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Molecular cloning of a pathogen/wound-inducible PR10 promoter from Pinus monticola and characterization in transgenic Arabidopsis plants

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Abstract

In Pinus monticola (Dougl. ex D. Don), the class ten pathogenesis-related (PR10) proteins comprise a family of multiple members differentially expressed upon pathogen infection and other environmental stresses. One of them, PmPR10-1.13, is studied here by investigating its transcriptional regulation in transgenic Arabidopsis plants. For functional analyses of the PmPR10-1.13 promoter, a 1,316-bp promoter fragment and three 5′ deletions were translationally fused to the ß-glucuronidase (GUS) reporter gene. The 1,316-bp promoter-driven GUS activity first appeared in hypocotyls and cotyledons in 2- to 3-day-old seedlings. As transgenic plants grew, GUS activity was detected strongly in apical meristems, next in stems and leaves. No GUS activity was detected in roots and in reproductive tissues of flower organs. In adult plants, the PmPR10-1.13 promoter-directed GUS expression was upregulated following pathogen infection and by wounding treatment, which generally mimic the endogenous expression pattern in western white pine. Promoter analysis of 5′ deletions demonstrated that two regions between −1,316 and −930, and between −309 and −100 were responsible for the wound responsiveness. By structural and functional comparisons with PmPR10-1.14 promoter, putative wound-responsive elements were potentially identified in the PmPR10-1.13 promoter. In conclusion, PmPR10-1.13 showed properties of a defence-responsive gene, being transcriptionally upregulated upon biotic and abiotic stresses.

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Abbreviations

4MU:

4-Methylumbelliferone

RT-PCR:

Reverse transcriptase-polymerase chain reaction

PR:

Pathogenesis-related

X-Glu:

5-Bromo-4-chloro-3-indolyl-β-D-glucuronic acid

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Acknowledgements

This research was funded by the Canadian Forest Service, Canadian Biotechnology Strategy, and Forestry Innovation Investment awarded to A.K.M.E. We thank R. Hunt, R. Sturrock, and H. Williams for thoughtful comments on the manuscript.

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Authors and Affiliations

  1. Pacific Forestry Centre, Canadian Forest Service, Natural Resources Canada, 506 West Burnside Road, Victoria, BC, V8Z 1M5, Canada

    Jun-Jun Liu, Abul K. M. Ekramoddoullah, Nina Piggott & Arezoo Zamani

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  1. Jun-Jun Liu
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  2. Abul K. M. Ekramoddoullah
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  3. Nina Piggott
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Correspondence to Abul K. M. Ekramoddoullah.

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Liu, JJ., Ekramoddoullah, A.K.M., Piggott, N. et al. Molecular cloning of a pathogen/wound-inducible PR10 promoter from Pinus monticola and characterization in transgenic Arabidopsis plants. Planta 221, 159–169 (2005). https://doi.org/10.1007/s00425-004-1428-x

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