MONOHYBRID AND DIHYBRID CROSSES IN DROSOPHILA MELANOGASTER

Genetics Class D
Study Program of Biology Education, Faculty of Teacher Training and Education, Jember University
Kalimantan Streat number 37, Kampus Tegalboto, Jember 68121

ABSTRACT
Among the subjects in the genetics practicum is to demonstrate medel I law and Mendel II law by
practicing monohybrid cross and dihybrid cross on fruit flies (Drosophila melanogaster) as test materials which
have many advantages including: easy to obtain, easy to maintain, low cost. maintenance, does not need a lot of
space, visible size, not dangerous, has many offspring, has a short life cycle, its life cycle ranges from 10-15
days, variable, simple chromosomes, has many mutants, and male flies do not cross. In this practicum, fruit flies
were crossed randomly with the prepared medium. Results have been successfully obtained in accordance with
Mendel's law, namely the monohybrid cross obtained F1 results with a phenotype ratio of 3: 1 and genotype 1:
2: 1.While in dihybrid cross, the phenotype ratio is 9: 3: 3: 1 and the genotype ratio is 1: 2: 1: 2: 4: 2: 1: 2: 1.
This result shows that the crossover was done well.
Keyword : Monohybri, dihybrid, mendel

INTRODUCTION mutations and produced mutants that were different

from normal conditions. The differences are mainly
Genetics is the study of how life is carried related to eye color, eye shape, and wing shape.
through time. It investigates how the wide diversity Some types of mutations in Drosophila
of organisms can be simplified into long strands of melanogaster that can be seen from the phenotype
four-letter code, and how this code is translated are eye color, eye shape, wing shape, and body
into twenty words. Gregor Mendel is often color mutations so that various strains of
considered the father of modern genetics, but his Drosophila melanogaster are known, including
great research on conspecific hybridization was not white (w), sepia (se), eyemissing (eym), curled
truly appreciated at the time (Edwards, 2016: 1). (cu), taxi (tx), dumpy (dp), and vestigial (vgl). The
Genetic inheritance is explaining how differences in phenotypes that appear are due to
characteristics can be inherited from parents to changes in the genotype from normal conditions. In
children and from generation to generation by addition to having an impact on the phenotype,
using fundamental laws of classical genetics or changes in the genotype will cause physiological
Mendelian genetics. Crosses or hybridization in changes such as the reproductive process (Taufika
biology is marriage between individuals or et al, 2020: 50).
populations that have genetic differences that have Monohybrid cross is a cross that only pays
the result of combined traits from their parents or attention to one different characteristic. For
recombination of genes in their offspring. (Awang- example, only paying attention to seed color or skin
Kanak et al, 2016: 284) color, monohybrid cross is a simple form of
If one parent has many alleles that are inheritance and is the basis for understanding the
expressed through a trait that appears to be a mechanism of inheritance. The parent individual is
dominant phenotype, then he will inherit these called P1 or parental generation (parent
traits while if the oldest one contains more generation), P1 offspring is called F1 or the first
recessive alleles then he will also pass these traits filial generation, and individuals resulting from
to his child so that when both parents inherit these marriage between F1 are called F2 or second filial
traits to the child then the nature of the child is not generation. Note that in each marriage One of the
the middle or intermediate character of the two traits does not appear in F1 but then reappears in F2
parents but the combination of the two is not an and with an almost constant ratio of 3 : 1 to explain
even mixture. Mendel also said that the traits this result Mendel introduced a term for an
carried by a gene are not influenced by the inheriting factor called the unit factor he assumed
experience of the individual who inherits the trait that These units act as basic units of traits that are
(Arumingtyas, 2016: 2) passed down from generation to generation so that
The fruit fly (Drosophila melanogaster) is it can determine the trait of each individual factor
known to have undergone many mutations that unit is now called an allele. Dihybrid cross is a
produce intraspecific genotypic variations called cross between 2 different properties that have a
strains. Apart from normal conditions (N), several phenotype ratio of 9: 3: 3: 1 (Irawan, 2011 : 29-31)
strains were found which were the result of
 Drosophila melanogaster, is an organism and memory and sleep (Cambridge University,
that is very useful for conducting research. It is 2014 : 217)
relatively easy to culture, has a conveniently short The life cycle of Drosophila melanogaster
lifespan, consists of mostly post-mitotic tissues reveals two peak periods : one encompassing the
(except for the reproductive system and a small part four initial hours of the embryonic development
of the gut), and has a fully sequenced genome and and one encompassing from the L3 larval stage
well-characterized genetics. This insect uses many onward. Drosophila melanogaster, as all
mitochondrial transporters homologous to the holometabolous insects, has an indirect
human carriers, as well as the same mechanisms for development with two active freeroaming phases,
transporting several metabolites from cytosol to the larva and the adult, and two inactive sessile
mitochondria and vice versa. Furthermore, it offers developmental phases, the embryo and the pupa.
