RATIFICATION PAGE

Complete report of genetics laboratory with the title “Maintenance

Medium for Fruit Flies (Drosophila melanogaster)”. Which made by:
name : Nisa Andini Sofyan
ID : 1614442010
class : Biology Education of ICP
group : V (five)
After checked by assistant and assistant coordinator, so this report accepted.

Makassar, September 2018

Assistant Coordinator Assistant

Sagita Cahyani
Paewa Panennungi, S.Pd
ID. 1514441007

Known,
Lecture of Responsibility

Hartati, S. Si, M. Si, Ph. D

ID : 19740405 200003 2 00
 CHAPTER I
INTRODUCTION

A. Background
The world around us is filled with variety of things. These things can be
classified as living things or unliving things. Unliving things like soil, air, rock,
water, or something made by human like chair, table, handphone. Then humans,
animals, plants we called it as living things, specifically organism. Organism do
activities like breathing, growth, making movement, capable to stimulate, etc.
On biology, organism is a group of organ systems, organs, tissues, and cells. So,
animals, plants, and humans includes organism. On the earth, there are humans,
animals, plants, bacteria, fungi, as a living things.
Everythings on this earth have a reason why them exist. One of reason is
to fullfill humans necessary, include fruit flies. Drosophilla melanogaster have
bben used to study genetics for over 100 years. Fruit flies are great to work with
in research setting because they are relatively easy to take care of, especially
compared to larger and more expensive organisms. Because you can get this fruit
flies free. Also, the fruit flies are small so this allows biologist or scientist to
keep up millions of them at one time. That’s support you to learn in genetics in
biology.
By studying biology, everyone will be able to know the reasons behind the
sudden changes happen. Everyone who have interest of biology, pay intention
for development courses. Biologist are answer their question by research. They
ar curious fellows. So they keep researching, experimenting, and and did more
trial-error things to get suitable answer. model organisms are species that are
studies to understand the biology of other oragnisms, often humans. For
example, in laboratory biologist or scientific need a representative as human
models to keep researching and laboratory utility while learn about genetics.
Genetics is one of sub material has been learned in biology. Genetics is
study of genes, heredity, chromosomes, and how the parental characteristic
delivered to their child. genetics arose out the identification of genes, the
fundamental units responsible for heredity. Genetics may be defined as the study
of genes at all levels, including human genetics by used fruit flies. Elizabeth said
that fruit flies share 75% of the genes that cause disease of humans. So biologist
or scientist can learn about human genetics by studying fruit fly genetics. In a
fruit flies lab, there are stacks and stacks fruit flies living in little vials. the only
care they need is to have their food changed every 10-14 days by transferring
them into new vials. They eat a pastey substance, called medium, that goes in
the bottom of vialsl can be kept at room temperature in laboratory. However, we
need suitable medium for better life span of fruit flies. A medium doesn’t
workout with a wrong recipe. Researcher investiged the recipe of suitable
medium for fruit flies or Drosophilla melanogaster.
Drosophilla melanogaster or fruit flies is a small, common fly found near
unripe and rotted fruit. It has been in use for over a century to study genetics and
behavior. Fruit flies are easily obtained from the wild and many biological
science companies carry variety of different mutations. Why Drosophilla
melanogaster? It has chromosomes large, undivided, allowing for easy
observation. The chromosomes of fruit flies gave scientists their first glimpse of
genetic variations between species and individuals. Because female fruit flies
often mate with more than one male during mating session, virgin fruit flies are
necessary for scientific study. using fruit flies that have previously mated may
result in mixes offspring, which skews experiment results. studying the genetic
makeip, transcription and replication of fruit fly can assist in better
understanding these processes in other eukaryotic orgainisms, such as humans.
So, the biologist or scientist usually culture the population of Drosophilla
melanogaster.
Medium or media is the first step in preparing culture vials by adding food
media. There are variety of types food avaible for the fruit flies; some require
cooked medium. Cooked medium with a jelly, bananas, fermipant, sugar, water,
and corns powder. This laboratory need us to learn how to make a medium for
fruit flies. Before we start to culture and do investigation DNA mutants for
Drosophilla melanogaster.
B. Purpose
1. To find out how to set medium for fruit flies (Drosophilla
melanogaster).
2. To understand how to make a medium for fruit flies (Drosophilla
melanogaster).
C. Benefits
This laboratory activity makes us learn about process, and technique. An
eggs cannot be a moths without process of growth. A better growth depends on
environment around it, like a good medium for developing fruit flies
(Drosophilla melanogaster), while understand how to set a medium.
 CHAPTER II
REVIEW OF LITERATURE

