RATIFICATION PAGE

Complete report of Plant Physiology observations with the title “ Medium

Maintanance of Fruit Flies (Drosophila melanogaster)”. Which made by:
Name : Yulianda
ID : 1614441010
Class : ICP Of Biology Education
Group : III (Three)
After checked by assistant and assistant coordinator, so this report accepted.

Makassar, 20th March 2018

Assistant Coordinator, Assistant,

Paewa Panennungi, S.Pd

Known,
Lecture of Responsibility

Dr. Ismail, M.S

ID. 19611231 198603 1 015
 CHAPTER I
INTRODUCTION

A. Background
Metamorphosis and pasting is a very important process in the life cycle, will
only form if the female larvae successfully metamorphose from male larvae to
benthic and attached to the substrate to then grow into new colonies as a
response signal received from the environment. Female larvae on spawning
type corals (broadcaster) generally metamorphoses a day to a week after
leaving the parent colony and undergoing fertilization.
Metamorphosis occurs when the larva undergoes physiological and
morphological changes permanently. The process of metamorphosis begins
when the planula larvae change to resemble a plate with oral and aboral
portions flattened and the septa will be formed fingerwise with the mouth as its
center. At this time the larvae form the cilia for movement and secreting the
sensor cell to detect the surface of the substrate. The larvae are reported to
crawl the surface of the substrate and select a suitable substrate for attaching
actively.In holometabolism, the larvae are very different from the adults.
Species of fruit fly, Drosophila melanogaster, a commonly harmless
common insect that is a fungus-eating creature that grows on the fruit. Fruit
flies are easy-to-breed insects. From one marriage alone can be produced
hundreds of offspring, and the new generation can be bred every two weeks.
This characteristic makes fruit flies an ideal organism for genetic studies.
The development begins after fertilization, which consists of two periods.
First, the embryonic period in the egg at the time of fertilization until the young
larvae hatch from the egg and this occurs within approximately 24 hours. At
times like this, larvae do not stop to eat. Metamorphosis in Drosophila includes
perfect metamorphosis, ie from eggs - instar larvae I - instar larvae II - instar
larvae III - pupa - imago. The development begins immediately after
fertilization, which consists of two periods. First, the embryonic period in the
egg at the time of fertilization until the young larvae hatch from the egg and
this occurs within approximately 24 hours. And at times like this, larvae do not
stop for food. The second period is the period after hatching of the egg and is
called postembrionic development which is divided into three stages, namely
larvae, pupa, and imago.
Drosophila larvae are white, segmented, worm-shaped, and dig with black
mouths near the head. For breathing on the trachea, there is a pair of spiracles
both of which are at the anterior and posterior ends. When the cuticle is not soft
anymore, young larvae periodically change skin to reach adult size. The old
cuticle is removed and the new integument is expanded with a high feeding
rate.Preadult tissue restriction (before adulthood) is called anlagen.
The main function of pupa is for the external development of anlagen to
adult form. Adults on Drosophila melanogaster in one lifecycle is about 9 days.
After coming out of the pupa, the fruit fly is still pale and the wings are not yet
stretched. Meanwhile, the female fly marries after 8 hours and will store sperm
in a very large number of male fruit flies.
During feeding, larvae create channels in the medium, and if there are many
channels then the growth of cultures can be said to be good.
When the Drosophila larvae form a pupa shell, the body is shortened, the
cuticle becomes hard and pigmented, without head and wings called instar
larvae 4. Pupa formation is characterized by the formation of heads, wing pads,
and legs. The puppets use cuticles in the third instar. At this stage of the pupa,
the larvae are inactive, and in this state, the larvae change into adult flies. Each
type of Drosophila melanogaster especially males have different arrangements
between types of one with the other. The period of development of Drosophila
melanogaster varies among other temperatures, generally all types of cold-
blooded.
Life cycle of Drosophila is very important to know because if we know it
we can provide appropriate treatment in its care. In addition, we can know the
right conditions for each phase. Metamorphosis of fruit flies (Drosophila
melanogaster) starts from the eggs of fertilization. Fruit flies have levels of
reproduction and the rate of production is also higher than other types of
 insects. This is due to their ability to perform very efficient reproduction is also
effective especially in the mating season. After fertilization, the fruit fly fly will
lay eggs. Based on this, so this lab is done for how to make fruit fly medium,
and can know the difference between male and female and life cycle of
Drosophila melanogaster.
B. Purpose
1. To know how to prepare cointainer medium maintanance of fruit flies
(Drosophila melanogaster).
2. To know how to made medium maintanance of fruit flies (Drosophila
melanogaster).
C. Benefit
After do this lab we can know how to prepare cointainer medium
maintanance of fruit flies and we can know how to made medium maintanance
of fruit flies.
 CHAPTER II
LITERATURE REVIEW

