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Optimizing TILLING and Ecotilling techniques for potato (Solanum tuberosum L)
- Source :
- BMC Research Notes, Vol 2, Iss 1, p 141 (2009), BMC Research Notes
- Publication Year :
- 2009
- Publisher :
- BMC, 2009.
-
Abstract
- Background The 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. Findings Three 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. Conclusion This 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.
- Subjects :
- TILLING
Medicine(all)
Soil salinity
Biochemistry, Genetics and Molecular Biology(all)
Mutant
fungi
lcsh:R
food and beverages
lcsh:Medicine
General Medicine
Biology
Solanum tuberosum
General Biochemistry, Genetics and Molecular Biology
Salinity
Horticulture
lcsh:Biology (General)
Botany
Technical Note
Cultivar
lcsh:Science (General)
lcsh:QH301-705.5
Gamma irradiation
lcsh:Q1-390
Subjects
Details
- Language :
- English
- ISSN :
- 17560500
- Volume :
- 2
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- BMC Research Notes
- Accession number :
- edsair.doi.dedup.....8697c6ffe0073b8af7f0dee3b967743a