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Molecular Cloning and Functional Analysis of an Ethylene Receptor Gene from Sugarcane (Saccharum spp.) by Hormone and Environmental Stresses

Ai-Qin Wang, Xing-Zhi Ye, Jing-Li Huang, Jun-Qi Niu, Ming Liu, Yong-Bao Pan, Li-Tao Yang, Yang-Rui Li
Sugar tech 2015 v.17 no.1 pp. 22-30
DNA primers, Saccharum, amino acid sequences, amino acids, complementary DNA, corn, drought, ethephon, ethylene, ethylene production, gene expression, gene expression regulation, genes, haploidy, internodes, leaves, molecular cloning, plant tissues, polypeptides, quantitative polymerase chain reaction, receptors, reverse transcriptase polymerase chain reaction, rice, signal transduction, sugarcane, temperature
Ethylene receptor (ethylene response sensor, ERS) is the primary component involving in the ethylene biosynthesis and ethylene signal transduction pathway. In the present study, a GZ-ERS gene encoding ERS was cloned from a sugarcane cv. YL17 (Saccharum spp.) using RT-PCR and ligation-mediated PCR with primers designed based on conserved amino acid sequences of ERS from rice and corn. The GZ-ERS gene was 2,243 base pairs in length and coded for a polypeptide of 633 amino acid residues. The amino acid sequence of GZ-ERS encoded protein was 97, 96 and 91 % identical to OS-ERS (AAX95525) of rice, ZmERS25 (AAR25567) of corn, and Sc-ERS (ACF60981) of a sugarcane cv. ROC22, respectively. Genomic Southern analysis using a GZ-ERS cDNA insert as the probe indicated the presence of a single copy of GZ-ERS gene per haploid sugarcane genome. Real-time qPCR analysis showed that GZ-ERS was widely expressed in the vegetative tissues of sugarcane, and its expression in maturing internodes and leaf tissues was higher than in immature tissues. Furthermore, the expression level of GZ-ERS was the highest in maturing leaves, but very low in immature internodes. The GZ-ERS expression level in maturing leaves was enhanced by ethephon treatment, but was reduced under low temperature, drought, and dark conditions. The results from this study may help understand the regulation of gene expression in sugarcane and its responses to plant growth regulator ethephon and environmental stresses. As the sugarcane ERS family has multiple members, other members need to be isolated and characterized in future studies.