Methodological aspects of increasing the intensity of callusogenesis and organogenesis of Linum usitatissimum L. in vitro

S. Mishchenko, H. M. Machulsky
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Abstract

Aim. Improving methods for increasing the efficiency of obtaining callus cultures and somaclones of flax (Linum usitatissimum L.) in vitro. Methods. Hypocotyl segments were cultured on Murashige and Skoog nutrient medium supplemented with sucrose (30 g/l) and phytohormones at various concentrations. Other conditions: photoperiod 16 hours, illuminance 2500 lx, relative humidity 60–80%, air temperature 22–24°C. Results. The ability to form callus and somatic embryogenesis of flax depends on the phytohormonal composition of the nutrient medium, the size of the explants and the distance between them. Conclusions. For intensive callus formation and somatic embryogenesis in vitro, the optimal concentrations of BAP (mg/l) can be expressed as 1.0 ≤ BAP ≤ 1.75; the optimal concentrations of BAP for the medium supplemented with NAA (0.05 mg/l) 0.5 ≤ BAP ≤ 2.0; the optimal concentration of NAA for the medium supplemented with BAP (1.0 mg/l) 0.025 ≤ NAA ≤ 0.150; and the optimal concentrations of IAA for the medium supplemented with BAP (1.0 mg/l) 0.05 ≤ IAA ≤ 0.50. Addition of 0.5 mg/l GA3 to the medium with NAA and BAP is effective. It is optimal to use hypocotyl explants 3–6 mm long and place them at a distance of 1.5–2.5 cm from each other. Organogenicity of callus is significantly reduced in the process of subculturing.
增加离体羊草愈伤组织和器官发生强度的方法学研究
的目标。提高亚麻愈伤组织培养和体细胞无性系体外培养效率的方法改进。方法。下胚轴在添加蔗糖(30 g/l)和不同浓度植物激素的Murashige和Skoog培养基上培养。其他条件:光周期16小时,照度2500 lx,相对湿度60-80%,空气温度22-24℃。结果。亚麻愈伤组织形成和体细胞胚胎发生的能力取决于营养培养基的激素成分、外植体的大小和外植体之间的距离。结论。对于愈伤组织形成和体胚发生,BAP的最佳表达浓度(mg/l)为1.0≤BAP≤1.75;添加NAA的培养基中BAP的最佳浓度(0.05 mg/l)为0.5≤BAP≤2.0;添加BAP的培养基中NAA的最佳浓度(1.0 mg/l)为0.025≤NAA≤0.150;添加BAP的培养基中IAA的最佳浓度为(1.0 mg/l) 0.05≤IAA≤0.50。在含有NAA和BAP的培养基中添加0.5 mg/l GA3是有效的。下胚轴外植体长3 ~ 6 mm,间距1.5 ~ 2.5 cm为最佳。继代培养过程中愈伤组织的有机发生性明显降低。
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