A novel AP2/ERF family transcription factor from Glycine soja, GsERF71, is a DNA binding protein that positively regulates alkaline stress tolerance in Arabidopsis
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Here we first found that GsERF71, an ERF factor from wild soybean could increase plant alkaline stress tolerance by up-regulating H+-ATPase and by modifing the accumulation of Auxin.
Alkaline soils are widely distributed all over the world and greatly limit plant growth and development. In our previous transcriptome analyses, we have identified several ERF (ethylene-responsive factor) genes that responded strongly to bicarbonate stress in the roots of wild soybean G07256 (Glycine soja). In this study, we cloned and functionally characterized one of the genes, GsERF71. When expressed in epidermal cells of onion, GsERF71 localized to the nucleus. It can activate the reporters in yeast cells, and the C-terminus of 170 amino acids is essential for its transactivation activity. Yeast one-hybrid and EMSA assays indicated that GsERF71 specifically binds to the cis-acting elements of the GCC-box, suggesting that GsERF71 may participate in the regulation of transcription of the relevant biotic and abiotic stress-related genes. Furthermore, transgenic Arabidopsis plants overexpressing GsERF71 showed significantly higher tolerance to bicarbonate stress generated by NaHCO3 or KHCO3 than the wild type (WT) plants, i.e., the transgenic plants had greener leaves, longer roots, higher total chlorophyll contents and lower MDA contents. qRT-PCR and rhizosphere acidification assays indicated that the expression level and activity of H+-ATPase (AHA2) were enhanced in the transgenic plants under alkaline stress. Further analysis indicated that the expression of auxin biosynthetic genes and IAA contents were altered to a lower extent in the roots of transgenic plants than WT plants under alkaline stress in a short-term. Together, our data suggest that GsERF71 enhances the tolerance to alkaline stress by up-regulating the expression levels of H+-ATPase and by modifying auxin accumulation in transgenic plants.
KeywordsBicarbonate stress ERF transcriptional factor Wild soybean Arabidopsis H+-ATPase Auxin
This work was supported by the National Natural Science Foundation of China (31670272 to XD, 31171578 to YZ, 31501331 to DZ), Heilongjiang Provincial Higher School Science and Technology Innovation Team Building Program (2011TD005), and the NEAU starting grant to XD.
Yang Yu and Yanming Zhu conceived the project; Yang Yu, Xiangbo Duan, Chao Chen, Lei Cao, Xuewei Song, Zaib_un Nisa, Jiyang Yu and Qiang Li performed the experiments; Pinghui Zhu, Dan Zhu, Jianying Du, Yu Song and Huiqing Li analyzed the data; Yang Yu wrote the manuscript; Beidong Liu, Kuide Yin and Xiaodong Ding revised the manuscript.
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