Skip to main content

Isolation and Functional Analysis of Effector Proteins of Magnaporthe oryzae

  • Protocol
  • First Online:
Magnaporthe oryzae

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2356))

Abstract

In nature, plants have evolved a myriad of preformed and induced defenses to protect themselves from microbes. Upon microbial infection, the recognition of the microbe-associated molecular patterns (MAMPs) by the pattern recognition receptors (PRRs) triggers the first stage of defense response (Dodds and Rathjen, Nat Rev Genet 11:539–548, 2010). However, in order to develop microbial delivery, effectors target PRRs for deregulating immune responses and facilitating host colonization (Thomma et al., Plant Cell 23:4–15, 2011). Here, we contribute a protocol for the screening system of Magnaporthe oryzae effectors and construct a fluorescent system to trace secretory proteins in the sheath infection samples. Using the tobacco rattle virus (TRV) system, the proteins including LysM, Chitin, Cutinase, and CFEM domains were selected and divided into two kinds according to the results of cell death induced or inhibited test in Nicotiana benthamiana. Then, candidate effectors can be deleted or overexpressed in M. oryzae. The barley or rice infection with M. oryzae, rice leaf sheath inoculation, and subcellular localization during infection can be performed to explore the functions of these effectors.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Dodds PN, Rathjen JP (2010) Plant immunity: towards an integrated view of plant-pathogen interactions. Nat Rev Genet 11:539–548

    Article  CAS  Google Scholar 

  2. Thomma BP, Nürnberger T, Joosten MH (2011) Of PAMPs and effectors: the blurred PTI-ETI dichotomy. Plant Cell 23:4–15

    Article  CAS  Google Scholar 

  3. Giraldo MC, Valent B (2013) Filamentous plant pathogen effectors in action. Nat Rev Microbiol 11:800–814

    Article  CAS  Google Scholar 

  4. Park CH, Chen S, Shirsekar G, Zhou B, Khang CH, Songkumarn P, Afzal AJ, Ning Y, Wang R, Bellizzi M, Valent B, Wang GL (2012) The Magnaporthe oryzae effector AvrPiz-t targets the RING E3 ubiquitin ligase APIP6 to suppress pathogen-associated molecular pattern-triggered immunity in rice. Plant Cell 24:4748–4762

    Article  CAS  Google Scholar 

  5. Park CH, Shirsekar G, Bellizzi M, Chen S, Songkumarn P, Xie X, Shi X, Ning Y, Zhou B, Suttiviriya P, Wang M, Umemura K, Wang GL (2016) The E3 Ligase APIP10 connects the effector AvrPiz-t to the NLR receptor Piz-t in rice. PLoS Pathog 12:e1005529

    Article  Google Scholar 

  6. Zhang S, Xu JR (2014) Effectors and effector delivery in Magnaporthe oryzae. PLoS Pathog 10:e1003826

    Article  Google Scholar 

  7. Giraldo MC, Dagdas YF, Gupta YK, Mentlak TA, Yi M, Martinez-Rocha AL, Saitoh H, Terauchi R, Talbot NJ, Valent B (2013) Two distinct secretion systems facilitate tissue invasion by the rice blast fungus Magnaporthe oryzae. Nat Commun 4:1996

    Article  Google Scholar 

  8. Jia Y, McAdams SA, Bryan GT, Hershey HP, Valent B (2014) Direct interaction of resistance gene and avirulence gene products confers rice blast resistance. EMBO J 19:4004–4014

    Article  Google Scholar 

  9. Kang S, Sweigard JA, Valent B (1995) The PWL host specificity gene family in the blast fungus Magnaporthe grisea. Mol Plant-Microbe Interact 8:939–948

    Article  CAS  Google Scholar 

  10. Mosquera G, Giraldo MC, Khang CH, Coughlan S, Valent B (2009) Interaction transcriptome analysis identifies Magnaporthe oryzae BAS1-4 as biotrophy-associated secreted proteins in rice blast disease. Plant Cell 21:1273–1290

    Article  CAS  Google Scholar 

  11. Peyyala R, Farman ML (2006) Magnaporthe oryzae isolates causing gray leaf spot of perennial ryegrass possess a functional copy of the AVR1-CO39 avirulence gene. Mol Plant Pathol 7:157–165

    Article  CAS  Google Scholar 

  12. Mentlak TA, Kombrink A, Shinya T, Ryder LS, Otomo I, Saitoh H, Terauchi R, Nishizawa Y, Shibuya N, Thomma BP, Talbot NJ (2012) Effector-mediated suppression of chitin-triggered immunity by Magnaporthe oryzae is necessary for rice blast disease. Plant Cell 24:322–335

    Article  CAS  Google Scholar 

  13. Antoniw, J.F. and White, R.F. (1986) Changes with time in the distribution of virus and PR protein around single local lesions of TMV infected tobacco. Plant Mol Biol 6:145–149

    Google Scholar 

  14. Li H, Lu J, Liu X, Zhang L, Lin F (2012) Vectors building and usage for gene knockout, protein expression and fluorescent fusion protein in the rice blast fungus. J Agric Biotechnol 20:94–104

    Google Scholar 

  15. Lacomme C (1989) Bax-induced cell death in tobacco is similar to the hypersensitive response. Proc Natl Acad Sci U S A 96:7956–7961

    Article  Google Scholar 

  16. Sweigard JA, Carroll AM, Kang S, Farrall L, Valent B (1995) Identification, cloning, and characterization of PWL2, a gene for host species specificity in the rice blast fungus. Plant Cell 7:1221–1233

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Xiao-Hong Liu .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Dai, MD., Li, Y., Sun, LX., Lin, FC., Liu, XH. (2021). Isolation and Functional Analysis of Effector Proteins of Magnaporthe oryzae . In: Jacob, S. (eds) Magnaporthe oryzae. Methods in Molecular Biology, vol 2356. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1613-0_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1613-0_16

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1612-3

  • Online ISBN: 978-1-0716-1613-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics