地锦体细胞胚胎发生研究

Somatic Embryogenesis of Parthenocissus tricuspidata

以地锦幼胚为外植体,接种在附加6 BA2mg·L-1及2,4 D0～4mg·L-1的改良B5培养基中诱导愈伤组织,愈伤诱发率0～100% 对诱导出的愈伤组织采用6 BA和2,4 D不同浓度组合的培养基进行继代,对能稳定继代的愈伤组织,以改良B5为基本培养基进行分化培养,采用L16(44)正交设计,研究6 BA、NAA、AC及LH四因素不同浓度组合对体细胞胚胎发生的影响。结果表明:适宜地锦幼胚体细胞胚胎发生的培养基为:愈伤组织诱导:改良B5+6 BA2 0mg·L-1+2,4 D2 0mg·L-1;愈伤组织继代:改良B5+6 BA2 0mg·L-1+2,4 D1 0mg·L-1;胚状体分化及萌发:改良B5基本培养基。胚状体诱发率33%,成苗率53%,移栽成活率85%。胚状体为外起源发生方式,经球形胚、心形胚、鱼雷形胚、子叶形胚等阶段发育成熟。

Calli were induced from immature embryos of Parthenocissus tricuspidata which were cultured in improved B5 medium supplemented with 2.0 mg·L^(-1) 6-BA and 0～4 mg·L^(-1) 2,4-D. The inducing frequency of callus ranged from 0 to 100%.Calli were subcultured in the same base medium with various combinations of 6-BA and 2,4-D. Somatic embryoids occurred from improved B5 medium without any regulators. Results showed that the suitable mediums for embryogenesis and plant regeneration from immature embryos of Parthenocissus tricuspidata were:(1)callus inducement:improved B5+6-BA 2.0 mg·L^(-1)+2,4-D 2.0 mg·L^(-1);(2) subculture medium:improved B5+6-BA 2.0 mg·L^(-1)+2,4-D 1.0 mg·L^(-1);(3)embryogenic and germination medium:improved B5.The embryogenic frequency was 33%,the germination frequency of embryoids was 53%,and the survival ratio after transplant was 85%. Embryoides occurred from exterior of callus and developed through the stages of globular embryoid,heart shaped embryoid,tin fish-shaped embryoid and cotyledon differentiation stage embryoid.