Marker Assisted Development and Characterization of Beta-Carotene Rice

Beta-carotene producing transformants were produced in the background of Nagdongbyeo , a Japonica rice cultivar. Introgression of the carotenoid locus in the transformant, PAC4-2 into the elite cultivar Ilpumbyeo was started. To initiate a backcrossing program, we surveyed 220 SSR markers and found that 38% of them were polymorphic between Ilpumbyeo as a recurrent parent and the PAC4-2 as a recipient parent. The selection strategy comprising foreground and background selection was employed. First, foreground selection was practiced in BC1, BC2, and BC3 generations using the transgene specific PCR-based marker in addition to visual scoring of the seed color. Marker-based background selection combined with phenotypic selection was employed from BC3F2 to BC3F4 generations. Blast search indicated that the transgene PAC4-2 was located between SSR markers, RM6 and RM482. 240 BC3F3 and 63 BC3F4 lines were evaluated for four agronomic traits including days to heading. Most of the lines were similar to Ilpumbyeo in agronomic traits evaluated. The percentage of PAC4-2 genome ranged from 4% to 21% with a mean of 12.5%, which was higher than the expected for an unselected BC3 backcross population. This could be explained by the fact that two genes for beta-carotene and the stripe virus resistance were targeted in this study. We selected 10 representative BC3F5 lines from 63 BC3F4 lines based on agronomic traits and carotenoids content. The selection strategy would be appropriate for the introgression of beta-carotene gene in a breeding program.