Department
of Agriculture
Bureau
of Agricultural Research
RDMIC Bldg.,
Elliptical Rd. Cor. Visayas Ave.
Phone Nos.:
(632) 928-8624 & 928-8505 ·
Fax: (632) 927-5691
DETAILED
PROPOSAL FORMAT
A. bASIC
iNFORMATION
1.
Title of the Project: Establishment of Rapid Propagation Techniques
for Seedless Breadfruit in Marinduque
2.
Proponent (s)
2.1. Name
and Signature: Carlos J. Andam
2.2. Designation: Executive Assistant for
External and Linkages
2.3. Institution: Marinduque State
College
2.4. Address: Tanza, Boac,
Marnduque
2.5. Telephone
Number (s): (042) – 332-2028/09184299794
2.6. Fax
Number: (042) –
332-2728
2.7. Email
Address: cjandam@yahoo.com
2.8. Name
and Signature: Marilyn A. Mayores
2.9. Designation: Research and Extension
Coordinator
2.10. Institution: Marinduque State
College
2.11. Address: Tanza, Boac,
Marnduque
2.12. Telephone
Number (s): (042) – 332-2028/09266109011
2.13. Fax
Number: (042) –
332-2728
2.14. Email
Address: ellen_mayores@yahoo.com
2.15. Name
and Signature: Harvey A. Dulay
2.16. Designation: Agricultural Technologist
2.17. Institution: Marinduque State
College
2.18. Address: Tanza, Boac,
Marnduque
2.19. Telephone
Number (s): (042) –
332-2028/09475760562
2.20. Fax
Number: (042) –
332-2728
3.
Implementing Agency
3.1. Lead
Agency: Marinduque
State College
3.2. Collaborating
Agency (s):
Department of Agriculture,
Bureau of Agricultural Research (source of fund)
Provincial Government of
Marinduque-Provincial Agricultural Office (Source of data & information,
Co-implementing agency)
4.
Project Duration: 2.5 years
5.
Project Location: MSC Tanza, Boac,
Marinduque
6.
Total Budget Requested
6.1. Budget Requested 3,540,200.00
6.2. Agency Counterpart
6.3. Other Sources
B. Technical
Description
1.
Rationale
Breadfruit
Artocarpus atilis) has long been an
important staple crop and a primary component of traditional agroforestry
systems in Oceania. This fruit when cooked at all maturity stages, is high in
carbohydrates, and is a good source of minerals and vitamins.(Ragone & Paull, 2006).
Currently,
this seedless breadfruit is underutilized and seemingly being neglected
commodity in the Philippines despite the enormous food uses as reported by many
literature.
The
initiative of Marinduque State College in developing breadfruit as a source of
flour to support local industries which are flour requiring for their raw
materials is in response to reducing our dependency on imported wheat or wheat
flour coupled with the recurring increases in its price in the country.
Since
this crop produces a seedless fruit, it is therefore being propagated
asexually. Vegetative propagation techniques are necessary for the propagation
of this fruit. However, because this commodity is not popularly known in the
country, production there is no commercial experience in massive production of
this crop.
With
this scenario, there is a need to establish the research and development for
the propagation of breadfruit in order to support the needed raw materials for
the production of breadfruit and breadfruit - based product and conserve
endemic breadfruit tree.
2.
Objectives
General
objective:
To develop rapid propagation techniques
for the production of quality planting materials of seedless breadfruit in
Marinduque.
Specific
objectives:
a.
Conduct benchmarking on how seedless breadfruit is being propagated in a
recognized institutions focusing on the conservation and sustainable production
of breadfruit.
b.
To evaluate different natural vegetative propagation techniques for breadfruit;
c.
To formulate optimized media for the in
vitro micro-propagation of breadfruit;
d.
Establish nursery for validating propagation technique which have high
potential for massively multiplying this seedless breadfruit in order to
support an intensified planting in the province.
3.
Review
of Literature
Breadfruit
is a rich source of carbohydrates, minerals and vitamins. Though it is widely
distributed, it has often categorized as underutilized and little attention has
been given to its large scale commercial cultivation due to limited knowledge
on planting materials, agronomic practices and production techniques. (Deivanai and Bhore, 2010)
Vegetative
propagation
This
seedless breadfruit is often propagated through the root shootlets, root
cutting, air layering (marcotting), budding and grafting into seedling root
stocks and by the use of in vitro
tissue culture technique. (Ragone 1988 and Fowness and Raynor 1993). However,
vegetative propagation depends on the local climate and water availability (Deivanai and Bhore, 2010).
