Random Amplified Polymorphic DNA Markers Are Useful for Purity Determination of Tomato Hybrids

Hybrid Primerz Sequence My Fx PH236 E03960 CCAGATGCAC + – E03900 CCAGATGCAC – + F101600 GGAAGCTTGG + – F10720 GGAAGCTTGG + – F10660 GGAAGCTTGG – + F08840 GGGATATCGG + – G16650 AGCGTCCTCC – + H091180 TGTAGCTGGG – + I021400 GGAGGAGAGG + – I09500 TGGAGAGCAG + – I09470 TGGAGAGCAG – + J171000 ACGCCAGTTC – + L11600 ACGATGAGCC – + L131450 ACCGCCTGCT – + PH254 G02 GGCACTGAGG + – Identifying breeding lines and determining hybrid purity are major quality requirements in plant breeding and seed production. Cultivar contaminations may affect yield, uniformity, and quality of the product. In addition, identifying cultivars is essential for proprietary reasons. Until recently, morphological markers have been used to identify cultivars and to determine hybrid purity. However, such tests are time consuming and expensive, and often insufficient characteristic morphological traits exist to allow for cultivar discrimination. Molecular markers such as isozymes and restriction fragment length polymorphisms (RFLP) also have been used for cultivar identification (Bunce et al., 1986; Tanksley and Jones, 1981). More recently, random amplified polymorphic DNA (RAPD) markers have been used for fingerprinting plant species (Hu and Quiros, 1991). Using RAPDs has gained popularity because 1) a small amount of DNA is required per reaction, 2) numerous oligonulcleotide primers allow for polymorphism screening, 3) each primer detects multiple bands, and 4) the technique is fast and simple, requiring no sophisticated equipment and no radioactive isotopes. In tomato (Lycopersicon esculentum L.), several molecular marker techniques have been used for cultivar identification. Alcohol dehydrogenase allozymes were used for testing hybrid purity by Tanksley and Jones (1981). Highly polymorphic telomeric sequences allowed the tomato cultivar distinction using pulsed field gel electrophoresis (Broun et al., 1992). An oligonucleotide repeat of GATA