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A soybean cyst nematode resistance gene points to a new mechanism of plant resistance to pathogens

Nature volume 492, pages 256–260 (2012)Cite this article

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Abstract

Soybean (Glycine max (L.) Merr.) is an important crop that provides a sustainable source of protein and oil worldwide. Soybean cyst nematode (Heterodera glycines Ichinohe) is a microscopic roundworm that feeds on the roots of soybean and is a major constraint to soybean production. This nematode causes more than US$1 billion in yield losses annually in the United States alone1, making it the most economically important pathogen on soybean. Although planting of resistant cultivars forms the core management strategy for this pathogen, nothing is known about the nature of resistance. Moreover, the increase in virulent populations of this parasite on most known resistance sources necessitates the development of novel approaches for control. Here we report the map-based cloning of a gene at the Rhg4 (for resistance to Heterodera glycines 4) locus, a major quantitative trait locus contributing to resistance to this pathogen. Mutation analysis, gene silencing and transgenic complementation confirm that the gene confers resistance. The gene encodes a serine hydroxymethyltransferase, an enzyme that is ubiquitous in nature and structurally conserved across kingdoms. The enzyme is responsible for interconversion of serine and glycine and is essential for cellular one-carbon metabolism. Alleles of Rhg4 conferring resistance or susceptibility differ by two genetic polymorphisms that alter a key regulatory property of the enzyme. Our discovery reveals an unprecedented plant resistance mechanism against a pathogen. The mechanistic knowledge of the resistance gene can be readily exploited to improve nematode resistance of soybean, an increasingly important global crop.

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Figure 1: Rhg4 positional cloning and functional validation of SHMT by TILLING.
Figure 2: Haplotypes identified at SHMT in 28 soybean lines.
Figure 3: Functional validation of SHMT by VIGS, RNAi and complementation.
Figure 4: Modelled structure and biochemical analysis of SHMT.

Accession codes

Primary accessions

GenBank/EMBL/DDBJ

Data deposits

All sequences have been deposited in GenBank/EMBL/DDBJ under the following accession numbers: Forrest SHMT full-length genomic DNA, JQ714083; Essex SHMT full-length genomic DNA, JQ714084; Forrest SHMT cDNA sequence, JQ714080; Essex SHMT cDNA sequence, JQ714079; Forrest SHMT TILLING mutant F6266 sequence, JQ714081; Forrest SHMT TILLING mutant F6756 sequence, JQ714082; Forrest SUB1 cDNA sequence, JQ762395; Essex SUB1 cDNA sequence, JQ62396; Forrest SUB1 promoter sequence, JQ762397; Essex SUB1 promoter sequence, JQ904711.

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Acknowledgements

We thank X. Yang for technical assistance with soybean hairy root propagation and M. Kroll for proofreading the manuscript. We thank R. Hussey, W. Gassmann, P. Gresshoff, A. Bendahmane, D. Xu and B. McClure for critical reading of the manuscript. We thank K. Sharma and S. Puthur for use of the HPLC facility and technical help. This work was supported by the Illinois-Missouri Biotechnology Alliance (project 2005-3 to M.G.M. and K.M.), United Soybean Board (project 0253 to K.M., S.C. and M.G.M.; project 3253 to K.M. and S.C.; project 2268 to K.M. and M.G.M; project 1251 to J.H. and S.A.W.), USDA-NIFA (grant 2006-35300-17195 to K.M.), the National Science Foundation Plant Genome Research Program (grant 0820642 to S.A.W., J.H., M.G.M. and T.J.B.), the National Science Foundation CAREER Program (DBI-0845196 to D.K.), Missouri Soybean Merchandising Council (project 258 to M.G.M.), Illinois Soybean Association, North Central Soybean Research Program, Iowa Soybean Association, and a Department of Education Graduate Assistance in Areas of National Need (GAANN) Fellowship (to S.D.W.).

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Author notes

  1. Shiming Liu and Pramod K. Kandoth: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Plant, Soil and Agricultural Systems, Southern Illinois University, Carbondale, 62901, Illinois, USA

    Shiming Liu, Aziz Jamai, Tarik El-Mellouki & Khalid Meksem

  2. Division of Plant Sciences, Christopher S. Bond Life Sciences Center and Interdisciplinary Plant Group, University of Missouri, Columbia, 65211, Missouri, USA

    Pramod K. Kandoth, Greg Yeckel, Robert Heinz, John Alden & Melissa G. Mitchum

  3. Department of Computer Science, Christopher S. Bond Life Sciences Center and Informatics Institute, University of Missouri, Columbia, 65211, Missouri, USA

    Samantha D. Warren & Dmitry Korkin

  4. Department of Plant Pathology and Microbiology, Iowa State University, Ames, 50011, Iowa, USA

    Chunling Yang, Parijat S. Juvale, John Hill, Thomas J. Baum & Steven A. Whitham

  5. Department of Agronomy, Iowa State University, Ames, 50011, Iowa, USA

    Silvia Cianzio

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Contributions

S.L. and P.K.K. contributed equally as first authors. K.M. and M.G.M. contributed equally as senior authors. S.L. carried out mapping and haplotyping studies. S.L., A.J. and T.E.-M. developed the TILLING population used in this study. S.C. developed two of the RIL populations used in this study. S.L. identified the TILLING mutants and conducted the haplotyping analysis. G.Y. and R.H. carried out SCN phenotyping of all RILs, TILLING mutants, and soybean lines. G.Y. collected leaf tissues for RIL and TILLING studies. P.K.K. collected leaf tissues haplotyping analyses. P.K.K. and S.L. confirmed the mutations, cloned the genes, and conducted sequence analyses. P.K.K., C.Y., R.H. and P.S.J. developed and carried out VIGS assays. P.K.K. conducted RNAi experiments. P.K.K. carried out the promoter analysis. P.K.K. and J.A. carried out the complementation analysis. S.D.W. and D.K. performed the computational analysis. P.K.K. performed the biochemical studies. J.H., S.A.W. and T.J.B. provided materials and advice for VIGS analysis. K.M., M.G.M. and D.K. designed the research, and together with S.L., P.K.K. and S.D.W. wrote the manuscript. All authors reviewed and commented on the manuscript.

Corresponding authors

Correspondence to Melissa G. Mitchum or Khalid Meksem.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1–4. (PDF 900 kb)

Supplementary Table 1

This file contains the haplotyping of soybean Pls at the Rhg4 and rhg1 locl. (XLSX 17 kb)

Supplementary Table 2

This file contains details of the primers used in this study. (XLSX 15 kb)

Supplementary Data 1

This file contains data relevant to this study. (TXT 569 kb)

A homology model of the Essex GmSHMT homodimer showing functional annotation of the Forrest polymorphisms.

Shown are each of the GmSHMT monomers with the ligand binding sites and Forrest mutations mapped onto their surface (we note that the contributing parts of the ligand binding sites are not identical between the two monomers). When two ligand binding sites overlap only the colored surface for one of the sites is shown. (MOV 27613 kb)

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Liu, S., Kandoth, P., Warren, S. et al. A soybean cyst nematode resistance gene points to a new mechanism of plant resistance to pathogens. Nature 492, 256–260 (2012). https://doi.org/10.1038/nature11651

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