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

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selected for their tolerance to 25 ppm of Juglone, but new screening for resistance to M. fijiensis in these plants still has to be confirmed through inoculation with the fungus, which is a very slow process. On the other hand, the results obtained by Lepoivre et al. (2002) confirm the possibility of selecting banana plants resistant to M. fijiensis metabolites but in their research this approach did not result in higher resistance to black leaf streak disease. Since the leaf discs of the resistant cultivar (in this case ‘FHIA- 01’) were less affected by juglone (a host-specific toxin) applied at 150 ppm, in contrast to susceptible cultivars, the leaf disc immersion method can be a simple and rapid method to measure the reaction for tolerance/resistance to juglone. A low extent of necrosis in susceptible irradiated cultivars was observed (selected ones); however, new studies to reconfirm tolerance/resistance to juglone on the selected material must be undertaken. Van Harten (1998) mentioned that in the case of selection with phytotoxins and culture filtrate appears to be more effective than the use of the pathogen itself. Lepoivre et al. (2002) reported that chloroplast is target site of juglone. When juglone was used, swelling chloroplasts were observed by electron microscopy in ethyl acetate crude extract (EaCE)-treated leaves. Upon observation, ‘Fougamou’ (a partially resistant cultivar) chloroplasts appeared to be less affected by juglone in contrast to ‘Grand Naine’. These results suggest that the chloroplast is one of the primary action sites of juglone. 3. 3. 5. Relative DNA content measuring by using flow cytometer Histograms of the relative DNA content of ‘Cavendish Enano’, ‘Williams’ and ‘FHIA-01’ are showing in Fig. 16. The relationships 41
etween the irradiation doses and the relative DNA content (Fig. 17) showed that the values from the three cultivars (‘Cavendish Enano’, ‘Williams’ and ‘Orito’) showed an imperceptible variation in all the applied doses. However, a perceptible variation was possible to observe in ‘FHIA-01’ when higher doses were applied. The variation of ‘FHIA-01’ was better observed when a frequency distribution of the relative DNA contents was analyzed (Fig. 18). The analysis detected reduction of the relative DNA content when the irradiation dose was increased. The lowest value was 1.947 (96% of donor), observed at 300 Gy. These results suggest that deletion of DNA was occurred when high doses were applied in ‘FHIA-01’, exhibiting a shift to the left, indicating loss of whole or part of chromosomes. Aneuploid is the mean having one or more complete chromosomes in excess of, or less than the normal haploid, diploid or polyploidy number characteristics of the species. Seventeen plants with lower DNA content were selected corresponding to the doses of 200 and 300 Gy. FAO/IAEA (2002) screened aneuploid mutants by flow cytometric analysis in irradiated banana plants. Aneupliod mutants can be detected through chromosome counts with rather time-consuming process. Results obtained by flow-citometry were compared to chromosome counting in meristem shoot-tip cells. It could be shown that flow cytometry is sensitive enough to detect aneuploidy in bananas. With such a sensitive and fast technique we are now screening routinely for aneuploid mutants in addition to screening for juglone resistance. Aneuploid mutants would be important tools for basic research in Musa. Dolezel et al. (2002) reported that flow cytometry have been used to determine ploidy levels of Musa accessions. Flow cytometric ploidy assay involved preparation of suspensions of intact nuclei from small amount of the leaf tissue and the analysis of fluorescence intensity 42
Page 1 and 2: Gamma Rays and Carbon Ion-Beams Irr
Page 3 and 4: Table of contents Chapter 1 1.1. In
Page 5 and 6: 4.2.1.1. Cultivars of banana……
Page 7 and 8: and ‘Valery’, which are exporte
Page 9 and 10: ‘Cavendish Enano’, ‘Williams
Page 11 and 12: Although it is now found only in Au
Page 13 and 14: varieties bred at the Jamaican prog
Page 15 and 16: such as ‘Paka’ (AA) and ‘Pisa
Page 17 and 18: Resistance (PR). In connection with
Page 19 and 20: large number of improved mutant cro
Page 21 and 22: which migrated faster (Rf = 0.44) t
Page 23 and 24: is juglone (5 hydroxy-1,4-napthoqui
Page 25 and 26: experiments involving various biolo
Page 27 and 28: Tanaka (1999) reported novel genes
Page 29 and 30: 2. 1. Materials and Methods Chapter
Page 31 and 32: 2. 1. 1. 4. FHIA-01 ‘FHIA-01’(M
Page 33 and 34: (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: ‘Williams’ (93%), the necrotic
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 and 86: placement among the plants (Fig. 30
Page 87 and 88: hours, photographs of the leaf-disc
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
Page 139 and 140:
A ‘Cavendish Enano’ plant with
Page 141 and 142:
I extend my gratitude also to Dr. K
Page 143 and 144:
Cohan, J, P., C. Abadie, K. Tomekp
Page 145 and 146:
Hase, Y., M. Yamaguchi and A. Tanak
Page 147 and 148:
Molina, G. C. and J. P. Krausz (198
Page 149 and 150:
Roux, N. S. (2001). Mutation induct
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