School Science Lessons
2024-12-11
(UNBiol3)
Genetics, Heredity
Please send comments to: j.elfick@uq.edu.au
Contents
9.1.0 Amino acids DNA codons
9.2.0 Nitrogenous bases
9.3.0 Mitosis and meiosis
9.4.0 DNA and RNA
9.6.0 Drosophila experiments, Mendel's Laws
9.7.0 Chromosomes
9.8.0 Evolution
9.9.0 Human genetics, heredity
9.1.0 Amino acids DNA codons
The code for a amino acid is a 3 letter codon.
Most amino acids have more than one codon, e.g. phenylalanine has TTT and TTC.
End of chain has 3 codons.
The first letter in a codon is in position 1,
The second letter in a codon is in position 2
The third letter in a codon is in position 3.
After position 3 is END of chain.

Alanine, DNA codons

Arginine, DNA codons

Asparagine, DNA codons

Aspartic acid, DNA codons

Cysteine, DNA codons

Glutamine, DNA codons

Glutamic acid, DNA codons

Glycine, DNA codons

Histidine, DNA codons

Isoleucine, DNA codons

Leucine, DNA codons

Lysine, DNA codons

Methionine, DNA codons

Phenylalanine, DNA codons

Proline, DNA codons

Serine, DNA codons

Threonine, DNA codons

Tryptophan, DNA codons

Tyrosine, DNA codons

Valine, DNA codons

Amino acids and their 64 codons
The genetic code is the relation between a DNA codon and an amino acid.
Note that some amino acids are coded by several codons, e.g. Alanine, and some by only two codons, e.g. Asparagine, and some by only one codon, e.g. Methionine.
Ala Alanine GCT, GCC, GCA, GCG
Arg Arginine CGT, CGC, CGA, CGG, AGA, AGG
Asn Asparagine AAT, AAC
Asp Aspartic acid GAT, GAC
Cys Cysteine TGT, TGC
Gin Glutamine CAA, CAG
Glu Glutamic acid GAA, GAG
Gly Glycine GGT, GGC, GGA, GGG
His Histidine CAT, CAT
Ile Isoleucine ATT, ATC, ATA
Leu Leucine TTA, TTG, CTT, CTC, CTA, CTG
Lys Lysine AAA, AAG
Met Methionine ATG
Phe Phenylalanine TTT, TTC
Pro Proline CCT, CCC, CCA, CCG
Ser Serine TCT, TCC, TCA, TCG
Thr Threonine ACT, ACC, ACA, ACG
Trp Tryptophan TGG
Tyr Tyrosine TAT, TAC
Val Valine GUU, GUC, GUA, GUG
END of chain TAA, TAG, TGA
Position 1 is called the 5' end
Position 3 is called the 3' end
AAA
AAC
AAG
AAT
ACA
ACC
ACG
ACT
AGA
AGG
ATA
ATC
ATG
ATT
CAA
CAG
CAT
CAT
CCA
CCC
CCG
CCT
CGA
CGC
CGG
CGT
CTA
CTC
CTG
CTT
GAA
GAC
GAG
GAT
GCA
GCC
GCG
GCT
GGA
GGC
GGG
GGT
GUA
GUC
GUG
GUU
TAA
TAC
TAG
TAT
TCA
TCC
TCG
TCT
TGA
TGC
TGG
TGT
TTA
TTC
TTG
TTT
9.2.0 Nitrogenous bases
A base, nitrogenous base, when linked to other molecules, forms a nucleotide in DNA or RNA.
The five nitrogenous bases found in DNA and RNA:
A Adenine, 6-aminopurine, vitamin B4, (C5H5N5).
G Guanine, 2-​Amino-​6-​hydroxypurine, (C5H5N5O).
C Cytosine, 4-​Amino-​2-​hydroxypyrimidine, (C4H5N3O).
T Thymine (only in DNA), 2, 4-​Dihydroxy-​5-​methylpyrimidine, 5-Methyluracil, (C5H6N2O2).
or
U Uracil (only in RNA), 2, 4-​Dihydroxypyrimidine, 2, 4-Pyrimidinediol, (C4H4N2O2).

9.3.0 Mitosis and meiosis
9.3.1 Mitosis
9.3.2 Mitosis in cells of onion root tip
9.3.3 Meiosis
9.3.4 Meiosis in grasshopper testes
9.3.5 Meiosis, Lilium

9.4.0 DNA and RNA
9.4.1 DNA and RNA
4.1.10 Genetic engineering, Safety in school science
9.4.2 Role of RNA
9.4.3 Structure of DNA
9.4.4 Complexity of the genome
See 16.3.2.8: Nucleosides, nucleic acids, DNA.
See diagram 16.3.2.8.1: DNA molecule.
See diagram 16.3.2.8.2: Ribose, deoxyribose, nucleotide.
See diagram 16.21.10: Purines.
See diagram 16.21.13: Pyrimidines.
Experiments:
9.4.5 Isolate DNA from cheek cells, (Experiment)
9.4.6 Isolate DNA from sweetbread, (calf thymus gland), (Experiment)
9.4.7 Isolate DNA from wheat germ, kiwi fruit, strawberries, (Experiment)
9.4.8 Isolate DNA from strawberries, (Experiment)
9.4.9 Isolate DNA from any living material, (Experiment)

9.6.0 Drosophila experiments, Mendel's laws
Experiments:
9.6.1 Fruit flies, (Drosophila melanogaster), cultures of fruit flies
9.6.2 Fruit fly strains
9.6.3 Drosophila experiments, Mendel's Laws, Cultures of fruit fly
9.6.4 Mendel's experiments
9.6.5 Mendel's First Law, Law of Uniformity, (inheritance of one pair of characteristics)
9.6.6 Mendel's First Law, Law of Uniformity (inheritance of two pairs of characteristics)
9.6.7 Mendel's Second Law, Law of Segregation
9.6.8 Mendel's Third Law, Law of Independent Assortment
9.6.9 Sex-linked inheritance, introduction of recessive characteristic by female
9.6.8 Sex-linked inheritance, introduction of recessive characteristic by male

9.7.0 Chromosomes
Chromosomes are structures in the cells composed of DNA and proteins that contain genes.
In the eukaryotes, which have a nucleus and mitochondria in the cells, they are usually linear structures.
All multicellular organisms are eukaryotes.
Experiments:
9.7.1 Genetics, lethal factors, (Experiment)
9.7.2 Giant chromosomes, (Experiment)
9.7.3 Salivary chromosomes
3.2 Chromosome stain, Aceto-orcein stain, microscopy stain
5.4.4 Chromosomes, Conception, sex of the baby

9.8.0 Evolution
9.8.1 Evidence for evolution
9.8.2 Natural selection
9.8.3 Position statements on the teaching of biological evolution

9.9.0 Human genetics
9.9.1 Human genetic inheritance
9.9.2 Down syndrome
9.9.3 Fingerprint variations
9.9.4 Human traits with simple inheritance patterns
9.9.5 PTC tasters and non-tasters
9.9.6 Unattached / attached earlobe.

