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The Eucalyptus grandisGenome Project: Genome and transcriptome resources for comparative analysis of woody plant biology
BMC Proceedings volume 5, Article number: I20 (2011)
The International Year of Forests - 2011 [http://www.un.org/en/events/iyof2011/] will be a milestone for forest tree genomics. The draft genome sequence of Eucalyptus grandis was released in January 2011 in the USA (Phytozome [http://www.phytozome.net]) and in Belgium (BOGAS, [http://bioinformatics.psb.ugent.be/webtools/bogas/]). The genome sequencing was funded by the US Department of Energy (DOE) and performed at the DOE Joint Genome Institute (JGI) in collaboration with members of the Eucalyptus Genome Network (EUCAGEN, [http://www.eucagen.org]) who contributed genetic materials, linkage maps, EST resources and bioinformatics support. The E. grandis genome together with that of Populus trichocarpaand other woody plant genomes recently completed (e.g. Vitis, Cacao, Prunus, Citrus and Malus)will provide excellent opportunities for comparative studies of the unique biology of woody plants. Eucalypts are currently the most widely grown hardwood fibre crop in the world and eucalypt breeding programs will benefit greatly from the new genomic resources. The reference genome sequence of Eucalyptus, a foundation tree genus in Australia comprising more than 70% of the native forest estate, will also offer important benefits for ecological and evolutionary biology studies. We report the sequencing, assembly and annotation of the E. grandis genome.
Genome sequencing and assembly
Whole-genome (8X) shotgun sequencing was performed for a partially inbred (S1), 17-year-old tree of E. grandis (est. genome size 640 Mbp, n = 11), BRASUZ1 (Suzano, Brazil). A total of 7.7 million Sanger reads (5.4 Gbp) were produced from plasmid, fosmid and BAC libraries. An inbred genotype was selected to circumvent perceived problems with the assembly of a highly heterozygous eucalypt genome. However, microsatellite genotyping showed that BRASUZ1 was much less homozygous than expected, with large parts of the genome remaining heterozygous presumably due to viability selection. This finding was confirmed during the assembly of the S1 genome - approximately 25% of the assembly occurred in two haplotypes of 3-4X coverage, while the remainder of the genome assembled into a single haplotype of 6-7X coverage. Linkage maps with over 2400 DArT and microsatellite markers were subsequently used as a framework for the assembly of 11 large chromosome scaffolds. The chromosome scaffolds contained 88% (605 Mbp) of the draft assembly, with the remainder of the assembly sequence (85 Mbp) in 4941 smaller scaffolds. Based on similarity searches with 1.6 million ESTs from BRASUZ1, it was estimated that 96% of expressed gene loci were included in the 11 chromosome assemblies.
Genome annotation was performed in parallel at the JGI and at the University of Ghent. Both annotation teams used ab initio and homology-based annotation approaches supported by over 4 million 454-FLX-Titanium ESTs produced by the JGI, as well as Sanger, 454 and Illumina EST data provided by collaborators. The two annotations revealed that the 11 chromosome scaffolds contain more than 90% of the predicted protein-coding loci (total 44,974 - JGI, 47,974 - UGent). More than 70% of the predicted genes had EST support and 9,961 (18%) alternatively spliced transcripts were detected. The two annotations are being compared and a joint annotation may be released for the main E. grandis genome paper.
The Vitis genome , representing an early diverging Rosid lineage (Vitales), was found to contain the ancient hexaploidization event shared by Rosids and Asterids, but none of the more recent genome duplications found in the Rosid lineages represented by Arabidopsis and Populus. A preliminary analysis performed at UGent of genome duplication in E. grandis (representing the Rosid order Myrtales) suggested that the Eucalyptus genome most likely contains one more recent duplication event, in addition to the paleohexaploidy event.
E. globulus is a temperate eucalypt with superior wood properties compared to E. grandis and is viewed as the premier eucalypt species for pulping. The two species occur in different sections (Maidenaria and Latoangulatae) of the subgenus Symphyomyrtus and their genome sizes differ substantially (E. globulus - 530 Mbp, E. grandis - 640 Mbp, ). The JGI has performed genome-wide resequencing (>30X Illumina PE) of an E. globulus clone (X46, Forestal Mininco, Chile). Approximately 75% of the E. globulus Illumina reads mapped to the E. grandis reference genome and sequence analysis in these regions revealed an average sequence divergence of 1.5% between the two genomes. Other eucalypt genomes currently being resequenced by collaborators will generate a valuable resource for studies of eucalypt genome evolution.
The large amount of transcriptome sequence data was produced the project includes 1.9 million xylem and leaf ESTs (454 reads) from BRASUZ1 and 2.1 million 454 reads from E. globulus (X46) xylem and leaf tissues. Together with other large 454 datasets (e.g. ) and Illumina mRNA-Seq data produced by collaborators, the Eucalyptus research community now have access to excellent transcriptome resources some of which are already available in integrated genome and transcriptome browsers (Eucspresso [http://eucspresso.bi.up.ac.za/]).
The E. grandis genome sequence will be the first reference for the Rosid order Myrtales and will be informative for comparative genomic studies within the Eudicots. It will also deliver powerful tools for the application of genomics in eucalypt breeding programs.
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Myburg, A., Grattapaglia, D., Tuskan, G. et al. The Eucalyptus grandisGenome Project: Genome and transcriptome resources for comparative analysis of woody plant biology. BMC Proc 5, I20 (2011) doi:10.1186/1753-6561-5-S7-I20
- Genome Duplication
- Joint Genome Institute
- Transcriptome Sequence Data
- Transcriptome Resource
- Chromosome Scaffold