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Two Different Banana NPR1-Like Coding Sequences Confer Similar Protection Against Pathogens in Arabidopsis

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Abstract

Differences between the coding sequences of two banana homologues of the Non-expressor of Pathogenesis-Related 1 (NPR1) genes (MNPR1A and MNPR1B) were investigated as a possible cause for the differential activity of the two genes. Each of the MNPR1 coding sequences were expressed under the control of the cauliflower mosaic virus 35S promoter/terminator sequences in the transgenic Arabidopsis npr1-2 mutant. These MNPR1-expressing plants were then exposed to either the biotrophic oomycete Hyaloperonospora arabidopsidis, the necrotrophic fungus Botrytis cinerea, or the hemi-biotrophic bacterial pathogen Pseudomonas syringae. Expression of either MNPR1A or MNPR1B increased Pathogenesis-related 1 (PR-1) transcription in a similar manner in mutant plants and reduced pathogen growth, restoring resistance of the Arabidopsis npr1-2 mutant plants to pathogens. Sequence differences between MNPR1A and MNPR1B coding sequences did not affect MNPR1activity, suggesting a possible role for the involvement of other regulatory sequences in differential MNPR1 gene expression.

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Acknowledgement

Our appreciation goes to Prof Nick Brewin, Dr Jun Fan and Dr Henk-jan Schoonbeeck (John Innes Centre, UK) and Dr Georgina Fabro (Sainsbury Laboratory, UK) for the provision of facilities and for technical advice throughout the course of this work, to the Rockefeller Foundation and the University of Pretoria for funding, and to Prof. Xinnian Dong (Duke University, USA) for donation of mutants, and to Prof David Lawlor for critical review of the manuscript.

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Authors and Affiliations

  1. Plant Science Department; Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria, 0083, South Africa

    Rosita Endah Yocgo, Karl Kunert & Rachel Chikwamba

  2. John Innes Centre, Norwich Research Park, Colney, NR4 7UH, UK

    Gary Creissen

  3. Council for Scientific and Industrial Research (CSIR), Biosciences Division, Building 20, P O Box 395, Pretoria, 0001, South Africa

    Rachel Chikwamba

Authors
  1. Rosita Endah Yocgo
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  2. Gary Creissen
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  3. Karl Kunert
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  4. Rachel Chikwamba
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Corresponding author

Correspondence to Rachel Chikwamba.

Additional information

Communicated by: Ray Ming

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Figure S1

Pictures of typical Botrytis lesions on Arabidopsis leaves 72 h post Botrytis infection. Leaves (six per plant) of eight different Arabidopsis genotypes were infected with a 2.5 x 105 spores/mL of an inoculum of B. cinerea B05.10. Pictures were taken 72 hpi to illustrate the lesions formed Arrows on the leaves indicate the point of lesion initiation. The experiment was repeated twice using independent sets of plant material treated in the same manner described above. (JPEG 59 kb)

Figure S2

Pictures of infected plants 48 h post Pseudomonas syringae infection. A single leaf of eight different Arabidopsis genotypes were infiltrated primarily with either a 10 mM MgCL2 solution or PstavrRPM1 (5x106 cfu.ml-1 in 10mM MgCl2) inoculum. This was followed 48 later by a secondary infiltration with a Pst-DC3000-Lux (5x105 cfu.ml-1 in 10mM MgCl2) inoculum of three additional leaves per plant. Pictures of infected plants were taken digitally 48 h after the secondary infiltration. Some of the PstDC3000-Lux infected leaves (red arrows) and PstavrRPM1-infected leaves (white arrows) are represented (A and B). The experiment was repeated twice using independent sets of plant material treated in the same manner described above. (JPEG 129 kb)

Table SI

Primer sequences used for amplification of AtPR-1 and MNPR1 genes. (DOCX 11 kb)

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Yocgo, R.E., Creissen, G., Kunert, K. et al. Two Different Banana NPR1-Like Coding Sequences Confer Similar Protection Against Pathogens in Arabidopsis . Tropical Plant Biol. 5, 309–316 (2012). https://doi.org/10.1007/s12042-012-9112-y

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