摘要
从甘蓝型油菜试管苗下胚轴和幼叶分离的原生质体,在含2.4—D0.8mg/l、NAA0.5mg/l、6—BA0.5mg/l的改良BG培养基中浅层液体和固体平板培养3～5天。下胚轴原生质体第一次分裂,频率达10％～25％,幼叶原生质体培养6天,分裂细胞为8％～20％,至15天形成32细胞的细胞团。愈伤组织在含NAA0.2mg/l、IBA0.1mg/l、6—BA1.0mg/l的MS分化培养基上培养3～4周,产生不定芽,并在附加IBA0.5mg/l、6—BA0.2mg/l的1/2MS培养基上培养2周后形成完整小植株。
Vigorous protoplasts were isolated From young leaves and hypocotyls of Brassica napus L. cv. Qing You 2, Wan You 15 and 425-2, and culturcd on a modified soft solid BG medium (solidified by 0.2% agarose) and liquid thin layer. The media were supplemented with 2,4-D 0.8mg/l, NAA 0.5 mg/l and 6-BA 0.5 mg/l. After 3-5 days of cultlure, 10-25% of the hypocotylar protoplasts underwent the frist division (with differences among the cultivars), and A division frequency of 10-18% in young leaf protoplasts was achieved at 6 days from the initial culture. Colonies composed of 16-32 cells were observed at 15 days. Microcalli 1mm in diameter were transferred to and subcultured on MS medium supplemented with 2,4-D 0.8 mg/l and 6-BA 0.5 mg/l for callus proliferation. Adventitious buds were fromed on the surface of the calli after 3-4 weeks of culture at 25±1℃ and in continuous light (2,500-3,000 1x) on the differentiating medium containing IBA 0.1 mg/l, NAA 0.2 mg/l and 6-BA 1.0 mg, 1. Developing buds were excised from the calli when they were 2.0 cm in height, and rooted in 1/2MS medium with IBA 0.5 mg/l and 6-BA 0.2 mg/l. Rooted plantlets were obtained within 2 weeks.
出处
《西南农业大学学报（自然科学版）》
CSCD
1993年第6期518-521,共4页
Journal of Southwest Agricultural University
关键词
油菜
原生质体培养
植株再生
Brassica olcracca
bird rape
protoplast cultures/ plantlef regeneration.
