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Expression of a bacterial chitosanase in rice plants improves disease resistance to the rice blast fungus Magnaporthe oryzae

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Abstract

Plant fungal pathogens change their cell wall components during the infection process to avoid degradation by host lytic enzymes, and conversion of the cell wall chitin to chitosan is likely to be one infection strategy of pathogens. Thus, introduction of chitosan-degradation activity into plants is expected to improve fungal disease resistance. Chitosanase has been found in bacteria and fungi, but not in higher plants. Here, we demonstrate that chitosanase, Cho1, from Bacillus circulans MH-K1 has antifungal activity against the rice blast fungus Magnaporthe oryzae. Introduction of the cho1 gene conferred chitosanase activity to rice cells. Transgenic rice plants expressing Cho1 designed to be localized in the apoplast showed increased resistance to M. oryzae accompanied by increased generation of hydrogen peroxide in the infected epidermal cells. These results strongly suggest that chitosan exists in the enzyme-accessible surface of M. oryzae during the infection process and that the enhancement of disease resistance is attributable to the antifungal activity of the secreted Cho1 and to increased elicitation of the host defense response.

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Abbreviations

BSA:

Bovine serum albumin

GlcN:

d-Glucosamine

GlcNAc:

N-Acetyl-d-glucosamine

PCR:

Polymerase chain reaction

RT-PCR:

Reverse transcription polymerase chain reaction

SDS-PAGE:

SDS-polyacrylamide gel electrophoresis

CBB:

Coomassie brilliant blue

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

TLC:

Thin-layer chromatography

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Acknowledgments

We thank S. Omiya, A. Kikuchi, A. Kimura, M. Tomita, and R. Tokishige of Chiba University for providing information on chitosanase, recombinant Cho1 protein, and technical advice for detecting chitosanase activity. We also thank M. Nishimura and S. Tanabe of the National Institute of Agrobiological Sciences for fruitful discussions on the alteration of cell wall composition of M. oryzae and for providing technical advice regarding M. oryzae inoculation, respectively; and E. Nakajima, H. Kurano, and K. Iwasaki for producing rice transformants. This work was supported by the Program for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry in Japan.

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Authors and Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8572, Japan

    Yusuke Kouzai & Eiichi Minami

  2. Genetically Modified Organism Research Center, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602, Japan

    Yusuke Kouzai, Susumu Mochizuki, Eiichi Minami & Yoko Nishizawa

  3. Department of Materials and Life Science, Faculty of Science and Technology, Shizuoka Institute of Science and Technology, Fukuroi, Shizuoka, 437-8555, Japan

    Akihiro Saito

  4. Graduate School of Science and Technology, Chiba University, Chiba, 263-0022, Japan

    Akikazu Ando

Authors
  1. Yusuke Kouzai
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  2. Susumu Mochizuki
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  3. Akihiro Saito
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  4. Akikazu Ando
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  5. Eiichi Minami
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  6. Yoko Nishizawa
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Corresponding author

Correspondence to Yoko Nishizawa.

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Communicated by K. Toriyama.

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Kouzai, Y., Mochizuki, S., Saito, A. et al. Expression of a bacterial chitosanase in rice plants improves disease resistance to the rice blast fungus Magnaporthe oryzae . Plant Cell Rep 31, 629–636 (2012). https://doi.org/10.1007/s00299-011-1179-7

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  • DOI: https://doi.org/10.1007/s00299-011-1179-7

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