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Cloning and Characterization of a GmSAGT1 Gene Whose Transcriptional Expression Is Associated with the Level of Resistance to Soybean Downy Mildew

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Abstract

We have cloned a GmSAGT1 gene coding for a bi-functional enzyme of serine/alanine glyoxylate aminotransferase from a soybean variety Zaofeng5 resistant to soybean downy mildew (SDM). The GmSAGT1 expressed in Escherichia coli exhibited serine glyoxylate aminotransferase (SGT) and alanine glyoxylate aminotransferase (AGT) activity that produced 20-fold more glycine than that of the control. The amount of hydrogen peroxide (H2O2) dramatically increased in E. coli expressing GmSAGT1, and bacterial growth was severely inhibited. Real-time polymerase chain reaction (RT-PCR) showed that GmSAGT1 could be detected in Zaofeng5 prior to salicylic acid (SA) treatment and increased by fivefold after SA induction. In contrast, it was not detectable in susceptible variety Heinong10 prior to and only slightly increased after SA induction. Fifteen soybean varieties belonging to highly resistant (HR), medium resistant (MR), resistant (R), susceptible (S), and highly susceptible (HS) category were used for real-time quantitative RT-PCR (qRT-PCR) analysis, which showed the average expression level of GmSAGT1 in HR group was 13 times of that in HS group. Northern blot analysis indicated that the expression level of GmSAGT1 in HS, S, MR, and R group was 5, 22, 39, and 65 % of that in HR, respectively. Transgenic tobacco expressing GmSAGT1 increased resistance to a fungal pathogen Rhizoctonia solani. Our results have demonstrated that higher expression of GmSAGT1 gene is closely correlated to increased SDM resistance.

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Abbreviations

AGT:

Alanine glyoxylate aminotransferase

DI:

Disease index

EST:

Expressed sequence tag

GOX:

Glycolate oxidase

H2O2 :

Hydrogen peroxide

HR:

Highly resistant

HS:

Highly susceptible

IPTG:

Isopropyl β-d-thiogalactoside

MDM:

Melon downy mildew

MR:

Medium resistant

MS:

Murashige and Skoog medium

PR proteins:

Pathogenic-related proteins

R gene:

Resistance gene

R:

Resistant

ROS:

Reactive oxygen species

S:

Susceptible

SA:

Salicylic acid

SAR:

Systemic acquired resistance

SDM:

Soybean downy mildew

SGT:

Serine glyoxylate aminotransferase

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Acknowledgments

This work was financially supported by the National High-tech R&D Program (2011AA100606) and “The development of new varieties of animals and crops” under the National High Priority R&D Program (2013ZX08009-003-005). We sincerely thank Prof. Lijuan Qiu in the National Germplasm Bank, China for kindly providing us seeds of soybean varieties.

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Authors and Affiliations

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Wenli Sun, Chao Yin, Mei Li, Huaguang Zhang, Shuangfeng Shi, Shirong Jia, Zhixing Wang & Yuhui Liu

  2. College of Plant Protection, Shenyang Agricultural University, Shenyang, 110161, China

    Wenli Sun, Yuanhua Wu & Qiao Wu

Authors
  1. Wenli Sun
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  2. Chao Yin
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  3. Mei Li
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  4. Huaguang Zhang
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  5. Shuangfeng Shi
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  6. Yuanhua Wu
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  7. Qiao Wu
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  8. Shirong Jia
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  9. Zhixing Wang
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  10. Yuhui Liu
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Corresponding authors

Correspondence to Zhixing Wang or Yuhui Liu.

Additional information

Sun WL and Yin C contributed equally to this work.

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Sun, W., Yin, C., Li, M. et al. Cloning and Characterization of a GmSAGT1 Gene Whose Transcriptional Expression Is Associated with the Level of Resistance to Soybean Downy Mildew. Plant Mol Biol Rep 33, 1232–1244 (2015). https://doi.org/10.1007/s11105-014-0818-x

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