9.4.1 DNA and RNA
1. DNA is the master molecule that carries all of the inherited characteristics (genes) of an individual in the form of chromosomes.
Each individual (such as a human) receives one haploid set of 23 chromosomes from their father's sperm and one haploid set of 23 chromosomes from their mother's egg.
The two sets come together at conception when the diploid zygote, (fertilized egg) is formed.
Each eukaryotic chromosome, (the chromosomes of algae, fungi, plants and animals), carries thousands of genes, about 100 000 functional genes per cell.
A chromosome is analogous to a high capacity storage disk, (DVD disk), while genes are analogous to files on this storage disk.
If a chromosome could be completely unravelled, it would reveal a long ladder-shaped DNA molecule that is coiled into helical spirals.
At intervals along this double helix, the DNA ladder is wrapped around small beads of protein called nucleosomes.
2. The uprights of the DNA ladder are alternating 5-carbon sugars, (deoxyribose), and phosphates.
3. The rungs of the ladder are nitrogenous base pairs: purine bases (adenine and guanine) and pyrimidine bases (cytosine and thymine), with adenine always pairing with thymine and guanine always pairing with cytosine.
The four nucleobases in the nucleic acid of DNA:
G - guanine, C5H5N5O
C - cytosine, C4H5N3O
A - adenine, C5H5N5
T - thymine, C5H6N2O2
The four nucleobases in the nucleic acid of RNA, thymine is replace by uracil, C4H4N2O2
4. DNA and RNA have nucleotide sub units consisting of a phosphate, a sugar and a base.
A nucleotide is a molecule that combines a base with a five carbon sugar, ribose or deoxyribose, + a phosphate group. and is the basis of the nucleic acid sequence.
Every base pair has four different arrangements: A-T, T-A, C-G and G-C that allows millions of different possible arrangements in a DNA molecule.
Tiny amounts of DNA can be cloned into millions of copies with the PCR technique (Polymerase Chain Reaction) to give enough DNA to sequence gels into banding patterns to represent different base pair sequences and determine genetic "fingerprints" for crime identification or show relationships among plant and animal species.
The results can be compared with gene sequences in gene bank databases.
9.4.2 Role of RNA
The central role of RNA in human evolution, development and cognition, by Professor John S. Mattick, Institute for Molecular Bioscience, University of Queensland
Bioinformatic, genomic and experimental evidence all suggest that we have misunderstood the genetic programming of complex organisms for the past 50 years, because of the assumption, largely true for the unicellular prokaryotes, but not for multicellular eukaryotes, that proteins have transacted most genetic information.
The extent of non-protein-coding DNA increases with increasing complexity, reaching 98.8% in humans, suggesting that much of the information required to program development may reside in these sequences. Their studies indicate that these RNAs form a massive hidden network of regulatory information that regulates epigenetic processes and directs the precise patterns of gene expression during growth and development.
It also appears that RNA is central to brain development, learning and memory.
What was dismissed as "junk" RNA, because it was not understood may hold the key to understanding human evolution, development and cognition, and your idiosyncrasies and susceptibilities to complex diseases.
Finally, the observation that some RNA-directed epigenetic changes can be inherited raises the intriguing question: "Has evolution learnt how to learn?"
9.4.3 Structure of DNA
The structure of DNA was first suggested by J. D. Watson and F. H. C. Crick in a paper published in "Nature", April 25, 1953,
"MOLECULAR STRUCTURE OF NUCLEIC ACIDS, A Structure for
Deoxyribose Nucleic Acid", based on the X-ray evidence from research done by Rosalind Franklin and M. Wilkins.
The basic unit of DNA, the nucleotide, contains a sugar molecule (the aldopentose deoxyribose) a phosphate molecule and an organic nitrogenous base, a purine [adenine (A) or guanine (G)] or a pyrimidine [cytosine (C) or thymine (T)].
So DNA consists of a long string of nucleotides.
The structure of DNA has two helical chains with each chain coiled around the same axis with the sequence of the atoms in the two chains running in opposite directions.
In the diagram, the two ribbons represent the two phosphate to sugar chains and the horizontal rods represent the pairs of bases (base pairs) connecting the chains.
Only specific pairs of bases can bond together to adenine with thymine, and guanine with cytosine.
The protein-coding sequence of a single human gene may contain more than 1000 base pairs.
There are more than 30 000 different human genes.
RNA is similar to a single strand of DNA except that uracil (U) substitutes for thymine (T).
A codon is the sequence of three consecutive nucleotides in messenger ribonucleic acid (mRNA), which controls the placing of a particular amino acid into a protein or acts as the start or termination signal of a particular protein synthesis.
As there are 64 possible codons, i.e. 4 × 4 × 4 and 20 amino acids, so more than one codon can code for a particular amino acid, e.g. the code for arginine is CGC and CGA and CGG.
Just before cell division, the DNA double helix unwinds to allow each strand to be copied to form a sort of mirror image, pairing A with T and G with C.
The result is two DNA molecules each containing one old strand and one new strand with base pair sequences the same as the original DNA.
9.4.4 Complexity of the genome
The genome is the fully heritable DNA sequence of an organism.
The "atlas" of the genome is the Encyclopaedia of DNA Elements (ENCODE).
Only about 1.2% of the DNA codes for proteins.
Transcription factors, clusters of proteins binding to specific sequences near the genes, control the activity of the genes, so the specific sequences are called regulatory switches or regulatory DNA.
A gene may be influenced by many switches.
However, much DNA seems to have has no function and some DNA may be redundant, or be the result of harmless, but useless mutations.
So much of the DNA may be non-essential, but switches are still being discovered.
One line or research is to show the associations between specific parts of the genome and genetic diseases.
There is no correlation between genome size and complexity of a species.
The interaction of genes with the environment is still not understood.
There is a complex interaction between multiple genes so the effect of multiple genes for a characteristic, e.g. height, is not simply additive as was once thought.
9.4.5 Isolate DNA from cheek cells
Warning! Some school systems do not allow any experiments using human cells or the cells of the students!
Experiment
Dissolve half a teaspoon of salt in half a cup of water.
Add a little dishwashing liquid to break up the cells and release the DNA.
Put 25 mL of water in your mouth, but do not swallow it.
Move the water around your cheeks vigorously for 30 seconds to remove some cheek cells.
Spit the water into a clean cup.
Add 5 mL of this fluid to a 20 mL test-tube.
Add 2.5 mL of the salt and dishwashing liquid solution.
Put a stopper on the test-tube and move the test-tube up and down 3 times gently so that the contents do not form froth.
This movement breaks up the hundreds of cheek cells to release the DNA from the nucleus.
Gently pour into the test-tube a teaspoonful of ice cold ethanol that has been in a freezer for hours before the experiment.
Watch the point where the two layers meet.
Note the strands of DNA forming as cloudy filaments stretching up into the top ethanol layer.
DNA is not soluble in ethanol, so when the ethanol meets the DNA solution, it starts to precipitate form a DNA salt.
Use a glass hook or plastic tie wire to remove strands of DNA by slowly dipping up and down through the two layers.
Then gently invert the test-tube several times until the alcohol is mixed and the precipitated DNA will look like a small ball of white thread.
9.4.6 Isolate DNA from sweetbread (calf thymus gland)
9.2.30 Tensides
Cells of the calf thymus gland possess large nuclei from which large amounts of DNA can be obtained.
Buy some freshly-killed sweetbread from a butcher.
Deep frozen samples also produce comparable results.
Each tensides molecule of the kitchen detergent has a hydrophilic and a lipophilic end.
When they are taken up, the field of charge of molecules of the lipoprotein membrane is shifted, and hydrolysis occurs.
The membranes of the cell and nucleus are destroyed and histone molecules split off from DNA strands.
After the addition of kitchen detergent, the suspension becomes clear and viscous.
The viscosity is caused by liberated DNA molecules.
1. To produce a thymus suspension, cut up five sweetbreads in a mortar using a pair of scissors.
Add clean washed sand to the tissue in the mortar and grind with the pestle.
Add 20 mL of tap water and continue grinding until the water is completely clouded.
The water is clouded by cells and abraded cell detritus suspended in the water.
Filter the suspension through gauze.
2. To liberate DNA, add five drops of kitchen detergent to the cell suspension and shake thoroughly until the suspension becomes clear and viscous.
3. To obtain DNA, cover the suspension with one and a half times the amount of cold methylated spirit solution or ethanol solution in a narrow glass beaker.
Use a glass rod to remove the DNA precipitating as whitish strands.
Rotate the rod slowly and carefully dip it up and down occasionally to mix the suspension and the alcohol.
This action will precipitate more nucleic acid.
If the strands slip from the rod, use forceps to retrieve them.
9.4.7 Isolate DNA from wheat germ, kiwi fruit
DNA occurs inside a nuclear membrane within a cell membrane.
To see DNA, destroy the membranes and get a large enough quantity to be visible.
1. Put wheat germ, or kiwi fruit liquidized in a food processor, in the test-tube to a depth of 1 cm.
Add 3 mL of warm water and a drop of kitchen detergent.
Plug the test-tube with cotton wool and roll it in the hands gently for three minutes.
Float 2 mL of cold methylated spirit on top of the mixture and leave to stand.
The cloudiness at the interface of the liquids is the DNA.
Use an opened "slide-on" paper-clip to lift out the threads of DNA.
9.4.8 Isolate DNA from strawberries
Use two large frozen strawberries in a plastic bag and cooled rubbing alcohol [(CH3)2CHOH, isopropyl alcohol, 2-propanol].
To prepare the extraction solution, slowly add 50 mL of kitchen detergent to 250 mL of water so that no froth forms.
Slowly add 10 g of table salt (sodium chloride), then mix the solution.
Add water to make up the solution to 500 mL.
Add 20 mL of the extraction solution to the strawberries and seal the plastic bag after expelling the air.
Press down on the plastic bag for 5 minutes to crush the strawberries to a pulp.
Put the plastic bag on crushed ice and press down on the plastic bag again.
Filter the mixture of pulped strawberries and extraction solution into a beaker.
Put 2 mL of the filtrate into a test-tube and slowly add the same volume of cold rubbing alcohol.
Use a glass rod to remove the fluffy strands of DNA from the alcohol layer of the mixture.
9.4.9 Isolate DNA from any living material
Use broccoli, split peas, spinach leaf, lettuce, onions or chicken liver.
Put in a kitchen blender one half cup (100 mL) of extraction material, one eighth teaspoon (less than 1 mL) of table salt and 1 cup (about 200 mL) of cold water.
Run the kitchen blender on high for 30 seconds or until there is a runny pea soup consistency.
Pour this blended material through a strainer into another container.
Add 2 tablespoons (about 30 ml) of liquid detergent to the strained mixture and swirl to mix.
Let the mixture stand for 5-10 minutes.
Transfer the mixture into test-tubes one third full.
Tilt the test-tube and slowly pour cold ethanol into the test-tube down the side so it forms a layer on top of the blended mixture.
Observe the DNA rising into the alcohol layer from the blended mixture.
Use an opened "slide-on" paper-clip to draw the DNA into the alcohol.
9.6.1 Fruit flies, (Drosophila melanogaster), cultures of fruit flies
See diagram 9.35.1: Male and female Drosophila.
A foreleg, male with sex comb,
B male and larger female, abdomens with different numbers of segments,
C number of fruit flies in a 250 mL jar,
D number of fruit flies in a 500 mL jar
See diagram 9.35.2: Growth of Drosophila populations.
The common fruit fly, Drosophila, is used in genetic studies.
It is easy to culture and reproduces rapidly, so it is suitable for population studies.
Put ripe fruit, e.g. a banana, in the bottom of a jar and fit a paper funnel with a hole in the end in the mouth of the jar.
Put the jar in the open.
When fruit flies have entered the jar, remove the funnel and plug it loosely with cotton wool.
The fruit flies should be both males and females.
The females are larger, with a broader more pointed abdomen with seven segments.
The males are smaller and have a black-tipped more rounded abdomen with only five segments.
Only males have a fringe of black bristles on the forelegs called the sex comb.
The females will soon lay eggs and larvae will hatch after two days.
Put a piece of absorbent paper in the jar for the larvae to crawl on when they are ready to pupate and change to adult insects.
Put newly-hatched flies in another jar to start a new generation.
Make daily counts of the population in the bottle.
When numbers become very large, stand a piece of graph paper in the jar to count the number of pupae on the grid.
Maintain the bottle as long as the fruit flies survive.
Investigate the relationship between density of fly population, food preferences, temperature, life span and population.
9.6.3 Drosophila experiments
See diagram 9.35.1: Male and female Drosophila.
1. Attract fruit flies by putting overripe fruit in an open container, e.g. a glass jar.
After trapping the fruit flies, transfer them to small containers containing fruit chunks, e.g. banana.
Put a slice of ripe fruit in the bottom of the container and make a paper funnel with a hole in the end to fit the mouth of the container.
Put the container in the open air.
When six or eight fruit flies have entered (including both males and females) remove the funnel and plug it loosely with cotton wool.
The females are larger, with a broader abdomen.
The males are smaller and have a black-tipped abdomen.
Soon eggs will be deposited, and in 2 or 3 days the larvae will hatch.
Put a piece of paper in the container for the larvae to crawl on when they are ready to pupate.
The adult insects will come from the pupae.
Put newly-hatched fruit flies in another container to start a new generation.
To study fruit fly cultures, cut a piece of graph paper and stand it upright in the container so that you can sample a large population in the bottle by counting the number of pupae on the grid.
Make daily counts of the eggs, larvae, pupae and young adults of the population in a bottle.
Draw a graph to show the increase in population with time.
Maintain the culture for as long as the flies continue to survive.
2. Use conical flasks for breeding.
Prepare the artificial diet the day before.
Add 2 tablespoons beet juice syrup to 1 litre water.
Heat the mixture and stir constantly.
Add semolina until a thick paste forms.
Prevent moulds forming by stirring in 1 spatula tip of Nipagin, (methylparaben, CH3(C6H4(OH)COO), per litre.
Pour the mixture into clean breeding flasks to a depth of 2 cm.
After cooling, add 5 drops of a viscous suspension of bakers' yeast in tap water.
Close the breeding flasks with cotton wool plugs.
Before the flies are inserted, absorb any liquid collected on the surface of the feeding mixture with strips of filter paper to prevent the flies sticking to the surface.
Transfer the flies by tapping the breeding flask on the palm of the hand so that the flies fall to the bottom.
Quickly remove the cotton wool plug and place a collecting tube with the same size neck as the breeding glass on top of it.
By lightly tapping and shaking the flask, you can get the flies to enter the collecting tube.
Separate the two flasks and seal with cotton wool plugs.
Apply diethyl ether to the plug of the collecting tube so that the flies are anaesthetized within 20 seconds.
Shake the flies out on a sheet of filter paper and separate the sexes.
Use 8 females and 15 males for each fresh breeding batch.
The males are smaller than the females and the shapes of the abdomens differ.
The abdomen of the female is larger, more pointed and has 4 or 5 black transverse rings.
The abdomen of the male is smaller, more rounded with a black tip and has only two transverse rings.
The male has a row of bristles on the first foot section of each front leg.
To prevent the fruit flies sticking to the feeding mixture, transfer them to the breeding flask in small cones made from filter paper.
Make the cone by twisting a piece of filter paper about 5 cm square around the end of a pencil.
If unfertilized females are required, remove all the fruit flies from a breeding container containing a lot of pupae on the point of hatching, and after 6 hours, collect the fruit flies that have hatched.
Since the males are unable to copulate until 8 hours after hatching, the females among them cannot be fertilized.
Larvae obtained in the following manner are most suitable for preparing giant chromosomes.
Breeding colonies should not be overpopulated so remove adults from the breeding glass after they have deposited their eggs.
When the larvae are half grown, add more drops of viscous yeast suspension to the container and remove the cultures to a cool place, 15oC.
Take the fully grown larvae that have crept up the glass wall just before pupation for use as specimens.
3. Drosophila medium is an artificial diet consisting of 20 g of agar, 135 g of sugar, 38 g of yeast, 0.12 g of Nipagin (10 g L- 1 Nipagin in 70% ethanol) made up in 1 L of water and incubated at 20oC.
(Nipagin, methyl paraben, preservative, CH3(C6H4(OH)COO)
4. Use conical flasks for breeding.
See diagram 9.35.3 : Make a transfer cone.
Prepare the artificial diet the day before.
Add 2 tablespoons beet juice syrup to 1 litre water.
Heat the mixture and stir constantly.
Add semolina until a thick paste forms.
Prevent moulds forming by stirring in 1 spatula tip per litre of nipagin, methyl paraben, CH3(C6H4(OH)COO).
Pour the mixture into clean breeding flasks to a depth of 2 cm.
After cooling, add 5 drops of a viscous suspension of bakers' yeast in tap water.
Close the breeding flasks with cotton wool plugs.
Before the flies are inserted, absorb any liquid collected on the surface of the feeding mixture with strips of filter paper to prevent the flies sticking to the surface.
Transfer the flies by tapping the breeding flask on the palm of the hand, so that the flies fall to the bottom.
Quickly remove the cotton wool plug and place a collecting tube with the same size neck as the breeding glass on top of it.
By lightly tapping and shaking the flask, you can get the flies to enter the collecting tube.
Separate the two flasks and seal with cotton wool plugs.
Apply diethyl ether to the plug of the collecting tube so that the flies are anaesthetized within 20 seconds.
Shake the flies out on a sheet of filter paper and separate the sexes.
Use 8 females and 15 males for each fresh breeding batch.
The males are smaller than the females and the shapes of the abdomens differ.
The abdomen of the female is larger, more pointed and has 4 or 5 black transverse rings.
The abdomen of the male is smaller, more rounded with a black tip and has only two transverse rings.
The male has a row of bristles on the first foot section of each front leg.
To prevent the fruit flies sticking to the feeding mixture, transfer them to the breeding flask in small cones made from filter paper.
Make the cone by twisting a piece of filter paper about 5 cm square around the end of a pencil.
If unfertilized females are required, remove all the fruit flies from a breeding container containing a lot of pupae on the point of hatching, and after 6 hours, collect the fruit flies that have hatched.
Since the males are unable to copulate until 8 hours after hatching, the females among them cannot be fertilized.
Larvae obtained in the following manner are most suitable for preparing giant chromosomes.
Breeding colonies should not be overpopulated so remove adults from the breeding glass after they have deposited their eggs.
When the larvae are half grown, add more drops of viscous yeast suspension to the container and remove the cultures to a cool place, 15oC.
Take the fully grown larvae that have crept up the glass wall just before pupation for use as specimens.
9.6.2 Fruit fly strains
Drosophila has 8 chromosomes.
Different varieties are called mutants, because they have developed by mutation of genes.
Describe the following strains: Normal wild, vestigial vg, ebony e, white w, and Curly Cy.
Anaesthetize the fruit flies with diethyl ether.
Put them on a sheet of filter paper and examine them with a 6 × magnifying glass or stereoscopic magnifier.
Differentiate between the strains by means of their physical characteristics, e.g. colour of body and eyes.
Compile a table of observations:
Table 9.6.2
Strain characteristic Normal, wild type
+		 vestigial
vg		 ebony
e		 white
w		 Curly
Cy
Colour of body .
 . . . .
Colour of eyes . . . . .

