Cloning and Functional Analysis of Expansin TaEXPA9 Homologs in Winter Wheat in Frigid Regions

Abstract

BackgroundLow temperature is an important factor that influences the ability of winter wheat to safely overwinter. Excessive low temperatures restrict the regrowth of winter wheat, thus decreasing agricultural output. Non-enzymatic expansins, which are related to plant growth, have been reported to respond to drought, salinity, and low temperature stress. We obtained an expansin gene, TaEXPA9, that is induced by low temperature from a transcriptome analysis of ‘Dongnong winter wheat no. 2’—a winter wheat with high cold hardiness—but the expression pattern and function of this gene were unknown. We therefore analyzed the expression patterns of TaEXPA9-A/B/D in D2 in response to different abiotic stresses and exogenous phytohormone treatments in different organs. The entire length of TaEXPA9-A/B/D was obtained, and green fluorescent labeling was used for subcellular localization analysis of TaEXPA9-A/B/D on onion epidermis. The 35S::TaEXPA9-A/B/D expression vector was constructed, and an overexpression transgenic Arabidopsis thaliana line was obtained to examine the effects of the homologs of this expansin on plant growth and low temperature stress resistance. ResultsThe results showed that TaEXPA9-A/B/D transcription significantly increased at 4°C low temperature stress, its expression level was higher in the roots, and TaEXPA9-A/B/D was localized to the cell wall. The roots were well-developed in the overexpression A. thaliana, and the growth-related markers and setting rate were better than in the wild-type. Recovery was stronger in the overexpression plants after frost stress. At 4°C low temperature stress, the antioxidant enzyme activity and osmoregulatory substance content in the TaEXPA9-A/B/D-overexpressing A. thaliana plants were significantly higher than in the wild-type plants, and the degree of membrane lipid peroxidation was lower. ConclusionsIn summary, TaEXPA9-A/B/D participates in the low-temperature stress response and may increase the scavenging of reactive oxygen species caused by low temperature stress through the protective enzyme system. Additionally, TaEXPA9-A/B/D can increase the levels of small molecular organic substances to resist osmotic stress caused by low temperature.