The explanation of heredity most widely in favor during the 1800s was the
“blending” hypothesis, the idea that genetic material contributed by the two parents
mixes in a manner analogous to the way blue and yellow paints blend to make green.
This hypothesis predicts that over many generations, a freely mating population
will give rise to a uniform population of individuals. However, our everyday
observations and the results of breeding experiments with animals and plants
contradict that prediction. The blending hypothesis also fails to explain other
phenomena of inheritance, such as traits reappearing after skipping a generation.
An alternative to the blending model is a “particulate” hypothesis of inheritance:
the gene idea. According to this model, parents pass on discrete heritable units
genes that retain their separate identities in offspring. An organism’s collection of
genes is more like a deck of cards than a pail of paint. Like playing cards, genes can
be shuffled and passed along, generation after generation. Modern genetics had its
genesis in an abbey garden, where a monk named Gregor Mendel documented a
particulate mechanism for inheritance (Campbell, 2010: 262).
In 1866, Gregor Mendel, an Austrian monk and a plant breeder, published
his findings on the method and the mathematics of inheritance in garden pea plants.
The passing of traits to the next generation is called inheritance, or heredity. Mendel
was successful insorting out the mystery of inheritance because of the organism he
chose for his study the pea plant. Pea plants are easy to grow and many are true-
breeding, meaning that they consistently produce offspring with only one form of a
trait. Pea plants usually reproduce by self-fertilization. A common occurrence in
many flowering plants, self-fertilization occurs when a male gamete within a flower
combines with a female gamete in the same flower. Mendel also discovered that
pea plants could easily be cross pollinated by hand. Mendel performed cross
pollination by transferring a male gamete from the flower of one pea plant to the
female reproductive organ in a flower of another pea plant. Mendel rigorously
followed various traits in the pea plants he bred. He analyzed the results of his
experiments and formed hypotheses concerning how the traits were inherited. The
study of genetics, which is the science of heredity, began with Mendel, who is
regarded as the father of genetics (Biggs et al, 2008 : 277).
He called these new plants hybrids because they received different genetic
information, or different alleles, for a trait from each parent. The results of these
studies made Mendel even more curious about how traits are inherited. Garden peas
are easy to breed for pure traits. An organism that always produces the same traits
generation after generation is called a purebred. He found that tall plants crossed
with short plants produced seeds that produced all tall plants. Whatever caused the
plants to be short had disappeared. Mendel called the tall form the dominant factor
because it dominated, or covered up, the short form. He called the form that seemed
to disappear the recessive factor. Today, these are called dominant alleles and
recessive alleles (Biggs et al, 2008: 128).
An organism with two of the same alleles for a particular trait is
homozygous for that trait with two different alleles for a particular trait is
heterozygous.The outward appearance of an organism does not always indicate
which pair of alleles is present. The organism’s allele pairs are called its genotype.
The observable characteristic or outward expression of an allele pair is called the
phenotype (Biggs et al, 2008: 279).
Drosophila melanogaster as one the insect that has the role of a very
important in the development of the science of genetics as well as diploid organisms
model made in the laboratory because of the small size, had short life cycle, the
number of offspring produced very much cheaper costs and her treatment.
Drosophilla melanogaster during this a genetic mutation making it known by a
wide variety of strains. One example is the strains of sepia and plum, which is
mutant D. Melanogaster. The mutant has certain genetic abnormalities in
chromosome thus causing the occurrence of differences Phenotype if dibandingka
with Drosophilla melanogaster type normal (Wild Type). Drosophilla
melanogaster type characteristics normal is characterized by red eye, eye
compound spherical rather ellips and eyes single (oceli) at the top of his head with
relative sizes smaller than compound eyes, body colorntanned yellow with coloured
rings. Black in the back part of the body. Body size Drosophilla melanogaster
ranged between 3-5mm. Drosophilla melanogaster wing is quite long and
transparent. The position of the wings begins from thoracic vein, the edges of the
wings (costal vein) has the two sections are interrupted near the her body. aristanya
are generally shaped hair and have the 7-12 branching (Hotimah et al, 2017:55).
The ability of laboratory populations of Drosophila melanogaster for
selection_induced rapid changes of these parameters makes this species a
convenient object for studying plasticity and evolution of the life cycle. The nature
of selection factors underlying lifespan evolution remains controversial. In some
cases, lifespan shortening may be interpreted as a useful adaptation increasing
evolutionary potential of a population,which is supported by natural selection,
group selection, or “second order selection for evolvability”. D. melanogaster that
existed in the mode of short life cycle (reproduction at the age of 2 weeks after the
egg stage) for 180 generations (~7 years), and in lines derived from them that were
for 2 years subjected to selection for late reproduction (the age of imago admitted
for reproduction was gradually increased from 28 to 70 days). Rose has shown that
