Bamal Deepak, Singh Anoop, Swami Nisha, Sehrawat Anita Rani
Department of Botany, Maharshi Dayanand University, Rohtak, 124001, India.
Appl Biochem Biotechnol. 2025 May 27. doi: 10.1007/s12010-025-05284-3.
This study investigated the potential of green-synthesized copper oxide nanoparticles (CuO NPs) to enhance biomass production and therapeutic metabolite yields in Alhagi maurorum, a medicinal plant of significant pharmaceutical value. CuO NPs were biosynthesized using A. maurorum leaf extract as a reducing and capping agent, with characterization confirmed via UV-Vis spectroscopy, FTIR, XRD, SEM, TEM, and zeta potential analysis. Nanoparticles ranged from 7-30 nm in size. Callus induction and proliferation were established using Murashige and Skoog (MS) media supplemented with varying concentrations (0-12 mg/L) of CuO NPs combined with plant growth regulators. Maximum callus fresh weight (9.02 mg in cotyledon and 8.46 mg in hypocotyl) was achieved in MS media containing 3.0 mg/L BAP, 0.1 mg/L NAA, and 0.50 mg/L kinetin without CuO NPs. However, CuO NPs significantly enhanced metabolite production in a dose-dependent manner. Analysis of variance revealed statistically significant differences (p=0.001) across all biochemical parameters tested, with high F-values for peroxidase activity (7,755.74), total flavonoids (5,195.02), and total soluble sugar (5,702.18). At 8 mg/L CuO NPs, callus cultures exhibited elevated levels of total free amino acids (12.49±0.023 mg/g DW) and total soluble protein (35.617±0.033 mg/g DW), while control samples produced higher starch (35.547±0.23 mg/g DW) and total soluble sugar (121.56±0.091 mg/g DW) content. Significantly, CuO NP-treated cultures demonstrated enhanced secondary metabolite synthesis, with maximum total phenolic compounds (156.477±0.167 mg/g DW GAE) and flavonoids (58.307±0.179 mg/g QE) at 8 and 10 mg/L CuO NPs, respectively. Antioxidant enzyme analysis revealed that cotyledon-derived callus exhibited peak activities at specific CuO NP concentrations: superoxide dismutase (84.5±0.254% inhibition) and glutathione reductase (0.75±0.006% inhibition) at 8 mg/L; peroxidase (3.137±0.009 U), catalase (77.35±0.152 U), and ascorbate peroxidase (0.43±0.006 mM/mg FW) at 10 mg/L. HPLC analysis confirmed the novel presence of lupeol, an anticancer compound, in regenerated roots. These findings demonstrate the potential of CuO NPs for enhancing therapeutic metabolite production in A. maurorum tissue culture while suggesting optimal concentration ranges (8-10 mg/L) for maximum benefits. Further research is necessary to elucidate the molecular mechanisms governing nanoparticle-plant interactions and to address potential health implications.
本研究调查了绿色合成的氧化铜纳米颗粒（CuO NPs）在增强具有重要药用价值的药用植物骆驼刺生物量生产和治疗性代谢产物产量方面的潜力。以骆驼刺叶提取物作为还原剂和封端剂生物合成了CuO NPs，并通过紫外可见光谱、傅里叶变换红外光谱、X射线衍射、扫描电子显微镜、透射电子显微镜和zeta电位分析对其进行了表征。纳米颗粒的尺寸范围为7 - 30纳米。使用添加了不同浓度（0 - 12毫克/升）CuO NPs并结合植物生长调节剂的Murashige和Skoog（MS）培养基建立愈伤组织诱导和增殖体系。在不含CuO NPs的含有3.0毫克/升苄氨基嘌呤、0.1毫克/升萘乙酸和o.50毫克/升激动素的MS培养基中，获得了最大愈伤组织鲜重（子叶中为9.02毫克，下胚轴中为8.46毫克）。然而，CuO NPs以剂量依赖的方式显著提高了代谢产物的产量。方差分析显示，在所有测试的生化参数中存在统计学上的显著差异（p = 0.001），过氧化物酶活性（7755.74）、总黄酮（5195.02）和总可溶性糖（5702.18）的F值较高。在8毫克/升CuO NPs时，愈伤组织培养物中总游离氨基酸（12.49±0.023毫克/克干重）和总可溶性蛋白（35.617±0.033毫克/克干重）水平升高，而对照样品产生了更高的淀粉（35.547±0.23毫克/克干重）和总可溶性糖（121.56±0.091毫克/克干重）含量。值得注意的是，经CuO NP处理的培养物表现出次生代谢产物合成增强，在8毫克/升和10毫克/升CuO NPs时，总酚类化合物（156.477±0.167毫克/克干重没食子酸当量）和黄酮类化合物（58.307±0.179毫克/克干重槲皮素当量）分别达到最大值。抗氧化酶分析表明，子叶来源的愈伤组织在特定CuO NP浓度下表现出峰值活性：8毫克/升时超氧化物歧化酶（84.5±0.254%抑制）和谷胱甘肽还原酶（0.75±0.006%抑制）；10毫克/升时过氧化物酶（3.137±0.009单位）、过氧化氢酶（77.35±0.152单位）和抗坏血酸过氧化物酶（0.43±0.006毫摩尔/毫克鲜重）。高效液相色谱分析证实，再生根中存在新型抗癌化合物羽扇豆醇。这些发现证明了CuO NPs在提高骆驼刺组织培养中治疗性代谢产物产量方面的潜力，同时表明了最大效益的最佳浓度范围（8 - 10毫克/升）。有必要进一步研究阐明纳米颗粒与植物相互作用的分子机制，并解决潜在的健康问题。