In plant physiology it is common to show the free energy contained in water

is in the form of potential water (Ψ). The definition of water potential is the energy
per unit of water volume, the water potential is directly proportional to its
temperature. The osmotic potential is a chemical potential caused by dissolved
matter. The osmotic potential always has a negative value, this is because it tends
to move across the semi permeable membrane from pure water to water containing
the solute (Resb, 2009).
The plant cell can have large turgor pressure because cells contain large
concentrations of solutes. These solutes attract water into the cells through a process
known as osmosis, which involves water flowing in through semipermeable
membranes that prevent the passage of solutes but not of water. The inflow of water
swells the cells until a hydrostatic pressure is reached at which no more water will
flow in. In cells bathed in fresh water, such as algal cells in a pond, this equilibrium
hydrostatic pressure is known as the osmotic pressure (π) of the cell contents, and
is commonly about 500 kPa or 0.5 MPa.This osmotic pressure can be measured
directly with an osmometer, or it can be calculated from the solute concentration in
the cell (C) from the van‘t Hoff relation: π=RTC(1) where R is the gas constant,

T is the absolute temperature (in degrees Kelvin) and C is the solute concentration
in Osmoles L-1. At 25 ºC, RT equals 2.5 litre-MPa per mole, and π is in units of
MPa. Hence a concentration of 200 mOsmoles L-1 has an osmotic pressure of 0.5
MPa (Hawkesbury, 2015).
However, land plants are different from algae in a pond. Their leaves are in
air, and the water in their cell walls, unlike the water in a pond, is not free. It has a
negative hydrostatic pressure (discussed further in the next section). Thus, for a
given osmotic pressure (π) within a cell, the hydrostatic pressure, P, will be lower
than if the cell were bathed in free water. This difference is known as the water
potential (ψ) of the cell. It is zero in an algal cell in fresh water, but it is always

negative in land plants. Its value is the difference between P and π, that is:

ψ=P−π(2) An alternative notation for equation (2) used commonly by plant

physiologists is: ψw=ψp+ψs(3) (Hawkesbury, 2015).

In this case, ψwis the total water potential, ψs is the solute potential and ψp

is the pressure potential. Thus ψs is equal, but opposite in sign, to π. The notion of
water potential can be applied to any sample of water, whether inside a cell, in the
cell wall, in xylem vessels, or in the soil. Water will flow from a sample with a high
water potential to one with a low water potential provided the samples are at the
same temperature and provided that no solutes move with the water. Water potential
thus defined is always zero or negative, for by convention it is zero in pure water at
atmospheric pressure (Hawkesbury, 2015).
Turgor-driven strain does not affect in situ measurements because of the
large bulk of tissue surrounding the implanted hygrometer. The small cylindrical
cavity cut for the hygrometer would minimize strain. Also, radial geometry assists
water flow and dissipation of potential gradients near the cavity. Finally, the tuber
tissue volume provides a large capacitance for water, minimizing the impact of the
coring on the tissue. During the measurements reported here, the excised tissue
samples were isothermal with or warmer than the hygrometer body, so thermal
errors were in the opposite direction to expansion errors. Tissue warming, as
indicated by the zo in the SC-10, ordered fresh > harvest > stored as expected for
respiratory heating (20). The size of respirational errors in y6 can be estimated from
zo in the SC-10. The zo underestimates the temperature gradient between the
reference and the tissue because the measuring junction is 0.43 of the distance
between them and because of divergent heat flow geometry (William, 1985).
Correcting for the distance of the measuring junction from the tissue and
recognizing that the measured zo was removed electronically before making the
dewpoint measurement, a minimum error of 0.04 MPa obtains for the fresh tubers.
This value is in good agreement with the displacement of the fresh tuber data from
the 1:1 line in Figure 4; the lower respiration rate of stored tubers did not cause a
similar displacement. This fresh tuber respirational error is also present in Figure 2.
As reviewed by Laties (16), respiration increased 3- to 5-fold immediately after
cutting the tissue and at least another 2-fold within 8 h (William, 1985).
The water status of potato tubers has been investigated on a few occasions.
Large, diurnal fluctuations in tuber water potential have been observed, with tuber
water potential approaching soil water potential at night, and decreasing
substantially during the day. Water stress decreased both the maximum nighttime
and minimum daytime water potentials. Associated changes in tuber osmotic
potential were not reported. Water, osmotic and calculated pressure potentials have
been determined for tubers at. Calculated pressure potentials were approximately
0.5 MPa in tubers having water potentials of −0.2 MPa and pressure potentials were
as small as −0.1 to 0 MPa in tubers having water potentials of −0.7 to −1.0 MPa.
Changes in tuber water and osmotic potential were observed for tubers in storage.
Water potentials decreased and solute concentrations increased with time in storage
. It was suggested that the observed changes in osmotic potential were caused, in
part, by an accumulation of free sugars or organic acids. Calculated tuber pressure
potentials decreased to approximately zero in storage for tubers that had been water
stressed prior to placing them into storage (Bland and Tanner 1986).
 CHAPTER III
OBSERVATION METHOD

