洋地黄是Nilgiris的重要植物之一,印度的克什米尔和大吉岭地区,属于车前草科,具有众所周知的药理学应用。在目前的调查中,正在探索一种间接枝条器官发生的体外培养技术;生化属性,抗氧化活性和代谢组学分析是通过使用非目标气相色谱-质谱(GC-MS)和超高效液相色谱与电子喷雾电离/四极杆飞行时间质谱(UPLC-ESI-QTOF-MS)方法进行的。最初,叶片外植体用于愈伤组织诱导和增殖,在MS上获得最大愈伤组织频率(94.44%)和新鲜生物量(4.9g),用8.8µMBAP(6-苄基氨基嘌呤)+0.9µM2,4-D(2,4-二氯苯氧基乙酸)强化,随后在相同的MS培养基上观察到芽形成(间接器官发生),芽诱导频率为83.33%。稍后,体内的生化和抗氧化潜力-,评估了体外生长的叶片和叶片衍生的愈伤组织。明显较高的总酚,类黄酮,DPPH(2,2-二苯基-1-吡啶酰肼),与田间生长的叶片相比,在体外生长的愈伤组织和叶片组织中发现了POD(过氧化物酶)和SOD(超氧化物歧化酶)活性。每种甲醇提取物的GC-MS分析(体内,体外衍生的叶子和叶子衍生的愈伤组织)显示存在超过75种生物活性化合物,即loliolide,污名,α-生育酚,角鲨烯,棕榈酸,亚油酸,β-Amyrin,菜油甾醇等.具有巨大的治疗重要性。以正离子和负离子化模式进行每种甲醇提取物的基于UPLC-MS的代谢物指纹分析。获得的结果揭示了田间和实验室生长组织中植物化学成分的变化,表明体外培养条件对植物组织的影响。检测到的植物化合物属于各种类别,如类黄酮,类固醇,萜类化合物,碳水化合物,单宁,木脂素等.鉴定的药用重要代谢物是20,22-二氢地高辛,洋地黄毒苷,芹菜素,木犀草素,Kaempferide,迷迭香酸,Nepitrin和其他人。本研究的结果表明,通过在较短的时间内生产具有商业价值的此类重要植物化合物,可以成功地将其体外培养物用于新药发现,而不会损害植物的自然种群。
Digitalis purpurea L. is one of the important plant species of Nilgiris, Kashmir and Darjeeling regions of India, belonging to the family Plantaginaceae, with well-known pharmacological applications. In the present investigation, an in vitro culture technique of indirect shoot organogenesis of D. purpurea is being explored; the
biochemical attributes, the antioxidant activities and the metabolomic analyses were made by utilizing untargeted Gas Chromatography-Mass Spectrometry (GC-MS) and Ultra Performance Liquid Chromatography coupled with electronspray ionization/quadrupole-time-of-flight-mass spectrometry (UPLC-ESI-QTOF-MS) approaches. Initially, the leaf explants were used for callus induction and proliferation and maximum callusing frequency (94.44%) and fresh biomass (4.9 g) were obtained on MS, fortified with 8.8 µM BAP (6-benzyl amino purine) + 0.9 µM 2,4-D (2,4-dichlorophenoxyacetic acid), subsequently shoot formation (indirect organogenesis) was noted on the same MS medium with a shoot induction frequency of 83.33%. Later on, the
biochemical and antioxidant potential of in vivo-, in vitro grown leaf and leaf derived callus were assessed. Significantly higher total phenol, flavonoid, DPPH (2,2-diphenyl-1-picrylhydrazyl), POD (peroxidase) and SOD (superoxide dismutase) activities were noticed in in vitro grown callus and leaf tissues compared with field grown leaf. The GC-MS analysis of each methanolic extract (in vivo-, in vitro derived leaf and leaf derived callus) displayed the presence of more than 75 bioactive compounds viz loliolide, stigmasterin, alpha-tocopherol, squalene, palmitic acid, linoleic acid, beta-amyrin, campesterol etc. possessing immense therapeutic importance. The UPLC-MS based metabolite fingerprinting of each methanolic extracts were conducted in both positive and negative ionization mode. The obtained results revealed variation in phytochemical composition in field - and laboratory grown tissues, indicating the impact of in vitro culture conditions on plant tissues. The detected phytocompounds belongs to various classes such as flavonoids, steroids, terpenoids, carbohydrates, tannins, lignans etc. The medicinally important metabolites identified were 20, 22-dihydrodigoxigenin, digoxigenin monodigitoxoside, apigenin, luteolin, kaempferide, rosmarinic acid, nepitrin and others. The results of the present study suggest that in vitro culture of D. purpurea could successfully be utilized for the novel drug discovery by producing such important phytocompounds of commercial interest in shorter duration without harming the plants\' natural population.