Topic 4 – Plant Structure and Function, Biodiversity and Conservation

The Plant Cell

Plant cell wall - cell wall may have primary wall only or primary and secondary.

Composition of Primary Cell Wall:-

Primary cell walls are borne when new cell plates are formed during cytokinesis and they are continuously synthesized and re-modelled during cell
expansion. It is generally a thin, flexible and extensible layer formed while the cell is growing.
• Plant cells are connected to each other by middle lamella which is made up of pectin (glue).
• The cell wall has two parts in it that are matrix and cellulose fibres.
• Cellulose microfibrils are arranged vertically in the matrix of primary wall to provide flexibility.
• The matrixc that holds together the microfibrils is composed of short, branched polysaccharides known as hemicelluloses and pectins. These
short polysaccharides bind both to the surface of the cellulose and to each other, and hold the cellulose microfibrils together.
• These microfibrils are bundles of about 60–70 cellulose molecules. The microfibrils are arranged parallel and stuck together with a polysaccharide
matrix.
• Pectins are also an important component of the middle lamella – the region found between the cell walls of adjacent cells. The pectins act as
cement and hold the cells together.
• The arrangement of the cellulose microfibrils within a matrix of hemicelluloses and pectin makes the cell wall very strong
• There are some places where no cell walls are laid, these are known as plasmodesmata – these are fluid-filled cytoplasmic connections.
• The areas of the cell wall where plasmodesmata occur are known as pit region.

Pit region: At some places the cell wall is thin because only the first layer of cellulose is deposited. The result is a pit field in the cell wall.
Plasmodesmata – Areas where there is no cell wall and these are cytoplasmic connections between two adjacent cells and are often

located in pit field (region), aiding the movement of substances between cells.
Pit – a pit region becomes a pit when secondary wall is laid on primary wall. A cell with secondary cell wall is dead.

Composition of Secondary Cell Wall:- it is laid on top of primary wall

• It mainly consists of cellulose and lignin. The cellulose microfibrils are arranged in a criss-cross manner (Different angles or mesh of microfibrils
in secondary cell wall for strength) and lignin holds the microfibrils together so that microfibrils remain parallel.
• Once the secondary wall is laid, the cell dies off as lignin is impermeable to water.
• Secondary cell walls are produced after cessation of cell expansion.
• It is not found in all cell types. Some cells, such as the conducting cells in xylem (xylem vessel and sclerenchyma fibres), possess a secondary
wall containing lignin, which strengthens and waterproofs the wall.
• Secondary cell walls are typically deposited evenly on the inside of the primary walls except for the pit areas in which no secondary walls are
present. Pits are used for solute transport and communication between two neighbouring cells.

Lignin- it is a class of complex organic polymers that form key structural materials in the support tissues of vascular plants
and some algae. Lignins are particularly important in the formation of cell walls, especially in wood and bark, because they
lend rigidity, waterproofing and do not rot easily.
Note: Xylem vessels and Sclerenchyma cells have secondary cell wall that consists of lignin.

The Structure of Cellulose

• It is the main structural molecule of plant cell walls.
• It is a polymer of β- glucose monomers joined together in very long chains.
• A condensation reaction between the –OH group on the first carbon of one glucose and the –OH on the fourth carbon of the adjacent glucose
links the two glucose molecules. Each alternate glucose is inverted to allow the 1,4-glycosidic bond to form therefore, the molecule becomes
unbranched.
• -OH groups project outside in each cellulose molecule thus the cellulose molecules lie parallel, bound together by hydrogen bonds.
• Hydrogen bonding prevents water entering the molecule. Cellulose is therefore resistant to enzyme hydrolysis which makes it an excellent
structural polysaccharide. This increases the tensile strength.
• 60 to 70 cellulose molecules connected by hydrogen bonds form a microfibril, which further increases the tensile strength.
• Some bacteria have enzymes to break down cellulose.
• The products can be used as an energy source. E.g. Ruminants & Termites
Question – Describe the adaptations of cellulose molecules to carry out its functions efficiently.

