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Phytomining involves bioharvesting of metals from high biomass crops grown in soil substrates. This process has the potential for economic exploitation of low-grade mineralized soils (e.g., gold), which are too poor for conventional mining of metals. There is a lot of evidence that supports the synthesis of gold nanoparticles in living plants. In this study, the bioreduction of Au ions to Au nanoparticles (AuNPs/GNPs) by three types of Iranian alfalfa plants (Nikshahri, Hamedani, and Yazdi) is presented. The effects of pH, time, and concentration of chloroauric acid (HAuCl₄) as significant factors in the bioreduction reaction were also studied. The AuNPs were characterized by UV-vis spectroscopy, atomic absorption spectrometry, scanning electron microscopy (SEM), nanoparticle analyzer, and X-ray diffractometer (XRD) analysis. The synthesized particles were crystalline in nature, and different size of these …