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A1 Cell Division

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A1 Cell Division

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Grant Misinzo
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© © All Rights Reserved
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Cell Division

AQA-A AQA-B CIE Edexcel OCR

Complete: Complete: Complete: Complete: Complete:


1-6, 11-12 1-6, 8-12 1-7, 11-12 1-6, 8-12 1-7, 11-12

Learning Objectives Applications of cloning (pages 155-161)


8. Recognise the role of mitosis in producing clones.
1. Compile your own glossary from the KEY WORDS
Define the term: totipotent. Describe the current and
displayed in bold type in the learning objectives below.
potential uses of cloning technology.
9. Explain the principles involved in reproducing plants
Mitosis and the cell cycle (pages 152-154) by vegetative propagation. Describe methods of
2. Using diagrams, describe the behaviour of vegetative propagation including plant tissue culture
chromosomes during a mitotic cell cycle in (micropropagation). Recognise some of the benefits
eukaryotes. Include reference to: mitosis, growth (G1 and disadvantages involved in plant cloning technology.
and G2), and DNA replication (S). 10. Recognise embryo splitting as a method by which
3. Recognise and describe the following events in mitosis: animals can be cloned and explain the principles
prophase, metaphase, anaphase, and telophase. involved. Contrast embryo splitting with the more recent
nuclear transfer technique. Explain the principles
4. EXTENSION: With respect to both plant and animal
of nuclear transfer and describe the benefits and
cells, understand the term cytokinesis, and distinguish
disadvantages involved with this technique.
between nuclear division and division of the cytoplasm.
5. Understand the role of mitosis in growth and repair,
and asexual reproduction. Recognise the importance The role of meiosis (page 151)
of daughter nuclei with chromosomes identical in
11. Contrast the final products of mitosis and meiosis.
number and type. Recognise cell division as a prelude
Explain what is meant by the terms: homologous
to cellular differentiation.
pairs (of chromosomes), haploid, diploid, reduction
6. Demonstrate appropriate staining techniques in the division. Explain why a reduction division is necessary
study of mitosis in plant material, e.g. root tip squash. before fertilisation in sexual reproduction.
7. Explain how carcinogens can upset the normal 12. Distinguish between the two divisions of meiosis:
controls regulating cell division. Define the terms: Meiosis I (reduction division) and Meiosis II.
cancer, tumour suppressor genes, oncogenes. List Recognise the main features of these stages.
factors that increase the chances of cancerous growth.

