Genetic purity analysis of hybrid broccoli (Brassica oleracea var. italica) seeds using RAPD PCR

Determination of genetic purity of F-1-hybrid seeds is a quality control requirement in the production of hybrid Brassica vegetable seeds. Hybrid varieties of these vegetable crops have arisen from a limited germplasm base, making discrimination of parental and hybrid lines very laborious and troublesome. The use of RAPD PCR for evaluating seed purity in a commercial F-1-hybrid broccoli (Brassica oleracea var. italica) cultivar is demonstrated. DNA from an F-1-hybrid and its parental lines was subjected to RAPD PCR analysis with 20 random decamer arbitrary primers. Two primers produced suitable male and female specific markers and were chosen for purity testing of the F-1-hybrid seeds. The level of sib (inbred seed, seed from self pollination of parental lines) contamination detected by RAPD analysis was comparable with the commonly used grow out trial, whereas isozyme analysis failed to differentiate between the parental lines. Our study clearly demonstrates that RAPD PCR is a useful tool for genetic purity testing of commercial F-1-hybrid broccoli seeds.