Matvieieva N A, Shakhovsky A M, Belokurova V B, Drobot K O
a Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine , Kyiv , Ukraine.
Prep Biochem Biotechnol. 2016 May 18;46(4):342-5. doi: 10.1080/10826068.2015.1031393.
An efficient and rapid protocol for the establishment of Artemisia tilesii "hairy" root culture is reported. Leaf explants of aseptically growing plants were cocultured with Agrobacterium rhizogenes A4 wild strain or A. rhizogenes carrying the plasmids with nptII and ifn-α2b genes. Root formation on the explants started in 5-6 days after their cocultivation with bacterial suspension. Prolongation of explant cultivation time on the medium without cefotaxime led to stimulation of root growth. The effects of sucrose concentration as well as of the levels of synthetic indole-3-butyric acid (IBA) and native growth regulator Emistim on the stimulation of A. tilesii "hairy" root growth were studied. Maximum stimulating effect both for the control and for transgenic roots was observed in case of root cultivation on the media supplemented with IBA-up to 7.95- and 9.1-fold biomass increase, respectively. Cultivation on the medium with 10 μl/L Emistime has also led to the control roots growth stimulation (up to 2.75-fold). Emistime at 5 μl/L concentration led to 5.46-fold mass increase in only one "hairy" root line. Higher sucrose content (40 g/L) stimulated growth of two hairy root lines but had no effect on growth of the control roots.
报道了一种高效快速建立蒙古沙蒿“毛状”根培养体系的方法。将无菌培养植株的叶片外植体与发根农杆菌A4野生菌株或携带nptII和ifn-α2b基因质粒的发根农杆菌共培养。外植体与细菌悬浮液共培养5-6天后开始生根。在外植体于不含头孢噻肟的培养基上培养时间延长后，根的生长受到刺激。研究了蔗糖浓度以及合成吲哚-3-丁酸（IBA）和天然生长调节剂Emistim水平对蒙古沙蒿“毛状”根生长的刺激作用。在添加IBA的培养基上培养根时，对照根和转基因根均观察到最大刺激效应，生物量分别增加7.95倍和9.1倍。在含有10μl/L Emistime的培养基上培养也导致对照根生长受到刺激（增加2.75倍）。5μl/L浓度的Emistime仅使一个“毛状”根系的生物量增加5.46倍。较高的蔗糖含量（40g/L）刺激了两个毛状根系的生长，但对对照根的生长没有影响。
