*Article* **EDTA and IAA Ameliorates Phytoextraction Potential and Growth of Sunflower by Mitigating Cu-Induced Morphological and Biochemical Injuries**

**Naila Shah 1,2, Muhammad Irshad 1,\*, Anwar Hussain 1, Muhammad Qadir 1, Waheed Murad 1, Asif Khan 3, Muhammad Awais 4, Abdulwahed Fahad Alrefaei <sup>5</sup> and Sajid Ali 6,\***


**Abstract:** As an essential micronutrient, copper is vital for normal growth and development of plants, however, its accumulation in soil exerts a severe negative impact on the agronomic characteristics and yield of the crop plants. Phytoextraction is a low-cost method for restoring soil fertility and avoiding losses due to heavy metal contamination. We found that using EDTA and IAA together improved sunflower hyperaccumulation capacity. Sunflowers were cultivated under various levels of Cu (0 (control), 25, 50, and 75 mg/kg of soil) and treated with EDTA alone or combined with IAA. The results revealed that the amended treatment significantly enhanced the absorption and accumulation of Cu in the sunflowers. Furthermore, the various doses of Cu significantly reduced the root and shoot growth of sunflowers in a concentration-dependent manner by impairing the chlorophyll content, hormones (indole 3-acetic acid, salicylic acid, and gibberellic acid), flavonoids, phenolics, and antioxidant response. The injurious effect of Cu was reduced by the addition of EDTA alone, and the supplementation of IAA led to a significant restoration of shoot growth (~70%) and root growth (~13%) as compared to the plant treated with Cu alone. Moreover, significantly higher levels of chlorophyll content, GA3, endogenous IAA, and flavonoids were recorded, indicating the effectiveness of the treatment in ameliorating plant health. The results also showed considerable restoration of the catalase and ascorbate peroxidase activities in plants treated with EDTA and IAA. These results are suggestive that application of EDTA and IAA enhances the Cu absorption potential of sunflower and increases its tolerance to copper, which may not only serve as a better technique for phytoextraction of Cu, but also to bring Cu contaminated soil under cultivation.

**Keywords:** Copper hyperaccumulation; stress mitigation; EDTA and IAA; sunflower
