GIANT CHROMOSOMES

Yolanda Anggraeni
190210103128
Genetics of Class D
Biology Education Study Program, Faculty of Teacher Training and Education, University of Jember
Kalimantan Street Number 37 Kampus Bumi Tegalboto Jember 68121
ohyoland31@gmail.com

ABSTRACT
A chromosome is a structure in a cell that is a long series of molecules consisting of one DNA
molecule and various proteins. Giant chromosome or giant chromosome is a chromosome that has more elongation
than the chromosome in general. The function of giant chromosomes is generally the same, namely to enhance the
metabolic activity of a cell. The purpose of this practicum is to observe the giant chromosomes in Drosophila
melanogaster and to find out the shape of the giant chromosomes found in Drosophila melanogaster. The giant
chromosomes found in the salivary glands of 3rd instar larvae of Drosophila melanogaster 3. The salivary glands
that have been found in 3rd instar larvae of Drosophila melanogaster are dripped with enough FAA until the color of
the salivary glands turns cloudy then clean the remaining FAA by filling it with blotting paper, then dripping with
acetocamin and closing the preparation with a cover slip and scan for giant chromosomes. Giant chromosomes look
long dangling. These giant chromosomes can be found in the nurse cells of the ovaries, follicular cells that surround
the oocytes, fat cells, intestinal cells and abdominal histoblasts. The difference is in the location of the swelling.

Key Words: Chromosomes, Giant Chromosomes, Politen, Drosophila melanogaster

D. melanogaster is a fruit fly that has been
used as the subject of genetic research since the early
20th century, which states that D. melanogaster is a
research subject that is very extensively used in the
field of genetics. The characteristics of these insects
which have a fast life cycle, have only a few
chromosomes, a small genome size, and have giant
chromosomes in their salivary glands make D.
melanogaster chosen by genetic researchers in their
research (Fauzi, A., & Corebima, AD 2016: 278) .
The fruit fly (Drosophila melanogaster) was
the first organism to be used by humans to study
genetic analysis and is now one of the best known
and widely used eukaryotes. The analysis is used to
understand transcription and replication. There is an
expert who studies fruit flies with his students,
namely Thomas. Thomas Hunt Morgan conducted
experiments on fruit flies at the Fly Room Lab,
University of Columbia in 1910 and found many
principles of heredity such as sex linkage, epistasis,
multiple and gene mapping. Drosophila
melanogaster is widely used as an animal in genetic
experiments for several reasons. According to James,
fruit flies only need simple and inexpensive
equipment, easy to care for, harmless, short life
cycles, male and female are easy to distinguish,
imago has giant chromosomes (polytenes) in the
salivary glands, the female is capable of producing
500 eggs, has 3 pairs of autosomes and a pair of
gonosomes, and the mutant variations are numerous.
The fruit fly genome sequence was completed and
published in 2000 (Suharsono, S., & Nuryadin, E.
2019: 114).
Drosophila melanogaster is an insect that
has a small number of chromosomes, which is 2n = 8.
Drosophila melanogaster has a short life cycle of
about 10-12 days, produces a lot of eggs every time a
female Drosophila melanogaster lays eggs, so it is
easy to care for and has many mutant characters. .
Drosophila melanogaster has three important pairs of
chromosomes, which has an XX / XY chromosome
system for the assignment of sex chromosomes, has
giant chromosomes in the salivary glands of its
larvae, and in male Drosophila melanogaster there is
no crossing over or crossing over when meiosis
occurs. Based on this explanation, it is necessary to
carry out practicum regarding monohybridization of
fruit flies (Drosophila melanogaster) to determine
the differences between male and female flies
(Aurora, M. E. M., & Susilawati, I. O. 2020: 264).