selection for late reproduction resulted in a significant increase in Drosophila
lifespan, and maximal fecundity moved to a later term. If, on the contrary, selection
is aimed at early reproduction (or increased mating frequency at an early age), it
leads to lifespan reduction and shifts reproductive effort towards younger age
(Yakovleva et al, 2016:1445).
The fruit fly model, Drosophila melanogaster, has played a central role in the
debate over the relative importance of calories vs. specific nutrients to LS. Early
DR studies on D. melanogaster focused almost exclusively on the role of calories
in extending, although it is now clear that many of these studies confounded the
effects of calories with those of specific nutrients. More recent work has targeted
the specific nutrients that extend LS, and there is good evidence to suggest that
protein (P) restriction is largely responsible for the extension in LS observed in
many DR studies. However, despite considerable research, we still know
surprisingly little about the exact nutrient (or combination of utrients) that extends
LS in D. melanogaster. A major obstacle in addressing these questions has been the
large diversity of different approaches used to restrict dietary intake in this species.
The most common way to implement DR in D. melanogaster has been to restrict
the intake of a diet of fixed composition by either limiting access to food or diluting
the diet with water or another bulking agent. Studies using this approach have
typically used only a small number of diets, have not precisely measured food or
nutrient intake, and have used yeast (Jensen et al, 2015:605).
Yeast, of course, is a complex food consisting of many nutrients besides
protein, including essential fatty acids, minerals, vitamins, and cholesterol (which
insects cannot synthesize). What feature of reduced dietary yeast relative to
carbohydrates modulates lifespan, and in particular is this protein or a specific
amino acid? Although caloric intake has dominated views of rodent DR, methionine
restriction alone extends lifespan in both mice and rats. In Drosophila, flies fed a
media with casein as the sole source of protein were long lived upon diets with
intermediate levels of protein although even this best casein-based concentration
produced suboptimal longevity relative to diets made with yeast. Likewise, flies
were longest lived when fed an intermediate level of methionine in a defined diet
with relatively low total amino acids. Interpreting these data is complicated because
the only tested low-methionine diet might reflect malnutrition while the only tested
high-methionine diet increased total food consumption (Tatar et al, 2015:3).
Based on mutation of chromosomes, no. 3 on a mutant chromosome
mutations and sepia No. 2 on the mutant prunes, then allegedly there is the
difference of some phenotypes when compared with Drosophila melanogaster the
type normally. Therefore the research needs to be done about morphological
description Drosophilla melanogaster strains with normal type sepia and plum
(Hotimah et al, 2017:55).
It is a culture medium ingredients that became the environment culture
Drosophila melanogaster, consisting of nutrient mix. General media contains
water, energy sources, nitrogen, sulfur, phosphates, oxygen, hydrogen and others,
such as medium papaya. Papaya is used as medium because papaya has content
excellent nutrition, among other things, provides energy quite high compared with
fruits another. Papaya is rich in minerals such as potassium, carbohydrates, fats,
proteins, phosphorus, iron and calories (Rahmawati et al, 2016:252).
In the manufacture of medium for fruit flies there are many recipes in
preparation of media culture for Drosophila melanogaster and in research centers
have special laboratories for their fruit flies. The Bloomington Stock Center in
Drosophilla Indiana University is one of the most popular Drosophilla research
where there are seven different types of culture media used on this research center
are in each medium has excess funds keep the fruit flies. In the year 1984 began to
use some medium for customers to maintenance types – types of other Drosophila
and some years later a new recipe is used because the quality of bananas and tape
are not uniform, so the need is felt to gain the medium is more solid and reliable.
This new recipe is a modification of existing recipes and adapted to conditions in
Indonesia. Maintenance of the Drosophila melanogaster preferably done using
media with a moisture content of at least 60%, protein rough 13.20%, and gross
energy 2263.89 calorie/gram (Hartati, 2018:1)
Based on Hartati (2018) as for recipes that are now used to make around 25-
30 bottles of medium are as follows: banana 600 grams; gelatin (swallow) 7 grams;
brown sugar/sugar 150 grams; 20 g Yeast; nipagin-moldex tegosept//7 mL; sorbic
acid 5 mL; aquadest-411 mL.
For the maintenance of a stock of Drosophila melanogaster can use various
range of medium. The medium used is leaven used as material because has excellent
nutritional yeast, among others, provide a high enough energy. Rich yeast minerals
such as potassium, magnesium, phosphorus, iron, and calcium. Droshopilla
melanogaster is a species of winged insects belonging to the order of Dipteral
(nation flies). Drosophilla melanogaster is popular because it is very easy to breed
only takes two weeks to complete the entire cycle of life, easy maintenance, and
has many variations of fenotif are relatively easily observed. Drosophilla
melanogaster egg amount issued will be decreased if a lack of food. Adult fruit flies
that food shortages will result the larvae are small. These larvae were able to form
a pupa, but often small-sized time failed to develop into a mature individual phases
(Safitri et al, 2017:49).
 CHAPTER III
MATERIALS AND METHODS