a variety of powerful molecular genetics methods The larval and adult phenotypes, especially their
for studying transporters, many of which would be morphology, arise primarily through the genetic,
difficult to test in mammalian models (Curcio et al, cellular and tissue interactions of embryonic and
2020: 1). pupal (metamorphosis) development, respectively
In heterogeneous populations, there are (Zamora et al, 2019 : 1476).
several genotypes formed by the process of Drosophila has different morphological
recombination and segregation. Genotypic characteristics between males and females. In
variation can be eliminated by studying cross lines males Has a smaller body size when compared to
that are homozygous for some genes, or F1 females. Has three segments on the abdomen and
offspring from uniform heterozygous crosses for all has a genital comb. Whereas in females the size is
parental genes. As for the complete elimination of relatively larger, has six segments in the abdomen
the environment is not possible despite all the and does not have a genital comb. Drosophila
efforts to make an identical environmental melanogaster is a winged, small animal. Therefore,
experiment for all populations. In plants, for observing the morphology of these animals can use
example, a small variation in soil quality or tools such as loops and microscopes. In Drosophila
exposure to sunlight will result in a slightly melanogaster, 4 pairs of chromosomes are found.
different environment. Sometimes even for plants In general, males and females are the same, but
that are close together with a high cross, the there is a slight difference, namely that on one of
drosophila still shows variations in its phenotype the male chromosomes there is a curve like a hook.
(for example, in body size) caused by Meanwhile, regarding the sexual age of females for
environmental differences between animals in the mating in Drosophila melanogaster, various
same culture bottle. Therefore, the properties that information is obtained. There are several opinions
are susceptible to small environmental influences which state how old Drosophila melanogaster
will never be uniform even on inbred lines (Hartl & female reaches sexual maturity (Suparman et al,
Ruvolo, 2012 : 663) 2018: 41-42).
Drosophilla melanogaster is popular
because it is very easy to breed, it only takes two RESEARCH METHODS
weeks to complete the entire life cycle, is easy to
maintain, and has many phenotypic variations that To make a cross on fruit flies, there are several
are relatively easy to observe. The number of things that need to be prepared in advance. The
Drosophilla melanogaster eggs released will first, making medium is done by mixing all the
decrease if there is a lack of food. An adult fruit fly ingredients that have been prepared with added
that lacks food will produce small larvae (Aurora et water then blending until completely smooth. Then
al, 2020: 263). cook until it boils and a little thick. Then, put the
Drosophila melanogaster is an ideal medium in the culture bottle, after the medium is
organism for behavioral analysis genetic systems, warm, sprinkle with 7 pearls. Wait for it to cool and
because a large number of genetically identical insert the pupation paper and cover with a sponge
individuals can be bred quickly and economically stopper.
under controlled environmental conditions. Flies After the medium is ready, experiments can
are not only able to accept extensive genetic be started, the first is by doing inoculation. Prepare
manipulation but also for neuroanatomic and a small hose whose end is closed with gauze. Then,
electrophysiological studies. They also show a insert it into a large tube. Then, the combined hose
variety of behaviors that can be measured using a then put it in a bottle in which there is already a
simple behavior paradigm, including olfactory fruit fly. Vacuum the flies using the tube until they
behavior, gistation, courtship and mating, foraging, enter the large tube. And close the end of a large
starting behavior, phototaxis, negative geotaxis, tube and transfer the flies to the culture bottles.
open field locomotion, aggression, flight, Before crossing, it is necessary to isolate
sensitivity to alcohol, circadian behavior, learning. virgin first on female flies. The way to do virgin
isolation is by taking flies on the tube and
removing adult flies (imago) from the curl bottle. maintenance is not too expensive. But we must be
Then, take it out and write down the date. After 8 diligent about changing the medium once every
hours, remove all the newly hatched flies from the two weeks. (4) It doesn't take much space, the
pupan and select the female. Usually flies will not mutant stock is usually maintained in jam jars. One
mate for at least 8-10 hours from the pupa. Thus jar of jam can contain about 300-500 adult flies.
