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Transcriptional response of Scots pine trees to Heterobasidion annosum infection: analyses of antimicrobial protein Sp-AMP2

ABSTRACT: Understanding of mechanisms of resistance of forest trees against microbial pathogens is an essential prerequisite for the development of sustainable forestry practices and for the improvement of commercially-grown trees via either conventional breeding or rational genetic engineering. We have studied the transcriptional response of Scots pine trees to Heterobasidion annosum infection under field conditions. By comparing responses of trees to wounding and to fungal inoculation we could identify a set of genes that were specifically responding to fungal infection. We have also investigated a contribution of Scots pine antimicrobial protein Sp-AMP2 to the host antimicrobial defense to evaluate the potential of Sp-AMP genes as molecular markers for resistance breeding.

ORGANISM(S): Pinus sylvestris

PROVIDER: GSE87881 | GEO | 2017/12/31

SECONDARY ACCESSION(S): PRJNA348098

REPOSITORIES: GEO

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Transcript profiling of Pinus sylvestris trees as a response to Heterobasidion annosum infection under field conditions

Project description:In conifer forests of Northern Europe, a pathogenic fungus Heterobasidion annosum attacks the roots of Scots pine and causes mortality. Trees with infection grow slower and produce less timber with reduced quality. Despite applied control methods, such as switching tree species to a non-host species, or stump treatment, root and butt rot continues to be a serious forest health problem. Disease resistance breeding is a less-applied control method which has potential to improve tree health. However, neither conifer genotypes with absolute resistance to Heterobasidion sp. nor robust selection markers for resistance breeding have been found. We studied the responses of various Scots pine genotypes to Heterobasidion annosum infection and mechanic damage in drained peatland. Stems and roots of mature naturally regenerated Scots pine trees growing in drained peatland were either artificially infected with H. annosum or wounded and inoculated with sterile inoculum. Untreated trees from the study sites served as controls. Responses of different Scots pine genotypes to pathogen infection as determined by lesion size were recorded from samples harvested four months after inoculation, and least susceptible and highly susceptible genotypes were selected from the study material. Analysis of terpenoids from both least susceptible and highly susceptible pine genotypes by gas chromatography coupled with mass spectrometry indicates that some monoterpenes and sesquiterpenes are differentially induced depending on the susceptibility level. Transcriptomic microarray analysis was therefore conducted with RNA from stems of the least susceptible and highly susceptible Scots pine genotypes. Gene expression data from cDNA microarray were analysed by comparisons between the treatments, and the genotypes with different resistance level. The aim of the study is to highlight transcripts specific to differing levels of susceptibility.

2019-02-01 | GSE66168 | GEO

Comparative transcriptome analysis of early interaction events in Scots pine root tissues following challenge (1dpi)

Project description:Comparative transcriptome analysis of early interaction events in Scots pine root tissues following challenge with a pathogenic, saprophytic or symbiotic fungus. Seedlings of P. sylvestris (19 days post germination) were transferred to wet, sterile filter paper on Petri-plates. Thereafter, the roots of the seedlings were inoculated with the mycelial homogenate of either Heterobasidion annosum (FP5, P-type) a pathogenic root rot fungus which attacks Norway spruce, Scots pine and broad leaf trees or Laccaria bicolor, an obligate ectomycorrhizal symbiont or Trichoderma aureoviride- an obligate saprotroph. Thereafter, incubated for 30 minutes, during which time some hyphae adhered to the roots. The inoculated seedlings (ten) were then transferred to another wet sterile filter paper placed on 1% water agar in Petri dishes. A second set of moist sterile filter paper was laid over the roots. The region of the Petri-dish containing the roots was covered with aluminium foil and the edges of the plate sealed with parafilm. The seedlings were then incubated for 24 hr under a photoperiod of 16h light at 20 ÂºC. Control seedlings were âinoculatedâ with sterile distilled water.

2010-05-27 | E-GEOD-2089 | biostudies-arrayexpress

Comparative transcriptome analysis of early interaction events in Scots pine root tissues following challenge (1dpi)

Project description:Comparative transcriptome analysis of early interaction events in Scots pine root tissues following challenge with a pathogenic, saprophytic or symbiotic fungus. Seedlings of P. sylvestris (19 days post germination) were transferred to wet, sterile filter paper on Petri-plates. Thereafter, the roots of the seedlings were inoculated with the mycelial homogenate of either Heterobasidion annosum (FP5, P-type) a pathogenic root rot fungus which attacks Norway spruce, Scots pine and broad leaf trees or Laccaria bicolor, an obligate ectomycorrhizal symbiont or Trichoderma aureoviride- an obligate saprotroph. Thereafter, incubated for 30 minutes, during which time some hyphae adhered to the roots. The inoculated seedlings (ten) were then transferred to another wet sterile filter paper placed on 1% water agar in Petri dishes. A second set of moist sterile filter paper was laid over the roots. The region of the Petri-dish containing the roots was covered with aluminium foil and the edges of the plate sealed with parafilm. The seedlings were then incubated for 24 hr under a photoperiod of 16h light at 20 ºC. Control seedlings were ‘inoculated’ with sterile distilled water. Keywords: other

2004-12-21 | GSE2089 | GEO

Heterobasidion annosum gene expression from different tissues and mycelium grown on different substrates and under different biotic and abiotic stresses