9.6.4 Mendel's experiments
Gregor Mendel (1822-1884) used garden peas, (Pisum sativum), for most of his experiments, because they have constant different characters, flowers of hybrids can be protected from all other pollen and hybrids and offspring can produce viable seeds.
In 1854 Mendel chose seven varieties for experimental work:
1. smooth vs. wrinkled peas,
2. yellow cotyledons with yellow peas vs. green cotyledons with green peas,
3. white seed coats with white flowers vs. grey seed coats with purple flowers,
4. smooth vs. constricted pods,
5. green vs. yellow pods,
6. axial vs. terminal flowers,
7. tall vs. dwarf plant.
Pisum satisum is self-pollinating before the flower bud opens, so pea varieties are naturally pure varieties.
It is easy to open the immature large flowers, remove the nine anthers and pollinate from another plant by using a small brush.
Each crossbreeding can go both ways, from plant 1 to plant 2 and from plant 2 to plant 1, reciprocal crossing.
He started experiments with true breeding plants, so their offspring would be identical to the parents and changes in the progeny must be due to crossbreeding.
In 1865 Mendel published his study of inheritance "Versuche über Pflanzen-Hybriden", (Experiments in Plant Hybridization), that includes three principles concerning the inheritance of traits when cross-breeding.
Seed shape and seed colour can be seen in the pods of the parent plants without waiting for the seeds to be planted and grown, so he concentrated on these traits.
The principle of uniformity is that if two plants that differ in one trait are crossed they will be uniform in the chosen trait, which will be either one of the parents' traits.
So when parent 1 is crossed with parent 2, all the individuals of the hybrid, the first filial generation (F1) will have the same characteristic for any pair of characteristics.
A unit of heredity is now called a gene and the list of different genes is called the genotype.
A gene is sequence of DNA that the cell uses to produce a protein.
A gene may produce an RNA molecule that alters the activity of another gene.
Pairs of genes occupying the same place on different chromosomes are called alleles, e.g. the genes for round or wrinkled ripe seeds are alleles.
An allele is a version of a gene and they differ by one or more genetic base pairs.
A gene may have many alleles
The expression of genes in an organism is called the phenotype.
If true breeding tall plants are crossed with true breeding short plants and tall is dominant and short is recessive, the offspring, the F1 generation, will all be tall.
Punnet square
In the following Punnet square, the two possible gametes of the tall plants are shown horizontally and the two possible gametes of the tall plants are shown vertically.
Table 9.6.3 Gametes of tall plants
Gametes
 T
 T
t
 Tt (tall)
 Tt (tall)
t
 Tt (tall)
 Tt (tall)
When an individual of the F1 generation is crossed with another individual of the same F1 generation, the resulting hybrids are members of the F2 generation.
The principle of segregation is that the individuals of the F2 generation are not uniform.
Hereditary traits occur in pairs, one of each pair being inherited from each parent.
During meiosis, when gametes are formed, each pair of alleles, e.g. tall seeds / short seeds in the parent cells segregate (separate) into different gametes.
So any gamete carries either the tall seeds or short seeds gene, but not both and not neither.
In the following Punnet square, the gametes produced by the two parents, shown horizontally and vertically, are either tall or short.
If tall is dominant and short is recessive, let T = tall and t = short.
The genotype of the offspring can be TT (tall) or tT (tall) or Tt (tall) or tt (short) so the ratio of phenotypes is 3 tall plants to 1 short plant.
This study of one allele is called a monohybrid cross.
Table 9.6.3.1 Gametes of two parents, tall or short
Gametes
 T
 t
T
 T T (tall)
 t T (tall)
t
 T t (tall)
 t t (short)
Similarly, the allele round or wrinkled seeds, round is dominant and wrinkled is recessive.
Let R = round and r = wrinkled.
The result of the monohybrid cross would be a ratio of 3 (round) to 1 (wrinkled) seeds.
The genotypes TT and tt are said to be homozygous.
The genotype Tt (or tT) is said to be heterozygous.
The principle of independent assortment is that each trait is inherited independently of the other traits, so new combinations of traits can occur that were not existing before.
The segregation of one pair of alleles is independent of the segregation of any other pair of alleles.
This principle is valid only for genes on different chromosomes.
For a dihybrid cross the offspring have 16 possible different genotypes and the ratio of phenotypes is:
9 (tall round),
3 (tall wrinkled),
3 (short round),
1 (short wrinkled).
Table 9.6.3.2 Dihybrid cross
Gametes
 TR
 Tr
 tR
 tr
TR
 TRTR (tall round)
 TrTR (tall round) tRTR (tall round) trTR (tall round)
Tr
 TRTr (tall round) TrTr (tall wrinkled)
 tRTr (tall round) trTr (tall wrinkled)
tR
 TRtR (tall round) TrtR (tall round) tRtR (short round)
 trtR (short round)
tr
 TRtr (tall round) Trtr (tall wrinkled) tRtr (short round) trtr (short wrinkled)
Later, geneticists interpreted Mendel's principles as "Mendel's three laws" while others referred to principle 2 and 3 as Mendel's First Law and Mendel's Second Law.
9.6.5 Mendel's First Law, Law of Uniformity (inheritance of one pair of characteristics)
According to Mendel's First Law, when homozygous strains are crossed that differ by one or more characteristics, the offspring in the first filial generation (F1) will all have the same characteristics.
This is called the law of uniformity.
To investigate the validity of Mendel's First Law, put 8 unfertilized female fruit flies, Normal wild strain (+) and 15 males, Dark, ebony strain (e) in each of two breeding flasks containing Drosophila medium.
Then do a reciprocal hybridization with 8 unfertilized females, Dark, ebony strain (e) and 15 males, Normal wild strain (+) in each of two breeding flasks containing the Drosophila medium.
Leave the four breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the body colour of the first filial generation (F1) offspring in the four breeding flasks.
Count the numbers of each sex after anaesthetizing them with diethyl ether.
Use a magnifying glass to distinguish between males and females.
Compare the body colour of the first filial generation (F1), fruit flies with the body colour of the parents.
Note the sex ratio.
The parent fruit flies are homozygous for the characteristics under investigation.
The genotypes are +/+ for fruit flies with normal body colour and e/e for fruit flies with dark body colour.
So the germ cells are + or +, and e or e.
Insert all possible combinations of the genes in the Punnet square diagram below.
Note whether the theoretical result agrees with the practical result of this experiment.
Table 9.6.4.1
. (+) male (+) male
(e) female .
 .
(e) female . .
. (e) male (e) male
(+) female . .
(+) female . .

