Optimizing TILLING and Ecotilling techniques for potato Solanum tuberosum LReport as inadecuate

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BMC Research Notes

, 2:141

First Online: 17 July 2009Received: 20 March 2009Accepted: 17 July 2009DOI: 10.1186-1756-0500-2-141

Cite this article as: Elias, R., Till, B.J., Mba, C. et al. BMC Res Notes 2009 2: 141. doi:10.1186-1756-0500-2-141


BackgroundThe TILLING and Ecotilling techniques for the discovery of nucleotide polymorphisms were applied to three potato Solanum tuberosum cultivars treated with gamma irradiation. The three mutant cultivars tested were previously shown to exhibit salinity tolerance, an important trait in countries like Syria where increasing soil salinity is affecting agricultural production.

FindingsThree gene-specific primer pairs were designed from BAC sequence to amplify ~1 to 1.5 kb of gene target. One of the three primer pairs amplified a single gene target. We used this primer pair to optimize enzymatic mismatch cleavage and fluorescence DNA detection for polymorphism discovery. We identified 15 putative nucleotide polymorphisms per kilobase. Nine discovered polymorphisms were unique to one of the three tetraploid cultivars tested.

ConclusionThis work shows the utility of enzymatic mismatch cleavage for TILLING and Ecotilling in different varieties of potato. The method allows for rapid germplasm characterization without the cost and high informatics load of DNA sequencing. It is also suitable for mutation discovery in high-throughput reverse genetic screens.

Electronic supplementary materialThe online version of this article doi:10.1186-1756-0500-2-141 contains supplementary material, which is available to authorized users.

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Author: Rana Elias - Bradley J Till - Chikelu Mba - Bassam Al-Safadi

Source: https://link.springer.com/

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