Project description:Illumina technology was used to generate mRNA profiles of a time course of Laccaria bicolor S238N and Populus tremula x alba 717-1B4 in vitro ectomycorrhizal development. Total RNA was extracted, TruSeq mRNA Stranded libraries were constructed and and sequenced in triplicates (2 x 150 bp Illumina HiSeq3000) at the Genotoul sequencing facilities (Toulouse, France). Raw reads were trimmed for low quality (quality score 0.05), Illumina adapters and sequences shorter than 15 nucleotides and aligned to the L. bicolor v2 reference transcripts available at the JGI database https://mycocosm.jgi.doe.gov/Lacbi2/Lacbi2.home.html using CLC Genomics Workbench v8. Overall design: mRNA profiles of a time course of Laccaria bicolor S238N and Populus tremula x alba 717-1B4 in vitro ectomycorrhizal development were generated by Illumina HiSeq 3000 (2x150bp). Three biological replicates were sequenced."
Project description:Complete Populus genome sequences are available for the nucleus (P. trichocarpa; section Tacamahaca) and for chloroplasts (seven species), but not for mitochondria. Here, we provide the complete genome sequences of the chloroplast and the mitochondrion for the clones P. tremula W52 and P. tremula x P. alba 717-1B4 (section Populus). The organization of the chloroplast genomes of both Populus clones is described. A phylogenetic tree constructed from all available complete chloroplast DNA sequences of Populus was not congruent with the assignment of the related species to different Populus sections. In total, 3,024 variable nucleotide positions were identified among all compared Populus chloroplast DNA sequences. The 5-prime part of the LSC from trnH to atpA showed the highest frequency of variations. The variable positions included 163 positions with SNPs allowing for differentiating the two clones with P. tremula chloroplast genomes (W52, 717-1B4) from the other seven Populus individuals. These potential P. tremula-specific SNPs were displayed as a whole-plastome barcode on the P. tremula W52 chloroplast DNA sequence. Three of these SNPs and one InDel in the trnH-psbA linker were successfully validated by Sanger sequencing in an extended set of Populus individuals. The complete mitochondrial genome sequence of P. tremula is the first in the family of Salicaceae. The mitochondrial genomes of the two clones are 783,442 bp (W52) and 783,513 bp (717-1B4) in size, structurally very similar and organized as single circles. DNA sequence regions with high similarity to the W52 chloroplast sequence account for about 2% of the W52 mitochondrial genome. The mean SNP frequency was found to be nearly six fold higher in the chloroplast than in the mitochondrial genome when comparing 717-1B4 with W52. The availability of the genomic information of all three DNA-containing cell organelles will allow a holistic approach in poplar molecular breeding in the future.
Project description:Illumina HiSeq technology was used to generate mRNA profiles from Populus tremula x alba INRA 717-1B4 roots treated with Methyl jasmonate. Samples were harvested after two weeks either from untreated control roots or from Methyl jasmonate treated roots. Paired-end (2X100bp) reads were generated and aligned to the Populus trichocarpa (http://www.phytozome.net/poplar.php) using CLC Genomics Workbench 6. mRNA profiles from Populus tremula x alba INRA 717-1B4 roots treated with Methyl jasmonate as well as from control roots were generated by paired-end (2X100bp) Illumina HiSeq sequencing. Four samples were sequenced per lane, two biological replicates per treatment.
Project description:The Poplar transcriptome was analyzed in mycorrhizal root tips in contact with Laccaria bicolor for 2 weeks. During mycorrhization the roots were treated with either 250µm ACC, 10nM JA or 500µM SA and compared to untreated mycorrhiza or control roots without contact to L. bicolor. In addition the poplar mutants 35S::PttACO1 and 35S::Atetr1 were used We performed 27 hybridizations (NimbleGen) with samples derived from Populus tremula x Populus alba clone 717-1B4 control roots, untreated mycorrhiza, SA-treated mycorrhiza, ACC-treated mycorrhiza and JA-treated mycorrhiza (3 biological replicates each) as well as Populus tremula x Populus tremuloides T89 control roots, mycorrhiza, 35S::PttACO1 mycorrhiza and 35S::Atetr1-1 mycorrhiza (3 biological replicates). All samples were labeled with Cy3.