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.
Similar content being viewed by others
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
References
Appert C, Zon J, Amrhein N (2003) Kinetic analysis of the inhibition of phenylalanine ammonia-lyase by 2-aminoindan-2-phosphonic acid and other phenylalanine analogues. Phytochemistry 62:415–422
Arnerup J, Lind M, Olson A, Stenlid J, Elfstrand M (2011) The pathogenic white-rot fungus Heterobasidion parviporum triggers non-specific defence responses in the bark of Norway spruce. Tree Physiol 31:1262–1272
Boller T, Felix G (2009) A renaissance of elicitors: perception of microbe-associated molecular patterns and danger signals by pattern-recognition receptors. Annu Rev Plant Biol 60:379–406
Chang SJ, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep 11:113–116
Chen Z, Zheng Z, Huang J, Lai Z, Fan B (2009) Biosynthesis of salicylic acid in plants. Plant Signal Behav 4:493–496
Danielsson M, Lunden K, Elfstrand M, Hu J, Zhao T, Arnerup J, Ihrmark K, Swedjemark G, Borg-Karlson AK, Stenlid J (2011) Chemical and transcriptional responses of Norway spruce genotypes with different susceptibility to Heterobasidion spp. infection. BMC Plant Biol 11:154
Deflorio G, Horgan G, Woodward S, Fossdal CG (2011) Gene expression profiles, phenolics and lignin of Sitka spruce bark and sapwood before and after wounding and inoculation with Heterobasidion annosum. Physiol Mol Plant P 75:180–187
Dempsey D, Vlot AC, Wildermuth MC, Klessig DF (2011) Salicylic acid biosynthesis and metabolism. In: Arabidopsis book, vol 9, p e0156
Farmer EE, Caldelari D, Pearce G, Walkersimmons K, Ryan CA (1994) Diethyldithiocarbamic acid inhibits the octadecanoid signaling pathway for the wound induction of proteinase-inhibitors in tomato leaves. Plant Physiol 106:337–342
Fossdal CG, Hietala AM, Kvaalen H, Solheim H (2006) Changes in host chitinase isoforms in relation to wounding and colonization by Heterobasidion annosum: early and strong defense response in 33-year-old resistant Norway spruce clone. Tree Physiol 26:169–177
Franceschi VR, Krekling T, Christiansen E (2002a) Application of methyl jasmonate on Picea abies (Pinaceae) stems induces defense-related responses in phloem and xylem. Am J Bot 89:578–586
Franceschi VR, Krekling T, Christiansen E (2002b) Application of methyl jasmonate on Picea abies (Pinaceae) stems induces defense-related responses in phloem and xylem. Amer J Bot 89:578–586
Franceschi VR, Krokene P, Christiansen E, Krekling T (2005) Anatomical and chemical defenses of conifer bark against bark beetles and other pests. New Phytol 167:353–375
Germain H, Lachance D, Pelletier G, Fossdal CG, Solheim H, Seguin A (2012) The expression pattern of the Picea glauca defensin1 promoter is maintained in Arabidopsis thaliana, indicating the conservation of signalling pathways between angiosperms and gymnosperms. J Exp Bot 63:785–795
Hietala AM, Kvaalen H, Schmidt A, Johnk N, Solheim H, Fossdal CG (2004) Temporal and spatial profiles of chitinase expression by Norway spruce in response to bark colonization by Heterobasidion annosum. Appl Environ Microb 70:3948–3953
Hudgins JW, Franceschi VR (2004) Methyl jasmonate-induced ethylene production is responsible for conifer phloem defense responses and reprogramming of stem cambial zone for traumatic resin duct formation. Plant Physiol 135:2134–2149
Hudgins JW, Christiansen E, Franceschi VR (2003) Methyl jasmonate induces changes mimicking anatomical defenses in diverse members of the Pinaceae. Tree Physiol 23:361–371
Ingestad T, Kähr M (1985) Nutrition and growth of coniferous seedlings at varied relative nitrogen addition rate. Physiol Plant 65:109–116
Katagiri F, Tsuda K (2010) Understanding the plant immune system. Mol Plant Microbe Interact 23:1531–1536
Keeling CI, Bohlmann J (2006) Diterpene resin acids in conifers. Phytochemistry 67:2415–2423
Knoth C, Eulgem T (2008) The oomycete response gene LURP1 is required for defense against Hyaloperonospora parasitica in Arabidopsis thaliana. Plant J 55:53–64
Kozlowski G, Buchala A, Metraux JP (1999) Methyl jasmonate protects Norway spruce [Picea abies (L.) Karst.] seedlings against Pythium ultimum Trow. Physiol Mol Plant P 55:53–58
Krekling T, Franceschi VR, Krokene P, Solheim H (2004) Differential anatomical response of Norway spruce stem tissues to sterile and fungus infected inoculations. Trees-Struct Funct 18:1–9