9.6.6 Mendel's First Law, Law of Uniformity (inheritance of two pairs of characteristics)
Put 8 unfertilized female fruit flies, Dark, ebony strain (e) and 15 male fruit flies, Stump-winged vestigial strain (vg), in each of two breeding flasks containing the Drosophila medium.
Leave the breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the body colour and wing shape of the first filial generation (F1) offspring in both breeding flasks.
Anaesthetize the fruit flies with diethyl ether.
Place them on a sheet of filter paper and examine them under a magnifying glass.
Compare their appearance with the parents.
The parent fruit flies that were crossbreeds possessed the following observable characteristics:
Table 9.6.4.2
.
 Body colour Wing shape
Male Normal wild (+) Stump-winged, vestigial (vg)
Female Dark, ebony (e) Normal wild (+)
The genotypes are the following:
male + / + vg / vg
female e / e + / +
To find what types of germ cells can be formed, insert all possible combinations of the genes in the Punnet square.
Note whether the theoretical results agree with the experimental results.
Draw a Punnet square and predict whether the result of the experiment would have been the same if both recessive characteristics were introduced by one parent in the hybridization.
Table 9.6.4.2.1
.
 male (+) male (vg)
female (e) (+) (e) (vg) (e)
female (+) (+) (+) (vg) (+)

