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杜仲愈伤组织诱导和植株再生体系的建立
王 征1, 伍江波1, 刘雪梅,等1
中南林业科技大学 风景园林学院
摘要:
【目的】研究在BA、NAA、IBA、2,4 D等激素作用下,杜仲愈伤组织诱导、愈伤组织诱导丛生芽及丛生芽诱导生根各阶段的最佳培养基配方,建立杜仲植株再生体系,为杜仲转基因研究提供基础材料。【方法】以杜仲成熟胚、幼嫩胚、叶片和茎段为外植体,使用BA、NAA、IBA、2,4 D等激素,配制成不同质量浓度的培养基,进行杜仲愈伤组织诱导,同时分析愈伤组织诱导丛生芽和丛生芽诱导生根培养基的最佳配方,建立杜仲植株的再生体系。【结果】杜仲愈伤组织诱导的培养基分别为:成熟胚MS+BA 2.0 mg/L+NAA 2.5 mg/L,诱导率77.8%;幼嫩胚MS+BA 0.5 mg/L+2,4-D 2.0 mg/L,诱导率88.9%;叶片MS+BA 0.5 mg/L+NAA 0.5 mg/L,诱导率100%;茎段MS+BA 2.0 mg/L+NAA 0.5 mg/L,诱导率为100%。愈伤组织诱导不定芽形成的适宜培养基为MS+BA 1.0 mg/L+NAA 0.05 mg/L,不定芽诱导率为88.9%,增殖系数为3±1;杜仲在1/4MS的基本培养基中生根最好,生根率达80%,激素的存在反而会抑制生根。【结论】杜仲成熟胚、幼嫩胚、叶片和茎段均可通过愈伤组织途径诱导不定芽的形成,并诱导生根,建立杜仲愈伤组织再生体系,但不定芽的增殖系数较低。
关键词:  杜仲  成熟胚  继代周期  不定芽  愈伤组织
DOI:
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基金项目:“十二五”农村领域国家科技计划项目(2012BAD21B0502)
Establishment of callus induction and regeneration system for Eucommia ulmoides Oliver
WANG Zheng,WU Jiang-bo,LIU Xue-mei,et al
Abstract:
【Objective】In order to provide basis for transgenic research on bark of Eucommia ulmoides Oliver,the paper established the Eommia ulmoides regeneration system by studying the optimal culture medium for induced callus,induced adventitious bud,and adventitious bud induced roots.【Method】Mature embryos,young embryos,leaf,and stem were used to find the influence of type and concentration of hormones (BA,NAA,IBA,and 2,4-D) on callus induction,subculture,and rooting of Eucommia ulmoides Oliver.【Result】All explants could induce callus,but different media were needed.Medium of mature embryo was MS+BA 2.0 mg/L+NAA 2.5 mg/L with induction rate of 77.8%.Medium of young embryos was MS+BA 0.5 mg/L+2,4-D 2.0 mg/L with induction rate of more than 88.9%.Medium of leaf blade was MS+BA 0.5 mg/L+NAA 0.5 mg/L with induction rate of 100%.Medium of stem section was MS+BA 2.0 mg/L+NAA 0.5 mg/L,with induction rate of 100%.Medium of adventitious bud from callus media was MS+BA 1.0 mg/L+NAA 0.05 mg/L with induction rate of 88.9%,growth rate of 3±1.Rooting medium was 1/4MS with rooting rate of 80%.【Conclusion】Mature embryo and immature embryo,leaf and stem section of Eucommia ulmoides can establish regeneration system through callus,but the k factor of adventitious bud was low.
Key words:  Eucommia ulmoides Oliver  mature embryos  subculture time  adventitious bud  callus