the flies obtained must be virgins. The way to get The bottles can be stored in one shelf. (5) Visible
virgin female flies is pupa isolation. Males are size, although only 2-5 mm in size, the morphology
recognized by the sex comb on the tarsals of the and characteristics of the mutants can be observed
two front legs. This sign is only visible in the 3- with the naked eye. (6) Harmless, these fruit flies
day-old cocoons, because the primentation of the do not transmit disease or act as intermediate hosts.
bristles occurs only one day before leaving the (7) Having many offspring, a female fly can
cocoons. This method of isolation will produce produce 500 eggs during her life. (8) Has a short
very good accuracy. Selection of virgin female flies life cycle. Its life cycle ranges from 10-15 days. (9)
should be stopped after the third day. Age of male Variable, this fly has a phenotype that is easily
flies does not matter, because male flies of all ages distinguished on eye color, feather shape, body
can be used for crossing. color and wing shape. (10) Simple chromosomes,
Crosses were carried out by taking 2 bottles only have 4 pairs of chromosomes, so it's easy to
of fly culture of normal type (wild), curly type count them. Besides that, in the salivary glands
(walikot wing) curved upward. Then, put 5 normal there is a giant and arrayed polytene chromosome
type female flies and 5 curly type male flies (Cu) in that is useful for recognizing the location of genes.
the medium that already contains the medium and (11) Has many mutants. History of biological
pupation paper. Then, cover the culture bottle with research shows that Drosophila melanogaster is the
a sponge. provides information about the type of most mutant-rich organisms. Therefore it is very
cross and the date of the cross (initial). store the appropriate to be used as an experimental animal in
culture bottles in the designated place. On day 7 genetics practicum. (12) Male flies do not cross,
remove all parental. Note when the flies first Drosophila melanogaster males do not cross. Thus
appeared. On day 10, sedate the flies and count the the cross displacement can be calculated more
number of male flies that appear. Then, precisely. In animals or in plants crossing over can
differentiate the sex of flies and count their occur in both parental.
numbers. There are several factors that affect the yield
and growth of Drosophila melanogaster. Namely
RESULT AND DISCUSSION the factors of reproductive ability, sex, life cycle
and their genetic suitability to be examined for the
Monohybrid cross is a cross that occurs results of the crossing. Other factors are factors that
from one different trait, where the filial (F1) of the come from outside or are called external factors,
cross will have a phenotype ratio of 3 : 1. such as temperature, nutrition, hygiene /
Meanwhile, a dihybrid cross is a cross that is sterilization and so on.
carried out on a similar individual which consists of There are several tools and materials used in
two different properties, where the filial (F1) of the this practical experiment. Among them are fruit
cross will have a phenotype ratio of 9 : 3: 3: 1 from flies of various ages, male flies can be used from
the result of the parent cross. various ages. Small brush, foam stopper, cover the
There are many advantages that can be culture bottle. Large and small hoses, for
obtained by using fruit flies (Drosophila vacuuming and picking up fruit flies and
melanogaster) for this crossing experiment, transferring flies to culture bottles. Gauze, to cover
namely: (1) Easy to obtain, Drosophila the end of a small hose. Pupation paper, to take the
melanogaster is an insect that is cosmofolite. In pupa from the medium. Jam bottles, as culture
Indonesia alone there are more than 600 types of bottles and medium placement. Banana and Tape
fruit flies from the Drossophilidae family. Thus, and Brown Sugar as ingredients in a ratio of 7: 2: 1,
not all fruit flies found are Drosophila for the maintenance of Drosophila melanogaster.
melanogaster species, but this species can be bred Pernipan, for the manufacture of a fruit fly medium
independently. (2) Easy to maintain, Drosophila which functions to provide air space in the
melanogaster is usually kept in a bottle containing medium.
banana or tape medium. In addition, it also requires To make a cross on fruit flies, there are
sterile conditions such as in microbiological several things that need to be prepared in advance.
