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Trehalose metabolism is activated upon chilling in grapevine and might participate in Burkholderia phytofirmans induced chilling tolerance

Abstract

During the last decade, there has been growing interest in the role of trehalose metabolism in tolerance to abiotic stress in higher plants, especially cold stress. So far, this metabolism has not yet been studied in Vitis vinifera L., despite the economic importance of this crop. The goal of this paper was to investigate the involvement of trehalose metabolism in the response of grapevine to chilling stress, and to compare the response in plants bacterised with Burkholderia phytofirmans strain PsJN, a plant growth-promoting rhizobacterium that confers grapevine chilling tolerance, with mock-inoculated plants. In silico analysis revealed that the V. vinifera L. genome contains genes encoding the enzymes responsible for trehalose synthesis and degradation. Transcript analysis showed that these genes were differentially expressed in various plant organs, and we also characterised their response to chilling. Both trehalose and trehalose 6-phosphate (T6P) were present in grapevine tissues and showed a distinct pattern of accumulation upon chilling. Our results suggest a role for T6P as the main active molecule in the metabolism upon chilling, with a possible link with sucrose metabolism. Furthermore, plants colonised by B. phytofirmans and cultivated at 26°C accumulated T6P and trehalose in stems and leaves at concentrations similar to non-bacterised plants exposed to chilling temperatures for 1 day. Overall, our data suggest that T6P and trehalose accumulate upon chilling stress in grapevine and might participate in the resistance to chilling stress conferred by B. phytofirmans.

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Abbreviations

HAD:

l-2-Haloacid dehalogenase

PGPR:

Plant growth-promoting rhizobacteria

SnRK1:

Sucrose-non-fermenting-1-related kinase 1

T6P:

Trehalose-6-phosphate

TPS:

Trehalose phosphate synthase

TPP:

Trehalose phosphate phosphatase

TRE:

Trehalase

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Acknowledgments

This work was supported by the Region Champagne-Ardennes. Vitis vinifera L. Chardonnay clone 7535 was a gift from Moët et Chandon. B. phytofirmans strain PsJN was kindly provided by Dr. Essaid Ait Barka. We thank Pr. Petronia Carillo for her help in optimisation of the trehalose assay for grapevine.

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Correspondence to Christophe Clément.

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Fig. S1

Phylogenetic tree of TPSs isoforms in V. vinifera L. and A. thaliana. A phylip phylogenetic tree was constructed using Clustwal W2—Phylogeny program. The neighbour-joining algorithm was used to construct tree from the distance matrix. Sequences were obtained from the NCBI Gene Bank. Tree was drawn in a radial form using TreeView software (http://www.taxonomy.zoology.gla.ac.uk/rod/treeview.html) (PPT 55 kb)

Fig. S2

Alignment of VvTPS1, AtTPS1, OsTPS1 and SlTPS1 protein sequences. Amino acid residues that are similar in all sequences are shaded in grey and the ones that are predicted to be important for substrate binding (according to Lunn 2007; Vandesteene et al. 2010) in catalytically active TPS are shaded in red. All of these residues are perfectly conserved in the four sequences (PPT 139 kb)

Fig. S3

Alignment of VvTPPA-F and TaTPP (from Thermoplasma acidophilum). Amino acid residues that are similar in all sequences are shaded in grey and the ones that are predicted to be important for substrate binding in catalytically active TPP enzymes (according to Lunn 2007) are shaded in red. All of these residues are perfectly conserved in the 6 VvTPP sequences (except one conservative substitution of Lys to Arg in four isoforms VvTPPA, VvTPPB, VvTPPC and VvTPPE) (PPT 105 kb)

Fig. S4

Relative transcript number of VvTPPs non-chilled (a) and chilled (b) grapevine plants. Transcript number was calculated using the standard curve method and normalized with the Ef1α gene as internal control. Results represent the mean (± SD) of transcript number analysed from three independent experiments. Plants were exposed to chilling during 9 h, a time point when all VvTPPs gene induction peaked. n.d not detected; ds: detected only in stems; dl : detected only in leaves (PPT 59 kb)

Fig. S5

Protein coding region of the V. vinifera L. cv. Chardonnay TPPA cDNA and the deduced protein sequence. Differences from the orthologous V. vinifera L. cv. Pinot Noir cDNA and protein are highlighted in red (DOC 25 kb)

Fig. S6

Correlation between T6P and sucrose in roots, stems and leaves of V. vinifera L. plants. Correlation was evaluated in all samples (a), in non-chilled samples (b), in chilled samples (c) and in bacterised samples (d). All R² values are significant (P < 0.05) (PPT 59.5 kb)

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Fernandez, O., Vandesteene, L., Feil, R. et al. Trehalose metabolism is activated upon chilling in grapevine and might participate in Burkholderia phytofirmans induced chilling tolerance. Planta 236, 355–369 (2012). https://doi.org/10.1007/s00425-012-1611-4

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Keywords

  • Burkholderia phytofirmans strain PsJN
  • Chilling
  • Trehalose
  • Trehalose 6-phosphate
  • Vitis vinifera L.