9.6.7 Mendel's Second Law, Law of Segregation
According to Mendel's Second Law, the characteristics of the parent generation recur in the second filial generation (F2) in a quite specific numerical ratio.
Investigate the segregation of the dominant recessive characteristic pair Normal wild strain / Dark, ebony strain (+ / e) of the fly in the second filial generation.
Put 8 female and 5 male fruit flies from one of the first filial generations (F1) from the previous experiment in each of two breeding flasks containing Drosophila medium.
Leave the breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the body colour of the second filial generation (F2) offspring.
Anaesthetize the fruit flies with diethyl ether and place them on a sheet of filter paper.
Note the numerical ratio of the body colours of the parent fruit flies, Normal wild strain (+) and Dark, ebony strain (e).
The genotype of the fruit flies of the first filial generation (F1) crossed in this experiment is + / e.
The germ cells they can form are (+) or (e).
Insert all possible combinations of the genes in a Punnet square.
Does the theoretical result obtained with the Punnet square agree with the practical result from the hybridization experiment?
Make a comparative table using both sets of results.
Calculate numerical values from the Punnet square and the numbers obtained by counting in the experiment.
Note the deviations.
Note whether all fruit flies having the same appearance have the same genetic constitution, i.e. do fruit flies with the same phenotype always have the same genotype.
Table 9.6.5
. male male
female .
 .
female . .

9.6.8 Mendel's Third Law, Law of Independent Assortment
According to Mendel's Second Law, the characteristics of the parent generation recur in an exact numerical ratio (subdivision ratio) in the second filial generation (F2), following the Law of Segregation.
If different pairs of characteristics (alleles), are not contained on the same chromosome, they are distributed (assorted), independently during the formation of the germ cells and freely recombine.
This process is called Mendel's Third Law, Law of Independent Assortment.
Investigate the free recombination of genes, Mendel's Third Law, in the fruit fly.
Place 8 female and 15 male fruit flies from the first filial (F1) generation of the previous experiment in each of two breeding flask containing Drosophila medium.
Leave the breeding flasks to stand at room temperature.
About 10 days after the first fruit flies have hatched, examine the offspring in both breeding flasks, the second filial generation (F2) for their body colour and wing shape.
For this purpose, anaesthetize the fruit flies with diethyl ether.
Put them on a sheet of filter paper and examine them under a magnifying glass.
Note the number of different characteristic types, phenotypes.
Count each of the different types.
Note what phenotype has occurred for the first time.
Normal wild (+) and Dark ebony (e).
In the previous experiment, you crossed Stump-wing, vestigial (vg), and Dark body, ebony (e), fruit flies of the parent generation.
Their genotypes were vg/vg and +/+ respectively, +/+ e/e.
The offspring, in accordance with their genotypes, +/vg, are normal-coloured and have normal-shaped wings, e/+.
These fruit flies were crossed in the present experiment.
Note what type of germ cells they have.
Insert all possible combinations of the genes in a Punnet square.
Determine the external characteristics of the fruit flies according to their respective genes.
Note which characteristic types (phenotypes) must occur and how often.
Note whether this theoretical result agrees with the practical result of the experiment.
Table 9.6.6
. male male male male
female . . . .
female . . . .
female . . . .
female . . . .

9.6.9 Sex-linked inheritance, introduction of a recessive characteristic by female
The female has the pair of chromosomes (X/X), but the male has the genotype (X/Y), so the genes on the X chromosome are distributed differently in hybridization from those on the other chromosomes.
Investigate the inheritance of the sex-linked recessive characteristic white (white-eyed, w) in the fly introduced by the female in hybridization.
Put 8 unfertilized female fruit flies, white-eyed strain (w) and 15 male, Normal wild strain (+) in each of two breeding flasks containing Drosophila medium.
Leave the breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the eye colour and sex of the first filial generation (F1) offspring in both breeding flasks.
Anaesthetize the fruit flies with diethyl ether.
Put them on a sheet of filter paper and examine them under a magnifying glass.
Record the results in a table as follows:
Table 9.6.7.1
. male female
White eyes . .
Normal-coloured (red) eyes . .
Note whether the result in accordance with Mendel's First Law.
The gene for eye colour is on the X chromosome.
The females have two X chromosomes.
In this experiment they are white-eyed and their genotype is w/w.
The males possess only one X chromosome.
In this experiment they have normal coloured (red) eyes and their genotype is +/y.
Note what type of germ cells can be formed and insert all possible combinations of the genes in a Punnet square that follows.
Determine the external characteristics and sex of the files according to their respective genes.
Note whether this theoretical result agrees with the result of the experiment.
Table 9.6.7.2
. male male
female . .
female . .

2. Put 8 females and 5 males of the first filial generation (F1) from the above experiment in each of two breeding flasks containing Drosophila medium.
Leave the breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the eye colour and sex of the second filial generation (F2) offspring in both breeding vessels.
Anaesthetize the fruit flies with diethyl ether, place them on a sheet of filter paper and examine them under a magnifying glass.
Count the files and record the results in the following table:
Table 9.6.7.3
. male female
white eyes . .
normal coloured (red) eyes . .
Note the genotype of both the males and females of the first filial generation.
Note the types of germ cells they can be formed.
Insert all possible combinations of genes in the Punnet square that follows.
Determine the external characteristics, phenotypes and sex of the fruit flies according to their respective genes.
Note whether the theoretical results agree with the practical result of the experiment.
Table 9.6.7.4
. male male
female . .
female . .

9.6.10 Sex-linked inheritance, introduction of a recessive characteristic by a male
1. In the preceding experiments you investigated the heredity process when a sex-linked recessive characteristic is introduced by the female during hybridization.
Investigate the heredity mechanism when this characteristic is introduced by the male when the sex-linked recessive characteristic white (white eyes, w) in the fly is introduced by the male.
Put 8 unfertilized female fruit flies, Normal wild strain (+) and 15 males, white strain (w) in each of two breeding flasks containing Drosophila medium.
Leave the breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the colour of the eyes of the first filial generation (F1) offspring in both breeding flasks.
Anaesthetize the fruit flies with diethyl ether.
Put them on a sheet of filter paper and examine them under a magnifying glass.
Note whether the result is in accordance with Mendel's First Law.
Note the genotypes of the parent fruit flies and the germ cells can they form.
Insert all possible combinations of the genes in the following Punnet square.
Determine the external characteristics, phenotypes, of the fruit flies according to their respective genes.
Note whether the theoretical result obtained with the Punnet square agrees with the practical result of the experiment.
Table 9.6.8.1
. male male
female . .
female . .

2. Put 8 females and 15 males of the first filial generation (F1) from the above experiment in each of two breeding flasks containing Drosophila medium.
Leave the breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the eye colour and sex of the second filial generation (F2) offspring in both breeding flasks.
Anaesthetize the fruit flies with diethyl ether.
Put them on a sheet of filter paper and examine them under a magnifying glass.
Count the fruit flies and record the results in the following table.
Table 9.6.8.2
. male female
white eyes . .
normal coloured (red) eyes . .
Note the ratio of red-eyed to white-eyed fruit flies.
Note how the eye colour is distributed between the sexes and the genotype of the fruit flies of the first filial generation.
Note what types of germ cells these fruit flies can form.
Insert all possible combinations of the genes in the following Punnet square.
Determine the external characteristics (phenotypes) and sex of the fruit flies according to their respective genes.
Note whether this theoretical result agrees with the practical result of the hybridization experiment.
Table 9.6.8.3
. male male
female . .
female . .

