표제지
목차
초록 12
I. 서언 15
II. 연구사 18
1. 감(Diospyros kaki, Thunb.)의 기원 18
2. 감 품종 식별 및 유연관계 분석 19
3. PCNA-type 선발을 위한 분자표지 개발 21
4. 감에서 배수체 개체 유기 및 배수체 품종 개발 22
III. 연구결과 24
1. GBS 분석을 이용한 감 품종 식별 SNP 선발 및 KASP assay 24
1.1. 서언 24
1.2. 재료 및 방법 25
1.3. 결과 및 고찰 33
2. HRM을 이용한 PCNA 선발 64
2.1. 서언 64
2.2. 재료 및 방범 65
2.3. 결과 및 고찰 67
3. 기내 콜히친 처리에 의한 염색체 배가 감 식물체 유기 73
3.1. 서언 73
3.2. 재료 및 방법 74
3.3. 결과 및 고찰 75
IV. 종합고찰 83
V. 인용문헌 87
Abstract 98
Table 1. Cultivars and breeding lines for GBS analysis. 27
Table 2. Barcode sequences for GBS library. 29
Table 3. Published D. kaki representative transcripts information. 32
Table 4. Summary of sequencing raw data. 37
Table 5. Summary of the GBS sequence data after alignment to the reference sequence. 37
Table 6. Statistics of raw sequence data of persimmon cultivars after demultiplexing. 37
Table 7. Statistics of trimmed data of persimmon cultivars after demultiplexing. 40
Table 8. Statistics of mapping data of persimmon cultivars. 43
Table 9. The SNPs detected among the persimmon genotypes. 46
Table 10. Summary of SNP filter process. 48
Table 11. Read depths and SNPs by persimmon accessions. 51
Table 12. Primer sequences of KASP markers. 55
Table 13. Validation of KASP markers and the list of persimmon cultivars genotyped. 60
Table 14. Sequences of primers newly designed in the 5R region 68
Table 15. List of the cultivars used to examine the applicability of the high-resolution melting analysis using associated markers. 70
Table 16. Fruit setting of cross combinations for embryo culture. 76
Table 17. Effect of the colchicine concentration on polyploidy and viability. 76
Table 18. Effect of the zeatin concentration and treatment methods on shoots growth. 77
Table 19. Effect of NAA and IBA concentration and treatment methods on roots growth. 79
Fig. 1. Diagram of GBS analysis. 30
Fig. 2. Workflow for GBS library construction. 34
Fig. 3. Total gDNA isolated from 95 persimmon samples for GBS library construction. 35
Fig. 4. Result of GBS library construction-Purification. Lane 1: GBS library-1 before purificaton; Lane 2: GBS library-2 before purification;... 36
Fig. 5. Bioanalyzer image of GBS library-QC report. 36
Fig. 6. Phylogenetic tree of 95 persimmon culitvars using 38,762 SNP loci. 49
Fig. 7. Genotyping result for persimmon cultivars using KASP assay technique. The scatter plots with axes X and Y represent allele... 59
Fig. 8. Conversion of the SNP markers into barcode after changing into digital signal of 0 and 1. 62
Fig. 9. Conversion of the SNP markers of 32 persimmon cultivars into barcode. 62
Fig. 10. Schematic representation of the AST and ast allele-linked regions isolated from genomic libraries of 'Nishimurawase' and 'Jiro'... 69
Fig. 11. PCR amplification patterns of the Hrm marker (A) and PCR amplication patterns of the AST marker (B). Lanes 1-8: pollination... 71
Fig. 12. High-resolution melting analysis of pollination constant non-astringent (PCNA) and non-PCNA cultivars using Hrml marker.... 72
Fig. 13. Flow cytometric histograms of relative fluorescence intensity in the nuclei. A: 2x is D. Lotus (2n=2x) and 6x is D. kaki (Fuyu), B: 6x... 77
Fig. 14. Effect of zeatin concentration and treatments on shoot growth. A: dipping in the 10 mM zeatin for 5 seconds and incubate at 1/2 MS... 78
Fig. 15. Effect of NAA and IBA concentrations and treatment methods on roots growth. A: Dipping in the 10 mM NAA + 5% DMSO for 5... 80
Fig. 16. Comparison of stomatal density and length of 12x and 6x guard cells. 12x: Doubled chromosomes, 6x: non-doubling chromosomesor... 82
Fig. 17. Microscopic images of 12x and 6x guard cells. A: Seedling of fuyuXtaishu that has doubled chromosomes(12x) by colchicine... 82