A. Time and Place

Day / Date : Thursday, 13rd September 2018
Time : 15.50 until 17.50 WITA
Place : Microbiology lab floor II West Faculty of Mathematics and
Natural Science University of Makassar
B. Tools and Materials
1. Tools
a. Blender (1 piece )
b. Beaker glass (6 piece )
c. Knife (2 pieces)
d. Stove (1 piece )
e. Jar of jam (5 pieces)
f. Measure glass of 100 mL (1 piece )
g. Rubber band (5 pieces)
2. Material
a. Banana (600 gram)
b. Brown Sugar (150 gram)
c. Yeast ( 20 gram)
d. Jelly ( 7 gram)
e. Aquadest ( 411 mL )
f. Nipagin ( 7 mL )
g. Sorbid acid ( 5 mL )
h. Plastic ( 5 pack )
i. Paper napkins ( 5 pieces )
C. Work Procedure
1. Preparation of container culture
a. Prepare all the tools you want to use namely bottle medium and cover;
b. Clean wash bottles before use;
 c. Put all the bottles that have already cleared to the plastic and then tie with a
rubber;
d. The bottle into the sterilization autoclave for 3 hours at a temperature of
121o C;
e. Remove the bottle from inside the autoclave;
f. The bottle is ready to use
2. The making of media culture
a. Prepare the tools and materials that will be used;
b. Weight banana, jelly, sugar, and yeast;
c. Blander the banana 600 gram;
d. Mix the banana, water, jelly, and sugar;
e. Cooked it until boiling;
f. Pour it into the bottle about 40 mL / 40 gram;
g. Oily nipagin on the cover;
h. Put a little bit yeast;
i. Put the tissue paper on the bottle;
j. Covered the bottle with plastic;
k. The medium is ready for use.
 CHAPTER IV
RESULT AND DISCUSSIONS

