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O-methyltransferase(s)-suppressed plants produce lower amounts of phenolic vir inducers and are less susceptible to Agrobacterium tumefaciens infection

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Abstract

The first step of Agrobacterium tumefaciens/plant interaction corresponds to the activation of a transduction pathway of the bacterium by plant exudate. Phenolic compounds rapidly secreted by wounded plant cells induce the expression of bacterial virulence (vir) genes; however, little is known about their biosynthesis in plant. Here we show that inoculation of an Agrobacterium tumefaciens virulent strain on orthodiphenol-O-methyltransferases-suppressed tobacco plants leads to significantly smaller tumors compared to control plants. These transgenic plants are inhibited for caffeic acid O-methyltransferase class I or II (OMT; EC 2.1.1.6) and/or caffeoyl-coenzyme A O-methyltransferase (CCoAOMT; EC 2.1.1.104) that are involved in monolignol biosynthesis. The significant decrease of tumor size could be suppressed by the pre-activation of bacterial virulence, before inoculation, using acetosyringone a known vir inducer. Total soluble phenolic amounts and cell wall composition analyzed by FT-IR analysis did not show significant differences between transgenic and control plants. The potential of phenolic extracts from control and OMT-suppressed plants to induce virulence was evaluated using an Agrobacterium tumefaciens reporter strain carrying a vir::LacZ gene fusion plasmid. Lower vir-inducing activities were recorded for plants that show inhibition to caffeic acid O-methyltransferase activity. HPLC analysis confirmed that the levels of several phenolic compounds were differently affected by wounding and/or by bacterial inoculation. Statistical correlations were established between tumor sizes, vir-inducing activities, O-methyltransferases proteins accumulations and the levels of various soluble phenolic compounds such as acetosyringone. These results demonstrate the role of the O-methyltransferases of the phenylpropanoid pathway in the early production of soluble Agrobacterium tumefaciens vir inducers.

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Abbreviations

CCoAOMT:

Caffeoyl-coenzyme A O-methyltransferase

COMTI:

Caffeic acid O-methyltransferase of class I

COMTII:

Caffeic acid O-methyltransferase of class II

FT-IR:

Fourier transformed-infrared

HPLC:

High-performance liquid chromatography

OMT:

Orthodiphenol-O-methyltransferases

Vir :

Virulence

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Acknowledgments

We are grateful to Professor F. Delmotte from LBLGC University of Orléans and Léon Otten from IBMP du CNRS of Strasbourg for providing Agrobacterium tumefaciens strains. We acknowledge Dr G. Pinçon, Dr F. Martz, Dr V. Toquin and Professor R. Atanassova for their previous works on OMT plants. We thank Professor E. Lainé and Professor Franck Brignolas from LBLGC for helpful discussions. We are grateful to A. Guichard for taking good care of tobacco plants and to Gilles Moreau France for technical assistance. This work was supported by the Ministère de l’Education Nationale, de l’Enseignement Supérieur et de la Recherche.

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Authors and Affiliations

  1. Institut de Biologie Moléculaire des Plantes du Centre National de la Recherche Scientifique, Université Louis Pasteur, 12, rue du Général Zimmer, 67084, Strasbourg Cedex, France

    Stéphane Maury, P. Geoffroy & M. Legrand

  2. Laboratoire de Biologie des Ligneux et des Grandes Cultures, UPRES EA1207, Faculté des Sciences, Université d’Orléans, rue de Chartres, BP 6759, 45067, Orléans Cedex 2, France

    Stéphane Maury & A. Delaunay

  3. Laboratoire de Phytotechnologie, UPRES EA3900, Faculté de Pharmacie d’Amiens, Université de Picardie Jules Verne, 1 rue des Louvels, 80037, Amiens Cedex 01, France

    F. Mesnard

  4. INRA Parois Végétales et Matériaux Fibreux UMR FARE, 2 esplanade Roland Garros, BP224, 51686, Reims, France

    D. Crônier & B. Chabbert

Authors
  1. Stéphane Maury
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  2. A. Delaunay
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  3. F. Mesnard
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  4. D. Crônier
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  5. B. Chabbert
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  6. P. Geoffroy
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  7. M. Legrand
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Correspondence to Stéphane Maury.

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425_2010_1230_MOESM1_ESM.pdf

FT-IR analysis of cell wall composition in transgenic stems after wounding or agroinfection. Average spectra were obtained from the cell wall analysis of eight replicates. C, control plants transformed with empty plasmid; COMTI, transgenic tobaccos suppressed for COMTI; COMTII, transgenic tobaccos suppressed for COMTII; CCoAOMT, transgenic tobaccos suppressed for CCoAOMT; dAS, transgenic tobaccos suppressed for COMTI and CCoAOMT. Unwounded (a), wounded and mock inoculated (b) or wounded and Agrobacterium tumefaciens infected transgenic plants (c) (Supplementary material 1 (PDF 74 kb)

425_2010_1230_MOESM2_ESM.pdf

Total soluble phenolic contents of transgenic stems after wounding or agroinfection. Total soluble phenolic compounds were extracted and quantified using Folin-Ciocalteu’s reagent. Extracts were prepared from 6-hours post-treatment tobacco stems of unwounded (white bars), wounded and mock inoculated (grey bars) or wounded and inoculated with a saturated culture of Agrobacterium tumefaciens (black bars) transgenic plants: C, control plants transformed with an empty plasmid; COMTI, transgenic tobaccos suppressed for COMTI; COMTII, transgenic tobaccos suppressed for COMTII; CCoAOMT, transgenic tobacco suppressed for CCoAOMT; dAS, transgenic tobacco suppressed for COMTI and CCoAOMT. Mean values ± SE (n=4) are shown. Two independent experiments were performed. In order to evaluate genotypic and treatment effects, two-way ANOVA analysis was performed: for each graph, g indicates the genotype effect, t the treatment effect and gxt the genotype by treatment interaction. Significant effects are indicated at * P ≤ 0.05 or ** P ≤ 0.01 (Supplementary material 2 (PDF 11 kb)

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Maury, S., Delaunay, A., Mesnard, F. et al. O-methyltransferase(s)-suppressed plants produce lower amounts of phenolic vir inducers and are less susceptible to Agrobacterium tumefaciens infection. Planta 232, 975–986 (2010). https://doi.org/10.1007/s00425-010-1230-x

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