■ The Cell Cycle and Mitosis Biol. Sci. Rev., ■ Human Cloning - Why Ban it? Biol. Sci. Rev.
14(4) April 2002, pp. 37-41. Cell growth and 11(4) March 1999, pp.8-9. The ethics of human
division, key stages in the cell cycle, and the cloning and the advantages to be gained from it.
Textbooks complex control over different stages of mitosis.
■ Rebels without a Cause New Scientist, 13 July
2002, (Inside Science). The causes of cancer: the Internet
See the ‘Textbook Reference Grid’ on uncontrolled division of cells that results in tumour
pages 8-9 for textbook page references formation. Breast cancer is a case example.
relating to material in this topic. ■ Dance of the Chromosomes Biol. Sci. Rev.,
11(2) Nov. 1998, pp. 11-14. Techniques to explore See pages 10-11 for details of how to access Bio
Supplementary Texts the role of chromosomes in the cell cycle help us Links from our web site: www.biozone.co.uk.
Additional details of these texts are provided in the to find out what happens when steps go wrong. From Bio Links, access sites under the topics:
introductory resources section (pp. 4-6): ■ Fast Tissue Culture Biol. Sci. Rev., 10(3) Jan.
GENERAL BIOLOGY ONLINE RESOURCES >
■ Adds, J. et al., 2000. Molecules and Cells, 1998, pp. 2-6. Techniques for plant propagation
(includes design for a tissue culture project). Online Textbooks and Lecture Notes: • S-Cool!
(NelsonThornes), pp. 69-76. A level biology revision guide • Learn.co.uk •
■ Jones, N., et al., 2001. Essentials of Genetics, ■ Human Cloning Biol. Sci. Rev. 11(3) Jan. 1999,
Mark Rothery’s biology web site … and others
(John Murray), pp. 9-25. pp. 7-9. Nuclear transfer and the ethics of the
issues surrounding human and livestock cloning. CELL BIOLOGY AND BIOCHEMISTRY: • Cell
& molecular biology online • Cell structure and
function web links … and others > Cell Division:
Periodicals TEACHER’S REFERENCE • Cell division: Binary fission and mitosis •
■ Out of Control - Unlocking the Genetic Mitosis in the onion root tip… and others
Secrets of Cancer Biol. Sci. Rev., 11(3) Jan. 1999, BIOTECHNOLOGY > Applications > Cloning
pp. 36-39. The control of cell division: oncogenes and Tissue Culture: • Conceiving a clone •
See page 6 for details of publishers of periodicals:
and their role in the development of cancer. Tissue culture in the classroom … and others
STUDENT’S REFERENCE ■ Cloning for Medicine Scientific American,
December 1998, pp. 30-35. An excellent article
■ To Divide or Not to Divide Biol. Sci. Rev.,
describing the techniques and applications of
11(4) March 1999, pp. 2-5. The cell cycle: cell
growth and stages of cell division and their control.
cloning technology, including nuclear transfer. Software and video resources are
■ Mechanisms of Meiosis Biol. Sci. Rev., ■ Into the Clone Zone New Scientist, 9 May provided on the Teacher Resource
15(4), April 2003, pp. 20-24. A clear and thorough 1998, pp. 25-30. Where will the breakthroughs in Handbook on CD-ROM
account of the events and mechanisms of meiosis. cloning and genetic engineering lead us?
151
Cell Division
The life cycle of diploid sexually reproducing organisms (such gametes for the purpose of sexual reproduction. The difference
as humans) is illustrated in the diagram below. Gametogenesis between meiosis in males and in females should be noted (see
is the process responsible for the production of male and female spermatogenesis and oogenesis in the box below).

Human embryos have cells which are rapidly dividing


by mitosis. The term somatic means 'body', so the Female Male
cell divisions are creating new body cells (as opposed embryo embryo
to gametes or sex cells). The 2N number refers to how 2N 2N
many whole sets of chromosomes are present in each
body cell. For a normal human embryo, all cells will
Many Many
have a 2N number of 46. Somatic cell
mitotic mitotic
production
divisions divisions
Adults still continue to produce somatic cells by mitosis
for cell replacement and growth. Blood cells are Female Male
replaced by the body at the astonishing rate of two adult adult
million per second, and a layer of skin cells is
constantly lost and replaced about every 28 days.
2N 2N

Gamete production begins at puberty, and lasts until Meiosis Gamete Meiosis
menopause for women, and indefinitely for men. production
Gametes are produced by the special type of cell
division, called meiosis, which reduces the

Cell Division
chromosome number to half. Human males produce
about 200 million sperm per day (whether they are
used or not), while females usually release a single Egg Sperm
egg only once a month. 1N Fertilisation 1N

Fertilisation involves fusion of the sperm and the A single set of


egg to produce a single cell called the zygote. This chromosomes
cell has all the genetic information to build a human
Zygote
body as well as maintain it (metabolism).
2N
Several
Spermatogenesis Somatic cell mitotic
production divisions
Sperm production: Meiotic division of spermatogonia
produces the male gametes. This process is called
spermatogenesis. The nucleus of the germ cell in the Embryo
male divides twice to produce four similar-sized sperm 2N A double set of
cells. Many organisms produce vast quantities of male chromosomes
gametes in this way (e.g. pollen and sperm).
Somatic cell Many
Oogenesis production mitotic
divisions
Egg production: In females, meiosis in the oogonium
produces the egg cell or ovum. Unlike gamete production
in males, the divison of the cytoplasm during oogenesis Adult
is unequal. Most of the cytoplasm and one of the four
nuclei form the egg cell or ovum. The remainder of the 2N
cytoplasm, plus the other three nuclei, form much
smaller polar bodies and are abortive (i.e. do not take
part in fertilisation and formation of the zygote).

