Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: January 19, 2000
Publication Date: N/A
Technical Abstract: Sequence characterized amplified regions (SCARs) are breeder-friendly allele specific molecular markers that are very useful in marker-assisted breeding programs. We previously identified two RAPD markers linked to the beta-carotene (B) gene and one AFLP marker, E-ACA/M-CTG-100, linked to the beta modifier (Mo-B) gene in tomato. Amplified products were cloned and sequenced. SCAR primer pairs, SCAR 18f/r and SCBC92f/r from the RAPD markers and SCE3M7f/r from the AFLP marker were designed based on the sequence information. The same sized fragment as their progenitor RAPD or AFLP markers was amplified, but failed to retain the polymorphism. To resolve this problem, the amplified SCAR products from the null phenotype parent for the two RAPD markers were cloned and sequenced. Sequences from both parents for each marker were used for identifying restriction site mutations. In the SCAR18f/r amplified DNA product, a RsaI site mutation on allele of the high beta-carotene parent was identified and a codominant SCBC792-779 marker was generated. Since the SCE3M7f/r amplified fragment was too small to be used for subsequent enzyme digestion, an alternate technique, inverse PCR, has been used to convert this to SCAR.