Limits...
Physical mapping and BAC-end sequence analysis provide initial insights into the flax (Linum usitatissimum L.) genome.

Ragupathy R, Rathinavelu R, Cloutier S - BMC Genomics (2011)

Bottom Line: Compared to other plant genomes, the proportion of rDNA was found to be very high whereas the proportion of known transposable elements was low.The SSRs identified from BES will be valuable in saturating existing linkage maps and for anchoring physical and genetic maps.The physical map and paired-end reads from BAC clones will also serve as scaffolds to build and validate the whole genome shotgun assembly.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cereal Research Centre, Agriculture and Agri-Food Canada, Winnipeg, MB, Canada.

ABSTRACT

Background: Flax (Linum usitatissimum L.) is an important source of oil rich in omega-3 fatty acids, which have proven health benefits and utility as an industrial raw material. Flax seeds also contain lignans which are associated with reducing the risk of certain types of cancer. Its bast fibres have broad industrial applications. However, genomic tools needed for molecular breeding were non existent. Hence a project, Total Utilization Flax GENomics (TUFGEN) was initiated. We report here the first genome-wide physical map of flax and the generation and analysis of BAC-end sequences (BES) from 43,776 clones, providing initial insights into the genome.

Results: The physical map consists of 416 contigs spanning ~368 Mb, assembled from 32,025 fingerprints, representing roughly 54.5% to 99.4% of the estimated haploid genome (370-675 Mb). The N50 size of the contigs was estimated to be ~1,494 kb. The longest contig was ~5,562 kb comprising 437 clones. There were 96 contigs containing more than 100 clones. Approximately 54.6 Mb representing 8-14.8% of the genome was obtained from 80,337 BES. Annotation revealed that a large part of the genome consists of ribosomal DNA (~13.8%), followed by known transposable elements at 6.1%. Furthermore, ~7.4% of sequence was identified to harbour novel repeat elements. Homology searches against flax-ESTs and NCBI-ESTs suggested that ~5.6% of the transcriptome is unique to flax. A total of 4064 putative genomic SSRs were identified and are being developed as novel markers for their use in molecular breeding.

Conclusion: The first genome-wide physical map of flax constructed with BAC clones provides a framework for accessing target loci with economic importance for marker development and positional cloning. Analysis of the BES has provided insights into the uniqueness of the flax genome. Compared to other plant genomes, the proportion of rDNA was found to be very high whereas the proportion of known transposable elements was low. The SSRs identified from BES will be valuable in saturating existing linkage maps and for anchoring physical and genetic maps. The physical map and paired-end reads from BAC clones will also serve as scaffolds to build and validate the whole genome shotgun assembly.

Show MeSH

Related in: MedlinePlus

Distribution of GO-slim annotations of gene products predicted from BAC-End sequences: A. Molecular functions; B. Biological processes and C. Cellular locations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3113786&req=5

Figure 4: Distribution of GO-slim annotations of gene products predicted from BAC-End sequences: A. Molecular functions; B. Biological processes and C. Cellular locations.

Mentions: Mapping of predicted proteins from 24,962 BES to the UniProt database yielded 45,380 GO annotations as a result of multiple associations of individual predicted proteins with multiple functions, processes or components (Additional file 5: Table S5). Corresponding plant GO-slim categories were obtained for all three independent GO components namely, molecular functions (Figure 4A; Additional file 6: Table S6), biological processes (Figure 4B; Additional file 7: Table S7) and cellular components (Figure 4C; Additional file 8: Table S8). The top four GO categories for molecular function were 'binding' (19%), 'transferase activity' (15%), 'catalytic activity' (13%) and 'hydrolase activity' (13%). Similarly, in the categorization of biological processes, protein signatures associated with 'metabolic processes' (23%), 'cellular processes' (20%) and 'biosynthetic processes' (8%) were predominant among annotations representing 44 processes in total. Approximately 1.7% of the catalogued proteins were assigned with roles in lipid metabolism/catabolism.