Vermicompost
Another
alternative source of nutritious inputs in the field is the casts produced by
species of worm. Wastes are converted into compost which contains natural
nutrients that will suffice needs of field crops. This cast which is termed as
vermicast or vermicompost contains higher percentage of both macro or
micronutrients than garden compost (ICRISAT 2006).
Root
cuttings
The
roots which are present slightly below the soil surface often grow and produce
a shoot, especially when it is cut or damaged (Deivanai and Bhore, 2010). Further,
the success of root cuttings ranged from 80 to 85% and the cuttings can be made
ready for transplanting from the propagating bed in 3 -5 months time (Gunarto
1992).
Air
layering
Air
layering of the young tree will produce advantageous roots earlier than the
matured tree (Coronel 1983). The success of this propagation technique for
breadfruit trees still depend on the age of the tree(Deivanai and Bhore, 2010).
Stem cuttings
Another
possible method for breadfruit propagation is through the stem cuttings.
Hamilton, Criley and Chia 1982 reported that 95% of the stem cutting produce
sufficient root in 10 weeks after planting under mist condition.
In vitro Tissue culture
At
present there are very limited information regarding in vitro propagation of breadfruit. However, information regarding in vitro propagation of jackfruit is
very useful (Deivanai and Bhore, 2010). Shoot
tips, auxiliary buds (nodal sectors) seed and zygotic embryos can be used to
initiate the in vitro propagation
(Bhore and Vaishana 2010). It was also suggested that explants taken from new
branches or from recently pruned branches initiates better shoot proliferation
(Wilkins 1991).
The
first thing to be considered in micropropagation is the sterilization procedure
of explants to be initiated. Explants will be soaked into 70% ethanol for 1
minute. After which, ethanol will be replaced with 10% bleach (5.25 % sodium
hypochlorite) and occasionally stir to exposure the explants completely with
the sterilant for 15 minutes. Wash the explants 3-5 times with sterile
distilled water (Murch et al. 2008).
According
to Tuia, Taylor and Ragone , undated, Woody Plant Medium (WPM) (Llyod and
McCown, 1980) with the addition of 2.5 mg/L of 6-benzylaminopurine (BAP) was
the optimal medium in terms of survival, root production and shoot growth of Artocarpus altilis. On the other hand,
it was also found that Murashige and Skoog (MS) media (Murashige and Skoog
1962) supplemented with myo-inositol 100 mg/L, glycine 2 mg/L, Thiamine HCL 0.1
mg/L and pyridoxine 0.5 mg/L with the addition of 2.5 mg/L of 6 –
benzylaminopurine (BAP) increase the nodal budding. The pH of the media will be
adjusted to 5.7-5.8 before it solidified with 7.8 g/L agar. 10 ml of medium
will be dispense into a culturing vessel prior to autoclaving at 121˚c at 15
psi for 20 minutes.
Cultures will be placed
in the growing room at a temperature of 25˚C ±2˚C and provided with a light
intensity of 2668 lux irradiance and daylength of 18 hours. For the duration of
4 months the survival, establishment in the tissue culture, plant height, root
development, contamination by fungi or bacteria and production of phenolic will
be recorded followed by subculturing into fresh media (Tuia, Taylor and Ragone
, undated).
Once
the plantlets attain the optimum height they should be taken out for hardening.
Roy and Hadiuzzaman 1991, reported that about 80% of the plantlet survive if
rooted plantlet in the culture tube will be transferred to earthenware pots
containing sterile sand, soil and humus in the ratio of 1:2:1 and covered with
a transparent plastic sheet.
4.
Methodology
The
initial move for this project is conduct benchmarking on recognized
international and local institutions focusing on Horticultural Research and
development such Postharvest Horticulture Training and Research Center (PHTRC)
based at the University of the Philippines, Los Baños and the Breadfruit Institution
of the National Tropical Botanical Garden (NTBG) in Hawaii.
Study
1. Evaluation of natural vegetative propagation techniques of seedless
breadfruit
Study
1.1 Propagation through Root cuttings
This study will focus on the potential
natural propagation techniques for seedless breadfruit. Since literatures
emphasised that breadfruit can be propagated through root cutting, initial
visit to the PHTRC – UPLB will be conducted to acquire rooting hormone for the
enhancement of the rooting of collected cuttings.
Step 1. Collection of root cutting
stock
Roots that are present slightly below
the soil surface which about 1 to 2.5
inches
(2.5 to 6.25 cm) thick size will be cut into 9 inches (22 cm) long. This will
serve as the root cuttings. Cuttings will be collected from the various donor
breadfruit trees identified in the province.
Collected
stocks will be subject to cleaning to remove residual soil. The end of the root
cutting will be dipped into potassium permanganate to coagulate latex (Deivanai and Bhore, 2010).
