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An agglutinating chitinase with two chitin-binding domains confers fungal protection in transgenic potato

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Abstract

Brassica juncea BjCHI1 is a unique chitinase with two chitin-binding domains. Here, we show that, unlike other chitinases, potato-expressed BjCHI1 shows hemagglutination ability. BjCHI1 expression in B. juncea seedlings is induced by Rhizoctonia solani infection, suggesting its protective role against this fungus. To verify this, transgenic potato (Solanum tuberosum L. cv. Desiree) plants expressing BjCHI1 generated by Agrobacterium-mediated transformation were challenged with R. solani. We also transformed potato with a cDNA encoding Hevea brasiliensis β-1,3-glucanase, designated HbGLU, and a pBI121-derivative that contains cDNAs encoding both BjCHI1 and HbGLU. In vitro fungal bioassays using Trichoderma viride showed that extracts from transgenic potato lines co-expressing BjCHI1 and HbGLU inhibited fungal growth better than extracts from transgenic potato expressing either BjCHI1 or HbGLU, suggesting a synergistic effect. Consistently, in vivo fungal bioassays with soil-borne R. solani on young transgenic potato plants indicated that the co-expressing plants showed healthier root development than untransformed plants or those that expressed either BjCHI1 or HbGLU. Light microscopy and transmission electron microscopy revealed abundant intact R. solani hyphae and monilioid cells in untransformed roots and disintegrated fungus in the BjCHI1-expressing and the BjCHI1 and HbGLU co-expressing plants. Observations of collapsed epidermal cells in the co-expressing potato roots suggest that these proteins effectively degrade the fungal cell wall, producing elicitors that initiate other defense responses causing epidermal cell collapse that ultimately restricts further fungal penetration.

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Abbreviations

BjCHI1 :

Brassica juncea chitinase 1

CaMV :

Cauliflower Mosaic Virus

HbGLU :

Hevea brasiliensis β-1,3-glucanase

MS :

Murashige and Skoog

PVDF :

Polyvinylidene difluoride

TEM :

Transmission electron microscopy

UDA :

Urtica dioica agglutinin

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Acknowledgments

We thank Prof. Z.P. Lin (Peking University) for wild-type potato Desiree, Prof. E.C. Pua (National University of Singapore) for B. juncea seeds, X.Z. Ouyang for technical assistance in microscopy, A. Mak for N-terminal sequencing, and W. Ho and P. Tam for helpful discussions. This project was supported by the Department of Botany (University of Hong Kong), the Biotechnology Research Institute (Hong Kong University of Science and Technology) and the Research Grants Council of Hong Kong (Plant and Fungal Biotechnology Area of Excellence). K.J.Z. (Postdoctoral Fellowship) and K.L.F. (Postgraduate Studentship) were supported by the University of Hong Kong.

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  1. Kai-Jun Zhao

    Present address: Institute of Crop Breeding and Cultivation, Chinese Academy of Agricultural Sciences, Beijing, China

  2. Zhu-Mei He

    Present address: Biotechnology Research Centre, Zhongshan University, Guangzhou, China

Authors and Affiliations

  1. Department of Botany, The University of Hong Kong, Pokfulam Road, Hong Kong, China

    Mee-Len Chye, Kai-Jun Zhao, Zhu-Mei He, Sathishkumar Ramalingam & King-Leung Fung

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  1. Mee-Len Chye
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  2. Kai-Jun Zhao
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Correspondence to Mee-Len Chye.

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Chye, ML., Zhao, KJ., He, ZM. et al. An agglutinating chitinase with two chitin-binding domains confers fungal protection in transgenic potato. Planta 220, 717–730 (2005). https://doi.org/10.1007/s00425-004-1391-6

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