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GmNFYA3, a target gene of miR169, is a positive regulator of plant tolerance to drought stress

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Abstract

Nuclear factor Y (NF-Y) is a heterotrimeric transcription factor composed of NF-YA, NF-YB and NF-YC proteins. In this study, we identified and characterized a gene, GmNFYA3, which encodes the NF-YA subunit of the NF-Y complex in soybeans (Glycine max L.). Real time RT-PCR analysis indicated that GmNFYA3 was induced by abscisic acid (ABA) and abiotic stresses, such as polyethylene glycol, NaCl and cold. Subcellular localization analysis suggested that GmNFYA3 may activate its specific targets in the nucleus. Histochemical β-glucuronidase (GUS) staining revealed that the expression of the GUS gene driven by the GmNFYA3 promoter occurred in various transgenic Arabidopsis tissues. Coexpression in Nicotiana benthamiana and 5′ RACE assays indicated that miR169 directs GmNFYA3 mRNA cleavage in vivo. Overexpression of GmNFYA3 resulted in Arabidopsis with reduced leaf water loss and enhanced drought tolerance. In addition, the transgenic Arabidopsis exhibited increased sensitivity to high salinity and exogenous ABA. Moreover, the transcript levels of ABA biosynthesis (ABA1, ABA2), ABA signaling (ABI1, ABI2) and stress-responsive genes, including RD29A and CBF3, were generally higher in GmNFYA3 plants than in wild-type controls under normal conditions. These results suggest that the GmNFYA3 gene functions in positive modulation of drought stress tolerance and has potential applications in molecular breeding to enhance drought tolerance in crops.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (2007AA10Z139).

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  1. The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Zhiyong Ni, Zheng Hu, Qiyan Jiang & Hui Zhang

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  1. Zhiyong Ni
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Correspondence to Hui Zhang.

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Supplementary material 1 (DOC 35 kb)

11103_2013_40_MOESM2_ESM.tif

Supplemental Figure 1. Drought-tolerant phenotype of 35S:GmNFYA3 plant. Two-week-old Col-0 and 35S:GmNFYA3 plants were transferred from MS plates to the same soil tray and grown for an additional week. Drought stress was imposed by withholding water for 13 days and was followed by a period of rewatering for 3 days. (TIFF 4273 kb)

11103_2013_40_MOESM3_ESM.tif

Supplemental Figure 2 35S:GmNFYA3 plants improve water use efficiency (WUE). WUE of wild-type and 35S:GmNFYA3 plants under water-sufficient conditions was calculated as shoot dry weight divided by total water loss over a period of 4 weeks (mean ± SD, n = 12). Asterisks indicate a significant difference (**P < 0.01) compared with the corresponding controls. (TIFF 92 kb)

11103_2013_40_MOESM4_ESM.tif

Supplemental Figure 3. Salt sensitivity of 35S:GmNFYA3. (A) Growth of the wild-type and 35S:GmNFYA3-1 line on media with or without 100 mM NaCl. Photographs were taken at 7 days after the transfer of 5-day-old seedlings from MS medium to plates containing 100 mM NaCl. Bar = 1.5 cm (B) Measurement of root lengths of plants. Root lengths were measured 7 days after the transfer of 5-day-old seedlings from MS medium to plates without or with 100 mM NaCl. At least three independent experiments were performed and similar results were obtained. Each data point represents the average root length of thirty seedlings ± SD, and asterisks indicate a significant difference (*P < 0.05) compared with the corresponding controls. (TIFF 1910 kb)

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Ni, Z., Hu, Z., Jiang, Q. et al. GmNFYA3, a target gene of miR169, is a positive regulator of plant tolerance to drought stress. Plant Mol Biol 82, 113–129 (2013). https://doi.org/10.1007/s11103-013-0040-5

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