/L 2,4-D showed optimal induction result compared with modest effect at the concentration of 5 mg/L and 15 mg/L.Both the treatments of NAA in darkness and NAA or 2,4-D under light failed to induce FELB structure.The effect of 2.0 mg/L BAP on FELB regeneration was the optimum.The results of morphological and histological experiments showed that the FELB was internally originated from tobacco.