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Compare the structures and functions of starch and cellulose.
 Cellulose
Insoluble polysaccharide formed from β –glucose monomers
1,4 glycosidic bonds form unbranched polysaccharide
Alternate β-glucose molecules are inverted through 180o
Hydrogen bonds form between -OH groups in neighbouring cellulose chains,
forming bundles called microfibrils
long several thousand (glucose) molecules
cellulose molecules are only of one type of chain
Starch
Insoluble polysaccharide formed from α-glucose monomers
Branched amylopectin with 1,4 and 1,6 glycosidic bonds and
helical amylose with 1,4 glycosidic bonds only
Glucose monomers are not inverted
Hydrogen bonds are present inside the molecule
Short several hundred glucose monomers
starch can be a mixture of two types of molecules

Functions of the Cell Wall

It gives shape to the cell It protects the cell from damage It allows the movement of substances into and out of cells

Plant organelles
Plastids – These are organelles with double membranes that can be divided into three:
Chloroplast Amyloplast Chromoplast
Amyloplast
• They are responsible for the synthesis and storage of starch granules, through the polymerization of glucose
• Amyloplasts also convert this starch back into sugar when the plant needs energy. Large numbers of amyloplasts can be found in fruit and in
underground storage tissues of some plants, such as in potato tubers.
• Amyloplasts are plastids, specifically leucoplasts. Plastids are a specialized class of cellular organelles that carry their own genome and are
believed to be descendants of cyanobacteria (blue-green algae) which formed a symbiotic relationship with the eukaryotic cell.
• Starch synthesis and storage also takes place in chloroplasts, a type of pigmented plastid involved in photosynthesis. Amyloplasts and
chloroplasts are closely related, and amyloplasts can turn into chloroplasts; this is for instance observed when potato tubers are exposed to light
and turn green
Amyloplast Chloroplast
• This organelle is formed from leucoplast. • Double membrane bound organelle
• Double membrane bound organelle • Many internal (thylakoid) membranes; some are arranged as stacks
• They do not have pigments called grana
• The main function of the amyloplast is to store starch. • Chlorophyll found on the thylakoid membranes
• Amyloplast stores starch in the form of amylopectin. • Fluid filled interior called the stroma that contains circular DNA, RNA,
Found in large numbers in areas of a plant that store starch, eg potato 70s ribosomes and enzymes
tubers • Starch grains may also be present in the stroma.
Function:-It makes food through Photosynthesis.
Compare the structure of a chloroplast and an amyloplast
Feature present Chloroplast Amyloplast
Double membrane
Starch
DNA
Ribosomes
Thylakoid

Vacuole
Structure:-Plant cells have permanent vacuoles which contain cell sap, surrounded by a single membrane called the tonoplast. It has water soluble
substances.
Function: -
• It collects wastes, salts and water.
• Regulates the process of absorption of water by osmosis
• Involved in cell turgidity
• It also stores enzymes that breakdown old organelles
Using ticks and crosses indicate which of the structures in Table 1 are found in each cell type.
Table 1
Structure Plant cell Animal cell
Chloroplast
Chromosome
Smooth endoplasmic reticulum
Amyloplast
Large central vacuole
Tonoplast
Cell wall
Nuclear membrane
Golgi apparatus
 Mitochondrion
Centriole

The Structure of a Plant Stem

On the outside of the bundle are sclerenchyma fibres. In a young dicotyledon, the vascular tissue is in bundles towards the outside of the stem. In
trees and shrubs these separate bundles merge to form a continuous ring as the plant ages. The xylem vessels carry water and inorganic ions up
through the stem. The phloem transports sugars made by photosynthesis in the leaves up and down the plant.
Functions of plant stem
• Gives support to the plant
• Holds the leaves in the best position for obtaining sunlight for photosynthesis
• Support the flowers in a way that maximizes the likelihood of pollination occurring
• Helps movement of materials about the plant
• Carry out a small amount of photosynthesis
There are three basic types of tissue found within plants. the general location of these three types: dermal tissue (epidermis), vascular tissue and
ground tissue.
Plant tissues can also be broadly classified into two: Meristematic and Permanent tissues
Parenchyma
 The cells of parenchyma are living.
 Parenchyma is made up of thin, uniformly thick wall formed
mainly of cellulose.
 The intercellular spaces are large.
 Parenchyma is present in all the organs of plants.
 It fills up the spaces between different tissues and thus acts
as a packing tissue, it also stores food.
 It can differentiate into collenchyma and sclerenchyma

Collenchyma

• The cells of collenchyma are living.

