油松合子胚愈伤组织诱导和悬浮细胞系的建立

Induction of Callus from the Zygotic Embryo of Pinus tabuliformis Carr and Establishment of Suspension Cell Line

摘要: 松属植物的离体培养一直是国内外的研究重点，油松(Pinus tabuliformis Carr)是我国特有的常绿裸子植物，具有丰富的经济价值。本研究以油松合子胚为研究对象，通过组织培养诱导出愈伤，多次继代得到稳定增殖的愈伤组织，继而液体培养得到油松悬浮细胞。结果表明，油松在DCR培养基上诱导愈伤组织效果较好，使用激素浓度为2.0 mg/L 2,4-D+ 0.63 mg/L 6-BA + 0.61 mg/L KT + 0.1 μM BR的培养基诱导率可达82.9%；继代培养基为DCR + 1.1 mg/L 2,4-D + 0.45 mg/L 6-BA + 0.43 mg/L KT，固体继代获得稳定增殖的愈伤，液体培养得到油松悬浮细胞系。该方法可以为油松的遗传转化研究提供材料，也为今后油松的组织培养再生成苗提供思路。

Abstract: In vitro culture of Pinus is always the priority of research all over the world. Pinus tabuliformis Carr is an evergreen gymnosperm unique to China, which has economic value. In this study, P. tabuliformis zygotic embryos were used as the research subject. The key skills of how callus induce from zygotic embryos and obtain stable suspension cells were explored, by means of tissue culture with different medium and hormones. The results showed that P. tabuliformis could induce callus effectively on DCR medium with 2.0 mg/L 2,4-D + 0.63 mg/L 6-BA + 0.61 mg/L KT + 0.1 μM BR hormones. The induction rate of the medium was up to 82.9%. The subculture medium was DCR + 1.1 mg/L 2,4-D + 0.45 mg/L 6-BA + 0.43 mg/L KT. Stable proliferation callus is obtained by subculture medium and P. tabuliformis suspension cell line is obtained by liquid culture. The method can provide materials for the genetic transformation research of P. tabuliformis and provide ways for the tissue culture regenerated seedlings of P. tabuliformis in the future.