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Gene-based analysis of Puccinia species and development of PCR-based marker to detect Puccinia striiformis f. sp. tritici causing yellow rust of wheat

  • Fungal Diseases
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Abstract

Stripe rust is considered as the current major rust disease affecting winter cereal production across the world. A quick, reliable PCR-based marker was developed here to detect, identify and rapidly monitor Puccinia striiformis f. sp. tritici (Pst) in wheat-growing areas. Three respective sets of primers, designed from β-tubulin, squalene monooxygenase and ketopantoate reductase genes selected from the full genome of Puccinia striiformis f. sp. tritici, amplified sequences of 239, 358 and 1518 bp, respectively, in Pst pathotypes. A fragment of 1518 bp unique to Pst pathotypes was amplified using primer set PstKeto F1_30/Pst KetoR1_1547 and distinguished the pathogen clearly from different Puccinia spp. and other fungal pathogens. The detection limit of the marker (KetoPstRA1500, accession no. KU240073) by conventional PCR assay was 10 pg. This marker could detect the pathogen in the host before symptom expression. The sensitivity and utility of the marker were further enhanced in a qPCR-based assay that was developed with a newly designed primer set PstKeto F1_1246/Pst KetoR1_1547, which amplified a product of 302 bp and detected as little as 10 fg of DNA. This PCR/qPCR based marker is suitable for studying cultivar resistance, which requires accurate quantification of the pathogen in diseased host tissue.

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Acknowledgements

Authors are highly thankful to the Indian Council of Agricultural Research, AMAAS project (NBAIM/AMAAS/2014-15/68), Mau Nath Bhanjan, (Uttar Pradesh), India, for partially funding this work. Authors are also thankful to ICAR-Indian Institute of Wheat and Barley Research, Regional Station Flowerdale Shimla, (Himachal Pradesh), India for providing the pure inocula of rust pathotypes and the Director of ICAR-Indian Agricultural Research Institute, (New Delhi), India for providing the facilities to carry out the research.

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Authors and Affiliations

  1. Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India

    Rashmi Aggarwal, Sapna Sharma, Sangeeta Gupta, Vaibhav K. Singh & Deepika Kulshreshtha

  2. ICAR-Indian Agricultural Research Institute, Regional Station Wellington, The Nilgiris, Tamilnadu, India

    C. Manjunatha

Authors
  1. Rashmi Aggarwal
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  2. Sapna Sharma
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  3. Sangeeta Gupta
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  4. C. Manjunatha
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  5. Vaibhav K. Singh
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  6. Deepika Kulshreshtha
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Correspondence to Rashmi Aggarwal.

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The authors declare that they have no conflict of interest.

Additional information

The nucleotide sequence data (Puccinia striiformis f. sp. tritici specific marker: KetoPstRA1500) reported in the manuscript is available in the NCBI database under accession number KU240073.

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Aggarwal, R., Sharma, S., Gupta, S. et al. Gene-based analysis of Puccinia species and development of PCR-based marker to detect Puccinia striiformis f. sp. tritici causing yellow rust of wheat. J Gen Plant Pathol 83, 205–215 (2017). https://doi.org/10.1007/s10327-017-0723-x

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  • DOI: https://doi.org/10.1007/s10327-017-0723-x

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