怀槐愈伤组织形成与异黄酮积累

目的 :考察怀槐愈伤组织形成与总异黄酮积累的关系。方法 :不同培养基诱导怀槐子叶和下胚轴产生愈伤组织 ,紫外分光光度术测定愈伤组织中总异黄酮并含量。结果 :NAA /BA及含 2 ,4-D组合有利子叶愈伤组织的诱导 ;MS ,N6 ,B5培养基适宜子叶形成愈伤组织 ,而SH较适宜下胚轴产生愈伤组织。愈伤组织具有异黄酮合成能力 ,含有NAA激素组合诱导的子叶愈伤组织总异黄酮含量较高 ,而 2 ,4-D/BA组合诱导的下胚轴愈伤组织总异黄酮含量较高 ,高浓度 2 ,4-D/KT组合抑制子叶和下胚轴愈伤组织积累异黄酮。愈伤组织根据发育的形态特征可分为 4种类型 ,以黄色、疏松、颗粒状的Ⅱ型愈伤组织中总异黄酮含量最高。结论 :培养基与激素组合影响愈伤组织的形成及发育形态特征 ,并反映在愈伤组织异黄酮合成差异上 ,获得的黄色、疏松颗粒状愈伤组织为后续的细胞液体悬浮培养及异黄酮合成代谢的调控研究奠定了基础。

Callus Formation and Its Isoflavonoid Accumulation in Maackia amurensis

OBJECTIVE: To obtain Maackia amurensis callus and investigate some factors influencing total isoflavonoid production in callus. METHOD: Different media were used for callus induction from cotyledonary and hypocotyl explants. Total isoflavonoid content was determined by UV spectrophotometer. RESULT: Medium types and hormone combinations influenced both callus formation and their developmental states. MS medium supplemented with NAA/BA or containing 2,4-D was suitable for callus induction from cotyledonary explants. Among the basal media tested, MS, N6 and B5 were suitable for cotyledonary callus formation and SH for hypocotyl callus formation. The formed callus could synthesize isoflavones. Media containing NAA were suitable for isoflavonoid production in cotyledonary callus and media containing 2,4-D/BA were suitable for hypocotyl callus, but 2,4-D/KT at high concentration inhibited the isoflavonoid accumulation in hypocotyl and coytledonary calli. According to the developmentally morphological characters, the formed callus could be classified into four types. II-type callus with yellow in color and friability in texture showed the highest accumulation of isoflavones. CONCLUSION: The influences of medium type and hormone combinations on Maackia amurensis callus formation are reflected in both developmentally morphological characters and isoflavonoid accumulation of calli. Yellow, friable callus induced from cotyledonary and hypocotyl explants in MS or N6 medium supplemented with 2,4-D and BA exhibit the optimum growth and isoflavonoid production.