It is said to be giant chromosome (polytene
chromosome) because it contains thousands of times
more DNA than ordinary chromosomes. Practicum
observation of polytene chromosomes using the
salivary glands of instar III larvae D. melanogaster,
because during the third instar larval phase, the
salivary gland chromosomes are very large, clearly
visible, and provide a clear picture of the effect of
ecdyson on gene expression. The polytene
chromosome consists of 4 equal long arms, 1 short
arm, chromosome, band, interband, and puff. The
highly condensed portion of the polytene
chromosome has multiple copies of the DNA
sequence but because it is in a condensed state and
does not express the genetic information encoded in
DNA, it is called heterochromatin. Heterochromatin
is dark in color because it is in the condensed state.
Heterochromatin is not active in transcription
because it does not contain active genes.
Euchromatin is the part that is not condensed and
appears light in color. This happens because the
euchromatin does not undergo compaction.
Euchromatin contains active genes and almost all of
the genes that are transcribed so that they become
active parts in replicating (Safilu, Amiruddin, Ahdiat
A., et all. 2019: 30-31).
It has been shown that translocation can
change chromosome structure both genetically and
morphologically. Chromosomes can become longer
or shorter, depending on the size of the translocated
pieces. Normally inversion does not change
chromosome length, but if inversion includes
centromere (pericentric). The position of the
centromere can be greatly altered Deletion or
duplication, if viable. sometimes it can be detected
cytologically by a change in chromosome size (or a
band pattern in the case of Drosophila giant
chromosomes). or the presence of "blooms" in paired
figures. Chromosomes with unequal arm lengths can
be converted into Isochromosomes having the same
arm length and genetically homologous to each other,
by abnormal transverse division of the centromere.
Drosophila's telocentric X chromosome can be
transformed into an “X-attached” form by
misdivision in the centromere (Elrod, Susan, &
William Stansfield. 2010. 160).
In certain eukaryotic organisms there arc
special tissues where the chromosomes are of special
structures not found in other cells of the same
organism. These chromosomes attain their largest
size in the nuclei of their respective cell. Hence these
arc also called giant chromosomes. The giant
chromosomes arc found in the suspensors of the
embryo of certain plant, cells of salivary' glands of
Drosophila and Chironomous. in the cells of
malphigian tubules, epithelium lining of gut of
Drosophila, in the cells of fat bodies of larval stage
of certain Diptera, oocyte nuclei of certain vertebrate,
and invertebrate. Special types of chromosome have
been classified into two categories, namely Polytene
chromosome and Lampbrush chromosome. Polytene
chromosomes are those giant chromosomes in which
DNA is replicated in such a way that the daughter
chromatids do not separate. In more details, polytene
is achieved by replication of the DNA many times
without nuclear division (endomitosis) and the
resulting daughter chromatids do not separate and
remain aligned side by side to form a giant multi-
stranded chromosome. Polytene chromosome first
provided the evidence that eukaryotic gene is
regulated at the level of RNA synthesis. These
chromosomes are the valuable material for the study
of gene regulation because their gene transcription
can be seen directly in the microscope. Polytene
chromosome differs from polyploidy, in which there
is also excess DNA per nucleus, but in which the new
chromosomes are separated from each other (Vega,
Leslie, & Bret White. 2019: 94-95).
Polytenization is a modification of
endoreduplication in which the replicated
homologous chromatids remain paired. The best
known polytene chromosomes are the giant
chromosomes of Dipteran salivary glands. Variants
of these typically giant chromosomes are found in
many other insect tissues. A polytene chromosome is
composed of partially uncoiled chromosome strands
paired side-by-side. At certain points, called
chromomeres, the strand is folded. The chromomeres
are aligned and form the visible bands in the polytene
chromosome. In the Diptera, each polytene
chromosome consists of the two homologues. The
stretched-out parts of the chromosome strands are the
interband regions, whereas each band is thought to
contain one gene. The chromosomes replicate time
and again, and a Dipteran salivary chromosome may
finally consist of as many as 16,000 units. During the
development of a tissue, certain bands specific for the
tissue form so-called puffs. Puffing involves the
unraveling of the coiled DNA of a particular
chromomere; RNA is being synthesized at the puffs,
and this demonstrates that the puffed genes are active
in specific tissues during specific periods in
development. Polytene chromosomes are not limited
to insects but are also found in certain specialized
plant cells, such as the synergids and antipods in the
embryo sac as well as in suspensor and endosperm
cells. Polytene chromosomes induced by culture
conditions in pea roots (ordinarily pea roots consist
of 2C and 4C cells). Interestingly, polytene
chromosomes have also been described in the giant
trophoblast cells of the rabbit, mouse, and rat. The
banded structure is usually much less clear in non-
Dipteran polytene chromosomes, since mitotic
chromosomes tend to pair only in Diptera. However,
good banding has been obtained in the bean
Phuseolus by growing the plants at low temperatures
(Therman, Eeva, & Millard Susman. 2010: 153).
