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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。
In order to establish the callus induction and somatic embryogenesis system and to provide an effective culture system for the molecular mechanism of somatic embryogenesis, the cotyledons and hypocotyls of mature and unripe zygotic embryos were used in this study The effects of sterilizing agent and its treatment time on the sterilizing effect of explants and the effect of hormone concentration on callus induction and somatic embryogenesis were discussed. The results showed that the optimal explants and disinfection time were 10% NaCl O for 1 min. MS1 / 2 (all the components were reduced by half) was used as the basic medium, NAA 3 mg / L + BA 1.5 mg / L is the best hormone combination inducing callus and somatic embryogenesis in immature embryos and immature embryos, the induction rates are 6.5% and 9.7% respectively; NAA 5 mg / L and BA 2 mg / L The best hormones for inducing callus and somatic embryogenesis from the mature cotyledons of H. ammodendron and the hypocotyls were induced with the induction rates of 29.3% and 19.5%, respectively. The results complement and break through the previous studies only obtained from embryonic leaves of A. sojae somatic embryogenesis and effective from the petiole only callus proliferation results, and to reveal the mechanism of somatic embryogenesis effective callus and somatic embryogenesis system The establishment of the foundation.