1. Describe the purpose of the following types of cell division:

(a) Mitosis:

(b) Meiosis:

2. Explain the significance of the zygote:

3. Describe the basic difference between the cell divisions involved in spermatogenesis and oogenesis:

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Mitosis and the Cell Cycle
Mitosis is part of the ‘cell cycle’ in which an existing cell (the associated with growth and repair of tissue, and it is the method
parent cell) divides into two new ones (the daughter cells). by which some organisms reproduce asexually. The example
Mitosis does not result in a change of chromosome numbers below illustrates the cell cycle in a plant cell. Note that in animal
(unlike meiosis): the daughter cells are identical to the parent cells, cytokinesis involves the formation of a constriction that
cell. Although mitosis is part of a continuous cell cycle, it is divides the cell in two. It is usually well underway by the end of
divided into stages (below). In plants and animals mitosis is telophase and does not involve the formation of a cell plate.

Centrosome, which Centrosomes in plant


A Interphase later forms the spindle, Early Prophase B Late Prophase cells lack centrioles. In
is also replicated. animal cells, centrioles
Nuclear are associated with the
Membrane centrosomes but their
exact role is unclear.

Cell enters mitosis


C

Nucleolus DNA is replicated to DNA continues condensing into Chromosomes continue Metaphase
form 2 chromatids chromosomes and the nuclear to coil up and appear as
membrane begins to dissolve double-chromatids
Interphase: Stages G1, S, G2

G2 Second Gap: the The mitotic spindle is


chromosomes formed to organise the
begin condensing chromosomes. The
Synthesis of S spindle consists of fibres
DNA to replicate M Mitosis: nuclear division made of microtubules
Cell Division

chromosomes and proteins.


Cytokinesis: division of the
C cytoplasm and separation of
First Gap: the cell
grows and develops. G1 the two cells. Cytokinesis is Anaphase
distinct from nuclear division.

Division of the cytoplasm Two new nuclei form. The cell


(cytokinesis) is complete. The plate forms across the midline The chromosomes
two daughter cells are now of the parent cell. This is where segregate, pulling
separate cells in their own right. the new cell wall will form. the chromatids apart

F Cytokinesis E Telophase Late Anaphase

1. The five photographs below were taken at various stages through the process of mitosis in a plant cell. They are not in any
particular order. Study the diagram above and determine the stage that each photograph represents (e.g. anaphase).

Photos: RCN

(a) (b) (c) (d) (e)


2. State two important changes that chromosomes must undergo before cell division can take place:

3. Briefly summarise the stages of the cell cycle by describing what is happening at the points (A-F) in the diagram above:

A.

B.

C.

D.

E.

F.

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Root Cell Development
In plants, cell division for growth (mitosis) is restricted to growing below illustrates the position and appearance of developing and
tips called meristematic tissue. These are located at the tips of growing cells in a plant root. Similar zones of development occur
every stem and root. This is unlike mitosis in a growing animal in the growing stem tips, which may give rise to specialised
where cell divisions can occur all over the body. The diagram structures such as leaves and flowers.

Zone of
specialisation

Root hair cell


Phloem cells

a Xylem vessel

Root tip

Cell Division
growing in

RCN
Zone of this direction
elongation

Zone of
cell division c

The root cap protects the growing tip of the


root immediately behind it. The cells in the
root cap undergo cell division to replace cells
that are rubbed off as the root grows.

