Gamma Rays and CarbonIon-Beams Irradiation for Mutation ...

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Stover’s scale modified by Gauhl (Orjeda, 1998) in which 0 = no symptoms; 1 = presence of spot (up to 10 spot, stage 4 of the Fouré scale ); 2 = less than 5% of the leaf affected; 3 = from 6 to 15% of the leaf affected; 4 = from 16 to 33% of the leaf affected; 5 = from 34 to 50% of the leaf affected; and 6 = more than 50% of the leaf affected. 4. 2. 2. 2. Juglone toxin screening Juglone (5-hydroxy-1,4-naphthoquinone) is one of the most active toxin among the seven toxins produced by Mycosphaerella fijinesis Morelet, that induce the formation of necrotic lesions of plants leaf cells (Strobel et al., 1993). This toxin was used in this experiment to screen young plants as an indicator of resistance/tolerance to black Sigatoka disease. Previous results using different doses of juglone (Reyes-Borja et al., 2005) found that 150 ppm of this toxin was a suitable dose to get differences among the resistant cultivars and irradiated plants. The leaf- discs (5 discs per plant) were taken from the second expanded leaf of both ion-beams irradiated cultivars ‘Williams’ and ‘Cavendish Enano’ from the field planted experiment. Leaf-discs samples of ‘FHIA-01’ (resistant cultivar to black Sigatoka) were obtained from sucker plants available in the banana collection of the Estación Experimental Tropical Pichilingue, Ecuador. One liter of solution containing 150 ppm of juglone was prepared using distilled water. An amount of 10 mL were dispensed into a Petri dish (90 mm) and the leaf-discs were immersed onto the solution. Leaves were rinsed with distilled water and 1.5 mm-diameter discs were removed using a cork borer (No. 8). Petri dishes with leaf-discs samples were kept on light and room temperature during 24 hours (Fig. 33). A total of 440 leaf discs were analyzed (87 plants x 5 leaf discs/plant). After 24 81
hours, photographs of the leaf-discs were taken by using a digital camera Minolta Dimage for necrotic areas percentage estimation. Photos were processed using the GIMP 2.2 software by selecting hand-drawn regions and their pixels. The percentage of necrotic area was calculated by the following formula: Pixels of the full disc - Pixels of the green area Disc necrotic area (%) = ---------------------------------------------------------- x 100 4. 3. Results and Discussion Pixels of the full disc 4. 3. 1. Assessment of banana explants in laboratory conditions 4. 3. 1. 1. Explants slicing conditioning for ion-beams irradiation The analysis of variance for regeneration rate (%) reported high significance for slicing methods. The Tukey analysis (P≦0.05), also reported significant differences between the slicing methods, but there are not significant differences among the cultivars. However, when the weight of the explants was analyzed, this analysis reported significant difference between cultivars. The Fig. 34 shows the relationships of the explants weight (A) and regeneration rate (B) with the two slicing methods (vertical and horizontal) in ‘Williams’ and ‘FHIA-01’, 13-days after culture. Vertically sliced cuttings showed the highest regeneration rate (%) in both cultivars as is shown in Fig. 35. Values of 60-70% of regeneration rate were obtained in ‘Williams’ and ‘FHIA-01’ using the vertical cutting in contrast to only 37-43% in horizontally sliced cuttings. The weight of the explants were 82
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Gamma Rays and Carbon Ion-Beams Irr
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Table of contents Chapter 1 1.1. In
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4.2.1.1. Cultivars of banana……
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and ‘Valery’, which are exporte
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‘Cavendish Enano’, ‘Williams
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Although it is now found only in Au
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varieties bred at the Jamaican prog
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such as ‘Paka’ (AA) and ‘Pisa
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Resistance (PR). In connection with
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large number of improved mutant cro
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which migrated faster (Rf = 0.44) t
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is juglone (5 hydroxy-1,4-napthoqui
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experiments involving various biolo
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Tanaka (1999) reported novel genes
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2. 1. Materials and Methods Chapter
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2. 1. 1. 4. FHIA-01 ‘FHIA-01’(M
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(Gamma room)” and “Carbon ion-b
Page 35 and 36: esulting from “Gamma Room” expe
Page 37 and 38: traits as female sterility and part
Page 39 and 40: 3. 2. 1. 3. Chronic irradiation ( 1
Page 41 and 42: Gy. ‘Williams’ and ‘Orito’
Page 43 and 44: a diploid clone (genomic constituti
Page 45 and 46: ‘Williams’ (93%), the necrotic
Page 47 and 48: etween the irradiation doses and th
Page 49 and 50: moment they are at the development
Page 51 and 52: A B C D Fig. 2. Cultivars of banana
Page 53 and 54: Weight of plantlets (g) Height of p
Page 55 and 56: Control 50 Gy 100 Gy 150 Gy Fig. 6.
