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The use of GFP-transformed isolates to study infection of banana with Fusarium oxysporum f. sp. cubense race 4

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Abstract

Fusarium oxysporum f. sp. cubense (Foc) is the causal pathogen of Fusarium wilt of banana. To understand infection of banana roots by Foc race 4, we developed a green fluorescent protein (GFP)-tagged transformant and studied pathogenesis using fluorescence microscopy and confocal laser scanning microscopy. The transformation was efficient, and GFP expression was stable for at least six subcultures with fluorescence clearly visible in both hyphae and spores. The transformed Foc isolate also retained its pathogenicity and growth pattern, which was similar to that of the wild type. The study showed that: (i) Foc race 4 was capable of invading the epidermal cells of banana roots directly; (ii) potential invasion sites include epidermal cells of root caps and elongation zone, and natural wounds in the lateral root base; (iii) in banana roots, fungal hyphae were able to penetrate cell walls directly to grow inside and outside cells; and (iv) fungal spores were produced in the root system and rhizome. To better understand the interaction between Foc race 4 and bananas, nine banana cultivars were inoculated with the GFP-transformed pathogen. Root exudates from these cultivars were collected and their effect on conidia of the GFP-tagged Foc race 4 was determined. Our results showed that roots of the Foc race 4-susceptible banana plants were well colonized with the pathogen, but not those of the Foc race 4-resistant cultivars. Root exudates from highly resistant cultivars inhibited the germination and growth of the Fusarium wilt pathogen; those of moderately resistant cultivars reduced spore germination and hyphal growth, whereas the susceptible cultivars did not affect fungal germination and growth. The results of this work demonstrated that GFP-tagged Foc race 4 isolates are an effective tool to study plant–fungus interactions that could potentially be used for evaluating resistance in banana to Foc race 4 by means of root colonization studies. Banana root exudates could potentially also be used to identify cultivars in the Chinese Banana Germplasm Collection with resistance to the Fusarium wilt pathogen.

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Abbreviations

Foc :

Fusarium oxysporum f. sp. cubense

GFP:

Green fluorescent protein

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Acknowledgments

This work was supported by the National Natural Science Fund (30971991), Guangdong Natural Science Fund (10151064001000007), the Commonwealth Industry (Agriculture) Specific Fund (200903049–10), and the International Collaborative Project between China and South Africa governments (2010DFA32470). The authors would also like to thank Dr Xin Zhang of the Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences (CATAS), for kindly helping to transform the plasmid pCT74 into the Foc race 4 isolates, and Dr. Altus Viljoen for valuable comments and suggestions during the preparation of the manuscript.

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Authors and Affiliations

  1. Institution of Fruit Tree Research, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, Guangdong Province, China

    Chunyu Li, Qingming Sun, Qian Ye, Ganjun Yi & Bingzhi Huang

  2. Fujian-Taiwan Horticulture Research Center, Fujian Academy of Agricultural Sciences, Zhangzhou, 363005, Fujian Province, China

    Shi Chen

  3. College of Life Science, South China Agricultural University, Guangzhou, 510640, Guangdong Province, China

    Cunwu Zuo

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  1. Chunyu Li
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  2. Shi Chen
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  3. Cunwu Zuo
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  4. Qingming Sun
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  5. Qian Ye
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  6. Ganjun Yi
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  7. Bingzhi Huang
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Correspondence to Ganjun Yi.

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Chunyu Li, Shi Chen and Cunwu Zuo contributed to the work equally.

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Li, C., Chen, S., Zuo, C. et al. The use of GFP-transformed isolates to study infection of banana with Fusarium oxysporum f. sp. cubense race 4. Eur J Plant Pathol 131, 327–340 (2011). https://doi.org/10.1007/s10658-011-9811-5

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