1. Briefly describe what is happening to the plant cells at each of the points labelled (a) to (c) in the diagram above:

(a)

(b)

(c)

2. The light micrograph (below) shows a section of the cells of an onion root tip, stained to show up the chromosomes.
(a) State the mitotic stage of the cell labelled A and explain your answer:

(b) State the mitotic stage just completed in the cells labelled B and explain:

(c) If, in this example, 250 cells were examined and 25 were found to be in the
process of mitosis, state the proportion of the cell cycle occupied by mitosis:
B
A

3. Identify the cells that divide and specialise when a tree increases its girth (diameter):

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The Genetic Origins of Cancer
Normal cells do not live forever. Under certain circumstances, agents capable of causing cancer. Roughly 90% of carcinogens
cells are programmed to die, particularly during development. are also mutagens, i.e. they damage DNA. Chronic exposure to
Cells that become damaged beyond repair will normally undergo carcinogens accelerates the rate at which dividing cells make
this programmed cell death (called apoptosis or cell suicide). errors. Susceptibility to cancer is also influenced by genetic
Cancer cells evade this control and become immortal, continuing make-up. Any one or a number of cancer-causing factors
to divide regardless of any damage incurred. Carcinogens are (including defective genes) may interact to induce cancer.

Cancer: Cells out of Control


Normal cell Mutations (defects) in one or two of the controlling
Cancerous transformation results from genes cause the formation of a benign tumour; a
changes in the genes controlling normal cell tumour that is nonmalignant (not growing in size). As
growth and division. The resulting cells the number of controlling genes with mutations
increases, so too does the loss of control until the cell
become immortal and no longer carry out their
becomes cancerous (a 'renegade cell').
functional role. Two types of gene are normally
involved in controlling the cell cycle: proto-
oncogenes, which start the cell division Tumour-suppressor genes
process and are essential for normal cell When damage occurs, the
development, and tumour-suppressor tumour-suppressor gene p53
commands other genes to
genes, which switch off cell division. In their
bring cell division to a halt.
normal form, both kinds of genes work as a Damaged
team, enabling the body to perform vital tasks DNA
Cell Division

such as repairing defective cells and replacing


dead ones. But mutations in these genes can DNA
disrupt these finely tuned checks and If the damage is too molecule
serious to repair, p53
balances. Proto-oncogenes, through mutation,
activates other genes
can give rise to oncogenes; genes that lead that cause the cell to Proto-oncogenes
to uncontrollable cell division. Mutations to self-destruct. If repairs are made, Genes that turn on cell division. The
tumour-suppressor genes initiate most human then p53 allows the mutated form or oncogene somehow
cancers. The best studied tumour-suppressor cell cycle to continue. leads to unregulated cell multiplication.
gene is p53, which encodes a protein that
halts the cell cycle so that DNA can be
repaired before division.

The bloated, lumpy shape is readily


distinguishable from a healthy cell,
Features of Cancer Cells Cancer cells can go on dividing which has a flat, scaly appearance.
indefinitely, if they have a
The diagram right shows a single lung cell continual supply of nutrients,
that has become cancerous. It no longer carries and are said to be immortal. Metabolism may be
out the role of a lung cell, and instead takes deranged and the cell
on a parasitic lifestyle, taking from the body Cancer cells may have ceases to function in a
what it needs in the way of nutrients and unusual numbers of constructive way.
contributing nothing in return. The rate of cell chromosomes.
division is greater than in normal cells in the Cancerous cells lose
their attachments to
same tissue because there is no resting phase
neighbouring cells.
between divisions.

