Glutathione Peroxidase cDNA Cloning and Expression in Soybean Roots under Heterodera glycines Infection

Abstract

A gene named GmGPX1 encoding glutathione peroxidase (GPX) was cloned and sequenced from soybean roots infested Heterodera glycines by RT-PCR (Reverse Transcription PCR), which is a crucial enzyme in plant cells regulating reactive oxygen species (ROX). The cDNA length of cloned gene was 693bp, flanked by a 5'-untranslated region of 7bp and a 3'-untranslated region of 185bp, containing six exons and five introns. Genomic DNA fragment was located at Glycine max chromosome 5. The open reading frame of cDNA which encodes a polypeptide of 166 amino acid residues and protein molecular weight was 18375.8Da, theoretical isoelectric point 6.59. The deduced amino acid sequence showed about 99% and 64% homology with G max putative PHGPX (XP_003532707.1) and Arabidopsis thaliana GPX (NP_564813.1), respectively. The expression profile of the GmGPX1 in Xiaoliheidou (G. max) under H. glycines infection, which generates oxidative stress, was analyzed. Real time PCR analysis revealed that the GmGPX1 mRNA levels were increased stabilized from 1.3 to 1.47 times after exposure to H. glycines from 12h to 48h, and reduced to 1.07 times at 72h comparing with non-inoculation control. These results suggest that the GmGPX1 gene is induced by H. glycines at early stage in soybean roots and play an important role in removing oxidative damage.

The Soybean Purple Acid Phosphatase GmPAP14 Predominantly Enhances External Phytate Utilization in Plants

“…Quantitative RT-PCR (qPCR) was performed with SYBR Green Mix (Takara, Japan) on a CFX96 (Bio-Rad, United States). GmPAP14 primers (5′-TCAAGCAGCCCCTTCATTAG-3′ and 5′-AGTTTTCCTTCGGCAATCTTC-3′) and primers for the housekeeping gene GmActin11 (5′-ATCTTGACTGAGCGTGG TTATTCC-3′ and 5′-GCTGGTCCTGGCTGTCTCC-3′) were used in qPCR ( Wang et al, 2012 ). Relative expression was calculated using the 2 -ΔΔC t method ( Livak and Schmittgen, 2001 ).…”

Cloning of a new glutathione peroxidase gene from tea plant (Camellia sinensis) and expression analysis under biotic and abiotic stresses

“…Glutathione peroxidase (GPX) is the general name for a family of isozymes that removes ROS using GSH as an electron donor (Margis et al 2008 ; Wang et al 2012 ). Generally, in plants GPXs localized at different cell organelles as mitochondria, chloroplast, endoplasmic reticulum/cytosol or secreted (Ramos et al 2009 ).…”