**3. Confirmation of Satt228 marker**

204 Soybean – Genetics and Novel Techniques for Yield Enhancement

Fig. 1. Polyacrylamide gels of protein extracted from parents and F2 seeds. P1

band (21.5 KDa).

and F2 population.

locus differed between populations 1 and 2.

(Jinpumkong2) and P2 (C242) are parents and arrow points to the Kunitz trypsin inhibitor

Of the 1,000 RAPD primers tested on two parents of population 1 (Jinpumkong2 and C242), approximately 12 % (124) primers produced polymorphic DNA fragment differences between the parents. Only 35 primers were identified as being polymorphic between bulked DNA samples with SKTI protein and bulked DNA samples with no SKTI protein. Of those 35 primers, only 16 also exhibited polymorphism between parents. Among 342 primer sets of AFLP analysis, only 10 primers were shown polymorphism between parents. Three SSR primers (Satt409, Satt228 and Satt429) among 35 primers selected were shown polymorphism between parents. Total 48 markes (35 RAPD, 10 AFLP, and 3 SSR) were used to obtain segregation data from 94 F2 individuals of population 1. Figure 2 represents some example of segregating DNA fragment for SSR markers (Satt228) in parents, bulked samples

Fig. 2. Patterns of segregating DNA fragment for SSR primer Satt228 in parents, bulked samples, and F2 population. P1 is Jinpumkong2 (*TiTi*) and P2 is C242 (*titi*). B1 is bulked of present kunitz trypsin inhibitor protein individuals and B2 is bulked of absent individuals. A genetic map was constructed from the 48 segregating DNA markers and *Ti* locus. A total 11 DNA markers (4 RAPD, 4 AFLP, and 3 SSR) and *Ti* locus was found to be genetically linked in population 1. Three SSR markers, Satt409, Satt228, and Satt429 linked with *Ti* locus within 10 cM (Figure 3). Satt228 marker was very tightly linked with *Ti* locus at 0 cM. Three SSR markers linked with *Ti* locus in population 1 were applied in population 2. Only two SSR markers, Satt228 and Satt409 were linked with *Ti* locus. Satt228 marker was linked with *Ti* locus in 3.7 cM distance (Figure 3). The order and map distances of SSR markers and *Ti*