1. Explain how cancerous cells differ from normal cells:

2. Explain how the cell cycle is normally controlled, including reference to the role of tumour-suppressor genes:

3. With reference to the role of oncogenes, explain how the normal controls over the cell cycle can be lost:

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Cloning by Embryo Splitting
Livestock breeds frequently produce only one individual per cells from early embryos in mice. They are also learning how
pregnancy and all individuals in a herd will have different to force stem cells to differentiate into different tissues. Such
traits. Cloning (by embryo splitting or other means) makes it techniques may make it possible to manufacture cells or replace
possible to produce high value herds with identical traits more tissues damaged by illness (e.g. muscular dystrophy or diabetes).
quickly. This technique also has applications in the medical Individually matched stem cells could be made by transferring the
field, for example, in the cloning of embryonic stem cells. Such nucleus from one of the patient’s cells into a human egg to create
applications demonstrate the advances made recently in cloning an embryo. The embryo would be allowed to develop only to a
technology. Some of the most ambitious medical projects now stage where stem cells could be separated and cultured from it.
being considered involve the production of universal human Although the embryo would consist of only a few hundred cells,
donor cells. Scientists know how to isolate undifferentiated stem there would be many ethical issues raised by this technique.

Dr. David Wells, AgResearch

Dr. David Wells, AgResearch


Livestock are selected for cloning on the basis of Cloned embryos immediately prior to implantation The individuals produced by embryo splitting have

Cell Division
desirable qualities such as wool, meat, or milk into a surrogate. These are at the blastocyst stage the same characteristics as the parents.
productivity. (a mass of cells that have begun to differentiate).

The cloning process


Stages in Embryo Splitting (forming the twin) Artificial zona
begins here pellucida

Egg

Sperm
Zona pellucida: a
Zona pellucida
coating that promotes
normal cell division.

Egg cells are removed At the first stage of The zona pellucida is An artificial zona is The cells continue to divide,
from an animal and development, one of removed with an enzyme added, allowing forming genetically identical
fertilised in a petri dish. these fertilised eggs and the two cells are development to proceed. embryos. These are implanted
divides in two. separated. into surrogates.

1. With respect to animals, explain what is meant by cloning:

2. Briefly describe the possible benefits to be gained from cloning the following:

(a) Stem cells for medical use:

(b) High milk yielding cows:

3. Suggest one reason why it would be undesirable to produce all livestock using embryo splitting:

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Cloning by Nuclear Transfer
Clones are genetically identical individuals produced from one techniques (below). In animal reproductive technology, cloning
parent. Cloning is not new; it has been used in plant breeding has facilitated the rapid production of genetically superior stock.
for years. The early methods of animal cloning involved embryo These animals may then be dispersed among commercial
splitting; a process that mimics the natural process of producing herds. The primary focus of the new cloning technologies is to
identical twins. In recent years clones have been produced from provide an economically viable way to rapidly produce transgenic
both embryonic and non-embryonic cells using nuclear transfer animals with very precise genetic modifications.

Creating Dolly Using Nuclear Transfer

PHOTO: Courtesy Roslin Institute ©


Dolly Dies
Dolly, the Finn Dorset lamb born at the Roslin Institute (near Edinburgh) in July
1996, was the first mammal to be cloned from non-embryonic cells. Nuclear Dolly the sheep was euthanased
transfer has been used successfully to clone cells from embryonic tissue, but (put to sleep) on February 14th,
Dolly was created from a fully differentiated udder cell from a six year old ewe. 2003 after examinations showed
she had developed progressive
This cell was made quiescent and then ‘tricked’ into re-entering an embryonic
lung disease. Dolly was six years
state. Dolly’s birth was a breakthrough, because it showed that the processes
old; half the normal life
leading to cell specialisation are not irreversible; even specialised cells can be
expectancy of sheep. Post
‘reprogrammed’ into an embryonic state. The steps involved in creating Dolly
mortem examinations showed that her demise was
are outlined below. While cloning seems relatively easy to achieve using this
due to a viral infection, not uncommon in older sheep,
method, Dolly’s early death (right) has raised concerns that the techniques
especially those housed inside. Despite the concerns
could have caused premature ageing. Although there is, as yet, no evidence of some scientists, there is no evidence that cloning
Cell Division

for this, the long term viability of animals cloned from non-embryonic cells has was a factor in Dolly contracting the disease.
still to be established.

