title: The mycorrhiza helper Pseudomonas fluorescens BBc6R8 has a specific priming effect on the growth, morphology and gene expression of the ectomycorrhizal fungus Laccaria bicolor S238N.
abstract: The mycorrhiza helper Pseudomonas fluorescens BBc6R8 promotes the presymbiotic survival and growth of the ectomycorrhizal fungus Laccaria bicolor S238N in the soil. An in vitro fungal-bacterial confrontation bioassay mimicking the promoting effects of the bacteria on fungal growth was set up to analyse the fungal morphological and transcriptional changes induced by the helper bacteria at three successive stages of the interaction. The specificity of the P. fluorescens BBc6R8 effect was assessed in comparison with six other rhizobacterial strains possessing mycorrhiza helper or pathogen antagonistic abilities. The helper BBc6R8 strain was the only strain to induce increases in the radial growth of the colony, hyphal apex density and branching angle. These morphological modifications were coupled with pleiotropic alterations of the fungal transcriptome, which varied throughout the interaction. Early stage-responsive genes were presumably involved in recognition processes and transcription regulation, while late stage-responsive genes encoded proteins of primary metabolism. Some of the responsive genes were partly specific to the interaction with P. fluorescens BBc6R8, whereas others were mutually regulated by different rhizobacteria. The results highlight the fact that the helper BBc6R8 strain has a specific priming effect on growth, morphology and gene expression of its fungal associate L. bicolor S238N.
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: A
authors: A
authors: Deveau
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: B
authors: B
authors: Palin
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: C
authors: C
authors: Delaruelle
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: M
authors: M
authors: Peter
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: A
authors: A
authors: Kohler
authors: INRA-ENGREF, UMR1092, Unité Dynamique des Systèmes Forestiers, 54042 Nancy, France.
authors: J C
authors: JC
authors: Pierrat
authors: INRA-Agrocampus Rennes, UMR1099 'Biologie des Organismes et des Populations appliquée à la Protection des Plantes', 35 653 Le Rheu Cedex, France.
authors: A
authors: A
authors: Sarniguet
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: J
authors: J
authors: Garbaye
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: F
authors: F
authors: Martin
authors: INRA, UMR1136 'Interactions Arbres/Micro-organismes', IFR 110 Centre de Nancy, 54280 Champenoux, France.
authors: P
authors: P
authors: Frey-Klett
mesh_terms: D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D025282:Mycelium / Q000254:growth & development; D038821:Mycorrhizae / Q000235:genetics / Q000502:physiology*; D011551:Pseudomonas fluorescens / Q000145:classification / Q000502:physiology*; D012331:RNA, Fungal / Q000235:genetics; D014158:Transcription, Genetic
chemical_list: D004271:DNA, Fungal; D005656:Fungal Proteins; D012331:RNA, Fungal
references: Akiyama K, Matsuzaki K-I, Hayashi H. 2005. Plant sesquiterpenes induce hyphal branching in arbuscular mycorrhizal fungi. Nature 435: 824-827.
references: Allard S, Utley R, Savard J, Clarke A, Grant P, Brandl C, Pillus L, Workman J, Côté J. 1999. NuA4, an essential transcription adaptor/histone H4 acetyltransferase complex containing Esa1p and the ATM-related cofactor Tra1p. EMBO Journal 18: 5108-5119.
references: Aspray T, Frey-Klett P, Jones JE, Whipps J, Garbay J, Bending G. 2006. Mycorrhization helper bacteria: a case of specificity for altering ectomycorrhiza architecture but not ectomycorrhiza formation. Mycorrhiza. 16: 533-541.
references: Becker D, Bagley S, Podila G. 1999. Effects of mycorrhizal-associated streptomycetes on growth of Laccaria bicolor, Cenococcum geophilum, and Armillaria species and on gene expression in Laccaria bicolor. Mycologia 91: 33-40.
references: Bending G. 2007. What are the mechanisms and specificity of mycorrhization helper bacteria? New Phytologist 174: 707-710.
references: Bertaux J, Schmid M, Chemidlin Prevost-Boure N, Churin JL, Hartmann A, Garbaye J, Frey-Klett P. 2003. In situ identification of intracellular bacteria related to Paenibacillus spp. in the mycelium of the ectomycorrhizal fungus Laccaria bicolor S238N. Applied and Environmental Microbiology 69: 4243-4248.
references: Besserer A, Puech-Pagès V, Kiefer P, Gomez-Roldan V, Jauneau A, Roy S, Portais J-C, Roux C, Bécard G, Séjalon-Delmas N. 2006. Strigolactones stimulate arbuscular mycorrhizal fungi by activating mitochondria. PLoS Biology 4: 1239-1247.
references: De Boer W, Leveau J, Kowalschuk G, Klein Gunnewiek P, Abeln E, Figge M, Sjollema K, Janse J, Van Veen J. 2004. Collimonas fungivorans General. Nov, sp. Nov, a chitinolytic soil bacterium with the ability to grow on living fungal hyphae. International Journal of Systematic and Evolutionnary Microbiology 54: 857-864.
