Optimizing TILLING and Ecotilling techniques for potato (Solanum tuberosum L).
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.


RESULTS
Three gene-specific primer pairs were designed from BAC sequence to amplify approximately 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.


CONCLUSIONS
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.
Elias
Rana
R
Atomic Energy Commission of Syria, PO. Box 6091, Damascus, Syria. relias79@yahoo.com
Till
Bradley J
BJ
Mba
Chikelu
C
Al-Safadi
Bassam
B