• The primary cell wall of this tissue cells is made up of cellulose and the walls (corners) of these cells show the thickening due to cellulose.
Such thickening is more prominent in the angular walls, which come in contact with other cells.
• Some have chloroplasts hence they photosynthesise
• There are less air spaces between cells
• This tissue renders elasticity and flexibility to the organs. Hence Collenchyma is found beneath the epidermis.
• Collenchyma is never found in the underground organs of the plants.
Parenchyma Collenchyma

Sclerenchyma
• They have primary and secondary cell walls
• The secondary wall is strongly thickened with lignin as a result the mature cells are dead and cells without protoplasts and the lumen is
very much reduced.
• They have simple pits in their walls.
• Sclerenchyma fibres and sclereids are examples.
• No air spaces between cells.
In woody plants, secondary growth of stems and roots occurs through the activity of two lateral meristems: the vascular cambium and the cork
cambium.

Xylem:- It is a vascular tissue which has most of its cells dead (almost 95% of the cells found are dead). It contains different kinds of cells, out of
which the vessel element is prominent.
Xylem Vessel Trachied Sclerenchyma fibre Parenchyma

 There are two types of xylem cells involved in the transport of water, tracheids and vessel elements.
 Both types of cell have cell walls thickened with lignin. Lignin strengthens the wall enabling it to provide support. It also waterproofs the cell so
that it can conduct water.
 However, the impermeable lignin isolates the protoplast and as a result, mature xylem cells are dead and hollow.
 Lignin is laid down in the wall in a variety of patterns. For example, the conducting units may show annular, spiral, pitted or reticulate
thickening.
 Vessels are arranged in chains. The end wall of each vessel element is perforated, often missing altogether. This arrangement of elongated cells
enables continuous flow of water from one element to the next in the series.
 Lateral movement of water is possible due to pits.

Functions of xylem:-
• It provides mechanical support to plant.
• It helps in the transportation of water and mineral ions within the plant.

Development of a xylem vessel

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Adaptations of xylem vessel:-

1. Long cylindrical hollow tube made of vessel elements therefore water and minerals can move easily.
2. Thick lignified wall to withstand high pressure therefore the plant is supported.
3. It has pits therefore water can move sideways
4. Cross walls are removed – This reduces obstruction for the movement of water
5. No cell contents – Less obstruction for water movement

Describe and explain how the structure of Sclerenchyma fibres and the structure of Xylem vessels are appropriate to their functions.
Sclerenchyma –
 • it has tough lignified cell wall – to aid in the mechanical support of the plant
Xylem –
• it has tough lignified cell wall – to aid in the mechanical support of the plant and also to withstand high pressure due to water movement
• no cross walls – no obstruction to moving water and minerals
• pits – lateral movement of water and minerals
Sclerenchyma fibre Xylem vessel
Short structures with tapered ends Long cylinders whose end walls have broken
Ends closed Ends open
Tough lignin present in walls Tough lignin present in walls
Gives mechanical support due to tough lignin Gives mechanical support due to tough lignin
No transport of materials Transport water and minerals
phloem
Phloem tissue transports the soluble products of photosynthesis, mainly as sucrose with some amino acids. It comprises two types of cell: sieve
tube elements and companion cells. These cells are well adapted for transporting sucrose and amino acids:
● mature sieve tube elements have many pores in their end walls (sieve plates), have lost most of their cytoplasm and what little cytoplasm is left
runs as strands from cell to cell via pores in the sieve plate
● mature companion cells retain their nucleus and have cytoplasm densely packed with organelles; plasmodesmata between the two cells enable
each companion cell to control the activities of its adjacent sieve tube element

Sieve tubes and sieve elements

• Phloem contains unique tube-like structures called sieve tubes. Unlike xylem vessels, sieve tubes are made of living cells.
• Figure above shows the structure of a sieve tube and its accompanying companion cells. A sieve tube is made up of many elongated sieve
elements (also known as sieve tube elements), joined end to end vertically to form a continuous tube.
• Each sieve element is a living cell. Like a ‘normal’ plant cell, a sieve element has a cellulose cell wall, a cell surface membrane and cytoplasm
containing endoplasmic reticulum and mitochondria. However, the amount of cytoplasm is very small and only forms a thin layer lining the inside
of the wall of the cell.
• There is no nucleus, nor are there any ribosomes. Perhaps the most striking feature of sieve elements is their end walls. Where the end walls of
two sieve elements meet, a sieve plate is formed. This is made up of the walls of both elements, perforated by large pores.
• These pores are easily visible with a good light microscope. In living phloem, the pores are open, presenting little barrier to the free flow of
liquids through them.

