Hydrogen Peroxide-Induced Oxidative Stress in Sugarcane and Response Expression Pattern of Stress-Responsive Genes Through Quantitative RT-PCR

Abstract

Economic production of sugarcane is highly affected by oxidative stress due to biotic and abiotic stress factors. The main objective of this study was to determine which concentration of hydrogen peroxide can induce the oxidative stress conditions in sugarcane and determine whether several key gene expression responses reported in other species are triggered in sugarcane. Sixty-day-old sugarcane plants were sprayed with 30% H2O2 (300, 500 and 1000 ppm) for three consecutive days. Controls were maintained by spraying with water. Quantitative RT-PCR was performed to quantify the gene expression levels. The PCR products were cloned and sequenced to confirm the identity of the gene amplified. The representative sequences of the stress-responsive genes were deposited in GenBank (Acc. No KX828698 for HSP gene, KX828699 for APX gene, KX828700 for NAC transcription factor and KX828701 for ERF transcription factor). The expression pattern of stress-responsive key genes (HSP, NAC, ERF, GST, MYBAS and CAT) showed higher expression levels within 48 h of H2O2 treatment at 500 and 1000 ppm concentration. However, MYBAS transcription factor and catalase genes showed higher expression levels at a higher concentration of H2O2 (1000 ppm) of H2O2 at 48 h of treatment. Ascorbate peroxidase gene was up-regulated at 1000 ppm of H2O2 at 72 h of treatment. NAC transcription factor, heat shock proteins, ethylene-responsive factor and glutathione S-transferase showed a significant increase in expression at 500 ppm concentration of H2O2 at 48 h. Based on the expression pattern levels, 500 ppm of hydrogen peroxide at 48 h can induce the oxidative stress in sugarcane. The expression levels indicated that HSP, NAC, ERF, GST and MYBAS genes might play a role in signal transduction pathways, and APX and CAT are involved in scavenging reactive oxygen species.