Functional analysis of two stress-related promoters in Medicago varia cultivar Xinmu No.1

doi:10.11686/cyxb2016071

Abstract

Abstract: MvP5CS and MvNHX1 genes of the Medicago varia cultivar Xinmu No.1 and their promoter sequences have been successfully cloned in previous work. Based on this research, in this study two plant expression vectors, each containing Mvp5cs and Mvnhx1 promoters with a GUS gene as the reporter, were constructed and two tobacco transgenic plants with the promoter obtained by Agrobacterium-mediated method. PCR analysis showed that the two promoters had been stably integrated into the tobacco genome and GUS-staining proved that they have the function of activating gene expression. We explored this function by measuring GUS activity under four different abiotic stresses: drought, salinity, abscisic acid (ABA) and gibberellin (GA). The results showed that Mvnhx1 and Mvp5cs responded to the four stresses and had significant differences from the CaMV35S promoter. The Mvnhx1 promoter was superior to Mvp5cs under salt and ABA stress. After 48 h treatments of 100 and 150 mmol/L NaCl stress, the GUS activity of Mvnhx1 was 2.03 times and 3.23 times that of Mvp5cs. Under treatments of 25 and 50 μmol/L ABA stress, Mvnhx1 activity was significantly higher than the other two promoters. Mvp5cs was superior to Mvnhx1 under drought and GA stress. The GUS activity of the Mvp5cs was 2.22 times that of Mvnhx1 after 36 h treatment of drought stress. The Mvp5cs promoter reached its maximum value, which was 1.79 times that of Mvnhx1, after 36 h treatment with 70 μmol/L GA stress.

CHAO Zhao-Xia, REN Yan-Ping, QIAN Jin, YAO Zheng-Pei, XU Lei, ZHANG Hua. Functional analysis of two stress-related promoters in Medicago varia cultivar Xinmu No.1[J]. Acta Prataculturae Sinica, 2017, 26(1): 131-141.

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