谷子转录因子SiNAC18通过ABA信号途径正向调控干旱条件下的种子萌发

doi:10.3864/j.issn.0578-1752.2017.16.002

Abstract

Abstract: 【Objective】 Foxtail millet (Setaria italica L.) has strong tolerance to drought stress. The objective of this research is to screen key regulatory factors affecting the germination process under drought conditions in plants through reverse genetics method, which will contribute for further research of regulation mechanism of seed germination under drought condition. 【Method】 Multiple sequence alignment of SiNAC18 protein sequences and millet homologous sequences were made by using ClustalX 2 and MEGA 5.05 softwares, and the phylogenetic tree was constructed. The expression patterns of SiNAC18 under the stress condition were analyzed using the real-time PCR method. SiNAC18 protein subcellular localization was analyzed by transient transfection method. Biological function of SiNAC18 was analyzed by using overexpression of SiNAC18 in Arabidopsis thaliana. The expression of downstream genes of SiNAC18 was analyzed by real-time PCR. 【Result】 The length of SiNAC18 is 1 074 bp encoding a hydrophilic protein with polypeptide of 357 amino acids, and its molecular weight is about 38.8 kD. Phylogenetic tree analysis indicated that SiNAC18 belongs to NAP subgroup of group I in the NAC transcription factors family and has the highest homology with Arabidopsis gene AtNAC29. The amino acid sequence alignment results show that the N-terminal of SiNAC18 and the highest homology transcription factors of SiNAC18 in other species, including rice, Arabidopsis thaliana, soybean and maize, has A, B, C, D and E five conserved domains. The C-terminal of the protein has a high degree of polymorphism, demonstrating that the N-terminal sequence of SiNAC18 is associated with its downstream promoter. Real-time PCR results showed that SiNAC18 were induced by drought (PEG), high salt (NaCl) and hydrogen peroxide (H₂O₂) treatment. Subcellular localization results showed that SiNAC18 protein is localized in the nucleus. Gene function analysis showed that in the ABA and PEG stress treatments, the germination rate of SiNAC18 transgenic Arabidopsis thaliana and wild type seeds was significantly different. Under normal growth conditions, germination rate of the wild type Arabidopsis WT and SiNAC18 transgenic Arabidopsis was the same, and when the concentration of PEG was increased to 10% and 15% on MS medium, the germination rate of SiNAC18 transgenic Arabidopsis was significantly higher than WT. Under the conditions of 2 and 5 μmol·L^-1of ABA treatment, the germination rate of SiNAC18 transgenic Arabidopsis was significantly lower than that of WT. Analysis results showed that the expression of downstream genes of ABA related genes AtRD29A, proline synthesis related genes AtP5CR and AtPRODH and peroxidase gene AtPRX34 in SiNAC18 transgenic plants was higher than that in WT, which suggesting that SiNAC18 affects the germination rate of transgenic plants under drought conditions by affecting the expression of those downstream genes. 【Conclusion】 NAC like transcription factor gene SiNAC18 positively regulates the germination of plants under drought conditions through ABA and oxidative stress response signaling pathway in foxtail millet.

Key words: foxtail millet (Setaria italic L.), NAC transcription factors, seed germination, drought stress, ABA signaling pathway

DOU YiNing, QIN YuHai, MIN DongHong, ZHANG XiaoHong, WANG ErHui, DIAO XianMin, JIA GuanQing, XU ZhaoShi, LI LianCheng, MA YouZhi, CHEN Ming. Transcription Factor SiNAC18 Positively Regulates Seed Germination Under Drought Stress Through ABA Signaling Pathway in Foxtail Millet (Setaria italic L.)[J].Scientia Agricultura Sinica, 2017, 50(16): 3071-3081.

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Transcription Factor SiNAC18 Positively Regulates Seed Germination Under Drought Stress Through ABA Signaling Pathway in Foxtail Millet (Setaria italic L.)

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