Abstract
Background Ascorbate peroxidase (APx) is a key enzyme of the antioxidant metabolism, catalyzing the decomposition of hydrogen peroxide (H2O2) in water, using ascorbate as an electron donor. The H2O2 is a reactive oxygen species (ROS) produced constantly by aerobic metabolism. Under biotic and abiotic stress the level of H2O2 increases and, in large quantities, can cause cellular damage. In rice, there are eight APx genes that encode products target to different subcellular compartments: cytosol, peroxisoma, mitochondria and chloroplast. OsAPx1 gene encodes a cytosolic isoform of APx. The study of promoters is an important tool that allows to analyze the overall expression pattern of genes in plants.
Highlights
Ascorbate peroxidase (APx) is a key enzyme of the antioxidant metabolism, catalyzing the decomposition of hydrogen peroxide (H2O2) in water, using ascorbate as an electron donor
The transformed calli were grown in selection medium with hygromycin, regenerated into plants, acclimatized in a greenhouse and the confirmation of transgene was verified by PCR using specific primers for the Hpt and Gus genes
Green tissues were incubated in 70% ethanol for chlorophyll discoloration
Summary
Ascorbate peroxidase (APx) is a key enzyme of the antioxidant metabolism, catalyzing the decomposition of hydrogen peroxide (H2O2) in water, using ascorbate as an electron donor. Methods A sequence of approximately 2kb preceding the translation initiation site of the OsAPx1 gene was isolated, cloned into pENTR vector and recombined in pHGWFS7 vector, whichallows the fusion of the promoter sequence with two report genes, Gfp and Gus, and confers resistence to hygromycin. The transformation of rice calli, originated from nipponbare cultivar seeds, was performed via Agrobacterium tumefaciens.
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