Project description:Transcript profiles of H. annosum from different tissues and mycelium grown on different substrates and under different stresses were analyzed. The array probes were designed from gene models taken from the Joint Genome Institute (JGI, Department of Energy) H. irregulare genome sequence version 1. One aim of this study was to compare gene expression profiles of H. annosum grown on different substrates and under different biotic and abiotic stresses. We performed hybridizations (NimbleGen) with samples derived from H. annosum grown in either liquid Malt Extract Medium at 20Â°C (three biological replicates), under NaCl stress (three biological replicates), under CaCl2 stress (three biological replicates), at 8Â°C (three biological replicates), at 27Â°C (three biological replicates), saprotrophic growth on Scots pine sapwood (three biological replicates), bark (three biological replicates) or heartwood (three biological replicates). Additional hybridizations were performed with RNA from necrotrophic growth of Heterobasidion on Scots pine xylem (three biological replicates), necrotrophic growth on Scots pine phloem (three biological replicates), under nutrient starvation (three biological replicates) or under oxidative stress (three biological replicates). The Heterobasidion irregulare custom-exon expression array (4 x 72K) manufactured by Roche NimbleGen Systems Limited (Madison, WI) (http://www.nimblegen.com/products/exp/index.html) contained five independent, non-identical, 60-mer probes per gene model coding sequence. For 12,199 of the 12,299 annotated protein-coding gene models, probes could be designed. For 19 gene models, no probes could be generated, and 81 gene models shared all five probes with other gene models. Included in the array were 916 random 60-mer control probes and labelling controls. For 2032 randomly chosen gene models, technical duplicates were included on the array.

2013-11-01 | E-GEOD-39805 | biostudies-arrayexpress

Scots pine (Pinus sylvestris) differentially interacts with Cenococcum geophilum and other simultaneously inoculated mycorrhizal (ECM) fungi

Project description:To identify specific gene networks induced in host roots by C. geophilum, we inoculated seedlings of Scots pine simultaneously with C. geophilum and either Suillus granulatus or Rhizopogon roseolus, two common ECM fungi associated to pines. We then measured the differential expression of Scots pine genes in the respective mycorrhizas using oligoarrays.

2016-08-01 | GSE80549 | GEO

Pinus sylvestris

Project description:Transcriptional response of Scots pine trees to Heterobasidion annosum infection: analyses of antimicrobial protein Sp-AMP2

| PRJNA348098 | ENA

Microarray assay of the genetic response of Picea abies to Heterobasidion annosum infection - Loop1

Project description:The hypothesis of the experiment is that infected trees of high resistance express a wider variety of resistance genes than infected trees of low resistance, and that the level of expression of these resistance genes differs between infected and healthy branches. Also, some genes highly expressed in the infected state not expressed in the healthy state may be in response to the wounding rather than the actual infection. By comparing these expressions to that of wounded, uninfected branches, this could also be clarified. Three different Picea abies clones, of increasing resistance to fungal attack, have been infected. For all clones, three rametes have been infected with Heterobasidion of 1 month, by means of cutting the cambium and inoculating a piece of fungus growing in agar in the wound. For the low and medium resistant clone, a fourth ramete has been cut and inoculated with agar not containing any fungal material.

2008-05-03 | E-GEOD-10058 | biostudies-arrayexpress

Microarray assay of the genetic response of Picea abies to Heterobasidion annosum infection - Loop2

2008-05-03 | E-GEOD-10059 | biostudies-arrayexpress

Comparative transcript profiling of interaction events in Pinus sylvestris root tissues following challenge (5dpi).

Project description:The response mechanisms, recognition and specificity of conifer trees during interaction with pathogenic, saprotrophic or symbiotic ectomycorrhizal fungus were investigated. The roots of Pinus sylvestris were challenged for five days with either Heterobasidion annosum (a pathogenic root rot fungus which attacks Norway spruce, Scots pine and broad leaf trees); Laccaria bicolor (an obligate ectomycorrhizal symbiont); or Trichoderma aureoviride (an obligate saprotroph). The gene expression data from cDNA micro-arrays consisting of 2176 Pinus taeda genes were analysed using 2-interconnected mixed linear model statistical approach. The result of the pairwise comparisons of the different treatments against un-inoculated control led to identification of genes specifically differentially expressed in the pathogenic, saprotrophic and symbiotic interactions. The results were compared with similar data obtained for two other interaction stages: 1 and 15 days post inoculation. The result of this comprehensive expression profiling will hopefully shed more light on the mechanistic basis for recognition and response of conifer trees to pathogenic and non-pathogenic fungi. Keywords: stress response

2006-07-31 | GSE5408 | GEO

Comparative transcript profiling of interaction events in Pinus sylvestris root tissues following challenge (15dpi).

Project description:The response mechanisms, recognition and specificity of conifer trees during interaction with pathogenic, saprotrophic or symbiotic ectomycorrhizal fungus were investigated. The roots of Pinus sylvestris were challenged for fifteen days with either Heterobasidion annosum (a pathogenic root rot fungus which attacks Norway spruce, Scots pine and broad leaf trees); Laccaria bicolor (an obligate ectomycorrhizal symbiont); or Trichoderma aureoviride (an obligate saprotroph). The gene expression data from cDNA micro-arrays consisting of 2176 Pinus taeda genes were analysed using 2-interconnected mixed linear model statistical approach. The result of the pairwise comparisons of the different treatments against un-inoculated control led to identification of genes specifically differentially expressed in the pathogenic, saprotrophic and symbiotic interactions. The results were compared with similar data obtained for two other interaction stages: 1 and 5 days post inoculation. The result of this comprehensive expression profiling will hopefully shed more light on the mechanistic basis for recognition and response of conifer trees to pathogenic and non-pathogenic fungi. Keywords: stress response

2006-07-31 | GSE5407 | GEO

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