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针对马尾松胚性细胞系增殖困难的问题,本研究设定了2因素3水平的处理,分析了增殖培养基中蔗糖与肌醇对马尾松胚性细胞系增殖的影响。研究结果表明,胚性培养物的增殖倍数在9个处理间存在极显著性差异(P<0.01),并初步选出马尾松胚性细胞系增殖倍数较高的3个培养基:5号(蔗糖20 g·L-1+肌醇1.0 g·L-1)、7号(蔗糖30 g·L-1+肌醇0.1 g·L-1)和8号(蔗糖30 g·L-1+肌醇1.0 g·L-1)。胚性细胞在以上3个培养基具有不同分化发育反应,其中培养基5号中,培养基胚性细胞发育较慢;培养基7号中,胚性细胞发育较快且能形成具有完整结构的正常早期体细胞胚;在培养基8号中,胚性细胞易分化形成结构不完整、形态不正常的早期体细胞胚。综合考虑胚性细胞系增殖倍数与胚性细胞分化发育两方面的因素,在增殖培养基中添加蔗糖30 g·L-1和肌醇0.1 g·L-1的组合更适合马尾松胚性细胞系的增殖。
In order to solve the problem of the proliferation of Massonsson embryogenic cell line, we set up a two-factor-three level treatment and analyzed the effect of sucrose and inositol in proliferation medium on the proliferation of the embryonic cell line. The results showed that there was a significant difference (P <0.01) in the multiplication ratio of embryogenic cultures among 9 treatments, and three mediums with higher multiplication ratio of embryo lines were selected: No.5 Sucrose 20 g · L-1 + inositol 1.0 g · L-1), No.7 (Sucrose 30 g · L-1 + Inositol 0.1 g · L -1) and No.8 Sucrose 30 g · L -1 + Inositol 1.0 g · L-1). Embryonic cells in the above three media have different developmental differentiation reaction, medium No. 5, medium embryonic cells slow development; medium No. 7, embryonic cells developed rapidly and can form a complete structure Normal early somatic embryos; medium No. 8, embryogenic cells easily differentiated to form an incomplete structure, abnormal early embryonic somatic embryos. Considering the multiplication of embryogenic cell lines and the differentiation and development of embryogenic cells, the combination of sucrose 30 g · L-1 and inositol 0.1 g · L-1 in the culture medium is more suitable for the production of Masson’s pine embryogenic cells Department of proliferation.