http://en.wikipedia.org/wiki/Picea_glaucaAnnotating the cytochrome P450CYP720B gene family in white spruceShaun Jackman201...
White sprucePicea glauca               http://en.wikipedia.org/wiki/Picea_glauca   2
Cytochrome P450✤   Enzymes involved in    biosynthesis and    metabolism✤   Catalyze the oxidation of    organic compounds...
CYP720B gene family                                          http://en.wikipedia.org/wiki/Mountain_pine_beetle✤   Conifer-...
CYP720B in sitka sprucePicea sitchensis✤   Twelve CYP720B genes    identified in sitka spruce                      http://e...
White spruce sequencing dataPicea glauca✤   RNA-seq of eight tissues:    bark, embryo, flush bud,    mature needle,    mega...
Identify white spruce transcripts✤   Assemble the white spruce RNA-seq data    usingTrinity (Grabherr et al. 2011) (assemb...
White spruce CYP720B transcripts                       Sitka spruce                       White spruce                    ...
Assemble the white spruce genome✤   Assemble the white spruce genome sequence using    ABySS (Simpson et al. 2009)✤   Asse...
Identify white spruce genes✤   Identify CYP720B genomic contigs by aligning the sitka    spruce and white spruce transcrip...
CYP720B4✤   Expressed in both sitka spruce and white spruce                                                      White spr...
CYP720B7✤   Expressed only in sitka spruce                                     White spruce                               ...
CYP720B15/16/17 related✤   White spruce transcripts align, but no sitka spruce transcripts                                ...
Cytochrome P450 CYP720B Genes      Sitka spruce   White spruce   White spruce          EST         RNA-seq     genome asse...
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Annotating the cytochrome P450 CYP720B gene family in white spruce

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Annotating the cytochrome P450 CYP720B gene family in white spruce

  1. 1. http://en.wikipedia.org/wiki/Picea_glaucaAnnotating the cytochrome P450CYP720B gene family in white spruceShaun Jackman2012-11-23 1
  2. 2. White sprucePicea glauca http://en.wikipedia.org/wiki/Picea_glauca 2
  3. 3. Cytochrome P450✤ Enzymes involved in biosynthesis and metabolism✤ Catalyze the oxidation of organic compounds✤ Contain a heme cofactor✤ Primarily membrane bound 3
  4. 4. CYP720B gene family http://en.wikipedia.org/wiki/Mountain_pine_beetle✤ Conifer-specific gene family✤ Involved in the biosynthesis of diterpene resin acids, important in conifer defence against insects and fungus http://en.wikipedia.org/wiki/Resin 4
  5. 5. CYP720B in sitka sprucePicea sitchensis✤ Twelve CYP720B genes identified in sitka spruce http://en.wikipedia.org/wiki/Picea_sitchensis✤ Full-length transcript sequences available✤ Use these sequences to identify orthologs in white spruce (Picea glauca) Katrin Geisler 5
  6. 6. White spruce sequencing dataPicea glauca✤ RNA-seq of eight tissues: bark, embryo, flush bud, mature needle, megagametophyte, seedling, xylem and young bud✤ Whole genome sequencing 24 lanes of Illumina HiSeq for 64-fold coverage Hamberger, Ohnishi et al. (2011) Plant Phys. 157: 1677-1695 6
  7. 7. Identify white spruce transcripts✤ Assemble the white spruce RNA-seq data usingTrinity (Grabherr et al. 2011) (assembled by Mack Yuen)✤ Identify the white spruce CYP720B transcripts by aligning the sitka spruce transcripts to the white spruce RNA-seq contigs using BWA-SW (Li and Durbin 2010)✤ Cluster the sitka spruce and white spruce transcripts using CLC Genomics Workbench 7
  8. 8. White spruce CYP720B transcripts Sitka spruce White spruce 8
  9. 9. Assemble the white spruce genome✤ Assemble the white spruce genome sequence using ABySS (Simpson et al. 2009)✤ Assembled 8.4 billion reads using 996 processors on 83 machines with an aggregate 4 TB of RAM✤ Assembled 18 Gbp in 5 million scaffolds larger than 500 bp with a scaffold N50 of 6 kbp 9
  10. 10. Identify white spruce genes✤ Identify CYP720B genomic contigs by aligning the sitka spruce and white spruce transcripts to the white spruce genome assembly using BWA-SW✤ Align the sitka spruce and white spruce transcripts to the CYP720B genome contigs using gmap (Wu and Watanabe 2005) 10
  11. 11. CYP720B4✤ Expressed in both sitka spruce and white spruce White spruce Sitka spruce 11
  12. 12. CYP720B7✤ Expressed only in sitka spruce White spruce Sitka spruce 12
  13. 13. CYP720B15/16/17 related✤ White spruce transcripts align, but no sitka spruce transcripts White spruce Sitka spruce 13
  14. 14. Cytochrome P450 CYP720B Genes Sitka spruce White spruce White spruce EST RNA-seq genome assembly CYP720B2 yes yes CYP720B4 yes yes CYP720B5 yes yes CYP720B7 no yes CYP720B8 no partial CYP720B9 no yes CYP720B10 no partial CYP720B12 yes yes CYP720B15 yes partial CYP720B16 no partial CYP720B17 no partial 14
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