Gupta Ruby, Pandey Pallavi, Singh Sailendra, Singh Dhananjay Kumar, Saxena Archana, Luqman Suaib, Bawankule Dnyaneshwar U, Banerjee Suchitra
Plant Biotechnology Department, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Lucknow, 226015, Uttar Pradesh, India.
Academy of Scientific and Innovative Research, CSIR-CIMAP Campus, Lucknow, 226015, Uttar Pradesh, India.
Protoplasma. 2016 Jul;253(4):1145-58. doi: 10.1007/s00709-015-0875-5. Epub 2015 Aug 28.
The present study reports the Agrobacterium rhizogenes-mediated hairy root induction in of an ethno-medicinally significant herb-Boerhaavia diffusa L., for elucidating the underlying competence regarding its biosynthetic (i.e. boeravinone B and eupalitin) and bioactivity (antibacterial, antioxidant and anti-inflammatory) potentials. Host plant-specific receptiveness towards A. rhizogenes strains and disparity in compatibility threshold of leaf and nodal explants were evident. Only leaf explants responded, attaining hairy root induction with the ATCC 15834 followed by A4 and SA79 strains in reducing order of transformation efficiency. The growth behaviours differed amongst independent rhizoclones, and two clones of A4 (RBH) and ATCC 15834 (RBT8) origin demonstrated higher growth potentials. Polymerase chain reaction amplification of rol genes confirmed their transformed nature. Optimization of the appropriate solvent and reverse phase high-performance liquid chromatography parameters relating to the targeted metabolite production in the selected RBH and RBT8 clones revealed higher accumulation of eupalitin with the RBH clone having the best result of 1.44 times greater yield over the control root. Compared to the selected rhizoclones, the control roots however showed higher boeravinone B content. Devising a modified "stirred-tank" reactor through equipping with marine impellers and ring spargers facilitated high-density RBH root biomass yield with 6.1-fold and 1.15-fold yield increment of the boeravinone B and eupalitin respectively compared to shake-flask cultures. Considering the control roots, the RBH clone revealed analogous antioxidant/antibacterial activities with improved anti-inflammatory potential. The hairy root mediated higher production of boeravinone B and eupalitin could be achieved for the first time in bioreactor.
本研究报道了发根农杆菌介导的民族药用重要草本植物——白花蛇舌草的毛状根诱导，以阐明其生物合成（即波拉维酮B和优帕利汀）和生物活性（抗菌、抗氧化和抗炎）潜力的潜在能力。宿主植物对发根农杆菌菌株的特异性接受度以及叶和节外植体在兼容性阈值上的差异是明显的。只有叶外植体有反应，按照转化效率从高到低的顺序，用ATCC 15834菌株继之以A4和SA79菌株诱导出毛状根。不同独立根克隆的生长行为不同，A4（RBH）和ATCC 15834（RBT8）来源的两个克隆表现出更高的生长潜力。rol基因的聚合酶链反应扩增证实了它们的转化性质。对所选RBH和RBT8克隆中与目标代谢物产生相关的合适溶剂和反相高效液相色谱参数进行优化，结果显示优帕利汀积累量更高，RBH克隆的产量比对照根高1.44倍，效果最佳。然而，与所选根克隆相比，对照根中的波拉维酮B含量更高。通过配备船用叶轮和环形气体分布器设计一种改进的“搅拌罐”反应器，与摇瓶培养相比，促进了高密度RBH根生物量的产量，波拉维酮B和优帕利汀的产量分别增加了6.1倍和1.15倍。与对照根相比，RBH克隆显示出类似的抗氧化/抗菌活性，且抗炎潜力有所提高。首次在生物反应器中实现了毛状根介导的波拉维酮B和优帕利汀的更高产量。