9.7.1 Genetics, lethal factors
After many mutations, the genotype may become altered to such an extent that the offspring are no longer viable.
Genes that mutate in this way are called lethal factors, e.g. (CY) mutant of the fruit fly.
These fruit flies have upwards curving wings.
The characteristic is dominant.
Fertilized egg cells in which (CY) it is homozygous do not develop.
Experiment
Put 8 unfertilized female and 15 male fruit flies, curly strain (Cy) in each of two breeding flasks containing Drosophila medium.
Leave the breeding flasks to stand at room temperature.
Ten days after the first fruit flies have hatched, note the shape of the wings of the offspring in each breeding container.
Anaesthetize the fruit flies with diethyl ether.
Put them on a sheet of filter paper and examine them under a magnifying glass.
Count the fruit flies with upwards curving wings and normal wings.
The dominant characteristic Curly homozygous fruit flies, genotype (CYCY) cannot exist so the hybrid fruit flies must have the genotype Cy/+.
Insert all possible combinations of the genes in the Punnet square.
Determine from the Punnet square what ratio there ought to be of fruit flies with upwards curving to normal wings in the first filial generation (F1).
Note whether this theoretical value corresponds to the practical result of the experiment.
Note the apparent deviation from Mendel's Laws and why it occurs.
Table 9.6.9
. male male
female . .
female . .

