Midterm Report for FNCA Mutation Breeding Sub-Project on Insect Resistance in Orchid
Evaluation Items

Indonesia

Malaysia

1. The selection of Starting
breeding Materials.
- Whether it wasappropriate or not.
2. The determination ofculture methodologies- Tissue culture ofclones, and Isolationand multiplication ofselected mutant clones.

D. ‘Sonia 17 Red’ and D. ‘Jayakarta’, were D. ‘Sonia 17 Red’ and D. ‘Jayakarta’ , D. mirbellianum,
selected for starting breeding materials.
were selected for starting breeding materials

3. Irradiation to Starting
Materials.
- The determination of
the methodologies, e.g.
optimum dose, radiation
sensitivity, etc.

PLBs of Dendrobium ‘Sonia 17 Red’ fromThailand were irradiated with acute gamma
irradiation at doses 0, 30, 70, 80, 90
and split dose 70 + 30 Gy. The cultures of 30, 70,
80 and 70 + 30 Gy showed better performance
although their growth was slower compared withuntreated cultures. The culture with the dose 90
Gy showed rosette and pale leaf color.
These works are conducted at the tissue culture
laboratory of CRDIRT and IOCRI.

D. ‘Sonia 17 Red’ PLBs were obtained fromThailand.
The PLBs of D. ‘Jayakarta’ were initiated on
modified Vacin and Went medium enriched withcharcoal.
This work was conducted at the tissue culture
laboratory of CRDIRT.

4. Selection of Mutants Screening techniques have not been established.
CloneswithInsectResistance.
- Establishment ofscreening technique.

ThailandD. ‘Sonia 17 Red’ and Dendrobium. ‘Sonia Earsakul’ wereselected for starting breeding materials

Young shoots from healthy plants were collected fromcommercial orchid nurseries for tissue culture.
-The lateral buds and terminal buds were excised from sterileshoots and cultured in liquid Vacin-Went medium.
-After 2 months under agitation condition, explants produced
mass of protocorm like bodies (PLBs). The PLBs were
It is difficult to categorize the vegetative stages of the multiplied in liquid medium and transflask to solid medium for
PLBs multiplication.(4 months)
proliferating protocorms after the irradiation treatment.
PLBs of Dendrobium ‘Sonia 17 Red’ from Thailand were The PLBs were irradiated with gamma rays;irradiated with acute gamma irradiation at doses 0, 30, 70, Phase I : Dendrobium ‘Sonia No.17 red’
Acute gamma rays 0,60,70,80,90,100 Gy
80, 90 Gy.
Dendrobium ‘Sonia Earsakul’High dose acute gamma rays 0,60,70,80,90,100D. ‘Jayakarta’ was irradiated with acute gamma ray at 30Gy
Gy and split doses (10 + 10 + 10 Gy). The dose was
Low acute gamma rays 0,2,4,6,8,10 Gy
selected based on data of previous study.
Split dose acute gamma rays 0,20,20+20,40 Gy
Chronic gamma rays 0 , 400, 800 Gy
D. mirbellianum protocorms were irradiated with acutegamma ray and ion beam. However it is difficult to Results: growth retardation was observed but it is not possible
accurately determine LD50. For ion beam irradiation the to determine the optimum dose from the current results.
doses used were, 0, 0.2, 0.4, 0.8, 1.0, 2.0, 4.0, 6.0, 8.0,
Phase II : Dendrobium ‘Sonia No.17 red’ and Dendrobium
10.0, 12, 15, 20, 30, and 50 Gy.
‘Sonia Earsakul’
Protocols for gamma and ion beam irradiation have been ・ In vitro seedlings
established for Dendrobium protocorms
Chronic gamma rays 50, 50+50, 100+50,100+100, 200+100
All of seedlings were grew in nursery for 6months.
PLBs
・Acute + Split dose chronic gamma rays
0+ 50 + 50
10+ 50 + 50
20+ 50 + 50
30+ 50 + 50
In vitro screening method has been developed. Selection of insect resistance mutant clones with natural
Preliminary results have shown that up to 28% of the infestation techniques.
irradiated plantlets and 14% of the control plantlets were Seedlings of each treatment were grown in nursery withoutable to survive after 3 months infestation with mites, application of insecticide and fungicide. During four months
however a better and more efficient screening method is under natural infestation of thrips, 0-17.4 % of seedlingdamaged by thrips and up to 67.27-92.31 % after 10 months ofstill needed.
the experiment.

D. ‘Sonia 17 Red’ and D. ‘Jayakarta’ PLBs were obtained
from Thailand and Indonesia respectively.
Standard orchid culture techniques are followed. For D.
mirbellianum the protocorms were initiated from selfedflower.

5. Testing and Evaluation The project has not reached field trial stage yet.
of Selected Mutants in thefield.
- The data of field trials
and estimate of theirability.
6. Other result and ripple
effects of the project, if
there are.

7. Opinion about the Mass rearing of thrips for the selection of tolerant
project (problems, ideas, mutants is needed. It is very difficult andremarks, proposal, etc.)
requires much effort and fund. Collaboration
with other research institutes and universities inconducting the research activities is needed due
to limited fund for the implementation of theproject.

The project has not reached field trial stage yet.

The thrips damaged and non-damaged seedlings were grown toflower in nursery.
The screening technique for selection of thrips tolerance on
flowers will be conducted under natural and artificialinfestation.

Training for junior researchers in mutagenesis technology, Workshop on plant resistance to insects was organized for
introduction of new technology (ion beam irradiation).
trainees from Malaysia and Thailand.
The technology and techniques developed can be extended A scientific TV programme was made on the nuclear
for the improvement of other important crops.
technology applications in orchid research in Thailand byJapanese.
A good awareness of the technology has been created in theorchid industry. The industry has shown great interests and at
least two companies have utilized the technology for orchidimprovement.
Mass rearing of thrips for the selection of tolerant mutants Mass rearing of thrips for the selection of tolerant mutants isis needed. It is very difficult and requires much effort and needed. It is very difficult and requires much time, effort andfund.
fund.
Lack of funding, three varieties are too many to work on at Additional funding is needed as the current funding is finishedat the end of October 2005.
one time.