The molecular timecourse of hybrid aspen/Laccaria bicolor ectomycorrhiza development

The aim of this temporal transcriptomics study is to identify key genes regulated in hybrid aspen root tips and the ectomycorrhizal fungus Laccaria bicolor during their interaction and in comparison to control roots or free living mycelium at various time points. It will reveal the dynamics of expression of our key genes within the pectin modulation and hormone pathways, as well as novel target genes in both organisms. Experiments have been performed under sterile condition and material has been analysed from 3, 7, 14, 21 and 28days of interaction from the following conditions: control roots without fungus (5 lateral root tips/sample), roots in interaction with the fungus (5 root tips/sample, note: these samples contain material from both fungus and plant) as well as free living mycelia as fungal control. For control roots and roots in interaction with the fungus, each 4 replicate samples were prepared per timepoint. For free living mycelia 3 replicate samples were prepared. This makes a total of 55 samples. RNA was extracted and turned into cDNA using smart-seq2 (Picelli et al. 2014, Nature Protocols) which hence was used at SciLifeLab for generating sequencing libraries using Rubicon Thruplex DNA-seq kit (for low amount of input material) and sequencing on one Illumina NovaSeq6000 S2 flowcell lane, 2x 150bp reads.