Cold-inducible expression of AZI1 and its function in improvement of freezing tolerance of Arabidopsis thaliana and Saccharomyces cerevisiae |
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Authors: | Xu Zhi-Yan Zhang Xin Schläppi Michael Xu Zi-Qin |
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Institution: | a Key Laboratory of Resource Biology and Biotechnology in Western China (Ministry of Education), Provincial Key Laboratory of Biotechnology, Institute of Life Sciences, Northwest University, Xi’an, Shaanxi 710069, People's Republic of China b Department of Biological Sciences, Marquette University, Milwaukee, WI 53233, USA |
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Abstract: | AZI1 (AZELAIC ACID INDUCED 1) of Arabidopsis thaliana could be induced by azelaic acid and was involved in priming of systemic plant immunity. In the present work, expression of AZI1 in response to low temperature was investigated via RNA gel blot analysis. AZI1 could be induced slowly by cold stress and more than 6 h treatment at 4 °C was required to detect an increase in mRNA abundance. However, the high expression state could not be maintained stably and would decline to basal level when the plants were transferred to room temperature. In order to clarify the function of AZI1 in resistance to abiotic stresses, overexpressing, RNA interference and T-DNA knockout lines of this gene were used in electrolyte leakage assays. Overexpression of AZI1 resulted in reduced electrolyte leakage during freezing damage. In contrast, AZI1 knockdown and knockout lines showed increased tendencies in cellular damage after freezing treatment. To further validate the potential resistance of AZI1 to low-temperature stress, Saccharomyces cerevisiae cells were transformed with pESC-AZI1 in which AZI1 was under the control of GAL1 promoter. Compared to yeast cells containing empty pESC-URA, the survival rate of yeast cells harboring AZI1 increased obviously after freezing treatment. All these results suggested that AZI1 might be multifunctional and associated with cold tolerance of Arabidopsis. |
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Keywords: | 8CM eight-cysteine-motif Col-0 Arabidopsis ecotype Columbia-0 El electrolyte leakage HyPRP hybrid proline-rich protein ICE1 inducer of CBF expression 1 LTP lipid transfer protein LTRE low temperature responsive element MS Murashige and Skoog medium PCR polymerase chain reaction PRD proline-rich domain SA salicylic acid TCA trichloroacetic acid Ws Arabidopsis ecotype Wassilewskija |
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