多变小冠花的组织培养和植株再生

Plant Regeneration of Coronilla varia L. from Callus Cultures via Embryogenesis and Organogenesis

本试验利用未完全展开的多变小冠花(Coronilla varia L.)子叶诱导成愈伤组织,通过体细胞胚胎发生和器官发生两条途径获得了再生植株。愈伤组织在含6-BA 0.5mg/L,KT 0.5mg/L,2,4-D2.0mg/L,CH400mg/L和Gln 225mg/L的MS培养基上培养2—3周后,转移到含6-BA 1.5mg/L,NAA 0.7mg/L,CH 400mg/L,Gln225mg/L的MS培养基上培养4周,再转到6-BA 0.5mg/L,KT 0.5mg/L,椰乳20ml/L,CH 400mg/L和Gln 150mg/L的MS培养基上,5周内形成胚状体。在1/2MS培养基上,胚状体萌发长成植株。愈伤组织在附加6-BA 1.0mg/L,NAA 0.5mg/L,KT 0.5mg/L,CH 400mg/L和Gln 225mg/L的MS培养基上,形成绿色瘤状组织。后者在含6-BA0.7mg/L,NAA 0.2mg/L,椰乳20ml/L,CH 400mg/L和Gln150mg/L的MS培养基上,诱导产生丛状不定芽,不定芽在生根培养基上诱导成完整植株。

Coronilla varia L. plants regenerated from callus cultures which were initiated from un-fully extended cotyledon segments inoculated on MS medium, with 6-BA 0.5 mg, KT 0.5 mg, 2,4-D 2.0 mg, CH 400 mg and Gln 225 mg per liter. Green areas of callus, produced in 2 or 3 weeks of culture, were selected for inductions of embryogenesis and organogenesis. Somatic embryoids formed from on and inside the calli tandemly cultured for 4 weeks on MS medium containing 6-BA 1.5 mg/l, NAA 0.7 mg/l, CH 400 mg/l and Gln 225 mg/l and 5 weeks on MS medium plus 6-BA 0.5 mg/l, KT 0.5 mg/l, coconut milk 20 ml/l, CH 400 mg/l and Gln 150 mg/l. On 1/2 MS medium, the embryoids germinated and plants regenerated. Organogenesis occured from the calli which contained green nodulars formed during the period culture of 2 to 4 weeks on MS medium with addition of 6-BA 1.0 mg/l, NAA 0.5 mg/l, KT 0.5 mg/l, CH 400 mg/l and Gln 225 mg/l. Each callus produced 6 throuh 24 shoots. Shoots produced and proliferated on MS medium supplemented with 6-BA 0.7 mg/l, NAA 0.2 mg/l, coconut milk 20 ml/l, CH 400 mg/I and Gin 150 mg/I, and rooted on rooting medium, MS plus 0.25 mg/l NAA.