experiments. However, it is better if all material The first, making medium is done by mixing all the
tools are sterilized first to avoid mold ingredients that have been prepared with added
contamination. Fungal growth can be prevented by water then blending until completely smooth. Then
giving sodium benzoate and tape for the cook until it boils and a little thick. Then, put the
fermentation process. (3) Low maintenance costs, medium in the culture bottle, after the medium is
because the food medium is very simple, the warm, sprinkle with 7 pearls. Wait for it to cool and
insert the pupation paper and cover with a sponge
stopper. N (Normal) : e (Black body)
After the medium is ready, experiments can 3 : 1
be started, the first is by doing inoculation. Prepare
a small hose whose end is closed with gauze. Then, The results were obtained by crossing the
insert it into a large tube. Then, the combined hose P1(Parent) between male fruit flies strain e and
then put it in a bottle in which there is already a female fruit flies strain N, the resulting gametes
fruit fly. Vacuum the flies using the tube until they were N and e which then formed F1(Filial) fruit
enter the large tube. And close the end of a large flies with genotype Ne (N Heterozygous). Then, to
tube and transfer the flies to the culture bottles. get the F2 result, a cross between F1 and F1 was
Before crossing, it is necessary to first isolate carried out which resulted in 4 puppies with
virgin virgin female flies. The way to do virgin gametes NN, Ne, Ne, ee. Those who have a normal
isolation is by taking flies on the tube and phenotype ratio and a black body are 3 : 1 and a
removing adult flies (imago) from the curl bottle. Genotype ratio are 1: 2: 1.
Then, take it out and write down the date. After 8 The second observation, dihybrid crosses
hours, remove all the newly hatched flies from the from male fruit flies of the ecl strain and female
pupan and select the female. Usually flies will not fruit flies of the N strain, with the following results:
mate for at least 8-10 hours from the pupa. Thus
the flies obtained must be virgins. The way to get P1 : Strain ecl >< Strain NN
virgin female flies is pupa isolation. Males are : eecc >< EECC
recognized by the sex comb on the tarsals of the G1 : e,c dan EC
two front legs. This sign is only visible in the 3-
F1 : EeCc (N Heterozygous)
day-old cocoons, because the primentation of the
bristles occurs only one day before leaving the
cocoons. This method of isolation will produce P2 : F1 >< F1
very good accuracy. Selection of virgin female : EeCc >< EeCc
flies should be stopped after the third day. Age of G2 : EC, Ec, eC, ec dan EC, Ec, eC, ec
male flies does not matter, because male flies of all F2 :
ages can be used for crossing.
Crosses were carried out by taking 2 bottles ♀♂ EC Ec eC ec
of fly culture of normal type (wild), curly type EC EECC EECc EeCC EeCc
(walikot wing) curved upward. Then, put 5 normal
Ec EECc EEcc EeCc Eecc
type female flies and 5 curly type male flies (Cu) in
the medium that already contains the medium and eC EeCC EeCc eeCC eeCc
pupation paper. Then, cover the culture bottle with
ec EeCc Eecc eeCc eecc
a sponge. provides information about the type of
cross and the date of the cross (initial). store the
culture bottles in the designated place. On day 7 E_C_ : E_cc : eeC_ : eecc
remove all parental. Note when the flies first 9 : 3 : 3 : 1
appeared. On day 10, sedate the flies and count the
number of male flies that appear. Then, E_C_ : (Normal)
differentiate the sex of flies and count their E_c_ : (Brownish yellow body, compound eyes
numbers. maroon / brownish)
The following are the results of observations e_C_ : (Black body, red compound eyes)
of monohybrid and dihybrid crosses that have been e_c_ : (Black body, compound eyes maroon /
carried out. The first observation, Monohybrid brownish)
crosses from male fruit flies strain e and female NB :
fruit flies strain N, with the following results : - ecl = Black body, compound eye color maroon /
brownish = eecc
P1 : Strain e >< Strain N - N = Brownish yellow body, red compound eyes =
G1 : N and e EECC
F1 : N Heterozygous
P2 : F1 >< F1 The results were obtained by crossing the P1
: Ne >< Ne between male fruit fly ecl strains and female fruit
G2 : N, e and N,e flies strain N, the resulting gametes were e, c and
F2 : NN, Ne, Ne, ee EC which then formed F1 fruit flies with the EeCc
(N Heterozygous) genotype. Then, to get the result
♀♂ N e of F2, a cross between F and F1 is carried out
N NN Ne which results in 16 tillers. Which has a phenotype
e Ne ee
ratio is 9: 3: 3: 1 and with a genotype ratio of 1: 2: Dolce. 2020. Drosophila melanogaster
1: 2: 4: 2: 1: 2: 1.
mitochondrial carriers: similarities and
There are several factors that can affect
the failure of crosses in Drosophila melanogaster. differences with the human carriers.
Among them are abnormalities in chromosomes or
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