W287 “Shrinking” the Giants: An innovative approach for de novo sequencing of conifer genomes

Date: Sunday, January 15, 2012
Time: 10:20 AM
Room: Sunrise
Konstantin V. Krutovsky , Department of Ecosystem Science and Management, Texas A&M University, College Station, TX
Iraida N. Tretyakova , Laboratory of Forest Genetics and Selection, V.N. Sukachev lnstitute of Forest, Krasnoyarsk, Russia
Irina V. Chubugina , Department of Genetics and Selection, Forest Protection Center, Krasnoyarsk, Russia
Natalja V. Oreshkova , Laboratory of Forest Genetics and Selection, V.N. Sukachev lnstitute of Forest, Krasnoyarsk, Russia
Craig S. Echt , Southern Institute of Forest Genetics, USDA Forest Service, Saucier, MS
Nurul Islam-Faridi , Department of Ecosystem Science and Management, Texas A&M University, College Station, TX
C. Dana Nelson , Southern Institute of Forest Genetics, USDA Forest Service, Saucier, MS
The gigantic genome size of conifers and high allelic variation impede their complete genome sequencing. To overcome the allelic variation problem we are developing and using haploid tissue cultures starting with very early-stage female gametophytic tissue. As a case study, we present data based on a haploid tissue culture generated from loblolly pine (Pinus taeda L.), the reference tree for pine genomics. The haploid condition of the genomic DNA was confirmed by genotyping SSR markers that are heterozygous in the diploid tissue of the parent tree. This is the first report of haploid tissue culture for loblolly pine. Similar results were obtained for Siberian larch (Larix sibirica Ledeb.) making this novel technique of reducing sequencing complexity by eliminating heterozygosity possible for the conifers.