水曲柳子叶和下胚轴愈伤组织和体胚的诱导

Callus Induction and Somatic Embryogenesis from Zygotic Cotyledons and Hypocotyls of Fraxinus mandshurica Rupr.

为了建立水曲柳愈伤组织诱导和体胚发生体系,并为体胚发生分子机理研究提供有效培养系统,本研究以水曲柳成熟合子胚和未成熟合子胚的子叶和下胚轴为外植体,探讨了灭菌剂及其处理时间对外植体灭菌效果的影响和激素浓度对愈伤组织诱导和体胚发生的影响。结果表明:外植体处理试剂和消毒时间是10%Na Cl O处理1 min最为适宜;以MS1/2(MS所有成分均减半)为基本培养基,NAA 3 mg/L+BA1.5 mg/L为水曲柳未成熟合子胚子叶和下胚轴诱导愈伤组织和体胚发生的最佳激素组合,诱导率分别为6.5%和9.7%;NAA 5 mg/L+BA 2 mg/L为水曲柳成熟合子胚子叶与胚轴诱导愈伤组织和体胚发生的最佳激素组合,诱导率分别为29.3%和19.5%。研究结果补充和突破了前人研究只从水曲柳合子胚子叶获得体胚发生和只从叶柄获得有效愈伤组织增殖的结果,并为揭示体胚发生机理有效愈伤组织和体胚发生系统的建立奠定了基础。

For the purpose of establishing callus induction and somatic embryogenesis system for Manchuian Ash(Fraxinus mandshurica Rupr.), and providing an effective culture procedure for researching into the molecular mechanism of somatic embryogenesis, cotyledons and hypocotyls from mature and immature zygotic embryos of F. mandshurica) were used as explants for investigating the sterilazation effects of different agents and their treatment time on explans and the induction effects of hormone combinations on callus induction and somatic embryogenesis. The results showed that the optimal combination of reagent and its disinfection time was 10%Na Cl O treating 1 min; by using MS1/2(all of the components in MS were in half strongth) as the basic medium, the optimal hormone combination for callus induction and somatic embryogenesis from cotyledon and hypocotyl of immature zygotic embryos of F. mandshurica was NAA 3 mg/L plus BA 1.5 mg/L, the induction rates were 6.5%and 9.7% respectively; whereas the optimal hormone combination for callus induction and somatic embryogenesis from cotyledon and hypocotyl of mature zygotic embryos of F. amndshurica was NAA 5 mg/L plus BA 2 mg/L,with the induction rates were 29.3% and 19.5% respectively. This result provided the implement and breakthrough of former research results that only obtained somatic embryogenesis from cotyledons and callus from leaf petiole for F. mandshurica, and laid a foundation of establishing the effective culture system for further study on revealingthe mechanism of somatic embryogenesis.