Molecular Breeding

, 37:151 | Cite as

Development and validation of breeder-friendly KASPar markers for er1, a powdery mildew resistance gene in pea (Pisum sativum L.)

  • Yu Ma
  • Clarice J. Coyne
  • Dorrie Main
  • Stefano Pavan
  • Suli Sun
  • Zhendong Zhu
  • Xuxiao Zong
  • José Leitão
  • Rebecca J. McGeeEmail author
Short Communication


Powdery mildew of pea is caused by Erysiphe pisi DC and is a serious threat to pea (Pisum sativum L.) production throughout much of the world. Development and utilization of genetic resistance to powdery mildew is considered an effective and sustainable strategy to manage this disease. One gene, er1, conferring powdery mildew resistance, was previously cloned and sequenced, and the functional markers for each resistance allele were reported. Allele-specific DNA markers are efficient and powerful tools to facilitate crop improvement and new cultivar development in breeding programs. However, extensive application of these markers is limited by gel-associated obstacles. In this study, eight breeder-friendly kompetitive allele-specific PCR (KASPar) markers were developed to overcome the problems of gel-based markers and increase the efficiency of genotypic screening. In order to identify additional pea germplasm with powdery mildew resistance, these KASPar markers were deployed and used to genotype a pea collection derived from the USDA pea single-plant (PSP) collection. Simultaneously, a phenotypic screening and a genotypic validation using the corresponding gel-based functional markers were conducted on the PSP collection. One pea accession, PI 142775, was identified by both phenotyping and genotyping to carry the allele er1-1 for powdery mildew resistance, indicating that the KASPar assay is an efficient and robust tool for breeding for powdery mildew resistance.


er1 KASPar assay Marker-assisted breeding Powdery mildew Pisum sativum 



The authors thank Drs. Shimna Sudheesh, Sukhjiwan Kaur, and John W. Foster from the Centre for AgriBioscience, Bundoora, Australia for their contributions in this research.

Funding information

The authors gratefully acknowledge the funding from the Washington State Department of Agriculture Specialty Crop Block Grant K1273 and USA Dry Pea and Lentil Council Research grant.