references: Bolwerk A, Lagopodi A, Wijfjes A, Lamers G, Chin A, -Woeng T, Lugtenberg B, Bloemberg G. 2003. Interactions in the tomato rhizosphere of two Pseudomonas biocontrol strains with the phytopathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersci. Molecular Plant-Microbe Interactions 16: 983-993.
references: Brown C, Howe L, Soussa K, Alley S, Carroza M, Tan S, Workman J. 2001. Recruitment of HAT complexes by direct activator interactions with the ATM-related Tra 1 subunit. Sciences 292: 2333-2337.
references: Brulé C, Frey-Klett P, Pierrat J-C, Courrier S, Gérard F, Lemoin M-C, Roussellet J-L, Sommer J, Garbaye J. 2001. Survival in the soil of the ectomycorrhizal fungus Laccaria bicolor and effect of a mycorrhiza helper Pseudomonas fluorescens. Soil Biology and Biochemistry 33: 1683-1694.
references: Chapon A, Guillerm A-Y, Delalande L, Lebreton L, Sarniguet A. 2002. Dominant colonisation of wheat roots by Pseudomonas fluorescens Pf29A and selection of the indigenous microflora in the presence of the take-all fungus. European Journal of Plant Pathology 108: 449-459.
references: Cosma MP. 2002. Ordered recruitment: gene-specific mechanism of transcription activation. Molecular Cell 10: 227-236.
references: Di Battista C, Selosse M-A, Bouchard D, Senström E, Le Tacon F. 1996. Variations in symbiotic efficiency, phenotypic charachters and ploidy level among different isolates of the ectomycorrhizal basidiomycete Laccaria bicolor strain S238. Mycological Research 100: 1315-1324.
references: Duplessis S, Courty PE, Tagu D, Martin F. 2005. Transcript patterns associated with ectomycorrhiza development in Eucalyptus globulus and Pisolithus microcarpus. New Phytologist 165: 599-611.
references: Duponnois R, Garbaye J. 1991. Effect of dual inoculation of Douglas fir with the ectomycorrhizal fungus Laccaria laccata and mycorrhization helper bacteria (MHB) in two bare-root forest nurseries. Plant Soil 138: 169-176.
references: Duponnois R, Plenchette C. 2003. A mycorrhizal helper bacterium enhances ectomycorrhizal and endomycorrhizal symbiosis of Australian Acacia species. Mycorrhiza 13: 85-91.
references: Founoune H, Duponnois R, Bâ AM, Sall S, Branget I, Lorquin J, Neyra M, Chotte JL. 2002. Mycorrhiza helper bacteria stimulate ectomycorrhizal symbiosis of Accacia holoserica with the Pisolithus albus. New Phytologist 153: 81-89.
references: Frey-Klett P, Garbaye J. 2005. Mycorrhiza helper bacteria: a promising model for the genomic analysis of fungal-bacterial interactions. New Phytologist 168: 4-8.
references: Frey-Klett P, Pierrat J-C, Garbaye J. 1997. Location and survival of mycorrhizal helper Pseudomonas fluorescens during establishment of ectomycorrhizal symbiosis between Laccaria bicolor and Douglas fir. Applied and Environmental Microbiology 63: 139-144.
references: Garbaye J. 1994. Mycorrhiza helper bacteria: a new dimension to the mycorrhizal symbiosis. New Phytologist 128: 197-210.
references: Gasch A, Werner-Washburne M. 2002. The genomics of yeast responses to environmental stress and starvation. Functional & Integrative Genomics 2: 181-192.
references: Hildebrandt U, Ouziad F, Marner F-J, Bothe H. 2006. The bacterium Paenibacillus validus stimulates growth of the arbuscular mycorrhizal fungus Glomus intraradices up to the formation of fertile spores. FEMS Microbiology Letters 254: 258-267.
references: Huh C, Aldrich J, Mottahedeh J, Kwon H, Johnson C, Mrsh R. 1998. Cloning and characterization of Physarum polycepharum tectonins. Homologues of limulus lectin L6. Journal of Biological Chemistry 273: 6565-6574.
references: Kohler A, Delaruelle C, Martin D, Encelot N, Martin F. 2003. The poplar root transcriptome: analysis of 7000 expressed sequence tags. FEBS Letters 542: 37-41.
references: Le Quéré A, Eriksen KA, Rajashekar B, Schütznedübel A, Canbäck B, Johansson T, Tunlid A. 2006. Screening for rapidly evolving genes in the ectomycorrhizal fungus Paxillus involutus using cDNA microarrays. Molecular Ecology 15: 535-550.
references: Livak K, Schmittgen D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods. 25: 402-408.
references: Maier A, Riedlinger J, Fiedler H-P, Hampp R. 2004. Actinomycetales bacteria from a spruce strand: characterization and effects on growth of root symbiotic, and plant parasitic soil fungi in dual culture. Mycological Progress 3: 129-136.
references: Martin F, Duplessis S, Ditengou F, Lagrange H, Voiblet C, Lapeyrie F. 2001. Developmental cross talking in the ectomycorrhizal symbiosis: signals and communication genes. New Phytologist 151: 145-154.