Step 2. Planting Media preparation
and planting of root cuttings
Plastic
bags or polyethylene bags containing compost or dried manure or combination
will be used as planting media. Root cutting will be planted horizontally into
the media. This will be placed into the shaded portion of the nursery and
watered daily. Regular monitoring and evaluation will be done after the setting
up.
Study 1.2. Propagation
through air layering or marcotting
Braches
stem with 5-15 cm in diameter that does not bear flower and fruit will be
selected for air layering. A strip of bark with about 3-5 cm (1.4 - 2 in) will
be removed from the branch using a sharp knife. Opened bark will be wrapped
with moistened sphagnum moss and covered with plastic. A piece of foil will be
wrapped around to minimize evaporation. Regular monitoring evaluation will be
conducted after the set up to observe the occurrence of roots.
To
evaluate the best location in the canopy for marcotting, three locations (below
the canopy, within the canopy and top of the canopy) will be considered. With the
recommended size of branch for marcotting, different sizes of branch will be
considered for evaluation which includes branches with the size of 1-2 cm; 3-4 cm; 5-6 cm; 7-8 cm and 9-10 cm in
diameter.
Branches
with grown roots will be cut and place into a plastic bags containing different
ratio of medium as shown below.
a.
pure vermicompost;
b.
pure soil;
c.
1:3 vermicompost : soil;
d.
1:1 vermicompost : soil;
e.
3:1 vermicompost : soil;
Study 1.3. Propagation
through the stem cuttings
Stem
with less leaf will be selected for this study, cuttings will be treated with
plant regulating hormone to induce rooting. The set up will be placed into a
mist condition. Different sizes of stem cutting will be collected which
includes stem with the size of (1-2 cm; 3-4 cm; 5-6 cm; 7-8 cm; 9-10 cm) in
diameter and will be sown into a plastic pot containing different ratio of
medium:
a.
Pure vermicompost
b.
Pure soil
c.
1:3 Vermicompost : soil
d,
1:1 Vermicompost : soil
e.
3:1 Vermicompost : soil
Study
2. In vitro propagation of breadfruit
This
study will focus on the micro propagation of seedless breadfruit. Tree identified
within the province will be the source of shoot tips to be grown in MS media
and WPM for experimentation added with different concentration of
6-benzylaminopurine (BAP)
Step
1. Collection and preparation of explants from donor trees
Shoot
tips with a size of 0.5 cm to 1.0 cm long containing meristem and 2-3
leaves
primordial (Deivanai and Bhore, 2010) will be
collected from the donor tree found in various locations in the province
considering the climate and elevation of the area of collection.
Explants
will be soaked into 70% ethanol for 1 minute. After which, ethanol will be
replaced with 10% bleach (5.25 % sodium hypochlorite) and occasionally stir to
exposure the explants completely with the sterilant for 15 minutes. Wash the explants
3-5 times with sterile distilled water (Murch et al. 2008).
Step
2. Initial culture of the Explants
After surface sterilization, shoot tips
will be initially culture into tube contain 10 ml of MS media added with
different concentrations of 6-benzylaminopurine (BAP). On the other hand, same
concentration BAP will be added to WPM. The performances of the explants will
be evaluated from the two types of media used. Each culture tubes will be treated
with the following 6- benzylaminopurine concentrations (2.5 mg/L; 5 mg/L; 7.5
mg/L and 10 mg/L). It will be replicated three times.
Cultures
will be placed in the growing room at a temperature of 25˚C ±2˚C and provided
with a light intensity of 2668 lux irradiance and day length of 18 hours. For
the duration of 4 months the survival, establishment in the tissue culture,
plant height, root development, contamination by fungi or bacteria and
production of phenolic will be recorded followed by subculturing into fresh
media (Tuia, Taylor and Ragone , undated).
Step
3. Sub culturing
After
the initial culturing the explants will be subcultured into a fresh medium for
further growth development, root development, and optimization of height prior
to hardening.
Step
4. Hardening and Nursery Establishment
Plantlets
with optimum height from the culture room will be transferred to natural
environment and acclimatize them to nature condition. Plantlets will be remove
into the culture vessel, wash with tap water to remove exist media and dried
with tissue paper.
After cleaning, plantlets will be sown
into a seedling tray containing sterilized sand, soil and humus in the ratio of
1:2:1 and place it the greenhouse. Regular monitoring and evaluation will be
done in order to supervise the growth and development of the plantlets.
Step 5. Outplanting of matured
planting materials
From the nursery, matured plantlets
will be planted in identified land area at Mogpog, Marinduque for field evaluation.
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