TY - JOUR
T1 - Advancing Eucalyptus genomics
T2 - Identification and sequencing of lignin biosynthesis genes from deep-coverage BAC libraries
AU - Paiva, Jorge A.P.
AU - Prat, Elisa
AU - Vautrin, Sonia
AU - Santos, Mauro D.
AU - San-Clemente, Hélène
AU - Brommonschenkel, Sérgio
AU - Fonseca, Paulo G.S.
AU - Grattapaglia, Dario
AU - Song, Xiang
AU - Ammiraju, Jetty S.S.
AU - Kudrna, David
AU - Wing, Rod A.
AU - Freitas, Ana T.
AU - Bergès, Hélène
AU - Grima-Pettenati, Jacqueline
N1 - Funding Information:
This work was partially supported by Fundação para a Ciência e Tecnologia (Portugal) (GenEglobwq project - PTDC/AGR-GPL/66564/2006). Authors also acknowledge the fundings from the ERA-PG EUCANET project n° ANR-06-ERAPG-10-03, from the Brazilian Ministry of Science and Technology (GENOLYPTUS project - FINEP grant 1755-01 and CNPq grant 520489/02-0) and a CNPq research fellowship to DG. The authors gratefully acknowledge Nathalie Ladouce for help on cDNA clones selection and amplification for probe preparation.
PY - 2011/3/4
Y1 - 2011/3/4
N2 - Background: Eucalyptus species are among the most planted hardwoods in the world because of their rapid growth, adaptability and valuable wood properties. The development and integration of genomic resources into breeding practice will be increasingly important in the decades to come. Bacterial artificial chromosome (BAC) libraries are key genomic tools that enable positional cloning of important traits, synteny evaluation, and the development of genome framework physical maps for genetic linkage and genome sequencing.Results: We describe the construction and characterization of two deep-coverage BAC libraries EG_Ba and EG_Bb obtained from nuclear DNA fragments of E. grandis (clone BRASUZ1) digested with HindIII and BstYI, respectively. Genome coverages of 17 and 15 haploid genome equivalents were estimated for EG_Ba and EG_Bb, respectively. Both libraries contained large inserts, with average sizes ranging from 135 Kb (Eg_Bb) to 157 Kb (Eg_Ba), very low extra-nuclear genome contamination providing a probability of finding a single copy gene ≥ 99.99%. Libraries were screened for the presence of several genes of interest via hybridizations to high-density BAC filters followed by PCR validation. Five selected BAC clones were sequenced and assembled using the Roche GS FLX technology providing the whole sequence of the E. grandis chloroplast genome, and complete genomic sequences of important lignin biosynthesis genes.Conclusions: The two E. grandis BAC libraries described in this study represent an important milestone for the advancement of Eucalyptus genomics and forest tree research. These BAC resources have a highly redundant genome coverage (> 15×), contain large average inserts and have a very low percentage of clones with organellar DNA or empty vectors. These publicly available BAC libraries are thus suitable for a broad range of applications in genetic and genomic research in Eucalyptus and possibly in related species of Myrtaceae, including genome sequencing, gene isolation, functional and comparative genomics. Because they have been constructed using the same tree (E. grandis BRASUZ1) whose full genome is being sequenced, they should prove instrumental for assembly and gap filling of the upcoming Eucalyptus reference genome sequence.
AB - Background: Eucalyptus species are among the most planted hardwoods in the world because of their rapid growth, adaptability and valuable wood properties. The development and integration of genomic resources into breeding practice will be increasingly important in the decades to come. Bacterial artificial chromosome (BAC) libraries are key genomic tools that enable positional cloning of important traits, synteny evaluation, and the development of genome framework physical maps for genetic linkage and genome sequencing.Results: We describe the construction and characterization of two deep-coverage BAC libraries EG_Ba and EG_Bb obtained from nuclear DNA fragments of E. grandis (clone BRASUZ1) digested with HindIII and BstYI, respectively. Genome coverages of 17 and 15 haploid genome equivalents were estimated for EG_Ba and EG_Bb, respectively. Both libraries contained large inserts, with average sizes ranging from 135 Kb (Eg_Bb) to 157 Kb (Eg_Ba), very low extra-nuclear genome contamination providing a probability of finding a single copy gene ≥ 99.99%. Libraries were screened for the presence of several genes of interest via hybridizations to high-density BAC filters followed by PCR validation. Five selected BAC clones were sequenced and assembled using the Roche GS FLX technology providing the whole sequence of the E. grandis chloroplast genome, and complete genomic sequences of important lignin biosynthesis genes.Conclusions: The two E. grandis BAC libraries described in this study represent an important milestone for the advancement of Eucalyptus genomics and forest tree research. These BAC resources have a highly redundant genome coverage (> 15×), contain large average inserts and have a very low percentage of clones with organellar DNA or empty vectors. These publicly available BAC libraries are thus suitable for a broad range of applications in genetic and genomic research in Eucalyptus and possibly in related species of Myrtaceae, including genome sequencing, gene isolation, functional and comparative genomics. Because they have been constructed using the same tree (E. grandis BRASUZ1) whose full genome is being sequenced, they should prove instrumental for assembly and gap filling of the upcoming Eucalyptus reference genome sequence.
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U2 - 10.1186/1471-2164-12-137
DO - 10.1186/1471-2164-12-137
M3 - Article
C2 - 21375742
AN - SCOPUS:79952202673
VL - 12
JO - BMC Genomics
JF - BMC Genomics
SN - 1471-2164
M1 - 137
ER -