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为建立白头翁再生体系,以白头翁主根的切段和试管苗叶片为外植体,比较2种外植体离体培养差异,探讨不同植物生长调节剂对不定芽和愈伤组织诱导、愈伤增殖和再分化及芽苗生根的影响。结果表明:叶片可以通过愈伤组织再分化和直接产生不定芽2种途径建立再生体系,而根只诱导出了愈伤组织,增殖培养中逐渐褐化死亡;在含6-BA培养基上,叶块死亡,而根只诱导出少量愈伤组织;叶片诱导不定芽的培养基为MS+TDZ 0.3 mg/L+NAA 0.1 mg/L;愈伤组织增殖的培养基为MS+TDZ 0.2 mg/L+2,4-D 0.2 mg/L;再分化时,需要转接到TDZ和NAA组合的培养基上,其中处理组合TDZ 0.3 mg/L+NAA0.1 mg/L的再分化率达100%;生根培养基为1/2MS+NAA 0.2 mg/L+IBA 0.2 mg/L+蔗糖20 g/L。不同外植体离体培养存在差异,叶片较根易培养;TDZ对白头翁叶片培养效果较好,2,4-D对愈伤组织的诱导能力较强,而NAA适合于不定芽分化,NAA与IBA组合使用生根效果较好。
In order to establish a regeneration system of Pulsatilla, the cuttings of main roots of Pulsatilla and test tube seedling leaves were used as explants. The differences of in vitro culture of two explants were compared to investigate the effects of different plant growth regulators on the induction of adventitious buds and callus, callus proliferation And redistribution and root sprouting effects. The results showed that the regeneration system could be established by two ways of callus re-differentiation and direct adventitious buds, but the callus was induced only by the roots and gradually browning in the proliferation culture. On the medium containing 6-BA, The leaves were dead while the root only induced a small amount of callus. The medium for inducing adventitious buds was MS + TDZ 0.3 mg / L + NAA 0.1 mg / L. The callus proliferation medium was MS + TDZ 0.2 mg / L + 2,4-D 0.2 mg / L; when redifferentiation, it needs to be transferred to the combination medium of TDZ and NAA, and the re-differentiation rate of TDZ 0.3 mg / L + NAA 0.1 mg / L was 100 The rooting medium was 1 / 2MS + NAA 0.2 mg / L + IBA 0.2 mg / L + sucrose 20 g / L. The explants cultured in vitro differed in their in vitro culture, and the leaves were easier to culture than the roots. TDZ had better effect on the leaves of Pulsatilla, 2,4-D had stronger ability of inducing callus, while NAA was suitable for the differentiation of adventitious buds, NAA and IBA combination of rooting effect is better.