Genome wide analysis of HMA gene family in Hydrangea macrophylla and characterization of HmHMA2 in response to aluminum stress

Abstract

Aluminum toxicity poses a significant threat to plant growth, especially in acidic soils. Heavy metal ATPases (HMAs) are crucial for transporting heavy metal ions across plant cell membranes, yet their role in Al³⁺ transport remains unexplored. This study identified eight HmHMA genes in the genome of Hydrangea macrophylla, categorizing them into two major clades based on phylogenetic relationships. These genes were found unevenly distributed across six chromosomes. Detailed analysis of their physicochemical properties, collinearity, and gene structure was conducted. RNA-seq and qRT-PCR analyses revealed that specific HmHMA genes, notably HmHMA2, were predominantly expressed in roots and flowers under Al³⁺ stress, indicating their potential role in Al³⁺ tolerance. HmHMA2 showed significant expression in roots, especially under Al³⁺ stress conditions, and when expressed in yeast cells, it conferred resistance to aluminum and zinc but increased sensitivity to cadmium. Overexpression of HmHMA2 in hydrangea leaf discs significantly improved Al³⁺ tolerance, reduced oxidative stress markers like hydrogen peroxide and malondialdehyde, and enhanced antioxidant enzyme activity such as SOD, POD and CAT compared to controls. These findings shed lights on the potential role of HmHMAs in Al transport and tolerance in H. macrophylla.