Polytene chromosomes develop from the
chromosomes of diploid nuclei by successive
duplication of each chromosomal element
(chromatid) without their segregation. The newly
formed chromatids remain associated lengthwise and
together form a cable-like structure, referred to as
polytene chromosomes. In Drosophila melanogaster,
they are 70-110 times longer than typical metaphase
chromosomes. Extent of coiling of the DNA and its
associated proteins varies along the linear axis of
each chromatid, thereby leading to the variation in
chromatin concentration and compaction. Regions
with high DNA content are known as chromomeres.
In every polytene chromosome, homologous
chromomeres align exactly alongside each other and
so they fuse to form a transverse band. Bands are
separated from each other by interchromomeric
fragments of chromosomes (interbands) (Zykova, et
all. 2018: 179).
Polytene chromosomes of Drosophila
melanogaster are a convenient model for studying
interphase chromosomes of eukaryotes. They are
giant in size in comparison with diploid cell
chromosomes and have a pattern of cross stripes
resulting from the ordered chromatid arrangement.
Each region of polytene chromosomes has a unique
banding pattern. Using the model of four chromatin
types that reveals domains of varying compaction
degrees, we were able to correlate the physical and
cytological maps of some polytene chromosome
regions and to show the main properties of genetic
and molecular organization of bands and interbands,
that we describe in this review. On the molecular
map of the genome, the interbands correspond to
decompacted aquamarine chromatin and 5’ ends of
ubiquitously active genes. Gray bands contain
lazurite and malachite chromatin, intermediate in the
level of compaction, and, mainly, coding parts of
genes. Dense black transcriptionally inactive bands
are enriched in ruby chromatin. Localization of
several dozens of interbands on the genome
molecular map allowed us to study in detail their
architecture according to the data of whole genome
projects. The distribution of proteins and regulatory
elements of the genome in the promoter regions of
genes localized in the interbands shows that these
parts of interbands are probably responsible for the
formation of open chromatin that is visualized in
polytene chromosomes as interbands. Thus, the
permanent genetic activity of interbands and gray
bands and the inactivity of genes in black bands are
the basis of the universal banding pattern in the
chromosomes of all Drosophila tissues. The smallest
fourth chromosome of Drosophila with an atypical
protein composition of chromatin is a special case.
Using the model of four chromatin states and
fluorescent in situ hybridization, its cytological map
was refined and the genomic coordinates of all bands
and interbands were determined. It was shown that,
in spite of the peculiarities of this chromosome, its
band organization in general corresponds to the rest
of the genome. Extremely long genes of different
Drosophila chromosomes do not fit the common
scheme, since they can occupy a series of alternating
bands and interbands (up to nine chromosomal
structures) formed by parts of these
(Sidorenko, et all. 2019: 148).
The polytene chromosomes are a convenient
object for the visual study of gene activity, spatial
and structural genome organization.
chromosomes are formed as a result of successive
rounds of endoreduplication of each of the diploid
chromosomes and function as the
chromosomes (Dyka, et all. 2016: 222)
Polytene chromosomes are formed when the
products of multiple rounds of S-phase remain tightly
associated to form one supersized chromosome. In
particular polytene chromosomes are found in a
specific subset of endopolyploid cells, which have
undergone a variant cell cycle known as the
endocycle. During endocycles, cells
successive genome replications without
intervening nuclear envelope breakdown. For more
on the regulation of endocycles, we refer the reader
to several recent reviews on the topic. Because the
number of endocycles can vary between tissues and
organisms, polytene chromosomes
impressive range of ploidy from those that are only
reduplicated a single time to chromosomes made of
many thousands or even millions of genome copies.