Krokene P, Nagy NE, Solheim H (2008) Methyl jasmonate and oxalic acid treatment of Norway spruce: anatomically based defense responses and increased resistance against fungal infection. Tree Physiol 28:29–35
Likar M, Regvar M (2008) Early defence reactions in Norway spruce seedlings inoculated with the mycorrhizal fungus Pisolithus tinctorius (Persoon) Coker & Couch and the pathogen Heterobasidion annosum (Fr.) Bref. Trees-Struct Funct 22:861–868
Martin D, Tholl D, Gershenzon J, Bohlmann J (2002) Methyl jasmonate induces traumatic resin ducts, terpenoid resin biosynthesis, and terpenoid accumulation in developing xylem of Norway spruce stems. Plant Physiol 129:1003–1018
Mauch-Mani B, Slusarenko AJ (1996) Production of salicylic acid precursors is a major function of phenylalanine ammonia-lyase in the resistance of arabidopsis to Peronospora parasitica. Plant Cell 8:203–212
Meuwly P, Molders W, Buchala A, Metraux JP (1995) Local and systemic biosynthesis of salicylic acid in infected cucumber plants. Plant Physiol 109:1107–1114
Nagy NE, Fossdal CG, Krokene P, Krekling T, Lönneborg A et al (2004) Induced responses to pathogen infection in Norway spruce phloem: changes in polyphenolic parenchyma cells, chalcone synthase transcript levels and peroxidase activity. Tree Physiology 24:505–515
Pallas J, Paiva NL, Lamb C, Dixon RA (1996) Tobacco plants epigenetically suppressed in phenylalanine ammonia-lyase expression do not develop systemic acquired resistance in response to infection by tobacco mosaic virus. Plant J 10:281–293
Palovaara J, Hakman I (2008) Conifer WOX-related homeodomain transcription factors, developmental consideration and expression dynamic of WOX2 during Picea abies somatic embryogenesis. Plant Mol Biol 66:533–549
Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST (c)) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30:10
Ralph SG, Yueh H, Friedmann M, Aeschliman D, Zeznik JA, Nelson CC, Butterfield YSN, Kirkpatrick R, Liu J, Jones SJM, Marra MA, Douglas CJ, Ritland K, Bohlmann J (2006) Conifer defence against insects: microarray gene expression profiling of Sitka spruce (Picea sitchensis) induced by mechanical wounding or feeding by spruce budworms (Choristoneura occidentalis) or white pine weevils (Pissodes strobi) reveals large-scale changes of the host transcriptome. Plant, Cell Environ 29:1545–1570
Rozen S, Skaletsky HJ (1993) Primer3. http://biotools.umassmed.edu/bioapps/primer3_www.cgi
Sato M, Tsuda K, Wang L, Coller J, Watanabe Y, Glazebrook J, Katagiri F (2010) Network modeling reveals prevalent negative regulatory relationships between signaling sectors in Arabidopsis immune signaling. PLoS Pathog 6:7
Sooriyaarachchi S, Jaber E, Covarrubias AS, Ubhayasekera W, Asiegbu FO, Mowbray SL (2011) Expression and beta-glucan binding properties of Scots pine (Pinus sylvestris L.) antimicrobial protein (Sp-AMP). Plant Mol Biol 77:33–45
Stenlid J (1985) Population structure of Heterobasidion annosum as determined by somatic incompatibility, sexual incompatibility and isoenzyme patterns. Can J Bot 63:2268–2273
Sun H, Paulin L, Alatalo E, Asiegbu FO (2011) Response of living tissues of Pinus sylvestris to the saprotrophic biocontrol fungus Phlebiopsis gigantea. Tree Physiol 31:438–451
Thaler JS, Humphrey PT, Whiteman NK (2012) Evolution of jasmonate and salicylate signal crosstalk. Trends Plant Sci 17:260–270
van Loon LC, Rep M, Pieterse CMJ (2006) Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol 44:135–162
Vestman D, Larsson E, Uddenberg D, Cairney J, Clapham D, Sundberg E, von Arnold S (2010) Important processes during differentiation and early development of somatic embryos of Norway spruce as revealed by changes in global gene expression. Tree Genet Genomes 7:347–362
Woodward S, Stenlid J, Karjalainen R, Hüttermann A (1998) Preface. In: Woodward S, Stenlid J, Karjalainen R, Hüttermann A (eds) Heterobasidion annosum: biology, ecology, impact and control. CAB International, London, pp xi–xii
Yaqoob N, Yakovlev IA, Krokene P, Kvaalen H, Solheim H, Fossdal CG (2012) Defence-related gene expression in bark and sapwood of Norway spruce in response to Heterobasidion parviporum and methyl jasmonate. Physiol Mol Plant P 77:10–16
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.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
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)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00425-012-1822-8