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

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4. 2. 1. 4. Post-irradiation management After the evaluation of the explants growth at the laboratory, each treatment/cultivar growing in the Erlenmeyer flask were propagated three times using the same composition of the liquid medium mentioned above, in order to increase the number of explants for nursery and field conditions experiments, that were conducted in Ecuador. They regenerated into shoot tips and then they were planted individually into a test tube containing 10 mL of MS solid medium. Additionally, 0.5 mg of activated carbon for rooting was added by applying on the solid medium surface. A total of 1707 rooted plantlets were sent to Ecuador (December, 2005) in autoclaved plastic bags. Processing plants for bagging is shown in Fig. 23. The numbers of plantlets per cultivar/doses were as follows: for ‘Williams’ in the doses of 0 (94), 0.5 (95), 1 (105), 2 (95), 4 (32), 8 (87), 16 (103), 32 (70), 64 (95) and 128 Gy (60). For ‘Cavendish Enano’ in the doses of 0 (55), 0.5 (95), 1 (95), 2 (100), 4 (80), 8 (40), 16 (80), 32 (53), 64 (53) and 128 Gy (100). The materials were found free of viruses such as Banana Bunchy Top Virus (BBTV), Banana Bract Mosaic Virus (BBMV) and Banana Streak Virus (BSV) by using ELISA virus indexing technique. They also were free of any other pest and disease including nematodes, Mycosphaerella fijiensis, Fusarium oxisporum, and so forth. The plantlets arrived in the Laboratory of Biotechnology of the Estación Experimental Tropical Pichilingue, Instituto Nacional Autónomo de Investigaciones Agropecuarias (INIAP), Ecuador. Immediately, the bags containing the plantlets were kept in the tissue culture room at 26 o C and light conditions for two days to recover photosynthesis. When arrived, the banana material showed injured probably occurred during transportation (Fig. 24). 77
4. 2. 2. Assessment of banana plantlets at the nursery and at field conditions A soil bed was constructed inside a greenhouse and the plantlets were transferred on a soil substrate (soil : vermiculite = 1:1) and covered with a plastic sheet to avoid dehydration (Fig. 25). Factors such as transportation conditions, disease attack, or some abnormal behavior caused by the irradiation could bring about the high mortality (Fig. 26). Surviving plants were transplanted to bags and covered by using a cotton sheet and spraying water for three times a day to avoid plantlets dehydration (Fig. 27). Only 87 plants survived the nursery acclimatization. Plants were kept in a nursery plot until May 2006 and the bags were changed for a bigger one to recover vigor (Fig. 28). Fortunately, plants from every dose for the two cultivars survived except in the dose of 2 Gy of the ‘Cavendish Enano’. Using these materials (height of plant around 35 cm) the experiments for black Sigatoka inoculation in nursery plot and later on at the field conditions were conducted. 4. 2. 2. 1. Disease development period (DDP-days) and infection index (II-%) Three banana leaves per plant were inoculated. Plants were about 30 cm height and the younger expanded leaf was marked as the first leaf for inoculation by a conidial solution. Second and third successive young emitted leaves were inoculated by fragments of the diseased banana leaves. Prior to inoculation of the first leaf, a solution containing a M. fijiensis conidial was prepared. The cultures were provided by the Laboratory of Pathology of the Estación Experimental Tropical Pichilingue, INIAP (Fig 29, A). The colonies for inoculation were cut in small pieces 78
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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
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 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: were recorded. From the results, th
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
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‘W 1 II 31’. In the case of ‘
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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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