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

ABSTRACT: 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

ORGANISM(S): Pinus sylvestris Pinus taeda

PROVIDER: GSE2089 | GEO | 2004/12/21

SECONDARY ACCESSION(S): PRJNA104755

REPOSITORIES: GEO

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Project description:A pair of nearly isogenic tomato (Solanum lycopersicum) lines were used. These are: Motelle (Mi/Mi), root-knot nematode resistant and Moneymaker (mi/mi) root-knot nematode susceptible. A potato aphid (red biotype), imported from France, was used for infestation. An equal volume of sterile 1% carboxymethyl cellulose in water was added to the cleaned nematodes to keep them in suspension. The inoculum, consisting of 75-100 J2s, was dispensed in 10 µl aliquots using a repeat pipetter. The inoculum was pipetted 2 mm beneath the root tips. Control seedlings were inoculated with 0.5% carboxymethyl cellulose. After the desired incubation time (24 h or 36 h), seedlings were taken out of the boxes and a few set aside to check nematode infection rate. In our hand, this system results in 50% infection rate. The rest of the seedlings were placed on a water-saturated filter paper and the roots aligned. About 1 cm of the infected root tip was excised with a blade and quickly frozen on a polycarbonate membrane-lined 50 mm diameter Petri dish floating on liquid nitrogen and stored at -80C. Plants were grown for 6 weeks in a growth chamber under 16 h light/8 h dark at 25C in UCMix/sand, irrigated with water or MiracleGroR solution, then moved to an insect growth room (25C) 4-6 days before aphid infestation. Individual tomato leaflets were infested with 30-40 mixed stages of the potato aphids using plastic leaf cages. Four leaflets and two plants of each genotype were infested. Two plants of each genotype were used as controls. After the desired infestation time (6, 12, 24 or 48 h), leaflets were excised from the plants with a blade and sprayed with 1% SDS to force aphids to remove their stylets from the leaf tissue. Aphids were carefully removed from the leaflets and quickly frozen in liquid nitrogen. Control leaflets were excised and sprayed with 1% SDS before freezing. Samples were stored at -80C. RNA was isolated using the hot phenol method. Keywords: Direct comparison

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

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