Companion cells
• Each sieve element has at least one companion cell lying close beside it.
• Companion cells have the structure of a ‘normal’ plant cell, with a cellulose cell wall, a cell surface membrane, cytoplasm, a small vacuole and a
nucleus.
• However, the number of mitochondria and ribosomes is rather larger than normal, and the cells are metabolically very active.
• Companion cells are very closely associated with their neighbouring sieve elements. In fact, they are regarded as a single functional unit.
Numerous plasmodesmata pass through their cell walls, making direct contact between the cytoplasm of the companion cell and that of the
sieve element.

The mass-flow hypothesis

Translocation occurs from a cell in which a solute is made, called a
source, to a cell in which it is stored or used, called a sink. Movement
from source to sink is explained by the mass-flow hypothesis, shown
in Figure

At the source
● Sucrose is produced in the mesophyll cells of the leaf during photosynthesis.
● Using active transport, these sugars are loaded into sieve tube elements near the source. This lowers the water potential of these sieve tube
elements (makes their water potential more negative), causing more water to enter the sieve tube elements by osmosis.
● Entry of water into the sieve tube elements increases the volume and, consequently, the hydrostatic pressure of their contents.
● Water that enters the sieve tube elements is replaced by water from xylem vessels near the mesophyll cells.

At the sink
● Sucrose is converted to starch, which is stored. Starch has no effect on the water potential of these cells.
● The removal of sucrose, however, raises the water potential of these cells (makes their water potential less negative), so that water moves out of
these cells by osmosis.
● Loss of water from the cells at the sink decreases the volume and, consequently, the hydrostatic pressure of their contents.
● Within sieve tube elements water and its dissolved sucrose moves down a pressure gradient from source to sink, i.e. mass flow occurs.

Evaluating the mass-flow hypothesis

As its name suggests, the mass-flow hypothesis has not yet been fully accepted by scientists. The results of experiments designed to test it have
shown the hypothesis has some strengths but also some weaknesses. Some Strengths of the mass-flow hypothesis:
● The contents of sieve tube elements (phloem sap) taken from a source have been found to have a higher concentration of sucrose than those
taken from a sink. This confirms that the water potential gradient could be formed.
● In infected plants, viruses can be seen to be transported in the phloem from well-illuminated leaves to roots but not from leaves that were in the
dark to roots. This suggests that translocation occurs only when sucrose is produced during photosynthesis.
● If a phloem tube element is punctured, such as by inserting a hypodermic needle, phloem sap oozes out, suggesting it is under pressure.
Some weaknesses of the mass-flow hypothesis:
● Organic solutes have been found to move around plants in different directions. If the mass-flow hypothesis is correct, they should all move down
a pressure gradient.
● Measurements show that translocation of sucrose and amino acids can occur at different rates within one phloem tube element. If the massflow
hypothesis is correct, all solutes ought to be translocated at the same rate.

Differences between the structure of xylem and that of phloem.

• Xylem cell walls contain cellulose and lignin but phloem cell walls contain cellulose but no lignin
• Xylem is hollow but phloem has cell contents
• Xylem has pits but phloem does not
• Xylem does not have companion cells but phloem has companion cells
• Xylem has no end walls but phloem tube has sieve plates

Strength of plant fibres

Plant fibres can be cellulose, sclerenchyma and xylem vessels. They are used for different purposes. Plant fibres can be used in these ways because
they are:
long and thin Flexible strong

How do we extract fibres from plants?

