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Contents
초록 (Abstract) 12
I. Introduction 15
II. Materials and Methods 19
1. Mapping population 19
2. Determination of genotype for Kunitz trypsin inhibitor protein 19
3. Genomic DNA extraction 20
4. RAPD analysis 21
5. AFLP analysis 22
6. SSR analysis 27
7. SKTti marker analysis 27
8. Bulked segregation analysis 29
9. Linkage analysis 29
10. Development of population for marker confirmation 30
11. Screening of germplasm with titi genotype by Satt228 marker 34
III. Results 36
1. Identification of DNA marker linked to the Ti locus 36
1-1. Segregation of KTI protein 36
1-2. Detection of polymorphic DNA markers 38
1-3. Molecular genetic map of Ti locus 49
2. Confirmation of Satt228 marker identified for MAS 53
2-1. Screening of titi germplasms by Satt228 marker 53
2-2. Confirmation of Ti genotype in 'Jinpumkong2' × C242 population by Satt228 Marker 56
2-3. Confirmation of Ti genotype in GS06 × C242 population by Satt228 marker 67
2-4. Confirmation of Ti genotype in 20M183 × C242 population by Satt228 Marker 72
2-5. Confirmation of Ti genotype in 'Seoritae' × C242 population by Satt228 Marker 77
2-6. Confirmation of Ti genotype in 'Hannamkong' × C242 population by Satt228 Marker 81
2-7. Cosegregation of Satt228 marker and Ti locus 85
IV. Discussion 87
V. Conclusions 98
VI. References 101
Appendixes 114
감사의 글 (Acknowledgement) 120
Table 1. Nucleotide sequences of adapters, primers and… 24
Table 2. Three hundreds forty two selective primers used for AFLP analysis 25
Table 3. Genotype of Ti locus and seed coat color… 31
Table 4. Origin, seed coat color and genotype for Ti locus of… 35
Table 5. Observed and expected segregation of F₂ seeds from… 37
Table 6. Nucleotide sequence of seven markers mapped with… 52
Table 7. Satt228 marker allele and Kunitz trypsin inhibitor (KTI)… 59
Table 8. Satt228 marker allele and Kunitz trypsin inhibitor (KTI)… 70
Table 9. Satt228 marker allele and Kunitz trypsin inhibitor (KTI)… 75
Table 10. Satt228 marker allele and Kunitz trypsin inhibitor… 80
Table 11. Satt228 marker allele and Kunitz trypsin inhibitor (KTI)… 84
Table 12. Genetic relationship between allele of Satt228 marker… 86
Fig. 1. Comparison of nucleotide sequence for KTi (presence of Kunitz… 28
Fig. 2. The diagram of four population development with lacking… 33
Fig. 3. Polyacrylamide gels of protein extracted from parents and… 36
Fig. 4. Pattern of fragments amplified using Operon primers with… 38
Fig. 5. Pattern of DNA fragment using OPAC12 primer… 39
Fig. 6. Pattern of segregating DNA fragment for RAPD (OPAR15)… 40
Fig. 7. Screening of parental polymorphism using AFLP marker with… 41
Fig. 8. Pattern of DNA fragment using AFLP primer… 42
Fig. 9. Segregation of fragment in F₂ population with EcoR I-CAT/Mse… 43
Fig. 10. Pattern of fragments amplified using SSR primers with… 44
Fig. 11. Pattern of DNA fragment using SSR primer… 45
Fig. 12. Pattern of segregating DNA fragment for SSR primer… 46
Fig. 13. Pattern of DNA fragment using SKTti primer… 47
Fig. 14. Pattern of segregating DNA fragment for SKTti marker… 48
Fig. 15. Molecular linkage map of Ti locus defined using population… 49
Fig. 16. Molecular linkage map of Ti locus defined using population… 50
Fig. 17 Molecular linkage map A2 (Cregan et al., 1999) of Ti locus… 51
Fig. 18. Pattern of polyacrylamide protein gel (A) and of genomic… 54
Fig. 19. Pattern of genomic DNA amplification by Satt228 marker… 57
Fig. 20. Pattern of genomic DNA amplification by Satt228 marker… 68
Fig. 21. Pattern of genomic DNA amplification by Satt228 marker… 73
Fig. 22. Pattern of genomic DNA amplification by Satt228 marker… 78
Fig. 23. Pattern of genomic DNA amplification by Satt228 marker… 82
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