The transcription factor HcERF4 confers salt and drought tolerance in kenaf (Hibiscus cannabinus L.)

Abstract

Ethylene response factors (ERF) are members of the APETALA2/ERF transcription factor family, and they play an important role in plant growth, development, and response to various environmental stresses. In this study, an ERF transcription factor HcERF4 was isolated and characterized from kenaf. The results showed HcERF4 contained an open reading frame of 702 bp, which comprised of single AP2 domain. Subcellular localization analysis revealed that the HcERF4 gene was localized in the nucleus and cytoplasm. Transactivation assays in yeast demonstrated that HcERF4 functions as a transcriptional activator. Its expression profile was investigated by real-time PCR analysis in different tissues and under salt as well as drought stress condition. The result revealed that HcERF4 could be markedly induced by salt and drought in a time and concentration dependent manner. Moreover, silencing of HcERF4 by using the VIGS technique, caused a significant growth inhibition under salt or drought stress condition, with lower plant height, stem diameter, and fresh weight, indicating HcERF4 plays key roles response abiotic stress. In addition, physiological index analyses showed that the contents of MDA, O2−, H2O2, and free proline significantly increased, while the activities of antioxidant enzymes were significantly restrained in HcERF4 silenced plants. Furthermore, the expression level of eight important genes involved in plant growth regulation, including HcERF3, HcNAC29, HcSOD, HcP5CS, HcbZIP43, HcWRKY31, HcMYB44, and HcCBP25 were investigated, and the result showed that all eight genes were highly expressed in the leaves of the wild kenaf, but were significantly downregulated in the leaves of the VIGS kenaf plants under drought and salt stress conditions; Consequently, the present results indicate HcERF4 plays a positive role in regulating salt and drought stress in kenaf.