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Development of a SCAR marker for molecular detection and diagnosis of Tilletia controversa Kühn, the causal fungus of wheat dwarf bunt

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Abstract

Tilletia controversa Kühn (TCK) is an important quarantine pathogen that causes wheat dwarf bunt and results in devastating damage to wheat production. The fungus is difficult to be distinguished from T. caries and T. laevis, which cause wheat common bunt, based on morphological, physiological and symptomatological characteristics of the pathogens. The traditional detection of the fungus can be a long and tedious process with poor accuracy. The inter-simple sequence repeat (ISSR) technique has been used for identifying molecular markers for detection of TCK. Of 28 ISSR primers screened, ISSR-859 amplified a specific 678 bp DNA fragment from all TCK isolates but not from any isolates of the common bunt fungi or other pathogenic fungi tested. Based on the fragment sequence, a pair of sequence characterized amplified region (SCAR) primers was designed, which amplified a 372 bp DNA fragment specifically in TCK. The SCAR marker was detected using as low as 1 ng template DNA of TCK, and was also detected using broken teliospores and DNA from asymptomatic wheat samples. We developed the SYBR Green I and TaqMan Green I and TaqMan real-time polymorphism chain reaction methods to detect TCK with the detection limit of 0.1 fg with asymptomatic wheat samples. Further work is needed to develop a rapid test kit for this pathogenic fungus using the designed specific primers.

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Acknowledgments

We appreciate **anming Chen (USDA-ARS and Washington State University, USA) for critical revising the paper before we submitted. We appreciate Blair J. Goates, UADA-ARS, National Small-Grains Germplasm Research Facility for providing T. controversa and T. caries. This work was supported by Bei**g Natural Science Foundation (Grant No. L140012), Multidisciplinary Cooperation Project of Bei**g Nova Program (Grant No. Z141105001814116), Bei**g Nova Program (Grant No. Z131105000413057) and Ministry of Agriculture, P. R. China (Grant No. CARS-03).

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Authors and Affiliations

  1. State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Bei**g, 100193, China

    Li Gao, Huixin Yu, Fei Gao, Taiguo Liu, Bo Liu & Wanquan Chen

  2. School of Life Science and Engineering, Southwest University of Science and Technology, Sichuan, 621000, China

    Huixin Yu & **aohui Kang

  3. College of Agricultural and Forestry Science and Technology, Hebei North University, Zhangjiakou, 075100, China

    Wensu Han

  4. Northeast Agriculture University, Harbin, 150030, Heilongjiang, China

    Fei Gao & Jiguo Gao

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  1. Li Gao
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  2. Huixin Yu
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  3. Wensu Han
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  4. Fei Gao
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  5. Taiguo Liu
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  6. Bo Liu
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  7. **aohui Kang
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  8. Jiguo Gao
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  9. Wanquan Chen
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Correspondence to Li Gao or Wanquan Chen.

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Gao, L., Yu, H., Han, W. et al. Development of a SCAR marker for molecular detection and diagnosis of Tilletia controversa Kühn, the causal fungus of wheat dwarf bunt. World J Microbiol Biotechnol 30, 3185–3195 (2014). https://doi.org/10.1007/s11274-014-1746-5

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  • DOI: https://doi.org/10.1007/s11274-014-1746-5

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