references: Melin P, Schnürer J, Wagner EGH. 2002. Proteome analysis of Aspergillus nidulans reveals proteins associated with the response to the antibiotic concanamycin A, produced by Streptomyces species. Molecular Genetics and Genomics 267: 695-702.
references: Morel M, Jacob C, Kohler A, Johansson T, Martin F, Chalot M, Brun A. 2005. Identification of genes differentially expressed in extraradical mycelium and ectomycorrhizal roots during Paxillus involutus-Betula pendula ectomycorrhizal symbiosis. Applied and Environmental Microbiology 71: 382-391.
references: Paschlewski R, Pachlewska J. 1974. Studies on symbiotic properties of mycorrhizal fungi of pine (Pinus sylvestris L.) with the aid of the method of mycorrhizal synthesis in pure cultures on agar. (Translated Into English by Royal Dzieciolowski). Warsaw, Poland: Forest Research Institute.
references: Peter M, Courty P-E, Kohler A, Delaruelle C, Martin D, Tagu D, Frey-Klett P, Duplessis S, Chalot M, Podila G, Martin F. 2003. Analysis of expressed sequenced tags from the ectomycorrhizal basidiomycetes Laccaria bicolor and Pisolithus macrocarpus. New Phytologist 159: 117-129.
references: Peterson L, Bonfante P. 1994. Comparative structure of vesicular-arbuscular mycorrhizas and ectomycorrhizas. Plant and Soil 159: 79-88.
references: Podila G, Zheng J, Balasubramanian S, Sundaram S, Hiremath S, Brand J, Hymes M. 2002. Fungal gene expression in early symbiotic interactions between Laccaria bicolor and red pine. Plant and Soil 244: 117-128.
references: Poole E, Bending G, Whipps J, Read D. 2001. Bacteria associated with Pinus sylvestris-Lactarius rufus ectomycorrhizas and their effects on mycorrhizal formation in vitro. New Phytologist 151: 743-751.
references: Pray-Grant M, Schieltz D, McM, Ahon S, Wood J, Cook R, Workman J, Yates J, IIIGrant P. 2002. The novel SLIK histone acetyltransferase complex functions in the yeast retrograde response pathway. Molecular and Cellular Biology 22: 8774-8786.
references: Requena N, Füller P, Franken P. 1999. Molecular charcterization of GmFOX2 an evolutionary highly conserved gene from the mycorrhizal fungus Glomus mosseae, down regulated during interaction with rhizobacteria. Molecular Plant-Microbe Interactions 12: 934-942.
references: Riedlinger J, Schrey S, Tarkka M, Hampp R, Kapur M, Fielder H-P. 2006. Auxofuran, a novel metabolite that stimulates the growth of fly agaric, is produced by the mycorrhiza helper bacterium Streptomyces strain AcH 505. Applied and Environmental Microbiology 72: 3350-3557.
references: Sambrook J, Fritsch E, Maniatis T. 1989. Molecular cloning: a laboratory manual, 2nd edn . Cold Spring Harbor, NY, USA: Cold Spring Harbor Laboratory Press.
references: Schrey S, Schellhammer M, Ecke M, Hampp R, Tarkka M. 2005. Mycorrhiza helper bacterium Streptomyces AcH 505 induces differential gene expression in the ectomycorrhizal fungus Amanita muscaria. New Phytologist 168: 205-216.
references: Sen RE-L, Nurmiaho-Lassila K, Haahtela et T, Korhonen. 1994. Attachment of Pseudomonas fluorescense trains to the cell walls of ectomycorrhizal fungi. In: Azcon-Aguilar C, Barea JM, ed. Mycorhizas in integrated systems from genes to plant development. European Commission EUR 16728 EN. Luxembourg, 661-664.
references: Smith S, Read D. 1997. Mycorrhizal symbiosis, 2nd edn . London, UK: Academic Press.
references: Sterner D, Belotserkovskaya R, Berger S. 2002. SALSA, a variant of yeast SAGA, contains truncated Spt7, which correlates with activated transcription. Proceedings of the National Academy of Sciences, USA 99: 11622-11627.
references: Ström K, Schnürer Melin P. 2005. Co-cultivation of antifungal Lactobacillus plantarum MiLAB 393 and Aspergillus nidulans, evaluation of effects on fungal growth and protein expression. FEMS Microbiology Letters 246: 119-124.
references: Tachibana C, Yoo J, Tagne J-B, Kacherovsky N, Lee T, Young E. 2005. Combined global localization analysis and transcriptome data to identify genes that are directly coregulated by Adr1 and Cat8. Molecular Cell and Biology 25: 2138-2146.
references: Wu J, Zhang N, Hayes A, Panoutsopoulou K, Oliver S. 2004. Global analysis of nutrient control of gene expression in Saccharomyces cerevisiae during growth. Proceedings of the National Academy of Sciences, USA 101: 3148-3153.
references: Xie X, Wilkinson H, Correa A, Lewis Z, Bell-Pederson D, Ebbole D. 2004. Transcriptional response to glucose starvation and functional analysis of a glucose transporter of Neurospora crassa. Fungal Genetics and Biology 41: 1104-1119.