Isolation, Cloning, and Heterologous Maintenance of a Class III Peroxidase Gene from

Plant growth, development, and defense reactions are greatly dependent on the massive multigene family of heme- containing glycoproteins called class III plant peroxidases (EC 1.11.1.7). These enzymes are of prime importance for resistant pathogens, lignification, suberization, and reactive oxygen species (ROS) metabolism. The isolation, amplification, cloning, and bacterial preservation of a peroxidase gene from Solanum tuberosum (potato), a globally important crop constantly under attack by biotic and abiotic stress factors, are described in this study. The CTAB method was used for the isolation of genomic DNA from young sprouting buds of potato tubers. Using gene-specific primers, a conserved region of the peroxidase gene was amplified by polymerase chain reaction (PCR). After being ligated into a plasmid vector, the amplified region was heat-shocked to produce Escherichia coli DH5a competent cells. The successful transformation was confirmed by biochemical screening and selection with ampicillin. A useful tool for gene conservation and subsequent applications in molecular breeding and plant genetic engineering is offered by the conservation of the peroxidase gene in a bacterial system. The significance of the peroxidase genes in enhancing plant resistance by genetic engineering and their importance as pathogenesis-related (PR- 9) proteins is highlighted in the study. This study enhances our understanding of the role of peroxidase in plants and supports future applications of transgenic technology to improve plant resistance.