Activity report from Indonesia - APRSAF

The 15 th Session of
The Asia Pacific Regional Space Agency Forum
(APRSAF)
Hanoi, 9 – 11 December 2008
National Institute of Aeronautics and Space (LAPAN)
INDONESIA

Feasibility Study Report
Banana Experiment
by Dr. Fenny Dwivany, et al. (ITB)

Proposed Joint Research on JEM
Dr. Fenny M. Dwivany of SITH-ITB propose research on “The effect of a
space environment on the expression of fruit ripening associated
genes”
Hypothesis: Factor that induce fruit ripening: Autocatalytic process
when ethylene is produced by fruit itself or produced by other sources
in the space environment that can induce fruit ripening.
Hypothesis: Factors that inhibit fruit ripening: Compared to conditions
on the earth at l g, lack of convective mixing in space may cause large
boundary layers of O 2 and CO 2 , at different levels, to surrounding plant
organs. Elevated CO 2 level can reduce ethylene whilst reduced O 2 level
can reduce ethylene synthesis.
However, whether environmental condition during experiment in
KIBO will change the ACS and ACO gene expression profile and
resulting in inhibition or enhancement of fruit ripening still need to be
studied.

Feasibility Study
Experiment done in Tsukuba Space Center of JAXA
(Japan) and in ITB (Indonesia), January 2008
Dr. Fenny Dwivany (PI), Dr. Rizkita R. Esyanti, and Ms.
Listya U. Karmawan

EXPERIMENT SCHEDULE AT JAXA
DATE ACTIVITY EQUIPMENT CONSUMABLES
14 January 2008 Samples handling
Lab inspection, clinostat and incubator setting, starting experiment using
clinostat,
Incubator
15 January 2008 Ethylene treatment for Japan’s banana Incubator Etephon
16 January 2008 as above
17 January 2008 1. Put banana in centrifuge
2. Take data/sample as day-0 for clinostat using Chiquita banana (sample
code: ACli, ACt)
Centrifuge
Clinostat
Liquid nitrogen
Plastic/aluminium foil
18 January 2008 1. Take data/sample for centrifuge (control, vertical g, horizontal g)
(photo, isolation of banana finger)
2. Put Sunpride Cavendish banana in clinostat (was not used as day-0
sample)
Centrifuge
Clinostat and incubator
Freezer -80 o C
Liquid nitrogen
Plastic/aluminium foil
19 January 2008 1.Take data/sample as day-1 for clinostat using Sunpride Cavendish
banana (sample code: Bcli1, BCt1)
(photo, isolation of banana finger)
2.RNA extraction solution preparation
Clinostat and incubator
Freezer -80 o C
Laboratory glass ware*
Liquid nitrogen
Plastic/aluminium foil
20 January 2008 1.Take data/sample as day-2 for clinostat Sunpride Cavendish banana
(sample code: BCli2, BCt2)
(photo, isolation of banana finger)
Clinostat and incubator,
Freezer -80 o C
Liquid nitrogen
Plastic/aluminium foil
21 January 2008 1.Take data/sample as day-3 for clinostat Sunpride Cavendish banana
(sample code: BCli3, BCt3)
(photo, banana finger);
2. RNA extraction from all samples
Clinostat and incubator,
Freezer -80 o C
RNA extraction equipment*
Liquid nitrogen
Plastic/aluminium foil
Chemicals for RNA extraction

Banana samples:
Banana from Indonesia: The green mature bananas have
been treated with ethylene gas for 24 hr at 14 o C. After
ethylene treatment, the fruits were then sealed with
plastics bag and vacuumed. It takes about 24 hr after sealed
with plastics bag to arrive at JAXA on 14 Jan 08. Banana
fruits were then stored at 14 o C incubator until experiment
was started on 16 Jan 08.
Banana from Japan: The bananas were stored at 14 o C
incubator until 15 Jan 08 and treated with ethephon.
Ethephon treatment was performed 24 hr at 14 o C until
experiment was started on 16 Jan 08.

Experiments
Two experiments were performed: using clinostat and centrifuge. The
control was conducted without any treatment.
Clinostat
The room for Clinostat was set at 25 o C. Bananas were placed on
styrofoam box and rotated at clinostat for one day (Chiquita banana
from Japan/Philippines) and for three days (Sunpride Cavendish
banana from Indonesia). Two banana fingers were collected everyday
for sunpride (at day1, day 2 and day 3), whilst Chiquita only at day 0
Centrifuge
Centrifuge was set at 19-20 o C, 45-50 % humidity and acceleration 49
m/sec or 5 g. Total 6-8 bananas (Chiquita banana) were placed on
centrifuge chamber for one day. The treatments were vertical gravity,
horizontal gravity, and control.