Supplementary material

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  1. Andersen JR, Lubberstedt T (2003) Functional markers in plants. Trends Plant Sci 8(11):554–560CrossRefPubMedGoogle Scholar
  2. Attanayake R, Glawe D, Dugan F, Chen W (2009) Erysiphe trifolii causing powdery mildew of lentil (Lens culinaris). Plant Dis 93:797–803CrossRefGoogle Scholar
  3. Attanayake R, Glawe D, McPhee K, Dugan F, Chen W (2010) Erysiphe trifolii—a newly recognized powdery mildew pathogen of pea. Plant Pathol 59(4):712–720CrossRefGoogle Scholar
  4. Boutet G, Carvalho SA, Falque M, Peterlongo P, Lhuillier E, Bouchez O, Lavaud C, Pilet-Nayel ML, Rivière N, Baranger A (2016) SNP discovery and genetic mapping using genotyping by sequencing of whole genome genomic DNA from a pea RIL population. BMC Genomics 17(1):121CrossRefPubMedPubMedCentralGoogle Scholar
  5. Dwight ZL, Palais R, Wittwer CT (2012) uAnalyze: web-based high-resolution DNA melting analysis with comparison to thermodynamic predictions. IEEE/ACM Trans Comput Biol Bioinf 9(6):1805–1811CrossRefGoogle Scholar
  6. Falloon R, Viljanen-Rollinson S, Coles G, Poff J (1995) Disease severity keys for powdery and downy mildews of pea, and powdery scab of potato. N Z J Crop Hortic Sci 23(1):31–37CrossRefGoogle Scholar
  7. Fondevilla S, Rubiales D (2012) Powdery mildew control in pea. A review. Agron Sustain Dev 32(2):401–409CrossRefGoogle Scholar
  8. Fondevilla S, Torres AM, Moreno MT, Rubiales D (2007) Identification of a new gene for resistance to powdery mildew in Pisum fulvum, a wild relative of pea. Breed Sci 57(2):181–184CrossRefGoogle Scholar
  9. Gupta P, Roy J, Prasad M (2001) Single nucleotide polymorphisms: a new paradigm for molecular marker technology and DNA polymorphism detection with emphasis on their use in plants. Curr Sci 80(4):524–535Google Scholar
  10. Harland S (1948) Inheritance of immunity to mildew in Peruvian forms of Pisum sativum. Heredity 2(Pt 2):263CrossRefPubMedGoogle Scholar
  11. Heringa R, van Norel A, Tazelaar M (1969) Resistance to powdery mildew (Erisyphe polygoni D.C.) in peas (Pisum sativum L.) Euphytica 18(2):163–169CrossRefGoogle Scholar
  12. Holdsworth WL, Mazourek M (2015) Development of user-friendly markers for the pvr1 and Bs3 disease resistance genes in pepper. Mol Breed 35(1):1–5CrossRefGoogle Scholar
  13. Humphry M, Reinstaedler A, Ivanov S, Bisseling T, Panstruga R (2011) Durable broad-spectrum powdery mildew resistance in pea er1 plants is conferred by natural loss-of-function mutations in PsMLO1. Mol Plant Pathol 12(9):866–878CrossRefPubMedGoogle Scholar
  14. Neelam K, Brown-Guedira G, Huang L (2013) Development and validation of a breeder-friendly KASPar marker for wheat leaf rust resistance locus Lr21. Mol Breed 31(1):233–237CrossRefGoogle Scholar
  15. Pavan S, Schiavulli A, Appiano M, Marcotrigiano AR, Cillo F, Visser RG, Bai Y, Lotti C, Ricciardi L (2011) Pea powdery mildew er1 resistance is associated to loss-of-function mutations at a MLO homologous locus. Theor Appl Genet 123(8):1425–1431CrossRefPubMedGoogle Scholar
  16. Pavan S, Schiavulli A, Appiano M, Miacola C, Visser RG, Bai Y, Lotti C, Ricciardi L (2013) Identification of a complete set of functional markers for the selection of er1 powdery mildew resistance in Pisum sativum L. Mol Breed 31(1):247–253CrossRefGoogle Scholar
  17. Rosas JE, Bonnecarrère V, Pérez de Vida F (2014) One-step, codominant detection of imidazolinone resistance mutations in weedy rice (Oryza sativa L.) Electron J Biotechnol 17(2):95–101CrossRefGoogle Scholar
  18. Rosso ML, Burleson SA, Maupin LM, Rainey KM (2011) Development of breeder-friendly markers for selection of MIPS1 mutations in soybean. Mol Breed 28(1):127–132CrossRefGoogle Scholar
  19. Santo T, Rashkova M, Alabaça C, Leitão J (2013) The ENU-induced powdery mildew resistant mutant pea (Pisum sativum L.) lines S (er1mut1) and F (er1mut2) harbour early stop codons in the PsMLO1 gene. Mol Breed 32(3):723–727CrossRefGoogle Scholar
  20. Semagn K, Babu R, Hearne S, Olsen M (2014) Single nucleotide polymorphism genotyping using Kompetitive Allele Specific PCR (KASP): overview of the technology and its application in crop improvement. Mol Breed 33(1):1–14CrossRefGoogle Scholar
  21. Singh B, Singh AK (2015) Marker-assisted plant breeding: principles and practices. Springer, pp 77–122Google Scholar
  22. Sudheesh S, Lombardi M, Leonforte A, Cogan NO, Materne M, Forster JW, Kaur S (2015) Consensus genetic map construction for field pea (Pisum sativum L.), trait dissection of biotic and abiotic stress tolerance and development of a diagnostic marker for the er1 powdery mildew resistance gene. Plant Mol Biol Report 33(5):1391–1403CrossRefGoogle Scholar
  23. Sun S, Deng D, Wang Z, Duan C, Wu X, Wang X, Zong X, Zhu Z (2016a) A novel er1 allele and the development and validation of its functional marker for breeding pea (Pisum sativum L.) resistance to powdery mildew. Theor Appl Genet 129(5):909–919CrossRefPubMedGoogle Scholar
  24. Sun S, Fu H, Wang Z, Duan C, Zong X, Zhu Z (2016b) Discovery of a novel er1 allele conferring powdery mildew resistance in Chinese pea (Pisum sativum L.) landraces. PLoS One 11(1):e0147624CrossRefPubMedPubMedCentralGoogle Scholar
  25. Sun S, Wang Z, Fu H, Duan C, Wang X, Zhu Z (2015) Resistance to powdery mildew in the pea cultivar Xucai 1 is conferred by the gene er1. The Crop Journal 3(6):489–499CrossRefGoogle Scholar
  26. Warkentin T, Rashid K, Xue A (1996) Fungicidal control of powdery mildew in field pea. Can J Plant Sci 76(4):933–935CrossRefGoogle Scholar
  27. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols: Guide Meth Appl 18:315–322Google Scholar

Copyright information

© This is a U.S. government work and its text is not subject to copyright protection in the United States; however, its text may be subject to foreign copyright protection 2017

Authors and Affiliations

  • Yu Ma
    • 1
  • Clarice J. Coyne
    • 2
  • Dorrie Main
    • 1
  • Stefano Pavan
    • 3
  • Suli Sun
    • 4
  • Zhendong Zhu
    • 4
  • Xuxiao Zong
    • 4
  • José Leitão
    • 5
  • Rebecca J. McGee
    • 6
    Email author
  1. 1.Department of HorticultureWashington State UniversityPullmanUSA
  2. 2.Plant Germplasm Introduction and TestingUSDA ARSPullmanUSA
  3. 3.Section of Genetics and Plant Breeding, Department of Soil, Plant and Food ScienceUniversity of BariBariItaly
  4. 4.National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop ScienceChinese Academy of Agricultural SciencesBeijingChina
  5. 5.Laboratory of Genomics and Genetic Improvement, BioFIG, FCTUniversidade do AlgarveFaroPortugal
  6. 6.Grain Legume Genetics and Physiology Research UnitUSDA ARSPullmanUSA

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