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Agrobacterium-mediated transformation as a useful tool for the molecular genetic study of the phytopathogen Curvularia lunata

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Abstract

In order to explore the molecular mechanisms of virulence and genetic variance of Curvularia lunata in maize, an ATMT (Agrobacterium tumefaciens-mediated transformation) system was established in order to create a wide range of insertional transformants of C. lunata. Our results showed that the germinating conidia were the ideal starting material for transformation. Based on our optimised transformation condition, the transformation efficiency of C. lunata with T-DNA was improved greatly, and the average transformation frequency was as high as 84 ± 5 transformants per 1 × 106 germlings. Southern blotting results of 39 randomly-selected transformants showed a unique hybridisation pattern and predominant single-copy insertions. An ATMT library containing approximate 3000 transformants was generated, and four types of transformants were screened in terms of growth rate, sporulation, mycelial pigmentation, and toxin production in vitro. This library will be used to identify genes involved in the virulence of the pathogen.

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Acknowledgements

This work is supported by the National Basic Research Programme of China (973 Programme, 2006 CB101901) and the National Science & Technology Supporting Programme of China (2006BAD08A06).

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Authors and Affiliations

  1. Department of Resources and Environmental Sciences, School of Agriculture and Biology, Shanghai Jiaotong University, 800 Dongchuan Road, Shanghai, 200240, People’s Republic of China

    Tong Liu, Lixing Liu, Xue Jiang, Kehe Fu, Feihong Zhou & Jie Chen

  2. Department of Biology and Chemical Engineering, Jindezhen Comprehensive College, Jiangxi, 333000, People’s Republic of China

    Jumei Hou

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  1. Tong Liu
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  2. Lixing Liu
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  3. Xue Jiang
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  5. Kehe Fu
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  7. Jie Chen
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Correspondence to Jie Chen.

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Liu, T., Liu, L., Jiang, X. et al. Agrobacterium-mediated transformation as a useful tool for the molecular genetic study of the phytopathogen Curvularia lunata. Eur J Plant Pathol 126, 363–371 (2010). https://doi.org/10.1007/s10658-009-9541-0

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  • DOI: https://doi.org/10.1007/s10658-009-9541-0

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