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Molecular characterization of three ethylene responsive element binding factor genes from cotton
Long-Guo Jin1, Hui Li, Jin-Yuan Liu
1Laboratory of Molecular Biology and MOE Laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
Journal of Integrative Plant Biology
|June 12, 2010
Summary
Three novel Ethylene-Responsive Factor (ERF) genes in cotton, GhERF2, GhERF3, and GhERF6, were identified. These genes are involved in regulating plant responses to ethylene, abscisic acid, and various environmental stresses.
Area of Science:
- Plant Molecular Biology
- Plant Physiology
Background:
- Ethylene-Responsive Factors (ERFs) play a crucial role in regulating plant gene expression.
- Understanding ERFs is key to deciphering plant responses to environmental stimuli.
Purpose of the Study:
- To isolate and characterize novel ERF genes from cotton (Gossypium hirsutum).
- To investigate the function and expression patterns of GhERF2, GhERF3, and GhERF6 in response to various stresses.
Main Methods:
- Isolation of novel ERF genes using rapid amplification of cDNA ends-polymerase chain reaction.
- Transient expression analysis with GhERF-green fluorescent protein fusions to determine protein localization.
- Yeast and tobacco cell assays to assess transcriptional activation and DNA binding.
- Semi-quantitative reverse transcription-polymerase chain reaction for expression analysis.
- Promoter analysis to identify stress-responsive elements.
Main Results:
- GhERF2, GhERF3, and GhERF6 proteins are localized in the nucleus and exhibit transcriptional activation activity in yeast.
- GhERF6 specifically binds to GCC boxes.
- GhERF2 and GhERF3 are constitutively expressed in all cotton organs, while GhERF6 is expressed in vegetative organs.
- Expression of all three genes is rapidly induced by ethylene, abscisic acid, salt, cold, and drought treatments.
- Promoter analysis revealed stress-inducible elements in the upstream regions of these genes.
Conclusions:
- GhERF2, GhERF3, and GhERF6 function as positive trans-acting factors in cotton's response to ethylene, abscisic acid, and multiple environmental stresses.
- These findings provide insights into the regulatory mechanisms of cotton's stress response pathways.
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