生姜离体叶片愈伤组织悬浮细胞系建立与植株再生

摘要

'Laiwu'giant ginger was taken the as material to study the induction of leaf-derived callus,establishment of cell suspension and plant generation in vitro ginger.The results showed that young leaves of in vitro ginger as explants were cultured on revised Murashige and Skoog medium supplemented with 2,4-D 1.0 mg/L,6-BA 0.5 mg/L and sucrose 30 g/L,and it resulted in callus with loose structure rapid growth.Obtained callus was cultured in modified liquid MS medium supplemented with 2,4-D 0.15 mg/L,6-BA 6.0 mg/L and sucrose 30 g/L,and shook at 25 ℃ under dark condition for 25-30 d,then good-diversity and rapid-growing cell suspension cultures were established.The optimal parameter of cell-suspension cultures were: 1.0 g-1.5 g initial inoculum,a sub-culture every 15 d,and 3/4 volume of updating liquid medium during subculture.Plantlet formation occurred when suspension cell was transferred to solid MS medium containing NAA 0.2 mg/L,6-BA 10.0 mg/L and sucrose 30 g/L.%以'莱芜大姜'为试材,研究了生姜离体叶片愈伤组织的诱导以及细胞悬浮系建立与植株再生.结果表明,以生姜试管苗叶片为外植体,接种到MS+1.0 mg/L 2,4-D+0.5 mg/L 6-BA+30 g/L蔗糖的培养基上,可有效诱导出生长迅速、质地疏松的愈伤组织.将获得的愈伤组织接种到MS+0.15 mg/L 2,4-D+6.0 mg/L 6-BA+30 g/L 蔗糖的液体培养基上,25 ℃ 黑暗条件下震荡培养25-30 d,可建立分散性好、生长迅速的悬浮细胞系,细胞悬浮系培养的适宜参数为:初始接种量为1.0-1.5 g,继代培养的适宜间隔期为15 d,继代培养液体培养基更新比例为3/4.将悬浮细胞接种到固体培养基MS+0.2 mg/L NAA+10.0 mg/L 6-BA+30 g/L 蔗糖上可获得再生植株.