Cytogenetics and Plant Biotechnology Laboratory, Department of Botany (DST-FIST and UGC-DRS Funded), Visva-Bharati (A Central University), Santiniketan, West Bengal, 731235, India.
Microbiology Laboratory, Department of Botany (DST-FIST and UGC-DRS Funded), Visva-Bharati (A Central University), Santiniketan, West Bengal, 731235, India.
J Biotechnol. 2020 Nov 10;323:238-245. doi: 10.1016/j.jbiotec.2020.09.002. Epub 2020 Sep 5.
An efficient genetic transfection technique has been established using A4 strain of Agrobacterium rhizogenes for the first time in a medicinally valuable plant Solanum erianthum D. Don. The explants were randomly pricked with sterile needle, inoculated with bacterial suspension. The infected leaves were then washed and transferred to MS basal medium fortified with cefotaxime for hairy root induction. A maximum transformation efficiency of 72 % has been recorded after two days of co-cultivation period. The transfer of rolA and rolB genes from the bacterium to the plant genome has been confirmed in five transformed hairy rootlines by standard Polymerase Chain Reaction technique. On the basis of growth analysis and secondary metabolite study two potential rhizoclones (A4-HR-A and A4-HR-B) were selected. Rhizoclone A4-HR-A can produce highest amount of alkaloid, phenolic and flavonoid, whereas A4-HR-B was observed to be highest tannin producer. Alkaloid like solasodine, commercially important for steroidal drug synthesis, was quantified from leaf and A4-HR-A clone by an improved High Performance Liquid Chromatography method. This showed a sustainable increase (1.33 fold) in production of solasodine in hairy rootline.
首次在一种药用价值高的植物——冬葵(Solanum erianthum D. Don)中,利用发根农杆菌 A4 菌株建立了一种高效的遗传转染技术。将外植体用无菌针随机刺破,接种细菌悬浮液。然后将感染的叶片洗净,转移到添加头孢噻肟的 MS 基本培养基上,诱导毛状根的形成。在共培养两天后,记录到最高 72%的转化效率。通过标准聚合酶链反应技术,已在五条转化的毛状根系中证实了 rolA 和 rolB 基因从细菌转移到植物基因组中。基于生长分析和次生代谢产物研究,选择了两个潜在的根克隆(A4-HR-A 和 A4-HR-B)。根克隆 A4-HR-A 可以产生最高量的生物碱、酚类和类黄酮,而 A4-HR-B 被观察到是最高的单宁酸产生者。通过改进的高效液相色谱法,从叶片和 A4-HR-A 克隆中定量了类似于商用药用甾体药物合成的生物碱——茄碱。这表明毛状根系中茄碱的产量可持续增加(1.33 倍)。