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Overexpression of GmSUMO2 gene confers increased abscisic acid sensitivity in transgenic soybean hairy roots

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Abstract

Small ubiquitin-like modifier (SUMO) participates in post-translational modification of various target proteins. SUMOylation is an important molecular regulatory mechanism for plants to respond to abiotic stress. In the present study, GmSUMO2 gene was isolated from soybean seedlings for further study because of the highest expression level among these six SUMO genes in soybean. qRT-PCR results showed that GmSUMO2 gene were detected in root, leaf, cotyledon, seed root, flower, pod and seed, with the highest transcription level in cotyledon. Moreover, GmSUMO2 gene was transcriptionally regulated by 200 mM NaCl, 42 °C, 25 μM abscisic acid (ABA) and 20% PEG6000 during the 24 h period of treatment. Besides, western blot analysis using AtSUMO1 antibody indicated that the free SUMO levels and SUMOylation dynamics were regulated by ABA stimulus. Functional analysis indicated that overexpression of GmSUMO2 gene in soybean hairy roots accentuated the sensitivity to exogenous ABA. Furthermore, the expression levels of ABI3, ABI5, SnRK1.1 and SnRK1.2 were differentially regulated by GmSUMO2 in transgenic soybean hairy roots. Overall, these results provided a preliminary understanding of molecular characterization, expression and function of GmSUMO2 in soybean.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (31971832) and Natural Science Foundation of Heilongjiang Province of China (YQ2019C006).

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Authors and Affiliations

  1. College of Life Science, Northeast Agricultural University, Harbin, 150030, China

    Jingsong Guo, Sibo Wang, Guixin Wang, Runfeng Lu, Yuxin Wang & Wei Ji

  2. Key Laboratory of Molecular Genetics, China National Tobacco Corporation, Guizhou Institute of Tobacco Science, Guiyang, 550083, China

    Yushuang Guo

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  1. Jingsong Guo
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Contributions

JG performed the analysis and laboratory assays and wrote the manuscript. SW and GW contributed to the bioinformatics analysis and preparation of all figures. RL, YW, and YG provide help in analysis of qRT-PCR and western blot. WJ conceived and designed the experiments, facilitated the project, and assisted in manuscript preparation. All authors read and approved the final manuscript.

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Correspondence to Wei Ji.

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11033_2020_5433_MOESM1_ESM.png

Fig. S1 Schematic representation of plant expression vector pTF101-6×His-GmSUMO2 and pTF101-6×His-GmSUMO2(H94R) used for overexpression in soybean hairy roots, containing GmSUMO2 or GmSUMO2(H94R) fused with 6×His at the 5’ terminal cloned at Xba I and Sac I sites driven by CaMV35S promoter. Supplementary file1 (PNG 11 kb)

11033_2020_5433_MOESM2_ESM.png

Fig. S2 Overexpression of GmSUMO2 decreased the average areas of roots under 25 μM ABA treatment. The data represented the mean ± SD of three independent experiments. Statistical significance was determined using ANOVA (**, P<0.01). Supplementary file2 (PNG 12 kb)

Table S1 Names and sequences of the primers. Supplementary file3 (DOCX 22 kb)

Table S2 Composition of plant culture medium. Supplementary file4 (DOCX 19 kb)

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Guo, J., Wang, S., Wang, G. et al. Overexpression of GmSUMO2 gene confers increased abscisic acid sensitivity in transgenic soybean hairy roots. Mol Biol Rep 47, 3475–3484 (2020). https://doi.org/10.1007/s11033-020-05433-3

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