Page 57 and 58: Control 50 Gy 100 Gy 150 Gy 200 Gy
Page 59 and 60: Survival rate (%) for LD50 Survival
Page 61 and 62: A B C D E F Fig. 12. A plant from
Page 63 and 64: B D AA E Fig 14. Banana meristems a
Page 65 and 66: A B C Orito Orito Orito Cavendish E
Page 67 and 68: No. plants No. plants No. plants No
Page 69 and 70: Circunference (cm) Plant height (cm
Page 71 and 72: Table 1. Differences of the plantle
Page 73 and 74: Table 3. Differences of the surviva
Page 75 and 76: Table 5. Selected materials reporti
Page 77 and 78: Table 8. Factor of effectiveness an
Page 79 and 80: deletion rather than point mutation
Page 81 and 82: were recorded. From the results, th
Page 83 and 84: 4. 2. 2. Assessment of banana plant
Page 85: placement among the plants (Fig. 30
Page 89 and 90: ‘Williams’ (B) was obtained by
Page 91 and 92: class limits values are clearly sep
Page 93 and 94: values in ‘Cavendish Enano’ ran
Page 95 and 96: Only a partial resistance was obser
Page 97 and 98: Sigatoka but also fruit quality, po
Page 99 and 100: A C Fig. 23. Banana plantlets packa
Page 101 and 102: A C Fig. 25. Acclimatization of the
Page 103 and 104: A C Fig. 27. Survival plantlets for
Page 105 and 106: A C Fig. 29. Mycosphaerella fijiens
Page 107 and 108: A C Fig. 31. Labor during transplan
Page 109 and 110: B A Fig. 33. Establishment of the j
Page 111 and 112: A B Fig. 35. Regenerated plantlets
Page 113 and 114: Survival rate (%) for LD50 Survival
Page 115 and 116: 0 0.5 1 2 4 Fig 39. Banana plantlet
Page 117 and 118: A B C Fig. 41. Regenerated explants
Page 119 and 120: Range of optimum doses (Gy) 16 15 1
Page 121 and 122: Percent of plants Percent of plants
Page 123 and 124: LDNA-% LDNA-% DDP-days 100 80 60 40
Page 125 and 126: Fig. 49. Hexaploid cells in ‘Cave
Page 127 and 128: Table 10. Infection index (II-%), d
Page 129 and 130: Table 12. DDP-days, II-% and LDNA-%
Page 131 and 132: Chapter 5 General Discussion Gamma
Page 133 and 134: FHIA-01 were not able to compare du
Page 135 and 136: ‘W 1 II 31’. In the case of ‘
Page 137 and 138:
Summary Both Gamma rays and Carbon
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A ‘Cavendish Enano’ plant with
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I extend my gratitude also to Dr. K
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Cohan, J, P., C. Abadie, K. Tomekp
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Hase, Y., M. Yamaguchi and A. Tanak
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Molina, G. C. and J. P. Krausz (198
Page 149 and 150:
Roux, N. S. (2001). Mutation induct
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