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Identification and characterization of Dicer-like genes in leaf rust pathogen (Puccinia triticina) of wheat

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Abstract

Puccinia triticina (P. triticina) is one of the most devastating fungal pathogens of wheat which causes significant annual yield loss to the crop. Understanding the gene regulatory mechanism of the biotrophic pathogen is one of the important aspects of host-pathogen interaction studies. Dicer-like genes are considered as important mediators of RNAi-based gene regulation. In this study, we report the presence of three Dicer-like genes (Pt-DCL1, Pt-DCL2, Pt-DCL3) in P. triticina genome identified through computational and biological analyses. Quantitative real-time PCR studies revealed an increase in the expression of these genes in germinating spore stages. Heterologous expression combined with mass spectrometry analysis of Pt-DCL2 confirmed the presence of a canonical Dicer-like gene in P. triticina. Phylogenetic analysis of the Pt-DCLs with the Dicer-like proteins from other organisms showed a distinct cluster of rust pathogens from the order Pucciniales. The results indicated a species-specific duplication of Dicer-like genes within the wheat rust pathogens. This study, for the first time, reports the presence of Dicer-dependent RNAi pathway in P. triticina that may play a role in gene regulatory mechanism of the pathogen during its development. Our study serves as a vital source of information for further RNAi-based molecular studies for better understanding and management of the wheat leaf rust disease.

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Funding

TRS is thankful to the SERB-Department of Science and Technology, Govt. of India, for JC Bose National Fellowship. HD is thankful to the University Grants Commission (UGC), New Delhi, for providing Junior Research Fellowship (JRF).

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Authors and Affiliations

  1. ICAR-National Institute for Plant Biotechnology (formerly ICAR-National Research Centre on Plant Biotechnology), Pusa Campus, New Delhi, 110012, India

    Himanshu Dubey, Kanti Kiran, Rajdeep Jaswal, Tapan Kumar Mondal, Neha Jain, N. K. Singh & Tilak Raj Sharma

  2. School of Biotechnology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India

    Himanshu Dubey & Arvind M. Kayastha

  3. Seri-Biotech Research Laboratory, Central Silk Board, Bangalore, Karnataka, 560035, India

    Himanshu Dubey

  4. ICAR-Indian Institute of Wheat and Barley Research, Regional Station, Flowerdale, Shimla, 171009, India

    Subhash C. Bhardwaj

  5. Indian Council of Agricultural Research, Krishi Bhawan, New Delhi, 110001, India

    Tilak Raj Sharma

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  1. Himanshu Dubey
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  2. Kanti Kiran
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  3. Rajdeep Jaswal
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  4. Subhash C. Bhardwaj
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  5. Tapan Kumar Mondal
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  6. Neha Jain
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  7. N. K. Singh
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  8. Arvind M. Kayastha
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  9. Tilak Raj Sharma
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Contributions

TRS conceived and designed the project, HD performed computational analysis and biological experiments, and KK and RJ helped in biological experiments. NJ and NKS helped in the MALDI-TOF analysis, SCB provided biological material. TKM provided input during manuscript writing. HD, KK, TRS, and AMK wrote the manuscript.

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Correspondence to Arvind M. Kayastha or Tilak Raj Sharma.

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Dubey, H., Kiran, K., Jaswal, R. et al. Identification and characterization of Dicer-like genes in leaf rust pathogen (Puccinia triticina) of wheat. Funct Integr Genomics 20, 711–721 (2020). https://doi.org/10.1007/s10142-020-00745-w

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