Zinc Oxide Nanoparticles Induce Genetic Transformation of Silybum marianum L. Mediated by Agrobacterium rhizogenes ATCC13332
DOI:
https://doi.org/10.15649/2346075X.5211Keywords:
Agrobacterium rhizogenes, milk thistle, nanoparticles, rolBAbstract
Introduction. Nanoparticles have recently garnered substantial interest across various scientific disciplines due to their novel physicochemical properties. These particles have found broad applications in medicine, agriculture, environmental sciences, and pharmaceutical technologies. Objectives. To investigate the effect of zinc oxide nanoparticles (ZnO NPs) on the transformation efficiency of the medicinal plant milk thistle (Silybum marianum L.) mediated by Agrobacterium rhizogenes. Materials and Methods. Explants derived from 20-day-old seedlings were directly injected with Agrobacterium rhizogenes ATCC13332 using a 0.5 mL insulin syringe. Treated explants were cultured on solidified Murashige and Skoog (MS) medium devoid of growth regulators. Results. Hairy roots were induced in 55.86% of leaf explants, marking the first visible indicator of successful genetic transformation. Callus induction from hairy roots reached a 100% success rate on MS2 medium (MS + 1.0 mg·L-¹ benzyladenine [BA] + 0.1 mg·L-¹ indole-3-butyric acid [IBA]). Polymerase chain reaction (PCR) analysis confirmed the presence of the rolB gene in the genomic DNA of transformed tissues. Shoot regeneration occurred during hairy root maintenance. Furthermore, supplementing MS medium with varying concentrations (25–300 μg/mL) of ZnO NPs significantly enhanced hairy root induction, achieving a maximum rate of 100% in leaf explants cultured on MS medium containing 150 μg/mL ZnO NPs. Conclusions. Zinc oxide nanoparticles effectively enhanced the genetic transformation frequency in S. marianum, demonstrating their potential as a nanobiotechnological tool in plant genetic engineering.
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