无患子优树叶片体细胞胚胎发生及植株再生的研究

摘要

In this paper, using leaf of S. mukorossi as explants and the suitable conditions for induction of embryogenic callus and somatic embryos, as well as plant regeneration were determined through multi-factor orthogonal combination. On the medium of MS supplemented with 1.5 mg·L-1 6-BA and 0.1 mg·L-1 NAA, the leaf of S. mukorossi could be induced of callus. After 21 days of culture, the induction rate of callus was up to 92.5%. On the medium of MS added with 1.0 mg·L-1 6-BA, 0.5 mg·L-1 2,4-D and 1.0 mg·L-1 NAA, the callus could be differentiated cotyledon successfully after three generation subculture. The differentiation rate was up to 33.4%. Early somatic embryonic can develop the mature somatic embryo on the medium of MS+1.0 mg·L-1 6-BA+0.1 mg·L-1 NAA+60g·L-1 sucrose after 21 days, the mature rate was up to 45%;The mature somatic embryogenesis can sprouting and rooting on the medium of 1/2 MS added with 1.0 mg·L-1 6-BA and 0.5 mg·L-1 IBA. After 30 days, the rate of rooting could be up to 68.2%.%以一株无患子优树嫩叶为外植体，采用多因素正交试验筛选其愈伤组织诱导、体胚分化及植株再生的适宜培养基组成。结果表明：6-BA 对无患子叶片愈伤组织诱导的影响显著，在添加1.0 mg·L-16-BA 和0.1 mg·L-1 NAA 的MS 培养基上培养21 d，愈伤组织诱导率达92.5%；愈伤组织在添加1.0 mg·L-16-BA、0.5 mg·L-12,4-D 和1.0 mg·L-1 NAA的MS培养基上经3次继代，可分化出体胚，分化率为33.4%；初分化体胚在MS+1.0 mgoL-16-BA+0.1 mg·L-1 NAA+60 g·L-1蔗糖的培养基上培养21 d后可转变为成熟体胚，成熟率45%；成熟体胚在添加1.0 mg·L-16-BA和0.5 mg·L-1 IBA 的1/2 MS培养基上可萌发生根，培养30 d后生根率为68.2%。