Plant Biotechnology Laboratory, Post Graduate Department of Botany, Ramakrishna Mission Vivekananda Centenary College, Rahara, Kolkata 700118, India.
J Biotechnol. 2024 Jun 10;388:59-71. doi: 10.1016/j.jbiotec.2024.04.010. Epub 2024 Apr 16.
Withania somnifera (L.) Dunal is an important indigenous medicinal plant with extensive pharmaceutical potential. The root is the main source of major bioactive compounds of this plant species including withanolides, withanine, phenolic acids, etc. Hairy root culture (HRC) is a crucial method for low-cost production of active compounds on a large scale. Four different Agrobacterium rhizogenes strains have been used for the hairy root induction. Maximum transformation efficiency (87.34 ± 2.13%) was achieved with A4 bacterial strain-mediated transformed culture. The genetic transformation was confirmed by using specific primers of seven different genes. Seven HR (Hairy root) lines were selected after screening 29 HR lines based on their fast growth rate and high accumulation of withanolides and phenolic acids content. Two biotic and three abiotic elicitors were applied to the elite root line to trigger more accumulation of withanolides and phenolic acids. While all the elicitors effectively increased withanolides and phenolic acids production, among the five different elicitors, salicylic acid (4.14 mg l) induced 11.49 -fold increase in withanolides (89.07 ± 2.75 mg g DW) and 5.34- fold increase in phenolic acids (83.69 ± 3.11 mg g DW) after 5 days of elicitation compared to the non-elicited culture (7.75 ± 0.63 mg g DW of withanolides and 15.66 ± 0.92 mg g DW of phenolic acids). These results suggest that elicitors can tremendously increase the biosynthesis of active compounds in this system; thus, the HRC of W. somnifera is cost-effective and can be efficiently used for the industrial production of withanolides and phenolic acids.
睡茄(Withania somnifera(L.)Dunal)是一种具有广泛药用潜力的重要本土药用植物。其根是该植物物种主要生物活性化合物的主要来源,包括醉茄内酯、醉茄胺、酚酸等。发根培养(HRC)是大规模低成本生产活性化合物的重要方法。已经使用了四种不同的根癌农杆菌菌株进行发根诱导。使用 A4 细菌菌株介导的转化培养,获得了最高的转化效率(87.34±2.13%)。通过使用七个不同基因的特异性引物,证实了遗传转化。在筛选了 29 个 HR 线后,根据其快速生长速度和高积累醉茄内酯和酚酸含量,选择了 7 个 HR(发根)线。向优良根线施加两种生物和三种非生物诱导剂,以引发更多醉茄内酯和酚酸的积累。虽然所有诱导剂都有效地增加了醉茄内酯和酚酸的产生,但在五种不同的诱导剂中,水杨酸(4.14mg/L)在 5 天的诱导后,诱导醉茄内酯(89.07±2.75mg/g DW)增加了 11.49 倍,酚酸(83.69±3.11mg/g DW)增加了 5.34 倍,与未诱导培养物(7.75±0.63mg/g DW 醉茄内酯和 15.66±0.92mg/g DW 酚酸)相比。这些结果表明,诱导剂可以极大地增加该系统中活性化合物的生物合成;因此,W. somnifera 的 HRC 具有成本效益,可有效用于醉茄内酯和酚酸的工业生产。