Molecular Markers Associated with Cold-Hardiness in Camellia

Sequence-characterized amplified region (SCAR) markers from expressed sequence tag-polymerase chain reaction (EST-PCR) and random amplified polymorphic DNA (RAPD) markers were developed with the goal to separate cold hardy Camellia's from non-cold hardy ones. A total of 28 cold hardy and non-cold hardy Camellia genotypes were evaluated using 11 EST-PCR and 60 RAPD primers. One EST-PCR primer pair based on a Camellia sinensis sequence (here referred to as CAME 13; forward, 5'-CGGGCAGGTACCTTCTGATA-3' and reverse, 5'-TGCAGAAGATGGAGATGCAG -3') and one RAPD primer (Operon A-12) appeared to have potential for discriminating between the cold hardy and non-cold hardy genotypes. SCAR primer pairs were designed based on the sequence of the C. japonica bands amplified by the CAME 13 and Operon A-12 primers. The ESTSCAR-19839 primer pair (forward, 5'-CGGGCAGGTACCTTCTGATA-3'; reverse, 5'-CCTCAGCCGTAGTTGCATTT-3') proved useful for identifying noncold hardy Camellia plants, whereas the fragment amplified by the RAPD-based SCAR primers appeared monomorphic.