High-Throughput Discovery of Mutations in Tef Semi-Dwarfing Genes by Next-Generation Sequencing Analysis |
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Authors: | Qihui Zhu Shavannor M. Smith Mulu Ayele Lixing Yang Ansuya Jogi Srinivasa R. Chaluvadi Jeffrey L. Bennetzen |
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Affiliation: | *Department of Genetics and;†Department of Plant Pathology, University of Georgia, Athens, Georgia 30602;‡Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47906;§Pioneer Hi-Bred International, Inc., Johnston, Iowa |
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Abstract: | Tef (Eragrostis tef) is a major cereal crop in Ethiopia. Lodging is the primary constraint to increasing productivity in this allotetraploid species, accounting for losses of ∼15–45% in yield each year. As a first step toward identifying semi-dwarf varieties that might have improved lodging resistance, an ∼6× fosmid library was constructed and used to identify both homeologues of the dw3 semi-dwarfing gene of Sorghum bicolor. An EMS mutagenized population, consisting of ∼21,210 tef plants, was planted and leaf materials were collected into 23 superpools. Two dwarfing candidate genes, homeologues of dw3 of sorghum and rht1 of wheat, were sequenced directly from each superpool with 454 technology, and 120 candidate mutations were identified. Out of 10 candidates tested, six independent mutations were validated by Sanger sequencing, including two predicted detrimental mutations in both dw3 homeologues with a potential to improve lodging resistance in tef through further breeding. This study demonstrates that high-throughput sequencing can identify potentially valuable mutations in under-studied plant species like tef and has provided mutant lines that can now be combined and tested in breeding programs for improved lodging resistance. |
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Keywords: | brachytic2 dw3 ethyl methanesulfonate (EMS) mutagenesis Eragrostis tef green revolution genes |
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