玉竹愈伤组织诱导及其细胞悬浮培养体系的建立

摘要

以玉竹叶片和根状茎为外植体,研究植物生长调节剂、碳源和pH对玉竹愈伤组织诱导的影响以及植物生长调节剂对愈伤组织增殖的影响,通过正交试验初步建立并优化玉竹愈伤细胞的悬浮培养体系.结果表明,叶片和根状茎愈伤组织诱导的最佳培养基均为MS+1.0 mg·L-1 6-BA+0.5 mg·L-1 NAA+30％蔗糖,培养基最佳pH5.6,叶片和根状茎愈伤组织增殖的最佳植物生长调节剂组合均为2.0 mg·L-1 6-BA+0.8 mg·L-1NAA,且根状茎愈伤组织的诱导率和增殖倍数均明显高于叶片.根状茎愈伤细胞悬浮培养的最佳接种量80g·L-1,最佳植物生长调节剂组合1.5 mg·L-16-BA+0.6 mg·L-1 NAA,细胞生长曲线与多糖含量变化曲线均呈“S”形,细胞最佳继代周期16 d,多糖含量在培养14 d时最高,培养基pH呈先下降后上升然后趋于稳定趋势.%Effects of hormone,carbon source,and pH on induction,as well as that of hormone on proliferation of callus tissues from Polygonatum odoratum were studied.Rhizomes and leaves of the plants were used as the explants in the experiment.In addition,the suspension culture for propagating the callus cells was optimized.The results showed that the optimal medium for the induction of rhizome and leaf calluses was MS+ 1.0 mg 6-BA · L-1 +0.5 mg NAA · L-1 +30％ sucrose at pH 5.6.The optimal hormone combination for the callus proliferation was 1.5 mg 6-BA · L-1 and 0.6 mg NAA · L-1.The rates of callus induction and proliferation of the rhizomes were significantly higher than those of the leaves.For the suspension culture on rhizome callus cells,the optimum inoculum rate was 60 g · L-1,and the hormone combination of 1.5 mg 6-BA · L-1 +0.6 mg NAA · L-1.The growth and polysaccharide content of the cells in the suspension exhibited an S-shape curve with an optimal subculture cycle of 16 d.The polysaccharide content in the cells peaked after 14 d cultivation,while the pH of the medium decreased initially followed by an increase,and leveled off subsequently.