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Functional complementation of rice blast resistance gene Pi-k h (Pi54) conferring resistance to diverse strains of Magnaporthe oryzae

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Abstract

Blast disease of rice, caused by Magnaporthe oryzae is an explosive disease that can spread rapidly in conducive conditions. R-gene mediated resistance offers an environmentally sustainable solution for management of this important disease of rice. We have earlier identified a unique R-gene of rice, on chromosome 11 of Oryza sativa ssp. indica cultivar Tetep. In this study we report functional validation of the Pi-k h (Pi54) gene using complementation assay. The blast resistance candidate gene Pi-k h (Pi54) was cloned into a plant transformation vector and the construct was used to transform a japonica cultivar of rice Taipei 309, which is susceptible to M. oryzae. Transgenic lines containing Pi-k h (Pi54) gene were found to confer high degree of resistance to diverse isolates of M. oryzae. The callose deposition was analyzed and compared between the transgenic and non-transgenic rice plants and widespread deposition was observed at the infection sites in plants showing incompatible interaction. Successful complementation of Pi-k h (Pi54) gene confirmed that the gene is responsible for resistance to M. oryzae in transgenic lines developed during this study. Expression analysis of the gene in resistant plants revealed that the gene is pathogen inducible in nature and is not expressed constitutively. Detection of callose deposition in resistant plants containing Pi-k h (Pi54) gene implicates its involvement in the initiation of defense response cascade.

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Abbreviations

Avr:

Avirulence

CTAB:

Cetyl trimethyl ammonium bromide

ETI:

Effector triggered immunity

hpi:

Hours post inoculation

RT-PCR:

Real time PCR

NBS-LRR:

Nucleotide binding site-leucine rich repeat

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Acknowledgements

Financial assistance received from the Department of Biotechnology, Govt. of India by TRS (Project code: BT/AB/FG-2(PH-II)/2009) is gratefully acknowledged. We also thank Dr. K.V. Prabhu, National Phytotron Facility, Indian Agricultural Research Institute for providing basic facilities for transgenic research. Authors are also thankful to Dr. Gautam Chawla, Department of Nematology, IARI for valuable suggestions on histo-chemical experiments.

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  1. National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi, 110012, India

    Amit Kumar Rai, Satya Pal Kumar, Santosh Kumar Gupta, Naveen Gautam, Nagendera Kumar Singh & Tilak Raj Sharma

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  1. Amit Kumar Rai
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  2. Satya Pal Kumar
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  3. Santosh Kumar Gupta
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  5. Nagendera Kumar Singh
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Correspondence to Tilak Raj Sharma.

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Rai, A.K., Kumar, S.P., Gupta, S.K. et al. Functional complementation of rice blast resistance gene Pi-k h (Pi54) conferring resistance to diverse strains of Magnaporthe oryzae . J. Plant Biochem. Biotechnol. 20, 55–65 (2011). https://doi.org/10.1007/s13562-010-0026-1

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