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The primary module in Norway spruce defence signalling against H. annosum s.l. seems to be jasmonate-mediated signalling without antagonism of salicylate-mediated signalling

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Abstract

A key tree species for the forest industry in Europe is Norway spruce [Picea abies (L.) Karst.]. One of its major diseases is stem and butt rot caused by Heterobasidion parviporum (Fr.) Niemelä & Korhonen, which causes extensive revenue losses every year. In this study, we investigated the parallel induction of Norway spruce genes presumably associated with salicylic acid- and jasmonic acid/ethylene-mediated signalling pathways previously observed in response to H. parviporum. Relative gene expression levels in bark samples of genes involved in the salicylic acid- and jasmonic acid/ethylene-mediated signalling pathways after wounding and inoculation with either the saprotrophic biocontrol fungus Phlebiopsis gigantea or with H. parviporum were analysed with quantitative PCR at the site of the wound and at two distal locations from the wound/inoculation site to evaluate their roles in the induced defence response to H. parviporum in Norway spruce. Treatment of Norway spruce seedlings with methylsalicylate, methyljasmonate and inhibitors of the jasmonic acid/ethylene signalling pathway, as well as the Phenylalanine ammonia lyase inhibitor 2-aminoindan-2-phosphonic acid were conducted to determine the responsiveness of genes characteristic of the different pathways to different hormonal stimuli. The data suggest that jasmonic acid-mediated signalling plays a central role in the induction of the genes analysed in this study irrespective of their responsiveness to salicylic acid. This may suggest that jasmonic acid-mediated signalling is the prioritized module in the Norway spruce defence signalling network against H. parviporum and that there seems to be no immediate antagonism between the modules in this interaction.

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Abbreviations

ACS:

ACC synthase

AIP:

2-Aminoindan-2-phosphonic acid

DIECA:

Diethyldithiocarbamic acid

ET:

Ethylene

ERF1:

Ethylene response factor 1

JA:

Jasmonic acid

JAZ1:

Jasmonate ZIM domain 1

LOX:

Lipoxygenase

LURP1:

Late up-regulated in response to Hyaloperonospora parasitica

MeJA:

Methyl jasmonate

MeSA:

Methyl salicylate

PAL:

Phenylalanine ammonia lyase

PR1:

Pathogensis related protein1

SA:

Salicylic acid

STS:

Silverthiosulfate

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Acknowledgments

Professor J. Zón at University of Wroclaw, Poland, and Dr. John H Grabber and Professor John Ralph at University of Wisconsin-Madison, USA are acknowledged for their generous gift of 2-aminoindan-2-phosphonic acid. We thank Dr. Rena Gadijeva for skilful assistance in the inoculation study and Diem Nguyen for reading the manuscript. The Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning and the Swedish Foundation for Strategic Research provided financial support for the study.

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Correspondence to Malin Elfstrand.

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425_2012_1822_MOESM1_ESM.xlsx

Suppl. Table S1 Primers used for quantitative PCR analysis and RACE. First column contain the name of the gene, second and third column the sequences of forward and reverse primers respectively. References for already published primers and Genbank accession numbers for newly designed primers are indicated. (XLSX 11 kb)

425_2012_1822_MOESM2_ESM.pptx

Suppl. Fig S2 BestKeeper data for the considered reference genes; phosphoglucomutase (Vestman et al. 2010), eukaryotic translation initiation factor 4A (elF4A) (Palovaara and Hakman 2008), alpha-Tubulin and Elongation factor 1-α (ELF1α) (Arnerup et al. 2011). The Ct values of PGmutase, elF4A, ELF1α and alpha-Tubulin was determined by qPCR in a representative subset of the treatments in the experiment. The resulting data was analysed with the BestKeeper software version 1(Pfaffl et al. 2004, Biotechnology Letters 26: 509-515). The Ct values of the four tested genes (a), Pearson correlation coefficient (r) (b) and Regression Analysis of reference genes vs. BestKeeper (c). Actin (isotig08740-RNA seq dataset published in Danielsson 2011) was also tested but the amplicon produced multiple products (data not shown). (PPTX 123 kb)

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Arnerup, J., Nemesio-Gorriz, M., Lundén, K. et al. The primary module in Norway spruce defence signalling against H. annosum s.l. seems to be jasmonate-mediated signalling without antagonism of salicylate-mediated signalling. Planta 237, 1037–1045 (2013). https://doi.org/10.1007/s00425-012-1822-8

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  • DOI: https://doi.org/10.1007/s00425-012-1822-8

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