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Isolation and molecular characterization of pathogenesis related PR2 gene and its promoter from Brassica juncea

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Biologia Plantarum

Abstract

Pathogenesis-related (PR) proteins play key roles in plant disease resistance. Here, we isolated and characterized pathogenesis-related PR2 gene encoding β-1,3-glucanase from Brassica juncea and named it BjPR2 (GenBank accession number DQ359125). The amino acid sequence of BjPR2 showed ~99 % similarity with β-1,3-glucanase of Brassica rapa, B. napus, and B. oleracea. BjPR2 transcription was rapidly increased after Alternaria brassicae infection, salicylic acid application, and wounding, but the induction was delayed in response to jasmonic acid. To investigate the transcriptional regulation of BjPR2 gene, its promoter was isolated. In silico analysis of BjPR2 promoter showed cis-regulatory elements upstream of TATA and CAAT boxes responsive to defense, hormones, wounding, and plant developmental stage. Homozygous Arabidopsis thaliana lines were developed with plasmid construct having β-glucuronidase (GUS) reporter gene driven by BjPR2 promoter. The analysis of GUS protein in Arabidopsis lines showed that BjPR2 promoter drived distinct pattern of pathogen inducible expression after fungal infection (Alternaria brassicae, Erysiphe orontii), phytohormones, and wounding. It also showed age dependent and organ specific expressions. BjPR2 promoter drove strong GUS activity in Arabidopsis seedlings and showed organ specific expression at the later growth stages (lateral organ junctions, leaf serrate, base of siliques, and receptacle). Due to stress-inducible and tissue specific nature, the BjPR2 promoter can serve as a potential candidate in genetic engineering.

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Abbreviations

Bj:

Brassica juncea

ET:

ethylene

GUS:

β-glucuronidase

HR:

hypersensitive response

JA:

jasmonic acid

ORF:

open reading frame

PR:

pathogenesis-related

SA:

salicylic acid

SAR:

systemic acquired resistance

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Authors and Affiliations

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

    S. Ali, S. Rawat, N. M. C. Nayanakantha, Z. A. Mir & A. Grover

  2. Department of Advanced Zoology and Biotechnology, Presidency College, Chennai, 600005, India

    S. Ali & M. Sultana

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

    N. Chandrashekar

  4. Department of Plant Biology and Biotechnology, Presidency College, Chennai, 600005, India

    A. Manoharan

Authors
  1. S. Ali
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  2. N. Chandrashekar
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  3. S. Rawat
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  4. N. M. C. Nayanakantha
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  5. Z. A. Mir
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  6. A. Manoharan
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  7. M. Sultana
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  8. A. Grover
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Correspondence to A. Grover.

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Acknowledgments: We gratefully acknowledge the Project Director, the National Research Centre on Plant Biotechnology Pusa Campus New Delhi for providing all the facilities required to complete this work.

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Ali, S., Chandrashekar, N., Rawat, S. et al. Isolation and molecular characterization of pathogenesis related PR2 gene and its promoter from Brassica juncea . Biol Plant 61, 763–773 (2017). https://doi.org/10.1007/s10535-017-0726-7

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  • DOI: https://doi.org/10.1007/s10535-017-0726-7

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