A. Date and Place

Day/Date : Wednesday, 14th March 2018
Time : 10:50 a.m. – 12:30a.m.
Place : Third floor of Biology Laboratory
B. Equipment and Materials
1. Tools
a. Test tube
b. Test tube rack
c. Spoit
d. Drop pipette
2. Materials
a. Potato
b. Sucrose
C. Work procedures
1. Chardakov’s method

Immersed the
Prepared the tool Measured potato 3 pcs
potato to the
and materials test tube along
3gr/pieces.
15 minute

Observed. Is the Putted the potato out Added

solution buoy /fly and added in plain methylen
/sink ? sucrose blue
2. Gravimetry’s method

Immersed the
potato to the Weighted the potato first
test tube along
15 minute

vgvgvg
Prepared the sucose
solution

Immersed the potato

to the test tube along
15 minute

Out the potato from sucrose

Weighted the potato
(It’s better to let potato dry)
and noted.
 CHAPTER IV
OBSERVATION RESULT AND DISCUSSION

A. Observation Result
1. Chardakov’s method
Concertration Indicator
No note
(M) position
1. 0,10 Sink Osmotic process
2. 0,15 Sink Osmotic process
3. 0,20 Sink Osmotic process
4. 0,25 Sink Osmotic process
5. 0,30 Sink Osmotic process
6. 0,40 Sink Osmotic process
7. 0,50 Sink Osmotic process

2. Gravimetric’s method
Concertration
No Initial weight Final weight
(M)
1. 0,10 1,499 1,601
2. 0,15 1,5 1,506
3. 0,20 1,5 1,564
4. 0,25 1,5 1,670
5. 0,30 1,533 1,589
6. 0,40 1, 433 1,711
7. 0,50 1,450 1,349
B. Graph of Gravimetric

Initial weight Final weight

1.711
1.8 1.601 1.564 1.607 1.589
1.499 1.506
1.5 1.5 1.5 1.533 1.549
1.6 1.411 1.45
1.4
weight (gram)

1.2
1
0.8
0.6
0.4
0.2
0
0 0.1 0.2 0.3 0.4 0.5 0.6
concentration (M)

C. Discussion
1. Chardakov’s method
The working principle of potency water measurements on potato
cells by Chardakov method are two solutions of the same type, volume,
and concentration are treated differently. The first solution is soaked in
pieces of potato so it is called the immersion solution and the second
solution is not treated so that it is called the control solution. The
immersion solution was allowed to stand for 15 minutes in the hope that
after that time the solution has become isotonic. The isotonic solution
means the concentration of the marinating solution and the potato
network has been the same.
Soaking solution then dyed methylen blue to facilitate the
observation of the movement of the immersion solution in the control
solution. However, when the submersion solution is down, the solution
is hypertonic to the potato tissue so that water from the solution enters
the potato.
Based on the results of the experiment, data obtained in the form of
a solution of potato tissue immersion that has been added methylen blue
sink after inserted into the control solution at a concentration of 0.15 m.
 So it can be assumed that the potato concentration is lower than the
concentration of sucrose. Its mean that potential water is lessen because
the solution is enter to the potato.
2. Gravimetric’s Method
The working principle of the Gravimetric method is to look at the
mass changes of the potato tissue before and after the soaking of the
sucrose solution. Reduced masses of potatoes after soaking in a sucrose
solution indicates that the water contained in the potato cells comes out
into the solution because potato water potency is higher than the water
potential of the immersion solution. Similarly, when the mass of
potatoes after soaking in the sucrose solution has increased, it can be
concluded that potato water potency is lower than the water potential of
the immersion solution so that water enters the potato cells. Whereas if
the masses of potato remain the same before and after soaked sucrose
solution then potato water potency water equal to potential of
submersion solution.
Based on observations, all potatoes at different concentrations of
0.1M; 015M; 0.2M; 0.25M; 0.3M; 0.4M and 0.5M have mass increases.
It means that potato water potency is lower than the water potential of
the immersion solution so that water enters the potato cells.

