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Protoplast Cell Death Assay to Study Magnaporthe oryzae AVR Gene Function in Rice

  • Hiroyuki Kanzaki
  • Kentaro Yoshida
  • Hiromasa Saitoh
  • Muluneh Tamiru
  • Ryohei TerauchiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1127)

Abstract

We describe a protocol for transient gene expression in rice protoplasts and its application to the study of Magnaporthe oryzae avirulence (AVR) gene function. In this assay the gene encoding the firefly luciferase protein is transfected into rice protoplasts by electroporation together with the candidate AVR genes. The luminescence can then be used to assess the viability of rice protoplasts. The hypersensitive response (HR) caused by the interaction between M. oryzae AVR and rice R genes can subsequently be monitored by recording the decrease in luminescence from the transfected cells.

Key words

Protoplast Electroporation Hypersensitive response (HR) Luciferase Rice blast Rice Monocotyledons 

Notes

Acknowledgements

This work was supported by the Ministry of Agriculture, Forestry, and Fisheries of Japan (Genomics for Agricultural Innovation PMI-0010) and the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN), Japan, and Grant-in-aid for Scientific Research from the Ministry of Education, Cultures, Sports and Technology, Japan to HS and RT (Grant-in-Aid for Scientific Research on Innovative Areas 23113009).

References

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Hiroyuki Kanzaki
    • 1
  • Kentaro Yoshida
    • 1
  • Hiromasa Saitoh
    • 1
  • Muluneh Tamiru
    • 1
  • Ryohei Terauchi
    • 1
    Email author
  1. 1.Iwate Biotechnology Research CenterKitakamiJapan

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