We also note that polytene chromosomes can form in
cells that undergo transient nuclear
breakdown without subsequent chromosome
separation (Stormo and Donald. 2017: 2).
METHODOLOGY
The first step is to make a medium for the
breeding of Drosophila melanogaster. By weighing
the banana 350. After that, cut the banana, and put it
in the blender. Then weigh tapai 150 g, cut the tapai,
and put it in the blender. After that weigh the brown
sugar, cut the brown sugar, and put it in the blender.
After all the ingredients to make the medium go into
the blender, then add enough water to the blender.
Blend all ingredients. After that, heat all the
ingredients that have been blended, put the medium
in the jam jar, cover the medium, and wait for it to
warm. After it is warm, put 7 points of farnipan and
put in the pupation paper and close the jar of jam.
Then the second step is to do the inoculation
process of Drosophila melanogaster by selecting
genes instar 3 larvae of Drosophila melanogaster.Then put
the larvae in a watch glass and given a sea of 0.9%
NaCl. After that, determine the head, tail and neck of
the larvae. Then put the larvae on a slide and
Polytene observed under a microscope. Then separate the head
from the tail, by placing a pin on the head and neck
then pulling it off. Look for luda glands that have a
interphase kidney-like shape with a transparent color. Separating
the salivary glands that have been found from the
attached fatty fat. After that, drip the salivary glands
that have been found with sufficient FAA, until the
color of the salivary glands turns cloudy. Then clean
the remaining FAA by sucking it with blotting paper,
then dripping it with acetocarmin. After that, cover
the preparation with a cover slip, and look under the
undergo microscope, and the last thing is to look for the giant
any chromosomes on the preparations that have been
made, and draw them.
DISCUSSION
The tools and materials in the “Giant
Chromosome Observation” practicum consist of a
cover an light microscope, watch glass, glass object, cover
glass, pipette, 3rd instar larvae of Drosophila
melanogaster, 0.9% NaCl solution, FAA solution,
acetocarmin, and pins. The light microspcope serves
as a tool to clearly see the giant chromosomes in fruit
envelope fly larvae, because the chromosomes cannot be seen
with the naked eye and require other visual
assistance, so here you can use a light microscope to
see clearly the giant chromosomes. The watch glass
here functions as a container for placing the larvae of
instar 3 Drosophila melanoster which will be given
0.9% NaCl solution. The glass object serves to place
the object to be observed under a light microscope.
Cover glass serves to cover glass objects that have
objects that will be observed under the microscope.
This cover glass aims to protect objects from outside
interference.
The pipette serves to take a solution and
then move it to another place. The 3rd instar larvae of
Drosophila melanogaster is used because the 3rd
instar larvae are the most mature larvae, where these
larvae will enter the pre-pupa stage and then become
adult flies. The use of the salivary glands of 3rd instar
larvae is also due to their transparent color making
them easy to isolate and also having many politent
chromosomes. 3rd instar larvae have met the
nutrients and how to eat enough so that they are used
as objects for observation of giant chromosomes.
The 0.9% NaCl solution functions so that
these 3rd instar larvae do not experience dryness. The
FAA solution is used to maintain the shape of the
cells so that these cells do not carry out the next
phase of division. Acetocarmin is used to provide
color so that the chromosomes can be seen easily.
The pin serves as a tool to separate the head and tail
of the 3rd instar fruit larvae.
The observation procedure this time begins
by selecting the 3rd instar larvae of Drosophila
melanogaster, which is then placed on the arva in the
watch glass and given 0.9% NaCl solution. NaCl or
this physiological salt serves to keep instar 3 larvae
from drying out during isolation and to keep the
bodies of these 3rd instar larvae from being
destroyed. After that can be determined the head, tail
and neck of the larvae. The next step is to place the
larvae on a glass object and observe it under a
microscope.