Donor cells taken from udder: Cells from the udder of Unfertilised egg has nucleus removed: In preparation for
1 2
a Finn Dorset ewe were cultured in low nutrient medium the nuclear transfer, an unfertilised egg cell was taken from
for a week. The nutrient deprived cells stopped dividing, a Scottish blackface ewe. Using micromanipulation techniques,
switched off their active genes, and became dormant. the nucleus containing the DNA, was removed. This left a
recipient egg cell with no nucleus, but an intact cytoplasm and
the cellular machinery for producing an embryo.

Egg cell Blunt “holding


Nucleus is sucked pipette”
up micropipette

Donor cell micropipette


Nucleus
of egg cell
First
electric pulse

Finn
Dorset ewe Donor cell with
nucleus intact A time delay improves
Second
the process by allowing electric pulse
Cells are fused: The two cells (the 3 as yet unknown factors
dormant donor cell and the recipient egg in the cytoplasm to
cell) were placed next to each other and activate the chromatin.
a gentle electric pulse causes them to
fuse together (like soap bubbles). Fused cells
Egg cell without
nucleus
PHOTO: Courtesy Roslin Institute ©

Blackface ewe
Cell division is triggered: A second electric
4
pulse triggers cellular activity and cell division,
effectively jump-starting the cell into
production of an embryo. This reaction can
also be triggered by chemical means.

Dolly

After six days, the resulting


5
embryo was surgically implanted
into the uterus of the surrogate
mother; another Scottish
blackface ewe. Of the hundreds
Birth: After a gestation of 148 days, the pregnant blackface of reconstructed eggs, only 29
6 successfully formed embryos,
ewe gave birth to Dolly, the Finn Dorset lamb that is
genetically identical to the original donor. and only Dolly survived to birth.

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All photos: © Dr. David Wells, AgResearch


Cultured
donor cell Injection
Holding pipette needle

Zona
pellucida

Egg cell with nucleus


already removed

100 µm

Embryo micromanipulation laboratory in Hamilton, New Zealand. Such labs use A single cultured cell is injected underneath the zona pellucida (the outer
sophisticated equipment to manipulate ova (monitor’s image is enlarged, right). membrane) and positioned next to the egg cell (step 3 of diagram on the left).

Donor
cow
Lady

Cell Division
Elsie

10 cloned calves

Adult cloning heralds a new chapter in the breeding of livestock. Traditional breeding methods are slow, unpredictable, Lady is the last surviving cow of the rare
and suffer from a time delay in waiting to see what the phenotype is like before breeding the next generation. Adult Enderby Island (south of NZ) cattle breed.
cloning methods now allow a rapid spread of valuable livestock into commercial use among farmers. It will also Adult cloning was used to produce her genetic
allow the livestock industry to respond rapidly to market changes in the demand for certain traits in livestock products. duplicate, Elsie (born 31 July 1998). This result
In New Zealand, 10 healthy clones were produced from a single cow (the differences in coat colour patterns arise represents the first demonstration of the use
from the random migration of pigment cells in early embryonic development). of adult cloning in animal conservation.

1. Explain what is meant by adult cloning (as it relates to nuclear transfer techniques involving adult animals):

2. Explain how each of the following events is controlled in the nuclear transfer process:

(a) The switching off of all genes in the donor cell:

(b) The fusion (combining) of donor cell with enucleated egg cell:

(c) The activation of the cloned cell into producing an embryo:

3. Describe two potential applications of nuclear transfer technology for the cloning of animals:

(a)

(b)

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Plant Propagation
Large numbers of flowering plants are able to reproduce as when uniformity of plant performance is desirable (e.g. in fruit
asexually by vegetative propagation. Humans exploit this trees), to multiply sterile or seedless species, or to propagate
ability and many successful strains of crop plants are hardly species with flowers in which the stamens have changed to
ever grown from seed. Plants can spread quickly by vegetative petals and there is no pollen produced. In general, artificial
means when conditions are favorable. By doing this they do not propagation is a more efficient way to multiply certain kinds
need to produce flowers, pollen or seeds; processes with large of plants because it produces a larger plant faster than one
energy costs to the plant. Vegetative propagation by artificial raised from seed and it avoids seed dormancy. New varieties
means (opposite) results in genetically identical plants (clones) can be developed by grafting, which combines the favorable
year after year. Clones are produced for several reasons, such characteristics of two existing varieties.

