TILLING to detect induced mutations in soybean

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TILLING to detect induced mutations in soybean

Please use this identifier to cite or link to this item: http://dx.doi.org/10.1186/1471-2229-8-9

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dc.contributor.author Cooper, Jennifer L
dc.contributor.author Till, Bradley J
dc.contributor.author Laport, Robert G
dc.contributor.author Darlow, Margaret C
dc.contributor.author Kleffner, Justin M
dc.contributor.author Jamai, Aziz
dc.contributor.author El-Mellouki, Tarik
dc.contributor.author Liu, Shiming
dc.contributor.author Ritchie, Rae
dc.contributor.author Nielsen, Niels
dc.contributor.author Bilyeu, Kristin D
dc.contributor.author Meksem, Khalid
dc.contributor.author Comai, Luca
dc.contributor.author Henikoff, Steven
dc.date.accessioned 2012-08-29T15:26:35Z
dc.date.available 2012-08-29T15:26:35Z
dc.date.issued 2008-01-24
dc.identifier.citation BMC Plant Biology. 2008 Jan 24;8(1):9
dc.identifier.uri http://dx.doi.org/10.1186/1471-2229-8-9
dc.identifier.uri http://hdl.handle.net/10355/15036
dc.description.abstract Abstract Background Soybean (Glycine max L. Merr.) is an important nitrogen-fixing crop that provides much of the world's protein and oil. However, the available tools for investigation of soybean gene function are limited. Nevertheless, chemical mutagenesis can be applied to soybean followed by screening for mutations in a target of interest using a strategy known as Targeting Induced Local Lesions IN Genomes (TILLING). We have applied TILLING to four mutagenized soybean populations, three of which were treated with ethyl methanesulfonate (EMS) and one with N-nitroso-N-methylurea (NMU). Results We screened seven targets in each population and discovered a total of 116 induced mutations. The NMU-treated population and one EMS mutagenized population had similar mutation density (~1/140 kb), while another EMS population had a mutation density of ~1/250 kb. The remaining population had a mutation density of ~1/550 kb. Because of soybean's polyploid history, PCR amplification of multiple targets could impede mutation discovery. Indeed, one set of primers tested in this study amplified more than a single target and produced low quality data. To address this problem, we removed an extraneous target by pretreating genomic DNA with a restriction enzyme. Digestion of the template eliminated amplification of the extraneous target and allowed the identification of four additional mutant alleles compared to untreated template. Conclusion The development of four independent populations with considerable mutation density, together with an additional method for screening closely related targets, indicates that soybean is a suitable organism for high-throughput mutation discovery even with its extensively duplicated genome.
dc.title TILLING to detect induced mutations in soybean
dc.type Journal Article
dc.date.updated 2012-08-29T15:26:35Z
dc.description.version Peer Reviewed
dc.language.rfc3066 en
dc.rights.holder Jennifer L Cooper et al.; licensee BioMed Central Ltd.


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