9.7.2 Giant chromosomes
See 3.3.0: Aceto-carmine, microscopy stain.
Experiment
Investigate the form and structure of the giant chromosomes in the salivary gland cells of the larva of the fruit fly.
Press carefully on the coverslip with the handle stem of a dissecting needle until the salivary glands disintegrate into individual cells.
Apply slightly more pressure to squash the cells so that the chromosomes are pressed out.
Use filter paper to draw off the acetic acid that has spread under the coverslip and examine the preparation under a microscope with preferably an oil immersion lens.
Use magnification not less than 40 X.
Put a larva of the fly in a large drop of carmine acetic acid on a microscope slide.
Hold down the larva by pressing the side of a dissecting needle horizontally across it, about one third of the way along from the end of the abdomen.
Pierce the exoskeleton by pressing the point of another dissecting needle at a slight angle from the horizontal into the larva between the second and third segment, i.e. directly behind the fauces.
Pull the head section of the larva forwards until it becomes detached from the rest of the body, pulling the organs attached to it out of the body.
Identify the salivary glands from by their club-like shape and glazed appearance.
Remove the attached fatty tissue.
Transfer the salivary glands to a fresh drop of carmine acetic acid on a second slide and place a coverslip over them.
After 2 minutes place a drop of 45% acetic acid on the edge of the coverslip and draw it across under the coverslip by placing the edge of a filter paper strip at the opposite edge of the glass.
The chromosomes in the salivary gland cells of the larvae of fruit flies are so large they are called giant chromosomes.
These chromosomes form by repeated longitudinal division of the chromatic thread without subsequent splitting of the products of the division.
Note their shape and structure.
9.7.3 Salivary chromosomes
See 3.2 Aceto-orcein stain, microscopy stain.
Some fly larvae, e.g. Sarcophagid blowflies, have very large salivary gland chromosomes that are easy to study.
Blowflies can be collected from uncooked meat left outdoors.
Put a larva in a drop of salt solution on a microscope slide.
Use two dissecting needles to hold the tail end with one needle and pierce the head with the other needle.
Slowly stretch the larva between the two needles until the head, mouth parts, digestive tube and salivary glands are pulled free.
Separate the fat cells, digestive tube and head from the salivary glands.
Use a microscope to observe a cell squash of the glands stained with aceto-orcein stain.
Observe the large banded chromosomes.
9.3.1 Mitosis
The fundamental reproductive process called cell division may be studied by selecting an appropriate tissue that is growing rapidly.
A good source of such cells is the root tip region of onions or other related plants.
Onion bulbs, garlic cloves or onion sets placed in an aerated water bath provide large quantities of material.
The cells of growing and developing organisms are constantly multiplying by cell division.
In unicellular organisms, daughter cells form, which separate after the division process is complete and continue to develop independently.
In multicellular organisms the cell mass is increased by cell division. The organisms grow.
In cells without a nucleus cell division takes place using a simple constriction process.
However, the division process in nucleated cells involves a complicated mechanism of division of the cell nucleus, during which operation the chromosomes are clearly visible and are distributed between the two daughter cells after longitudinal splitting.
This process is called indirect nuclear division or mitosis.
Observe the different phases of indirect nuclear division, mitosis.
In the onion root tip, before mitosis starts, interphase, granules in the nucleus have the staining reaction of chromatin that consists of DNA and proteins, mostly histones.
During interphase DNA replication occurs so the chromosome appearing in prophase will have two identical sister chromatids.
Prophase:
The loose coils of the chromatin network condenses to become the 16 chromosomes of the onion cell, double structures with two identical chromatids joined by a centromere.
The nuclear membrane and nucleoli disappear.
Prophase occupies about two thirds of the time taken for mitosis.
Metaphase:
Curved microtubules, fibres, form a spindle outside the nucleus.
The centromeres move to the central plane of the spindle, metaphase plate.
The centromeres split in two.
Anaphase:
The identical chromatids move to the opposite poles of the spindle to leave two identical sets of chromosomes and a new nuclear membrane appears around each set.
Spindle fibres do not pull the chromatids to the poles of the dividing cell.
Each chromatid (composed of DNA + protein) has, among its proteins, a motor protein that crawls along the spindle fibre, dragging the whole chromatid along with it.
In the trailing fibre behind this protein, the spindle fibre is disassembled and disappears from microscopic view.
The spindle fibre itself is nonmotile and acts only like a railroad track as a path for the motor protein to travel on.
The chromatids are now chromosomes.
Telophase:
Nucleoli reappear.
The chromosomes return to being chromatin granules.
Cytokinesis is the division of the cytoplasm between the new nuclei and the formation of new cell walls.
Mitotic cell division is the basis of growth and tissue repair in multicellular organisms.
Mitosis keeps the chromosome number constant.
9.3.2 Mitosis in cells of onion root tip
See diagram 9.107.1: Mitosis.
See diagram 9.3.69: Mitosis in onion root tip cells.
See diagram 2.26: Drawing stain across specimen under coverslip.
In the onion root tip, before mitosis starts, interphase, granules in the nucleus have the staining reactions of chromatin that consists of DNA and proteins, mostly histones.
During interphase DNA replication occurs so the chromosome appearing in prophase will have two identical sister chromatids.
Experiment
1. Put an onion or shallot in moist absorbent paper in a warm place to obtain roots.
Cut off 1 cm lengths from the ends of roots and fix them in a solution of 1 part glacial acetic acid to 3 parts 95% alcohol.
Leave for 24 hours.
Put a piece of root in a drop of aceto-carmine on a slide.
Cut off 3 mm of the tip and discard the rest.
Gently warm over a lamp (light) bulb.
Place a coverslip over the drop of stain and apply gentle pressure to separate the cells.
These cells will show stages in mitosis.
2. In the morning, cut 5 mm from the end of a growing root of onion or pea.
Cut the piece of root twice longitudinally.
Put the longitudinal sections in a drop of carmine acetic acid on a microscope slide.
Cover with a coverslip and heat to boiling point over a small flame by moving the slide backwards and forwards to prevent excessive heating in one place.
Put a drop of 2% acetic acid at the side of the coverslip and draw it across under the coverslip with absorbent paper on the opposite side.
Press down on the coverslip with a scalpel handle to squash the cells.
Examine the cells under high power.
Look for dividing nuclei at different stages of development, prophase, metaphase, anaphase and telophase.
Count the number of chromosomes, e.g. onion 16 chromosomes and pea 14 chromosomes.
Find the different stages and count the number of each stage.
Use this information to estimate the relative lengths of time for each stage during mitosis.
The process takes about 2 hours with prophase taking two thirds of the time.
Repeat the experiment with root tip specimens taken at different times.
3. Cut off the white root tips of healthy specimens.
Cut a 3 mm cylinder from the end of a root.
Put it in a drop of aceto-carmine stain on a microscope slide.
Cut up the onion tip with a razor blade until the pieces are extremely small.
Cover the preparation with a cover glass.
With a piece of folded paper towelling over your thumb for protection, gently squash the pieces of root tip by pressing on the coverslip with a rolling motion.
Do not allow the cover glass to slide.
Then examine the preparation with the low power of a microscope.
Look for dark stained threadlike bodies.
These bodies are chromosomes or mitotic figures.
Find the find various types or stages and count the number of various stages.
From this information estimate the relative lengths of time that the various stages are present in a reproducing cell.
4. Plant an onion or shallot in moist absorbent paper in a warm place to obtain roots.
Cut off 1 cm lengths from the ends of roots and fix them in a solution of 1 part glacial acetic acid to 3 parts 95% alcohol.
Leave for 24 hours.
Put a piece of root in a drop of aceto-carmine on a slide.
Cut off 3 mm of the tip and discard the rest.
Gently warm over a spirit lamp.
Place a coverslip over the drop of stain and apply gentle pressure to separate the cells.
These cells will show stages in mitosis.
5. Put onion root tips in 1 mL orcein in a watch glass.
Heat over a spirit lamp for 1 minute or until the tips are soft.
Scrape the tips over a microscope slide to make an even smear.
Add drops of glycerine and a coverslip.
6. Examine a prepared slide showing cells in various stages of mitotic division, e.g. a stained longitudinal section through the root tip of the onion.
9.3.3 Meiosis
See diagram 9.108.1: Meiosis 1, Prophase 1.
See diagram 9.108.2: Meiosis 2. Metaphase, Anaphase Telophase.
See diagram: Meiosis
First division
Prophase I, Leptotene
The chromosomes appear like a long string with of beads, called chromomeres.
Prophase I, Zygotene
The homologous chromosomes from the male and female gametes lie closely alongside each other forming shorter and thicker paired structures, so appearing to halve the number of chromosomes.
They are attached to the inside wall of the nucleus in animals, but form a tangled heap in plants.
Prophase I, Pachytene
When the pairing complete and the chromosomes appear as "thick" strings.
The two chromosomes are referred to as a "bivalent, " while the same structure viewed as four chromatids is known as a "tetrad."
Prophase I, Diplotene
Each chromosome of a pair splits into two chromatids along their length except at the centromeres.
As the chromosomes separate, they remain attached at chiasmata where recombination of the genes occurs.
Prophase I, Diakinesis
The homologous chromosomes repel each other and continue contracting.
The nucleolus and nuclear membrane disappear.
Metaphase, I
The bipolar spindle appears and the paired chromosomes (tetrads) line up on the metaphase plate and attach to the spindle fibres with the centromeres.
Anaphase, I
Each homologue moves toward opposite poles so halving the number of chromosomes.
This process is the reduction division that characterizes meiosis.
Telophase, I
The chromosomes may become surrounded by a nuclear membrane, but usually the second division starts when the chromatids of each chromosome separate and move to the opposite pole of the spindle.
Second division, II
This division is similar to normal mitosis, except that the number of chromosomes is n not 2n.
The four n nuclei develop nuclear membranes during the telophase of the second division to produce a tetrad of pollen grains, but only one n nucleus develops in the ovule.
9.3.4 Meiosis in grasshopper testes
Meiosis can be seen in grasshopper testes or in (Ascaris lumbricoides), where the eggs remain dormant until fertilized.
During fertilization, the male and female gametes fuse to form a zygote, so the number of chromosomes doubles, n chromosomes from the male gamete and n chromosomes from the female gamete.
The 2n cells in the sex organs undergo meiosis to halve the number of chromosomes to n chromosomes in the male gamete or female gamete.
9.3.5 Meiosis, Lilium
See diagram 9.98.6: Fertilization in Lilium
See diagram 9.98.11 Lilium Megaspore mother cell, Prophase 1
See diagram 9.98.11 Lilium Megaspore mother cell, Telophase 1
See diagram 9.98.12a Lilium After meiosis
See diagram 9.98.12b Lilium After first mitosis, After second mitosis
Lilium has a superior ovary above the attachment of the anthers.
The ovary becomes a three-celled capsule.
The swollen base part of the carpels, gynoecium, contains six ovules.
The ovule contains the female gamete and after fertilization it becomes the seed.
The ovule has tissue, nucellus, surrounded by protective layers and is attached to the ovary by the funicle.
The region of the ovule where the nucellus joins the outer integuments is the chalaza.
Inside the ovule is the embryo sac, megaspore, containing the megaspore mother cell, megasporocyte, that contains a nucleus with 2 N, diploid, chromosomes.
Meiosis
By the process of meiosis the megaspore mother cell will produce four N, haploid, megaspores, which will develop into the gametophytes.
The four nuclei produced by meiosis migrate within the embryo sac, one nucleus to the micropyle end of the embryo sac and the other three to the chalaza end.
A large central vacuole forms in the embryo sac.
First mitosis
At the chalaza end of the embryo sac, the chromosomes from the three nuclei condense around a common spindle to form two nuclei with 3N chromosomes.
At the micropyle end of the embryo sac, the single N nucleus divides to form two nuclei with n chromosomes.
Second mitosis
At the chalaza end of the embryo sac, one nucleus aborts, but the other nucleus divides to form a 3N polar nucleus and antipodal cell.
At the micropyle end of the embryo sac, one nucleus forms two n synergids while the other nucleus forms the N egg nucleus, and the micropyle end N polar nucleus.
Double fertilization
At the chalaza end of the embryo sac, triple fusion forms a 5 N primary endosperm nucleus from the 3N polar nucleus + 1N male sperm nucleus from sperm 1 + 1 M micropyle end polar nucleus, 3 + 1 + 1 = 5N.
At the micropyle end of the embryo sac, a male n nucleus from sperm 2 with an n female nucleus to form a 2n zygote.
After fertilization
The primary endosperm nucleus divides to form the endosperm.
The zygote divides to form a base area and globular pro-embryo.
9.9.4 Human genetic inheritance
Members of a family are always interested in "family resemblance", how members of the same family look alike.
The resemblance is due to traits inherited from parents to offspring.
The "expression of traits" (phenotype), depends on an individual's "genetic make-up" (genotype).
The DNA in sex cells is called the germline and contrasts with the DNA in the other cells of the body, the somatic cells, which cannot pass on their DNA to any offspring.
Human genes are carried on 23 pairs of chromosomes.
One set of 23 chromosomes is inherited from the mother, and the other set of 23 chromosomes is inherited from the father.
About 10 000 genes are carried on 44 of the 46 chromosomes.
The other two chromosomes are the X and the Y chromosome.
Females have two X chromosomes and males have an X and a Y chromosome.
The X chromosome is similar to the other chromosomes, but the Y chromosome is much smaller with only about 30 working genes that control maleness.
In the human male, the 23rd "pair" of chromosomes, i.e. XY, has only one copy of the genes on the X-chromosome, inherited from the mother, and only one copy of genes on the Y-chromosome, inherited from the father.
Each trait may be controlled by a single pair of genes, i.e. one of each gene pair (allele) inherited from each parent, or controlled by multiple pairs of alleles, i.e. many gene pairs inherited from each parent.
Some traits are high frequency traits.
Some traits depend on the sex of a person.
For example, index finger (second finger) shorter than ring finger (fourth finger), is the dominant phenotype only in males, but index finger longer than ring finger is the dominant phenotype in females.
Human DNA contains about 25 000 genes.
However, we also have some genes in the cell mitochondria.
Mitochondrial DNA (mtDNA, mDNA) controls the Kreb's cycle in basic metabolism.
Sperm have a few mitochondria in the middle piece, but the ovum has about a quarter of a million mitochondria.
After fertilization, the mitochondria from the sperm all die.
So we inherit only mitochondria genes from the ovum of the mother, giving us the same mDNA as your mother had.
Mitochondrial DNA forms in a circle of 6, 589 DNA bases.
Mitochondrial DNA mutates much faster than nuclear DNA, so that almost everyone has different mitochondrial DNA, but relatively similar nuclear DNA.
Some mutations of mitochondrial DNA can cause serious problems, e.g. exercise intolerance syndrome, Alzheimer's disease, diabetes.
9.9.1 Human traits with simple inheritance patterns
Ethics of teaching about simple inheritance patterns.
Some secondary school students and their parents have become distressed when a simple inheritance pattern test, e.g. tongue rolling, provides evidence that the secondary school student is not the biological child of the parents.
Usually the student was adopted when young, but the parents had never told the student about this matter.
The teacher should not encourage students to draw conclusion about inheritance tests applied to the student's own family.
Some traits may not be due to a single pair of dominant or recessive genes.
For example, brown eye genes are dominant over green eye genes and both are dominant over blue eye genes.
However, eye colour is caused by many genes that control the colour and the amount of melanin pigments in the iris.
The child of both parents with light colour eyes may have dark colour eyes due to mutations and other changes in the chromosomes.
Eyes may be darker after exposure to light and mutations can cause unusual eye colours.
Similarly, hair colour is caused by multiple genes and may change with age.
Investigate the sameness and difference between your parents and you for the above genetic characteristics.
Perhaps all your siblings and one of your parents have the same unusual genetic traits, because this characteristic "runs in the family".
Some people have an unattached earlobe on one ear and attached earlobe on the other ear
Table 9.9.1
.
 Dominant Recessive
1.1
 Unattached earlobe, free earlobes Attached earlobe (to the face)
1.2
 Big toe shorter than second toe Big toe longer than second toe
1.3
 Widow's peak (downwards, V-shaped hair peak) No widow's peak
1.4
 Darwin's point on the ear, skin flap
 No Darwin's point skin flap
1.5
 Hair on middle segment of any finger
 No hair on any middle segments of fingers
1.6
 Straight thumb, cannot bend thumb back 50o "Hitchhiker's thumb"
1.7
 Little finger bends towards 4th finger, crooked pinkie
 Little finger straight, pinkie straight
1.8
 Facial dimples, cheek dimples
 No facial dimples when you smile
1.9
 Freckles
 No freckles
1.10
 Can roll tongue to form U-shape Cannot roll tongue
1.11
 Hair whorl clockwise Hair whorl anticlockwise
1.12
 Can taste PTC Cannot taste PTC, non-taster
1.13
 Left thumb on top of interlocking fingers
 Right thumb on top of interlocking fingers

