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目的:研究6-BA,萘乙酸(NAA),碳源,pH对罗汉果悬浮培养细胞生长的影响,探讨罗汉果细胞悬浮培养的最适条件。方法:考察1个培养周期内罗汉果悬浮细胞的鲜重和干重,绘制生长曲线和生长速率曲线;采用正交试验设计对6-BA,NAA,碳源,pH 4因素3水平的组合进行优化。结果:罗汉果悬浮细胞培养的生长周期约为21 d,0～6 d为细胞的延滞期,7～21 d为对数期,稳定期较短,第22天进入到衰亡期,在第21天时细胞鲜、干质量达到最大,分别为693.0,416.0 g.L-1,最大生长速率为0.040 g.L-1.d-1。NAA是影响罗汉果悬浮细胞生长的关键因素,其影响效应依次为NAA>6-BA>pH>碳源。悬浮细胞生长的适宜培养基为MS+1.0 mg.L-16-BA+0.02 mg.L-1NAA,4.0%蔗糖为碳源,初始培养基pH 5.5～6.0。结论:通过组织培养方式可以成功建立罗汉果细胞悬浮培养体系。
OBJECTIVE: To study the effects of 6-BA, NAA, carbon source and pH on the growth of Siraitia grosvenori in suspension culture, and to explore the optimal conditions for the suspension culture of Siraitia grosvenori. Methods: The fresh weight and dry weight of the suspension cells of Lo Han Guo during one culture period were investigated. The growth curve and growth rate curve were drawn. The orthogonal design was used to optimize the combination of 6-BA, NAA, carbon source and pH 4 factor 3 . Results: The growth period of Lo Han Guo suspension cells culture was about 21 days. The lag phase was 0-6 days, and the logarithmic phase was from 7 to 21 days. The stability period was short and reached the decay stage on the 22nd day. On the 21st day, The fresh and dry weight of cells reached 693.0 and 416.0 gL-1, respectively. The maximum growth rate was 0.040 gL-1.d-1. NAA is the key factor affecting the growth of suspension cells of Luo Han Guo, and its effect is NAA> 6-BA> pH> carbon source. The suitable medium for the growth of suspension cells was MS + 1.0 mg.L-16-BA + 0.02 mg.L-1NAA, 4.0% sucrose as carbon source and initial medium pH 5.5-6.0. Conclusion: The cell suspension culture system of Luohanguo can be successfully established through tissue culture.