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用组织培养结合辐射诱变的方法进行药用植物阳春砂的育种,对于加快其育种速度、实现高产优质具有重要意义。本研究通过不同浓度的6-BA和NAA配比筛选阳春砂组织培养不定芽的最适培养基,再以组培不定芽为材料,设置不同辐射剂量,进行60Co-γ射线辐射诱变;采用ISSR分子标记技术,对部分诱变材料进行遗传变异分析。结果表明,添加4.0mg/L 6-BA和0.1mg/L NAA的MS培养基不定芽诱导率和出芽倍数最高,分别达到91.4%和2.44;用于阳春砂组培不定芽γ射线辐射的合适剂量为16Gy;ISSR分析结果显示,16Gy辐照获得的2个植株AV16-1和AV16-2在遗传上发生了明显变异,与对照的遗传相似系数分别为0.549和0.563。本研究建立了组织培养结合辐射诱变培育阳春砂新种质的方法,为基于ISSR分子标记技术的诱变育种提供了参考。
Tissue culture combined with radiation mutagenesis of medicinal plants Yang spring sand breeding, to speed up its breeding speed, to achieve high yield and quality is of great significance. In this study, different concentrations of 6-BA and NAA ratio screening of adventitious buds of spring sand culture optimum culture medium, and then adventitious buds of tissue culture as the material, set different doses of radiation, 60Co-γ ray irradiation mutation; using ISSR molecular marker technology, genetic mutation analysis of some mutagenic materials. The results showed that MS medium supplemented with 4.0 mg / L 6-BA and 0.1 mg / L NAA had the highest adventitious bud induction rate and sprouting times, reaching 91.4% and 2.44, respectively. The result of ISSR analysis showed that the genetic diversity of two plants AV16-1 and AV16-2 obtained by 16Gy irradiation was significantly different, and the genetic similarity coefficients with the control were 0.549 and 0.563, respectively. In this study, we established a method of tissue culture and radiation mutagenesis to cultivate a new germplasm of Amomum villosum, providing a reference for mutation breeding based on ISSR molecular marker technology.