The above steps have been carried out, then
the head with the tail is separated. This can be done
by placing a pin on the head and neck of the larva,
then pulling it off. After that, look for the salivary
glands of the 3rd instar larvae, where these salivary
glands have a kidney-like shape with a transparent
color. Then, the salivary glands that have been found
are separated from the adhered fats.
The next step is the salivary glands that have
been found dripping with sufficient FAA solution,
until the color of the salivary glands turns cloudy and
cleaning the remaining FAA solution by sucking it
with suction paper, then dripping with acetocarmin,
then the preparation is closed with a cover glass and
then observation can be made. under a light
microscope. The observation looks for giant
chromosomes on the preparations that have been
made and then drawn.
The definition of a giant chromosome or
also called a polytene chromosome is a chromosome
that has more elongation than the chromosomes in
general. The function of giant chromosomes is
generally the same, namely to enhance the metabolic
activity of cells and regulate all work systems in
these cells. Giant chromosomes can be found in the
salivary glands of Drosophila melanogaster. The
giant chromosomes in fruit flies are found in the
larval stage of the fruit fly prepupa. In addition to the
salivary glands, these giant chromosomes can be
found on nurse cells in the ovaries, follicular cells
that surround the oocytes, fat cells, intestinal cells
and abdominal histoblasts. The difference is in the
location of the swelling.
The definite characteristic of this giant
chromosome is that it is longer and thicker than
normal chromosomes. Another characteristic feature
is that there are stripes of dark bands and light bands
that are regularly alternated. The bright band on this
giant chromosome is euchromatin with loosely
wound while the dark band is heterochromatin with a
dense coil and can undergo condensation.
The process of formation of giant
chromosomes is due to the endomitosis process in
which the chromosome strands replicate continuously
without nuclear division. In this process, large and
long chromosomes are formed like ribbons or what is
commonly known as polytene chromosomes. This
giant chromosome has many copies of genes that do
not separate from one another or in the sense that
they remain attached to each other so that in one cell
there are copies of information from several genes in
the chromosome. However, when in the repeated
endoreplication stages on the chromosomes, there are
parts that do not divide maximally, namely the
central spherical area or often known as the
centromere. As a result, the centromeres of these
crosmosomes are joined together into a solid
formation known as the centrosenter.
CONCLUSION
chromosome or also called a polytene
chromosome is a chromosome that has more
elongation than the chromosomes in general. The
function of giant chromosomes is generally the same,
namely to enhance the metabolic activity of cells and
regulate all work systems in these cells. Giant
chromosomes can be found in the salivary glands of
Drosophila melanogaster. The giant chromosomes in Stormo, B. M., & Fox, D. T. 2017. Polyteny: still a
fruit flies are found in the larval stage of the fruit fly giant player in chromosome research.
prepupa. In addition to the salivary glands, these Chromosome Research, (3-4): 2.
giant chromosomes can be found on nurse cells in the Suharsono, S., & Nuryadin, E. 2019. Pengaruh suhu
ovaries, follicular cells that surround the oocytes, fat terhadap siklus hidup Lalat Buah (Drosophila
cells, intestinal cells and abdominal histoblasts. The melanogaster). Jurnal Penelitian Biologi. (2):
difference is in the location of the swelling. 114.
The definite characteristic of this giant Therman, Eeva, & Millard Susman. 2010. Human
chromosome is that it is longer and thicker than Chromosomes (Structure, Behavior, and
normal chromosomes. Another characteristic feature Effects, Third Edition). New York: Henry
is that there are stripes of dark bands and light bands Krell.
that are regularly alternated. The bright band on this Vega, Leslie, & Bret White. 2019. Fundamental of
giant chromosome is euchromatin with loosely Genetics. United Kingdom: ED – Tech Press.
wound while the dark band is heterochromatin with a Zykova, T. Y., Levitsky, V. G., Belyaeva, E. S., &
dense coil and can undergo condensation. Zhimulev, I. F. 2018. Polytene chromosomes–
a portrait of functional organization of the
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