Physical mapping and BAC-end sequence analysis provide initial insights into the flax (Linum usitatissimum L.) genome.

Ragupathy R, Rathinavelu R, Cloutier S - BMC Genomics (2011)

Distribution of GO-slim annotations of gene products predicted from BAC-End sequences: A. Molecular functions; B. Biological processes and C. Cellular locations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3113786&req=5

Figure 4: Distribution of GO-slim annotations of gene products predicted from BAC-End sequences: A. Molecular functions; B. Biological processes and C. Cellular locations.
Mentions: Mapping of predicted proteins from 24,962 BES to the UniProt database yielded 45,380 GO annotations as a result of multiple associations of individual predicted proteins with multiple functions, processes or components (Additional file 5: Table S5). Corresponding plant GO-slim categories were obtained for all three independent GO components namely, molecular functions (Figure 4A; Additional file 6: Table S6), biological processes (Figure 4B; Additional file 7: Table S7) and cellular components (Figure 4C; Additional file 8: Table S8). The top four GO categories for molecular function were 'binding' (19%), 'transferase activity' (15%), 'catalytic activity' (13%) and 'hydrolase activity' (13%). Similarly, in the categorization of biological processes, protein signatures associated with 'metabolic processes' (23%), 'cellular processes' (20%) and 'biosynthetic processes' (8%) were predominant among annotations representing 44 processes in total. Approximately 1.7% of the catalogued proteins were assigned with roles in lipid metabolism/catabolism.

Bottom Line: Compared to other plant genomes, the proportion of rDNA was found to be very high whereas the proportion of known transposable elements was low.The SSRs identified from BES will be valuable in saturating existing linkage maps and for anchoring physical and genetic maps.The physical map and paired-end reads from BAC clones will also serve as scaffolds to build and validate the whole genome shotgun assembly.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cereal Research Centre, Agriculture and Agri-Food Canada, Winnipeg, MB, Canada.

ABSTRACT

Background: Flax (Linum usitatissimum L.) is an important source of oil rich in omega-3 fatty acids, which have proven health benefits and utility as an industrial raw material. Flax seeds also contain lignans which are associated with reducing the risk of certain types of cancer. Its bast fibres have broad industrial applications. However, genomic tools needed for molecular breeding were non existent. Hence a project, Total Utilization Flax GENomics (TUFGEN) was initiated. We report here the first genome-wide physical map of flax and the generation and analysis of BAC-end sequences (BES) from 43,776 clones, providing initial insights into the genome.

Results: The physical map consists of 416 contigs spanning ~368 Mb, assembled from 32,025 fingerprints, representing roughly 54.5% to 99.4% of the estimated haploid genome (370-675 Mb). The N50 size of the contigs was estimated to be ~1,494 kb. The longest contig was ~5,562 kb comprising 437 clones. There were 96 contigs containing more than 100 clones. Approximately 54.6 Mb representing 8-14.8% of the genome was obtained from 80,337 BES. Annotation revealed that a large part of the genome consists of ribosomal DNA (~13.8%), followed by known transposable elements at 6.1%. Furthermore, ~7.4% of sequence was identified to harbour novel repeat elements. Homology searches against flax-ESTs and NCBI-ESTs suggested that ~5.6% of the transcriptome is unique to flax. A total of 4064 putative genomic SSRs were identified and are being developed as novel markers for their use in molecular breeding.

Conclusion: The first genome-wide physical map of flax constructed with BAC clones provides a framework for accessing target loci with economic importance for marker development and positional cloning. Analysis of the BES has provided insights into the uniqueness of the flax genome. Compared to other plant genomes, the proportion of rDNA was found to be very high whereas the proportion of known transposable elements was low. The SSRs identified from BES will be valuable in saturating existing linkage maps and for anchoring physical and genetic maps. The physical map and paired-end reads from BAC clones will also serve as scaffolds to build and validate the whole genome shotgun assembly.

Show MeSH
Related in: MedlinePlus