Examples of Natural Vegetative Structures

Bulb Rhizome Stem tuber

Base of fleshy Creeping, underground


foliage leaf is a horizontal stem Developing
Outer scale containing stored food
food store tuber
leaves
Tuber: the tip of an
underground stem
swollen with food
Cell Division

Stem

Roots arise directly


Roots Roots grow out from the
from the stem
old tuber (parent plant)

Runners Suckers Root tuber


Remains of Remains of old
parent plant parent plant

New shoot

Suckers form
Runners spread in new plants
all directions giving
rise to new plants
Root tubers form when roots become swollen
with food. In spring, each bud uses the stored
food to produce a new plant.

Garlic propagates via a bulb. Axillary Ginger is a rhizome from which new Tussock grasses propagate from The roots growing out of this potato
buds (called cloves) form beside the growth will eventually lead to the horizontal stems that grow from buds will form new tubers and eventually
existing bulb. formation of independent plants. at the tip of the rhizomes. independent plants.

1. Explain what is meant by vegetative propagation:

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Artificial Vegetative Propagation

Cutting Grafting (see photo series below)


Cutting is a method of propagation where a vegetative structure Grafting is a procedure by which the structures of two or more
is removed from a parent plant and grown as a new individual. plants are joined. Typically a twig section (scion) from one plant
Cuttings are successfully used to propagate herbaceous plants, is joined to the shoot of another (the rootstock). Grafting is used
but can be used on woody plants with the use of hormones that for many fruit and landscape trees because it avoids juvenility,
promote root growth. and the special properties of the rootstock and the scion are
able to be incorporated in the same plant.

Scion

Root stock

1 A leaf and axial bud is cut 2 The cutting is placed in a 1 A scion is prepared by taking a 2 The graft is covered in wax to
from the parent stock growth medium containing cutting. The scion is then grafted prevent infection and held
rooting hormones. to another plant (root stock). together with twine or raffia.

Cell Division
Root stock

Scion
Scion

Incision into
parent plant

A scion is removed from the parent Scion being grafted onto the stem of The graft is sealed and covered to The graft is then labelled for future
plant prior to grafting. the root stock. prevent water loss and infection. reference and monitoring.

2. Explain how plants benefit by reproducing vegetatively:

3. Discuss how humans have benefited from the vegetative propagation of plants. Include reference to both biological and
economic benefits:

4. Distinguish between cutting and grafting and suggest when each might be used:

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Plant Tissue Culture
Plant tissue culture, or micropropagation, is a method used for possible because differentiated plant cells have the potential to give
cloning plants. It is used widely for the rapid multiplication of rise to all the cells of an adult plant. It has considerable advantages
commercially important plant species with superior genotypes, as over traditional methods of plant propagation (see table below), but it
well as in the recovery programmes for endangered plant species. is very labour intensive. In addition, the optimal conditions for growth
Plant productivity and quality may be rapidly improved, and resistance and regeneration must be determined and plants propagated in
to disease, pollutants, and insects increased. Continued culture of a this way may be genetically unstable or infertile, with chromosomes
limited number of cloned varieties leads to a change in the genetic structurally altered or in unusual numbers. The success of tissue
composition of the population (genetic variation is reduced). New culture is affected by factors such as selection of explant material,
genetic stock may be introduced into cloned lines periodically to the composition of the culturing media, plant hormone levels, lighting,
prevent this reduction in genetic diversity. Micro-propagation is and temperature.

Stock plants are kept as free from


1 Advantages of Tissue Culture
pests and pathogens as possible.
• Possible to create large numbers of clones from a single
seed or explant.

