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Establishing genomic tools and resources for Guizotia abyssinica (L.f.) Cass.-the development of a library of expressed sequence tags, microsatellite loci, and the sequencing of its chloroplast genome
Authors:Dempewolf Hannes  Kane Nolan C  Ostevik Katherine L  Geleta Mulatu  Barker Michael S  Lai Zhao  Stewart Megan L  Bekele Endashaw  Engels Johannes M M  Cronk Quentin C B  Rieseberg Loren H
Institution:The Biodiversity Research Centre and Department of Botany, 3529-6270 University Blvd, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4 Department of Plant Breeding and Biotechnology, Swedish University of Agricultural Sciences, Box 101, SE-230 53 Alnarp, Sweden Department of Biology, Indiana University, Bloomington, IN 47405, USA Addis Ababa University, PO Box 1176, Addis Ababa, Ethiopia Bioversity International, 00057 Maccarese, Rome, Italy Centre for Plant Research, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4.
Abstract:We present an EST library, chloroplast genome sequence, and nuclear microsatellite markers that were developed for the semi-domesticated oilseed crop noug (Guizotia abyssinica) from Ethiopia. The EST library consists of 25 711 Sanger reads, assembled into 17 538 contigs and singletons, of which 4781 were functionally annotated using the Arabidopsis Information Resource (TAIR). The age distribution of duplicated genes in the EST library shows evidence of two paleopolyploidizations—a pattern that noug shares with several other species in the Heliantheae tribe (Compositae family). From the EST library, we selected 43 microsatellites and then designed and tested primers for their amplification. The number of microsatellite alleles varied between 2 and 10 (average 4.67), and the average observed and expected heterozygosities were 0.49 and 0.54, respectively. The chloroplast genome was sequenced de novo using Illumina’s sequencing technology and completed with traditional Sanger sequencing. No large re-arrangements were found between the noug and sunflower chloroplast genomes, but 1.4% of sites have indels and 1.8% show sequence divergence between the two species. We identified 34 tRNAs, 4 rRNA sequences, and 80 coding sequences, including one region (trnH-psbA) with 15% sequence divergence between noug and sunflower that may be particularly useful for phylogeographic studies in noug and its wild relatives.
Keywords:chloroplast genome  EST library  genetic resources  Guizotia abyssinica  microsatellites  paleopolyploidy
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