표제지
목차
Abstract 9
Ⅰ. 서론 10
Ⅱ. 재료 및 방법 12
1. 키위 검은점무늬병의 병징 및 병원균 분리 12
1) 키위 검은점무늬병의 병징 12
2) 키위 검은점무늬병균 분리 12
3) 키위 검은점무늬병균의 병원성검정 15
2. 키위 검은점무늬병균의 동정 16
1) 균학적 동정 16
2) 분자생물학적 동정 16
3. 키위 검은점무늬병균의 실내 화학약제 선발 17
4. 키위 검은점무늬병균에 대한 길항세균의 항균 효과 18
1) 길항세균의 선발 및 동정 18
2) 길항세균의 화학약제에 대한 감수성 조사 19
3) 길항세균이 키위 검은점무늬병균 포자 발아에 미치는 영향 19
4) 길항세균의 효소활성 및 항균물질 생합성 유전자 분석 20
5) 길항세균의 최적 배양 온도와 pH 21
6) 길항세균의 길항력을 증가시키는 탄소원과 질소원 선발 21
7) 길항세균의 키위 과피 정착 농도 조사 21
6. 키위 검은점무늬병 방제를 위한 현장적용 평가 22
1) 봉지씌우기 방제효과 22
2) 화학 약제와 길항세균 및 봉지 씌우기 방제효과 22
Ⅲ. 결과 24
1. 키위 검은점무늬병 병원균 분리 24
2. 키위 검은점무늬병균의 병원성검정 25
3. 키위 검은점무늬병균의 동정 26
1) 균학적 동정 26
2) 분자생물학적 동정 28
4. 키위 검은점무늬병균이 실내 화학약제 선발 31
5. 키위 검은점무늬병균에 대한 길항세균의 항균 효과 32
1) 길항세균의 선발 및 동정 32
2) 길항세균의 화학약제에 대한 감수성 조사 33
3) 길항세균이 키위 검은점무늬병균 포자 발아에 미치는 영향 34
4) 길항세균의 효소활성 및 항균물질 생합성 유전자 분석 35
5) 길항세균의 최적 배양 온도와 pH 36
6) 길항세균의 길항력을 증가시키는 탄소원과 질소원 37
7) 길항세균의 키위 과피 정착 농도 조사 38
6. 키위 검은점무늬병 방제를 위한 현장적용 평가 39
1) 봉지씌우기 방제효과 39
2) 화학약제와 길항세균 처리 후 봉지씌우기 방제효과 40
Ⅳ. 고찰 41
Ⅴ. 요약 44
Ⅵ. 인용문헌 45
Table 1. List of fungicides used in this study 17
Table 2. List of antagonistic bacteria used in this study isolated from pericarp of kiwifruit 19
Table 3. Characteristics of primers used for the detection of genes involve in biosynthesis of anti biotics genes of Bacillomycin D, Fengycin, Iturin A, Surfactin, and Zwitermicin A 20
Table 4. Spray schedules of fungicides and antagonistic bacteria to control black spot on kiwifruit with covering paper bag 23
Table 5. Comparison of morphological characteristics of the present isolates with Colletotrichum fioriniae previously described 26
Table 6. Comparison of morphological characteristics of the present isolates with Diaporthe eres previously described 27
Table 7. Sensitivity of 2 types fungi to various fungicides on potato dextrose agar 31
Table 8. Antifungal activity of the selected antagonistic bacteria against Colletotrichum fioriniae and Diaporthe eres on potato dextrose agar 32
Fig. 1. Typical symptom of black spot of naturally infected 'Sweet gold' kiwifruit (A). Symptom of bulging spots on skin (B, E), Symptom of freckle spots on skin (C, F),... 13
Fig. 2. Symptom of black spot on kiwifruits on various varieties. 'Sweet gold' (A), 'Haegeum' (B), 'Gold one' (C), 'Hongyang' (D), Healthy exterior of 'Sweet gold' (E), Healthy interior of... 14
Fig. 3. Typical symptoms of orchard and low temperature storage of kiwifruit. 24
Fig. 4. Spore mass formed on the black spotted kiwifruit. (A) Pericarp with black spot on WA, (B) Spore mass of Colletotrichum sp., (C) Spore mass of Phomopsis sp.. 24
Fig. 5. Pathogenicity test of black spot on gold kiwifruit cv. Haegeum by artificial inoculation. In vitro and in vivo typical symptom of pericarp inoculated with Colletotrichum sp. (A, B) and... 25
Fig. 6. Morphology of Colletotrichum fioriniae SG-K2. (A, B) Colony of Colletotrichum fioriniae SG-K2 incubated for 7 days after on potato dextrose agar, (C) Conidia mass, (D) Conidia, (E)... 26
Fig. 7. Morphology of Diaporthe eres SG-K5. (A, B) Colony of Diaporthe eres SG-K5 incubated for 7 days after on potato dextrose agar, (C) Conidia mass, (D) α-conidia, (E) β... 27
Fig. 8. Phylogenetic tree of Colletotrichum fioriniae SG-K2 using maximum likelihood method based on ITS gene sequences. The numbers at the nodes were the percentages of bootstrap on 1,00... 29
Fig. 9. Phylogenetic tree of Diaporthe eres SG-K5 using maximum likelihood method based on beta-tubulin gene sequences. The numbers at the nodes were the percentages of bootstrap on... 30
Fig. 10. Phylogenetic tree of Bacillus velezensis H18 using maximum likelihood method based on 16s rRNA gene sequences. The numbers at the nodes were the percentages of bootstrap... 33
Fig. 11. Sensitivity of Bacillus velezensis H18 to two fungicides on nutrient agar. (C) Control. 33
Fig. 12. Comparison of the inhibition of the Bacillus velezensis H18 cell suspension and fungicide against Colletotrichum fioriniae SG-K2 (A), Diaporthe eres SG-K5 (B). Scale bars... 34
Fig. 13. Enzyme activities of Bacillus velezensis H18. (A) CMC agar medium, (B) Amylase agar medium, (C) Proteinase agar medium, (D) Solubility phosphate agar medium. 35
Fig. 14. PCR detection of antifungal activating gene corresponding to Iturin A (itu A), Surfactin (srf A). M: 1 kbp ladder. 35
Fig. 15. Effect of initial temperature (A) and pH (B) on cell growth of Bacillus velezensis H18 in nutrient broth for mass culture of antagonistic bacteria. 36
Fig. 16. Carbon sources (A) and nitrogen sources (B) that increase the antibacterial activity of antagonistic bacteria against 2 types fungi on potato dextrose agar. 37
Fig. 17. Population densities of antagonistic bacteria Bacillus velezensis H18. 38
Fig. 18. Incidences of black spot on kiwifruits covered with and without bag. 39
Fig. 19. Control efficacies of black spot on 'Sweet gold' kiwifruit orchard in Goseong (A) and Goheung (B). 1, 8, 15: Tebuconazole SC + Bag, 2, 9, 16; Bacillus velezensis H18 SC +... 40