A. Result
Medium of fruit flies
(Drosophilla melanogaster) Note

1. Plastic cover
1
2. Rubber
2
3
3. Jar of jam

4 4. Napkin
5. Yeast
5
6. Batter
6

Table 1. Parts of Media Culture

Source: Practicum Documentations

Source: https://www.sheffield.ac.uk/bateson/fly-facility/fly-food
Table 2. Medium Picture Compare
B. Discussion
For culture fruit flies (Drosophila melanogaster) to investigate DNA
genetics, we need to develop this species by make a viels media culture. After
did experiment in Biology Laboratory, we got result observation as like shown
in result table. First, table is describe the container of medium. Making a
container by use a bottle jar and modified lid. As laboratory instruction, before
use the container we need to sterilize it before. Alfreedo (2017), a technician in
Abdulaziz University Hospital said sterilization in laboratory is important to
minimize the growth of organisms and transmission of one individual to another.
In environment the use of disinfection techniques decreases the growth of
bacteria on surfaces, which leads to decrease in transmission of organisms
among population. So the focuss in our labs about developing the growth of
Drosophila melanogaster.
The parts of medium container includes the body and the lid. The body by
using bottle jar as a main container and place to keep the yeast. Then the lid
contains plastic, rubber as a lid to close the container perfectly. The detail is on
table 1. Next to the medium culture, the yeast, the viels, and how to make it.
Based on observation, we combine the materials of banana 600 grams; gelatin
(swallow) 7 grams; brown sugar/sugar 150 grams; 20 g Yeast; nipagin-moldex
tegosept//7 mL; sorbic acid 5 mL; aquadest-411 mL follow the instruction on
modul.
As for the function of each material to be used, first banana as food of furit
flies. Fruit flies are attracted to ripened or fermenting banana (fruit). Fruit flies
lay their eggs on the surface of fermenting foods, so the banana is the main
material to attrached the fruit flies. After banana, next it aren sugar. That has
function to give a sweet taste on the medium. The combination of banana and
sugar automatically serves as carbohydrate. Gelatin is used to keep the density
of dough, which is where the gelatin was galakton a complex carbohydrate,
which is extracted from the algae gelidium genus marin. Then, nipagin serves as
an antifungal in other words prevent the growth of mildew on the medium.
Sorbic acid serves as a preservative medium on fruit flies. And paper napkins to
soak up the excess water on medium and also as a place of fruit flies lay their
eggs. The rubber band function to bind to the plastic bottle cover jam. Yeast
dough is used to inflate and aquadest is used as a solvent.
By following the instruction, we find out the recipe for media culture of
fruit flies. How to manufacture medium manufacture of fruit flies (Drosophila
melanogaster) expressed in conformity with research experts who concluded
that fruit flies requires various elements of macro or micro, among others, in the
form of carbohydrates, proteins, vitamins and minerals. Carbohydrates and water
is a source of energy for living fruit fly activities, both in growth and
development as well as a reproduction. Based on literature by Tatar, yeast, of
course, is a complex food consisting of many nutrients besides protein, including
essential fatty acids, minerals, vitamins, and cholesterol (which insects cannot
synthesize).
During the lab, we did something little wrong. We were late to place a
dough into the container of medium. It should be quick while put in the dough
into the container, cause the dough is reaaly fast to dense. And once the dough
dense before place to the container, it can be lumpy. We cannot lost of attention
while did this experiment. Caused a good medium for a good culture.
But, we can use this medium and continue our activity to the next
laboratory about how to keep the frui flies, developing, and breeding in the viels
we made before.
 CHAPTER V
CLOSING

A. Conclusion
Drosophila melanogaster needs a media for culture. This labs activity,
make us know how to make a suitable medium for culturing fruit flies. A
medium recipe takes from combine the fruit (bananas), sugar, water, powder to
dense the dough, gelatin, and sorbic acid. This medium as a manipulate place
for Drosophila melanogaster to growth and develop.

B. Suggestion
Keep focuss and pay attention while during the laboratoy activity. Cause a
little mistake can takes a high risk in the result. Also, don’t forget to understand
the instruction fellow before do a laboratory. Cause biologist is all about learn
and focuss.
 REFRENCES

Biggs,et,al. 2008. Glencoe Science : Biology, New York, : Mcgrow.

Campbell, dkk. 2008. Biologi Jilid 1. Penerbit : Erlangga
Hartati, Irawan F. 2018. Genetics Modul Based on Scientific Way. Makassar:
Universitas Negeri Makassar.
Hotimah et al. 2017. Deskripsi Morfologi Drosophila melanogaster Normal, Starin,
dan Plum. Jember: Universitas Jember.
Jensen et al. 2015. Sex-specific Effects of Protein and Carbohydrate Intake on
Reproduction but not Lifespan in Drosophila melanogaster. UK: Aging
Cells published by Anatomical Society.
Rahmawati, et al. 2016. Pengaruh Tembakau dalam Medium Kultur terhadap
Jumlah Turunan Lalat Buah (Drosophila melanogaster). Banda Aceh:
FTK UIN Ar-Raniry.
Ramadani et al. 2016. Pemanfaatan Drosophila melanogaster sebagai Organisme
Model untk Mempelajari Pengaruh Faktor Ligkungan terhadap Ekspresi
Sifat Makhluk Hidup pada Perkuliahan Genetika. Malang: Universitas
Negeri Malang.
Safitri D, Bachtiar S. 2017. Pengaruh Penambahan Ragi pada Media terhadap
Perkembangbiakan Drosophila melanogaster. Malang: Universitas
Negeri Malang.
Tatar et al. 2014. Nutrient Control of Drosophila Longevity. United States: NIH
Public Access in Trends Endocrinol Metab.
Yakovleva, et al. 2016. Adaptation of Drosophila melanogaster to Unfavorable
Growth Medium Affects Lifespan and Age-Related Fecundity. Moscow:
Biochemistry Pleiades Publishing.