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Transgenic lines of melon (Cucumis melo L. var. makuwa cv. ‘Silver Light’) expressing antifungal protein and chitinase genes exhibit enhanced resistance to fungal pathogens

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Abstract

The oriental melon (Cucumis melo L. var. makuwa cv. ‘Silver Light’) is an important fruit crop in the tropical and subtropical regions. However, oriental melon production is severely decreased by fungal diseases. In this study, antifungal protein (AFP) and chitinase (CHI) fusion genes were introduced into oriental melons to control fungal diseases caused by Rhizoctonia solani and Fusarium oxysporum. Transformation of oriental melon (Cucumis melo L. var. makuwa cv. ‘Silver Light’) with Agrobacterium tumefaciens strain LBA4404 containing antifungal protein (AFP) and chitinase (CHI) fusion genes under the control of the cauliflower mosaic virus (CaMV) 35S promoter and neomycin phosphotransferase (nptII) gene as a selectable marker was performed. Cotyledon explants of oriental melon were inoculated by Agrobacterium suspensions with pBI121–AFP–CHI and cultured in a regeneration medium. After regeneration, genomic DNA polymerase chain reaction (PCR) was conducted to confirm the presence of putative transgenic shoots. Southern blot analysis confirmed that the AFP–CHI fusion gene was incorporated into the genomic DNA of the PCR-positive lines. RT-PCR analysis showed that the AFP–CHI fusion gene was expressed in the individual transgenic lines. Western blot analysis revealed the accumulation of CHI protein in leaves. A segregation analysis of the T1 generation confirmed the inheritance of the transgene. Our results demonstrated that the AFP–CHI fusion gene was effective in protecting the transgenic melon plants against fungal disease caused by Rhizoctonia solani and Fusarium oxysporum.

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Acknowledgments

This work was supported by the National Science Council of Taiwan, Republic of China (98-2321-B- 005-009-MY3).

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Authors and Affiliations

  1. Department of Food Science and Biotechnology, National Chung Hsing University, Taichung, Taiwan, ROC

    Ismail Bezirganoglu & Tony J. Fang

  2. Department of Entomology, National Chung Hsing University, Taichung, Taiwan, ROC

    Shaw-Yhi Hwang

  3. Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan, ROC

    Jei-Fu Shaw

  4. Department of Biological Science and Technology, I-Shou University, Kaohsiung, Taiwan, ROC

    Jei-Fu Shaw

  5. Department of Nutrition, China Medical University, Taichung, Taiwan, ROC

    Tony J. Fang

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  1. Ismail Bezirganoglu
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  2. Shaw-Yhi Hwang
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  3. Tony J. Fang
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  4. Jei-Fu Shaw
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Correspondence to Tony J. Fang or Jei-Fu Shaw.

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Bezirganoglu, I., Hwang, SY., Fang, T.J. et al. Transgenic lines of melon (Cucumis melo L. var. makuwa cv. ‘Silver Light’) expressing antifungal protein and chitinase genes exhibit enhanced resistance to fungal pathogens. Plant Cell Tiss Organ Cult 112, 227–237 (2013). https://doi.org/10.1007/s11240-012-0227-5

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  • DOI: https://doi.org/10.1007/s11240-012-0227-5

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