理化条件对新疆紫草毛状根培养及紫草素含量的影响

以发根农杆菌诱导的新疆紫草毛状根为试验材料,采用二阶段培养法,分别考察了NH4NO3、培养基、转速、温度、蔗糖对毛状根生长的影响及培养方式、酸水解酪蛋白对毛状根生产的影响。结果表明:毛状根在MS无铵液体培养基中生长速度较MS快了近1倍,第15天时毛状根的生长量达峰值,增殖8倍;在SH无铵培养基、转速为(120±5)r/min、温度为(25±1)℃时毛状根增殖倍数最大,为9.34倍。蔗糖质量浓度对毛状根的生长增殖影响不大;液体培养方式的紫草素及其衍生物含量(1.05%)远远高于固体培养(0.33%),是后者的3倍;添加0.5g/L的酸水解酪蛋白的其紫草素及衍生物含量为1.29%,较对照(1.17%)增加了10.3%。本研究为今后利用新疆紫草毛状根规模化生产提供了理论依据。

Effects of physical and chemical conditions on culture of Arnebia euchroma (Royle) Johnst hairy roots and content of shikonin and its derivatives

Using the Arnebia euchroma(Royle) Johnst hairy roots induced by Agrobacterium rhizogenes as a testing material,the culture system with two-stage culture method of Arnebia euchroma(Royle) Johnst hairy roots was established for the first time.Factors influencing hairy roots growth such as NH4NO3,different media,speed,temperatures,content of sucrose and factors on hairy roots producing such as method of culture,casein hydrolysate were investigated.The results showed that under the conditions which hairy roots inoculated with MS(no NH4+) liquid medium,the biological production of hairy roots proliferation was doubled that of MS solid medium,and it increased by 8 times cultivated every 15 d.Under the conditions which hairy roots inoculated with 50 mL SH(no NH4+) liquid medium,at(25±1) ℃,(120±5) r/min,the biological production of hairy roots proliferation increased by 9.34 times.The content of sucrose was not critical for hairy roots proliferation.The content of hairy roots shikonin from solid culture and the growth stage could be promoted from about 0.33% to 1.05%.Adding 0.5 g/L casein hydrolysate into the hairy roots,the biological production of hairy roots proliferation could be promoted from about 1.17% to 1.29%,providing a theoretical basis for cultivating large-scale production by applying Arnebia euchroma(Royle) Johnst hairy roots.