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Isolation and expression analysis of defensin gene and its promoter from Brassica juncea

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Abstract

The full-length pathogen-inducible plant defensin gene encoding 80 amino acid sequences of pathogen-related gene PR12 (Bjdefensin) was isolated from Brassica juncea. Conserved domain (CD) search reveals its similarity with gamma thionin and knottins families of plant antimicrobial peptides. The protein blast reveals more than 80% of its similarity with defensin-like protein of Brassica rapa and Camelina sativa. Gene expression studies revealed that the transcript levels of Bjdefensin gene increases significantly upon Alternaria infection, jasmonic acid and wounding treatments but was not induced by salicylic acid. To further study the stress-inducible regulation of Bjdefensin gene, its promoter (2.5 kb) was isolated and cloned upstream of GUS gene in pORER2 vector. In silico studies of Bjdefensin promoter showed many important conserved cis-elements, responsive to biotic and abiotic stresses. Histochemical GUS assay showed pathogen-inducible expression of Bjdefensin promoter after fungal infection. Bjdefensin promoter was also induced by Jasmonic acid and wounding.

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Abbreviations

SA:

Salicylic acid

JA:

Jasmonic acid

ET:

Ethylene

SAR:

Systemic acquired resistance

ISR:

Induced systemic resistance

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Acknowledgements

Author is thankful to Director NRCPB for all possible help and financial support during the tenure of the work. This study was funded by NRCPB. The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. National Research Centre on Plant Biotechnology, Pusa Campus, New Delhi, 110012, India

    Sandhya Rawat, Sajad Ali, N. N. Chamil Nayankantha, N. Chandrashekar & Anita Grover

  2. Fakir Mohan University, Vyasa Vihar, Balasore, Orissa, 756020, India

    Sandhya Rawat & Bhabatosh Mittra

  3. Division of Microbiology, CCUBGA, Indian Agricultural Research Institute, New Delhi, 110012, India

    N. Chandrashekar

  4. Plant Science Department, Rubber Research Institute of Sri Lanka Dartonfield, Agalawatta, Sri Lanka

    N. N. Chamil Nayankantha

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  1. Sandhya Rawat
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  2. Sajad Ali
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Correspondence to Anita Grover.

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Rawat, S., Ali, S., Nayankantha, N.N.C. et al. Isolation and expression analysis of defensin gene and its promoter from Brassica juncea . J Plant Dis Prot 124, 591–600 (2017). https://doi.org/10.1007/s41348-017-0103-y

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  • DOI: https://doi.org/10.1007/s41348-017-0103-y

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