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Transgenic Sugar Beet Expressing a Bacterial Mannitol-1-Phosphate Dehydrogenase (mtlD) Gene Shows Enhanced Resistance to Fungal Pathogens

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Abstract

The E. coli mtlD gene encoding mannitol-1-phosphate dehydrogenase has been shown to be involved in the plant response to abiotic stresses such as salinity, drought and chilling. Nevertheless, in this study transgenic sugar beet lines containing the mtlD gene under control of a stress-inducible rd29A promoter were evaluated for their potential of resistance against three fungi including Alternaria alternata, Botrytis cinerea and Cercospora beticola. Southern blot analysis revealed the low copy number (1-3 copies) integration of the transgene in the transgenic lines. Expression profiling by semi-quantitative-RT-PCR analysis showed different levels of cold-inducible expression of mtlD in independent T1 transformants. Mannitol content quantified by HPLC analysis ranged from 1.8 to 3.7 µmol g−1 dry weight in different transgenic lines. Detached leaf bioassay showed that lower transgene expression levels in the transgenic line mt-LS4-32 under non-stressed conditions reduced the disease severity (DS) of A. alternata and B. cinerea by 49 to 87 %, respectively. At the whole-plant level, under non-stressed conditions, the transgenic line showed better performance to A. alternata with a delay of fungal symptom appearance. Following exposure to low temperature (4 °C), the transgenic line showed no fungal infection after 14 days of inoculation. Transgenic lines inoculated with C. beticola significantly (P < 0.001) showed higher resistance than the non-transformed control plants. DS in transgenic lines expressing mtlD gene was zero and 0.05 % compared to 25 % for the control. These results indicate that the mtlD gene can be used as a target to improve plant tolerance to both abiotic and biotic stresses.

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Acknowledgments

The authors thank the Bioscience and Biotechnology Institute of Urmia University for providing the necessary laboratory facilities. The authors would also like to thank Dr. Y. Ghosta, Dep. of Plant Protection, University of Urmia, Urmia, Iran, for providing the fungal isolates A. alternata and B. cinerea.

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Authors and Affiliations

  1. Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

    Azadeh Goudarzi & Naser Safaie

  2. Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Urmia, Nasloo Campus, P.O. Box 165, Urmia, Iran

    Morad Jafari & Setare Mohammad Jafari

  3. Department of Agricultural Biotechnology, Bioscience and Biotechnology Institute, University of Urmia, Urmia, Iran

    Morad Jafari & Setare Mohammad Jafari

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  1. Azadeh Goudarzi
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Goudarzi, A., Jafari, M., Safaie, N. et al. Transgenic Sugar Beet Expressing a Bacterial Mannitol-1-Phosphate Dehydrogenase (mtlD) Gene Shows Enhanced Resistance to Fungal Pathogens. Sugar Tech 18, 192–203 (2016). https://doi.org/10.1007/s12355-015-0379-9

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