Attachment IV-1

Improvement of Banana for FUSARIUM Wilt Resistanceand High Fruit Quality through Mutation Induction
Rusli Ibrahim, Norazlina Noordin, Edrina Azlan & Azhar Mohamad
Malaysian Nuclear Agency (Nuklear Malaysia)
Agrotechnology & Biosciences Division
Bangi, 43000 Kajang, Selangor, Malaysia
Abstract
Popular local banana cultivar called Berangan (Musa spp. AAA) was selected as thestarting material for mutation induction using gamma rays. Radiosensitivity test was carriedout by irradiating meristem tips obtained from suckers with a series of gamma ray doses of0, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100Gy. Data for radiosensitivity test was determinedby 3 parameters, such as 1) Plant height (shoot height), 2) Percentage of survival ofirradiated explants and 3) Multiplication or Growth Rate. Based on the percentage survivalof irradiated explants, LD50 and LD100 obtained for cultivar Berangan were 50Gy and 80Gy
respectively. Using selected effective doses of 20, 30, and 40Gy, all in vitro shoots derived
from irradiated meristems were subcultured from M1V1 upto M1V5 and rooted. Theserooted in vitro plantlets were used to screen for tolerance/resistance against Fusarium wilt
disease. Four artificial screening techniques for Fusarium wilt disease had been developed,
such as 1) Dipping of in vitro shoots in Fusarium oxysporioum suspension (10 6 spores/ml)
for 1-2 hrs and later transferred to sterile sand media in the greenhouse, 2) Double tray
method by inoculating in vitro plantlets that had been planted in sand media, 3) Nursery
screening method, whereby rooted in vitro plantlets that had been hardened for 4-8 weeks
were transferred to media that had been pre-inoculated with Fusarium oxysporioum sporesuspension (106 spores/ml) for 2 weeks and 4) Direct field screening, whereby rooted
plantlets that had been dipped in Fusarium oxysporioum suspension (106 spores/ml) for 1-2
hrs were planted directly in the field. Based on the results obtained after screening oftreated plants using artificial inoculation of Fusarium oxysporioum in the field, threepotential mutants had been identified, such as 1) Tolerant/resistant mutant, 2) High yielding
mutant and 3) Early flowering mutant. These mutants will be further screened in the field
for multi-location trials.

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Keywords: Artificial inoculation, gamma irradiation, Fusarium wilt, in vitro mutagenesis,
meristems.

Attachment IV-2

Agreement and Request for the Extension of Banana Subproject

Conclusions
1.

2.
3.
4.

All participating countries such as Philippines, Vietnam and Malaysia have successfully
achieved the objectives of the project by identifying promising mutant lines which are
disease resistant (BBTV, Fusarium and Nematode).
Achievement from this project will be published as proceedings and protocols forirradiation and screening for Fusarium, BBTV and Nematode.
As a new member of this sub-project of banana, Bangladesh can adopt all the developed
disease screening methods by other participating countries.
The protocol for anther culture developed by Bangladesh can be adopted by other membercountries.

Recommendations
All participating countries agreed to extend this project until 2010 as Phase II towards achieving thefinal goal of obtaining resistant new varieties of banana based on the following reasons:Bangladesh- will adopt mutation breeding technologies which has been established by other participating
countries (Philippines, Vietnam, Indonesia, Malaysia) since Bangladesh just started the program
last year in 2007
- Bangladesh has established a good protocol for anther culture which can be adopted by otherparticipating countries
Malaysia- already identified 3 promising mutant lines of variety ‘Berangan’ banana which are resistant toFusarium wilt with improved agronomic characters such as early flowering, dwarf and highyielding- will be screened for multi location trials at 3 sites for 2 cropping season (in collaboration withcommercial plantations)- mutants will be registered and micro-propagated for tissue culture plantlet production
Philippines- already identified 5 promising mutant lines of variety ‘Lakatan’ which are resistant to BBTV andwill screen for multi location trials (farmer’s plot) for 2 cropping seasons- mutants will be registered for plant breeder’s right as new varieties- mutants will be propagated for commercialization
Vietnam- already identified 2 promising mutant lines of variety ‘Tay’ banana which are resistant toFusarium wilt
- will be screened for multi location trials at 2 sites (north and central)- mutants will be propagated for production