A nursery experiment was conducted during the period from 2-7-2022 to 14-9-2023, aiming to evaluate the effects of foliar application of Spirulina extract (SE) at two concentrations (5 and 10 mL/L) and nano-micronutrients of iron (NFe) and zinc (NZn) at two concentrations (2 and 4 mL/L) and their interactions on growth and nutritional status of Nemaguard peach, comparing with untreated control (spray with distilled water only). Results showed that the binary interaction between Spirulina fertilization and nano-microelements, the combination SE1 + (NF2+NZ2) treatment, gave the highest significant increase in seedling height and diameter, total chlorophyll leaf content, fresh and dry weight of leaves, potassium, magnesium, and zinc. Meanwhile, the combination SE2 + (NF2+NZ2) treatment significantly increased the number of branches and leaves/seedlings and leaf area, as well as the leaf content of total carbohydrates, nitrogen, phosphorus, and iron. Nano-Fe/Zn and Spirulina extract also increased the activity of microbes in the rhizosphere of seedlings compared to the control treatment. It could be concluded that applying Spirulina extract and nano-micronutrients (iron and zinc) to Nemaguard rootstock seedlings is an effective way to improve vegetative growth and leaf mineral content, promote seedling growth, and shorten the time needed to reach the appropriate size to be grafted onto.