Many important traits have been tagged allowing plant breeders to apply marker assisted selection (MAS) in rice. PCR itself is simple to set up, and requires little hands-on time. However, a crucial limiting step of MAS programs is the reliable and efficient extraction of DNA which can be performed on thousands of individuals. In this study, We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract DNA from leaf tissues for suitable MAS in rice. We followed the standard 2% CTAB extraction method in all the procedure. In addition we used the 1.2 ml 8-strip tube instead of 1.5 ml E-tubes to fit the 8-multichannel pipette and employ the 96 well plate to use the swing bucket centrifuge. Our modified CTAB DNA extraction method offers several advantages with respect to traditional and simple methods. 1) adult leaf samples collected in paddy field are applicable. 2) 96 leaf samples can be homogenized only one-time by using tungsten carbonate bead and 96well block. 3) semiautomatic loading method using 8-multichannel pipette from DNA extraction to electrophoresis of PCR products. 4) our system can extract about 400 leaf samples per day by only one technicion. Therefore, this method could be useful for marker assisted breeding in rice.

In rice, tillering is an important trait determining yield. To study tillering at the agricultural and molecular aspects, we have examined a spontaneous rice mutant that showed reduction in the number of culms. The mutant was derived from a F6 line of the cross of Junambyeo*4 / IR72. It could produce, on average, 4 tillers per hill in the paddy field while wild-type plants usually have 15. Except the reduced culm numbers, they also show pale green phenotypes. The phenotypes of this mutant were co-segregated as the monogenic Mendelian ratio (χ2 = 0.002, p = 0.969). In order to locate a gene responsible for the rcn phenotype, the mutant with the japonica genetic background was crossed with Milyang21 of the indica background. Bulked segregant analysis was used for rapid determination of chromosomal location. Three SSR markers (RM551, RM8213, and RM16467) on chromosome 4 were genetically associated with the mutant phenotype. Each of the 217 F2 plants was genotyped with simple sequence length polymorphisms. The data showed that RM16572 on chromosome 4 was the closest marker that showed perfect co-segregation among the F2 population. We suggest the new rcn gene studied here name as rcn10t because there was no report which exhibit a rcn phenotype with a pleiotropic effect of pale green (chlorophyll deficiency), and mapped at same position on chromosome 4.