• Selection of desirable traits is possible directly from the


culturing setup (in vitro), decreasing the amount of space
required for field trials.

• Reproduction of plants is possible without having to wait


Explant (in this for the onset of seed production.
case an axial bud)
• Rapid propagation is possible for species that have long
Cell Division

generation times, low levels of seed production, or seeds


2 Small pieces are cut (excised) that do not readily germinate.
from the plant. These pieces,
• Enables the preservation of pollen and cell collections
called explants, may be stem
from which plants may be propagated (like a seed bank).
tissue with nodes, flower buds,
leaves or tiny sections of shoot • Allows the international exchange of sterilised plant
tip meristems. materials (eliminating the need for quarantine).

• Helps to eliminate plant diseases through careful stock


3 The surfaces of the explants selection and sterile techniques during propagation.
are sterilised using solutions
such as sodium hypochlorite. • Overcomes seasonal restrictions for germination.

• Enables cold storage of large numbers of viable plants


in a small space.

4 The explants are transferred


to a culture vessel under
7 New shoots that develop are removed from the
sterile conditions.
explant and placed on new culture medium. The
process is repeated every few weeks so that a
few plants can give rise to millions of plants.

5 Incubation of culture vessels:


Duration: 3-9 weeks
Temperature: 15-30°C
Light regime: 10-14 hours per day
NOTE: Different kinds of hormones
in culture media produce different
growth responses. By changing
the relative levels of several plant 6 An undifferentiated Growth medium: Contains nutrients
hormones, the formation of callus, mass of cells known and growth regulators (plant hormones
roots and shoots can be initiated. as a callus develops. such as auxins, gibberellins and
cytokinins) set in an agar gel.
8 Tissue culture plants must be
acclimatised in special
glasshouses before they can
be planted outside. 9 Plant cell culture: If the callus
is suspended in a liquid nutrient
medium and broken up
mechanically into individual cells
it forms a plant cell culture that
can be maintained indefinitely.

© Biozone International 1998-2004


Photocopying Prohibited
Code: A 2
161
Micropropagation of the Tasmanian blackwood tree (Acacia melanoxylon)

Photos: Barry O’Brien


Leaf buds

Culture medium
Callus

Greening and formation of leaf buds on a Normal shoots with juvenile leaves growing from a callus on media. Seedling with juvenile foliage 6 months
callus growing on culturing medium. They appear identical to those produced directly from seeds. after transfer to greenhouse.

Micropropagation is increasingly used in conjunction with genetic culture allows the multiple propagation of trees with desirable traits

Cell Division
engineering to propagate transgenic plants. Genetic engineering (e.g. uniform timber colour). Tissue culture could also facilitate
and micropropagation achieve similar results to conventional solutions to other problems that cannot be solved by forestry
selective breeding but more precisely, quickly, and independently management. When combined with genetic engineering
of growing season. The Tasmanian blackwood (above) provides (introduction of new genes into the plant) problems of pest and
a good example of a plant suited to this type of manipulation. It is herbicide susceptibility may be resolved. Genetic engineering may
a versatile hardwood tree now being extensively trialled in some also be used to introduce a gene for male sterility, thereby stopping
countries as a replacement for tropical hardwoods. The timber is pollen production. This would improve the efficiency of conventional
of high quality, but genetic variations between individual trees breeding programmes by preventing self-pollination of flowers (the
invariably lead to differences in timber quality and colour. Tissue manual removal of stamens is difficult and very labour intensive).
Information courtesy of Raewyn Poole, University of Waikato (Unpublished Msc. thesis).

1. Explain the general purpose of tissue culturing plants:

2. (a) Explain what a callus is:

(b) Explain how a callus may be stimulated to initiate root and shoot formation:

3. Discuss the advantages and disadvantages of micropropagation compared with traditional propagation methods:

4. Describe a potential problem with micropropagation in terms of long term ability to adapt to environmental changes:

© Biozone International 1998-2004


Photocopying Prohibited
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