Karyanti, Devit Purwoko and Teuku Tajuddin

Biotech Center, BPPT

June 25, 2008
 Introduction
Indonesia has large
biodiversity for plant
producing oil.

One of the potential

plant is Jatropha curcas.
The seed contains
about 25-35% oil.
 Developing
Jatropha curcas
is one alternative
solution for the
availability of
biodiesel, land
conservation, as
well as the
opportunity for
increasing the
farmer’s income.
 Obstacles
The cultivation of Jatropha
curcas in Indonesia is short
of high quality clones.

There are huge variation in

seed yield capacity and oil
concentration in the seeds
among the seedling used
for cultivation or
plantation.
 Obstacles
Fruits of Jatropha in one brunch ripe at
different time. It causes difficulty during
harvesting, that farmer has to select ripening
fruits only. This makes harvesting time slow
and high cost.

Uniformity of
maturity will reduce
harvesting cost.
 The plants propagated by seeds are highly
heterozygous. Thus, variations in genetic
characters are to be expected.
In vegetative
propagation, unlike
generative method,
superior traits or
characters of
mother plants with
respect to yield, oil
quality and disease
resistances are
conserved in their
progenies.
On going Research in Biotech Center
Mother plants collection with specific
characters
In vitro & ex vitro propagation
DNA finger printing method to characterize
the mother plants
Developing the F2 or
backcross population
based on the selected
mother plants
Construct the linkage
mapping using DNA
markers
Ex vitro propagation
Mother plants collected in Biotech Center
 Jatropha from Gunung Kidul, Yogyakarta, grows well
in dry stone-soil, leaf dark-green in color and
resistance to salt.
 Jatropha from Bengkulu and Central Sulawesi, have
high concentration of seed oil content (37-39%).
 Jatropha from Sukabumi, West Java, and Mataram,
NTB have superior high seed yield (more than 20 fruit
per brunch).
 Jatropha from Bogor, West Java, has characteristic in
uniform fruits maturity.
 Jatropha from NTB with vigor, fast growing tree and
wide leaf, however very low seed yield (2-3 fruit per
brunch).
 And from other areas . . . .
Artificial crossing on
selected mother
plants
Purposes of in vitro research
 In order to supply the high quality seedling
in large amount, the micro-propagation
methods for this plant were studied.
 Increasing the multiplication rate of in
vitro culture
 METHODOLOGY
Leaf Explants
1

5 1 2 3

4 5
Culture Condition
 The leaves explants were obtained from the
2-weeks germinated seedling grown on agar
under sterile condition.
 Leaf explants of Jatropha curcas were
cultured in vitro for 8 weeks
 Room temperature: 27-29oC.
 Light intensity: 1500 Lux
 Photoperiod: 12 hour/day.
Experiment 1: Plant growth Regulator BAP & NAA

NAA BAP (ppm)

(ppm) 0 0.2 0.4 2.0 4.0

0 A B C D E

0.2 F G H I J
Experiment 2: Plant growth Regulator BAP & Kinetin

5 10 15 20 25 30
BAP 1 2 3 4 5 6
(ppm)

5 10 15 20 25 30
Kinetin 7 8 9 10 11 12
(ppm)
RESULTS AND DISCUSSION
Experiment 1 – Growth of Callus
NAA BAP (ppm)
(ppm) 0 0.2 0.4 2.0 4.0
0 – ++ +++++ +++ ++++
+++ ++++++ ++++++ ++++++ ++++++
0.2 (Rooting)

NAA BAP (ppm)

(ppm) 0 0.2 0.4 2.0 4.0

0.2
 F5

Root formation at the 5th section of leaf explants

on F treatment (0.2 ppm NAA)
Adventitious Shoot Formation
Average and Total number of shoots

NAA BAP (ppm)

(ppm) 0 0.2 0.4 2 4
1 1.2 7.2
0 0 0
(5) (6) (36)

0.2 0 0 0 0 0
 B3 B4 B5

Shoots were induced on leaves explants

no. 3, 4 and 5 of B treatment (BAP 0.2 ppm)
 E2 E3

D3 E5

Shoot were induced on leaf explants of

treatments D (2 ppm BAP) and E (4 ppm BAP),
8 week after culture
Media for shoot
elongation on lower BAP
 Experiment 2 – Growth of Callus
BAP
(ppm)
5 10 15 20 25 30
Size +++++ ++ +++ + + +
Soft Semi
Formation Solid Friable Solid Solid
friable friable
Greenish
Color Green Green White White Green
white
Averg No.
1.7 (5) 0 0 0 0 0
of Shoot
Length of
2 mm – – – – –
Shoot
Hormone BAP
5 ppm 10 ppm 15 ppm

20 ppm 25 ppm 30 ppm

 Experiment 2 – Growth of Callus

Kinetin
5 10 15 20 25 30
(ppm)
Size +++ ++++ +++++ ++++ – –
Semi
Formation Solid Friable Friable – –
solid
Color Green Green Green Green – –
Averg No.
0 0 2.7 (8) 3.3 (10) 0 0
of Shoot
Length of
– – 10 mm 15 mm – –
Shoot
Hormone Kinetin
5 ppm 10 ppm 15 ppm

20 ppm 25 ppm 30 ppm

Conclusions
1. The base part of the
leaves gave the best
result for shoot
induction

2. Media contains 4-5 ppm BAP, as well as

15-20 ppm Kinetin, produce higher shoots
formation
3. Media for shoot elongation is best in lower
BAP