Experiments
Gas sampling
Sampling of gas was performed
by placing one finger of banana
from each time point into glass
jar for 1 hr. The gas was then
collected using syringe. The
collected gas was measured for
O 2 and CO 2 content (ethylene
content was determined later).
Sampling of gasses from the glass jar.

Observation
Observation was made visually and using digital camera.
The color of banana fruits from day 1 to day 3 showed a gradual change
from green to yellow (data not shown).
Interestingly, each banana fruit treated in clinostat showed a sudden
increase of respiration when observed for 1 hour inside the gas chamber.
It was shown by the presence of more water vapor on the wall of the jar
compare to that in the control . Increase in respiration will lead to the
synthesis of more ethylene (auto catalytic), which will then expected to
cause an earlier maturation process compare to that in the control.
Sample treated in centrifuge was not observed for gas generation.
However, in the future, gas observation for this treatment should also
be conducted.

Observation: Day 1
A
B
Sunpride Cavendish Banana sample at day 1, after clinostat treatment for 16 hr
A. control (BCt1) B. treatment (BCli1)

Observation: Day 2
A
B
Water vapor
Sunpride Cavendish Banana sample at day 2, after clinostat treatment for 16 hr
A. control (BCt2) B. treatment (BCli2)

Observation: Day 3
A
B
Water vapor
Sunpride Cavendish Banana sample at day 3, after clinostat treatment for 16 hour
A. control (BCt3) B. treatment (BCli3)

Sample isolation and RNA extraction
Banana fingers were collected and cut into small cubes.
Immediately after cut, cubes were frozen in liquid nitrogen
and placed in -80 o C freezer. Samples were then proceed for
RNA isolation until step 8 (LiCl incubation at 4 o C) and
transported to Indonesa.
The samples were then proceed immediately after arrive in
Indonesia (stored for about 7 hr in 4 o C after arrive, 0.30 am-
7.30 am). The RNA extraction was finished on 24 Jan 08.
The quality of RNA was analyzed using spectrophotometer
and agarose gel. However, the ratios of A260/280 were around
1.0. In addition, electrophoresis result showed that RNA from
sample was degraded. Next time, RNA isolation should be
finished on site to minimize RNA degradation

Experiment at ITB
25S rRNA
18S rRNA
Banana fruit ripening process (day 0-day 6)
RNA isolation from banana pulp during fruit
ripening (day 1-day 7).

Experiment at ITB
(A)
200 bp
(B)
100 bp
400 bp
Electroferogram of RT-PCR results using the MaACO1 and MaACS1 gene specific
primers and three reference genes. The samples were from ripening banana’s pulp
which were harvested on day 1, day, 3, day 5 and day 7. (A) 1-4 (MaACS1 primer) and
(B) 1-4 (MaACO primer), 5-8 (Ma18S rRNA primer), L (100 bp DNA ladder), 9-12
(MaActin primer), 13-16 (MaGAPDH primer).

Experiment at ITB
Figure 3.4 Normalized MaACS and MaACO genes fold expression

Experiment at ITB
CO 2
production and CO 2
consumption during experiment

ISS Kids in Bahasa Indonesia

Future Plan

Banana Experiment
It should be considered whether a longer experiment can be
performed using centrifuge in various g forces in the future.
Clinostat experiment should be performed at least for 2-7 days
in the future
Sample treated in centrifuge was not observed for gas generation.
However, in the future, gas observation for this treatment should
also be conducted.
Video camera is needed to observe the formation of water vapor
on the wall the glass jar
A more practical, yet accurate, methods to measure fluctuation
of gasses (O2, CO2, and ethylene) during observation are needed
(such as probes connected to digital gas analyzer).
Next time, RNA isolation should be finished on site to minimize
RNA degradation

External payload utilization
Potential research field: SEDA, MAXI,
SMILES
Cooperative researches using KIBO EF
will be promoted in each research
community.
JAXA inform how to access each
research team.

Proposed Joint Research:
Solar Activities Influence
on Space Environment
in Satellite Orbit

Proposed Joint Research:
JEM Meteoroid Detection compared to
Meteor Wind Radar Detection

14000
Meteor Flux per Day
12000
10000
8000
6000
4000
2000
0
2002.8 2003 2003.3 2003.5 2003.8 2004 2004.3 2004.5 2004.8 2005 2005.3 2005.5 2005.8 2006 2006.3 2006.5 2006.8 2007
Year
Blue: Meteor Flux Red: Model of flux based on ecliptic and solar activity variation)
Meteor Wind Radar at Kototabang, Indonesia
From MWR detection,
there is an indication
that meteor influx
depend solar activity.
Is that Interplanetary
variation or
atmospheric variation

That is all....
Thank you