CHAPTER V
 CLOSING

A. Conclusion
When the cell is placed in a hypertonic solution then the liquid cell
will come out. In a hypotonic environment fluid from outside the cell will
enter the cell, whereas in the isotonic environment there will be no change.
When the solution enter in to potato, it can be concluded that potato water
potency is lower than the water potential of the immersion solution, this
event named osmotic process.
B. Suggestion
1. Always working together with members of group for the results that we
want to accomplish can be done as expected.
2. Be careful when putting the reaction tube in a rack tube. Put the in safe
place, keep the solution in a test tube does not spill.
 BIBLIOGRAPHY

Am. J. Pot Res. 2009. Tuber Water and Pressure Potentials Decrease and Sucrose
Contents Increase in Response to Moderate Drought and Heat Stress. DOI
10.1007/s12230-009-9109-8, Potato Association of America 2009.

Bland and Tanner. 1986. Plant Physiology. Institut de Sciences Ve’ge’tales, Centre
National de la Recherche Scientifiqueb, France.

Hawkesbury. 2015. Australian Centre for International Agricultural Research. The

Univerity of Queensland, Australia.

William L. Bland,dkk.1985. Measurement of the Water Potential of Stored Potato

Tubers. Department of Soil Science, University of Wisconsin-Madison,
Madison, Wisconsin 53706 (C.B.T.)
ada banyak resep dalam persiapan media kultur untuk d. melanogaster dan
laboratorium berbeda atau pusat penelitian memiliki media kultur yang mereka
formulasikan sendiri untuk lalat buah mereka. the bloomington drosophila stock center
di universitas indiana adalah salah satu pusat penelitian drosophila terpopuler. tujuh
jenis medium memiliki kelebihan dalam memelihara lalat buah.

untuk pemeliharaan stok drosophila melanogaster dapat digunakan bermacam-macam

medium. medium yang mula-mula dipergunakan adalah campuran antara pisang ambon
dan tape ketela pohon dengan perbandingan 6:1. medium tersebut dipakai selama lebih
dari 15 tahun. Pada tahun 1984 mulai digunakan beberapa medium yang dicobakan
dapat pula pemeliharaan jenis-jenis drosophila lainnya dan beberapa tahun terakhir ini
telah digunakan resep yang baru. Hal ini disebabkan oleh karena kualitas pisang dan
tape yang digunakan tidak pernah seragam, sehingga dirasakan perlu untuk
memperoleh medium yang lebih padat dan dapat diandalkan. Resep baru yang
digunakan merupakan modifikasi resep yang telah ada dan disesuaikan dengan kondisi
untuk Indonesia.

Alat dan bahan :

1. Alat
a. Botol kultur
b. Tutup busa
c. Kuas kecil
d. Gelas beaker
e. Gelas ukur 100 ml
f. Blender
g. Kertas serbet
h. Plastic pembungkus
i. Karet gelang
2. Bahan
a. Pisang ambon
b. Zat pencegah jamur (nipagin/tegosept/moldex)
c. Ragi
d. Agar-agar
e. Gula merah

Cara kerja :

1. Campur gula merah dengan aquades dan masak sampai mendidih !

2. Blenderlah pisang yang ranum hingga lumat !
3. Campurlah pisang yang sudah di blender ke dalam air, agar-agar dan gula yang
sedang mendidih dan diaduk sampai rata diatas api yang kecil dan didiamkan
sekitar 15 menit sehingga piang ikut menjadi matang !
4. Biarkan adonan mendidih sekitar 15 menit !
5. Oleskan anti jamur yang telah dipersiapkan sebelumnya pada botol dan
tutupnya !
6. Tuangkan dengan segera sekitar 40 ml/40 gr adonan yang masih panas dalam
botol biakan yang sudah disterilkan terlebih dahulu!
7. Letakkan kertas serbet yang telah dilipat sedemikian rupa pada medium!
8. Taburkan sedikit ragi pada medium yang sudah memadat !
9. Gunakan botol biakan apabila medium sudah padat !

CATATAN :

Medium yang tidak terpakai dapat disimpan dalam lemari es, tetapi harus
dipergunakan dalam batas waktu satu minggu. Setelah satu minggu biasanya
medium mengering dan sudah menjadi kurang baik bagi perkembangan drosophila
melanogaster.