落叶型冬青雌、雄株茎段的组织培养技术

Micropropagation of Ilex verticillata axillary buds from male and female seedlings

【目的】建立落叶型冬青(Ilex verticillata L.)雌雄株的组织培养快繁体系。【方法】以落叶型冬青优良雌株和雄株的幼嫩茎段为材料,通过添加不同浓度的6-BA、IBA、NAA、氯吡苯脲(CPPU)等植物生长调节剂,对外植体消毒时间、诱导培养基、增殖及生根培养基进行了优化。【结果】落叶型冬青雌株、雄株茎段外植体腋芽诱导最好的培养基分别为MS+0.5 mg/L 6-BA+0.2 mg/L IBA和MS+0.5 mg/L 6-BA+0.5 mg/L IBA,诱导率分别为93.3%和83.3%;雌株和雄株丛芽增殖效果最好的培养基分别是MS+1.0 mg/L CPPU+0.2 mg/L NAA和MS+1.0mg/L CPPU+0.1 mg/L NAA,增殖系数分别达3.48和1.25;落叶型冬青在1/2 MS+0.4 mg/L NAA培养基上雌、雄株的生根率分别为26.7%和20.0%;添加一种促根剂后,雌、雄株的生根率可达66.7%和46.7%。【结论】该组织培养体系特别是雄株组培体系的建立可为落叶型冬青苗木的快速繁殖提供技术支持。

[ Objective ] Establish a micropropagation system for male and female of llex verticillata L. plant （Winterberry）. [ Method]In this work, we optimized the culture mediums of axillary bud induction, clustered bud proliferation and shoot rooting by using semi-lignified buds of male and female seedling and adding different regulators such as 6-benzylaminopu- fine （ 6-BA）, indole-3-bntyric acid （ IBA）, naphthaleneaeetic acid （NAA）, and N- （2-chloro-4-pyridyl） -N＇-phenylurea （CPPU） in vitro. [ Result ] The optimal method for sterilization of explants was treatment with 70% alcohol for 30 s, and 0. 1% HgC12 for 8 min; the optimal media for axillary bud induction of female and male explants were MS＋0. 5 mg/L 6- BA＋O. 2 mg/L IBA and MS ＋ 0. 5 mg/L 6-BA ＋ 0. 5 mg/L IBA, and this treatment resulted in axillary bud germination rates up to 93.3% and 83.3%, respectively. Furthermore, the optimal media for female and male multiple shoot clumps were MS ＋ 1.0 mg/L CPPU ＋ 0.2 mg/L NAA and MS ＋ 1.0 mg/L CPPU ＋ 0.1 mg/L NAA, this treatment resulted in proliferation rates up to 3.48 and 1.25, respectively. The shoots were induced to form roots on a 1/2MS ＋ 0.4 mg/L NAA medium, and the rooting rates of females and males were only 26.7% and 20.0%, which increased to 66.7% and 46. 7%, respectively, when a rooting agent was added to the medium. [ Conclusion] The establishment of this tissue culture system, especially the male plant tissue culture, provides technical support for the rapid propagation of llex verticiUata species.