9.9.5 PTC tasters and non-tasters
See 1.13: Phenylthiocarbamide, PTC, phenylthiourea, PTU, C7H8N2S.
(Thiocarbamide, CH4N2S, NH2CSNH2, thiourea)
Be careful! Phenylthiourea (PTC, PTU, phenyl thiocarbamide, n-phenylthiourea, structural formula: C6H6NHCSNH2) is very toxic and may be fatal if swallowed.
1. The ability to taste phenylthiourea (bitter) and other compounds with an N-C= group is an autosomal dominant trait, i.e. the defective gene is dominant and will be inherited by half the offspring of either sex.
Phenylthiourea tasters detect saccharin, potassium chloride (KCl) and caffeine as more bitter.
2. Put a piece of water-soaked absorbent paper on the tongue and roll it around the tongue for 10 seconds.
Repeat with absorbent paper soaked in PTC (phenylthiourea).
Be careful! Do not swallow the PTC.
Rinse the mouth out with water.
Did you notice a bitter taste?
9.9.6 Unattached / attached earlobe
See diagram 9.234: 1. Darwin's point, 2, Unattached earlobe, 3. Widow's peak.
Investigate the sameness and difference between your parents and you for the above genetic characteristics.
Perhaps all your siblings and one of your parents have the same unusual genetic traits, because this characteristic "runs in the family".
Some people have an unattached earlobe on one ear and attached earlobe on the other ear.
9.9.3 Fingerprint variations
See diagram 9.233: Finger lengths.
Look at the tiny ridges on the pad of your fingers.
When these are inked and placed on paper, a fingerprint is made.
Put the stamp pad near the edge of a table and, in turn, roll your fingers over the pad.
Each time, make a fingerprint on your notebook.
Label the fingerprint with the number of the finger and whether it is the right or left hand.
9.9.2 Down syndrome
The Down syndrome occurs in all races, cultures and histories.
The syndrome was described by Langdon Down in 1866.
However, the occurrence of 47 chromosomes, because of extra chromosome 21 in children with Down Syndrome was by Lejeune, J. et al in 1959, ""Etudes des chromosomes somatiques de neuf enfants mongoliens." ("Study of somatic chromosomes of 9 mongolian children.")
Children with Down Syndrome were formerly called "Mongoloid".
Modern medicine and training techniques to modify incompetent motor behaviour allow persons with Down syndrome to live at home and contribute to the local community.
Persons with Down syndrome do not have identical "trisomy 21" chromosome aberrations, so their behaviour varies.
9.8.1 Evidence for evolution
1. Carbon dating and radioactive clocks used to date fossils and rocks
2. Comparative anatomy, e.g. forelimbs of animals, fossils of fishes and amphibians
3. Comparative embryology, e.g. similar structures at early stages
4. Comparative biochemistry, e.g. DNA and RNA in all living organisms
5. Animal and plant breeding, e.g. pigeon breeds
6. Geographical isolation, flora and fauna of islands or remote continents, Galapagos tortoises, kangaroos, New Zealand flightless birds
The theory of biological evolution is a working hypothesis, based upon scientific evidence, which tries to explain observed changes to living things over time.
Biological evolution is a process that results in heritable changes in a population spread over many generations.
Biological evolution can be more precisely defined as any change in the frequency of alleles within a gene pool from one generation to the next.
Key principles of biological evolution
1. Descent with modification
This principle describes the process by which species of living things can undergo modification over time, with such change sometimes resulting in the formation of new, separate species.
All species on Earth have descended from other species, and a single, common ancestor lies at the base of the evolutionary tree.
9.8.2 Natural selection
Natural selection is a process in which the differential adaptation of organisms to their environment selects those traits that will be passed on (by reproduction), with greater frequency from one generation to the next.
Natural selection acts on individuals, but it is populations that evolve.
Populations change through the process of natural selection.
1. Individuals in a population of a particular species show inherited variation, i.e. differences between individuals resulting from them having different alleles of a gene and different combinations of alleles of different genes.
2. In most species of organisms, far more new individuals are produced each generation than survive to maturity, with only a small number of individuals surviving to produce offspring.
3. In a particular habitat subjected to environmental change, the inherited variation between individuals can result in some individuals of a species, by chance, having better characteristics to allow them to survive to maturity and reproduce.
These better-adapted individuals consequently produce more offspring than the less well-adapted individuals.
4. Some of the offspring of these better-adapted parents will inherit the beneficial characteristics, giving them a better chance to survive to maturity and reproduce, and to pass on the beneficial characteristics to some of their offspring.
With each new generation there will be a higher proportion of individuals with the beneficial characteristics, eventually leading to a whole population better adapted to the changed environmental factors.
Natural selection may be defined as differential reproductive success (fitness) of genotypes in a gene pool.
In the context of evolution, "fitness' refers to the average reproductive output of a genotype in a gene pool.
9.8.3 Position statements on the teaching of biological evolution
1. American Association of Physics Teachers [AAPT].
"We believe in teaching that science is a process that examines all of the evidence relevant to an issue and tests alternative hypotheses.
For this reason, we do not endorse teaching the "evidence against evolution", because currently no such scientific evidence exists.
Nor can we condone teaching "scientific creationism", "intelligent design", or other non-scientific viewpoints as valid scientific theories.
These beliefs ignore the important connections among empirical data and fail to provide testable hypotheses.
They should not be a part of the science curriculum"
2. Australian Academy of Science [AAS].
"Australian Academy of Science sees no objection to the teaching of creationism in schools as part of a course in dogmatic or comparative religion, or in some other non-scientific context.
There are no grounds, however, for requiring that creationism and intelligent design be taught as part of a science course."
3. National Science Teachers Association [NSTA], Australia.
"Science teachers should not advocate any religious interpretations of the natural world and should be non-judgmental about the personal beliefs of students.
Policy makers and administrators should not mandate policies requiring the teaching of "creation science" or related concepts, such as so-called "intelligent design", "abrupt appearance", and "arguments against evolution".
However, the professional responsibility of science teachers and curriculum specialists to provide students with quality science education should not be compromised by censorship, pseudoscience, inconsistencies, faulty scholarship, political influences, or unconstitutional mandates".