To obtain fibres we must take the plant apart. This can be done mechanically by pulling out the fibres or by digesting the surrounding tissue.
Fortunately for us, cellulose – and particularly cellulose combined with lignin – is very resistant to chemical and enzymic degradation, whilst the
polysaccharides that hold the fibres together can be dissolved away.
The more lignin there is present, the harder it is to separate fibres. So to produce fibre pulp from trees, caustic alkali (NaOH) is required. In some
traditional processes the stems are piled in heaps, allowing bacteria and fungi to do decomposition to remove soft parts that hold fibres together.
This process is called ‘retting’.
During soaking, bacteria and fungi break down the soft tissues of the stems leaving the cellulose intact. It is then relatively easy to remove the
cellulose-rich fibres.
Extracting fibres from mature nettle stems
Safety
▪ Wear eye protection and gloves when handling the unretted nettles to avoid being stung.
▪ Wash your hands after handling the soaked fibres.
▪ Wear gauntlet gloves when ‘harvesting’ the nettles.
▪ Take care when using a scalpel or razor blade
▪ Wear rubber gloves when immersing the nettles in water to avoid stings
Procedure for extracting fibres
▪ Remove the leaves and any flowers from stems of mature stinging nettles.
▪ Place the stems in a bowl/bucket of water so that they are completely submerged. The stems are soaked for at least a week; leave them
outdoors because they are very smelly.
▪ Remove the stems from the water. Wash the stems to remove the softened tissue and then dry the remaining fibres. The outside cuticle
and epidermal layer will rub away and the central pith will be left when you peel away the fibres.
▪ These ‘fibres’ are made up of the vascular tissue; they contain both the xylem vessels and the sclerenchyma fibres.
Strength - Strength can be defined as the maximum stress a material can withstand without failing (breaking).
Tensile strength - Tensile strength is the maximum stress caused by a pulling force that a material can withstand without failing. Tensile strength =
force/cross sectional area

Factors that affect tensile strength of a plant fibre

Cross sectional area Length of fibre Dryness Type of fibre
Compression strength - Compression strength is the maximum stress caused by a pushing force that a material can withstand without crushing.
Devise an experiment to test the strength of the fibres on different concentrations of NaOH
The diagram below shows the equipment used to find the force needed to break each fibre

It is possible that adding alkali will increase the tensile strength of fibres so that they can be used in new materials.
• Thirty fibres of same species, length and cross sectional are were soaked in distilled water and the breaking force of each fibre was measured
using a force meter in newtons. This is done by gradually increasing the force until the fibre breaks, and record the mass.
• The breaking force (N) was then converted into tensile strength (Pa) and recorded. Repeat the experiment with other fibres of the same species
and calculate the mean tensile strength – to increase reliability.
• This was repeated with fibres soaked in different concentrations of NaOH.
• Must make sure other variables are constant – temperature, size of each individual mass used.

Safety precautions: wear goggles to protect eyes and make sure the area where weights will fall is clear. Boots must be worn.

Plant mineral uptake

▪ Some of the epidermal cells in younger roots are modified in a way that increases the root surface area in contact with the soil and the soil
solution found in it. These cells have their outer wall extended to form a thin-walled tubular structure that penetrates the spaces between
adjacent soil particles.
▪ Such cells are called root hair cells. Root hairs have very thin, cellulose walls that are readily permeable to water and dissolved mineral ions. The
cell sap of root hairs and the water found between soil particles contain dissolved solutes, but the cells have a lower water potential than the
solution outside in the soil.
▪ The cell surface membrane, just inside the cell wall, is selectively permeable. Osmosis takes place and water passes down the water potential
gradient from the soil solution into the root hair cell vacuole. The cell surface membrane includes transporter proteins in its structure enabling,
at the same time, the uptake of ions by facilitated diffusion and, when the uptake is against a concentration gradient, active transport.

Element Obtained Functions Deficiency symptoms

N as NO3− or Constituent of proteins, nucleic acids, vitamins, some plant growth substances Stunted growth, older leaves turn
NH4+ and chlorophyll. yellow.
Mg Mg2+ Constituent of chlorophyll, as magnesium pectate in middle lamella of the cell Older leaves develop yellow areas
wall, activation of some plant enzymes and growth slows down
Ca Ca2+ as calcium pectate in middle lamella which holds plant cells together, it is also The growing points die back and
important in the permeability of the cell membrane the young leaves are crinkly.

Factors affecting the uptake of mineral ions

1. Availability of oxygen
2. pH
3. Temperature

Temperature. Absorption of mineral salt is affected by change in temperature. In general, an increase in

temperature results increase in the absorption of salts up to a certain optimum level. At very high
temperature the absorption is considerably inhibited. The inhibition might be due to denaturation of proteins
which are directly or indirectly involved in mineral salt absorption. The change in temperature also affects the
process of diffusion. The rate of diffusion depends upon the kinetic energy of diffusing molecules or ions
which, in turn, dependent upon temperature.
The pH of the soil affects the availability of inorganic ions to plants. The chart below shows the relative availability of inorganic ions needed by
plants, at different soil pH values. The thickness of each line shows the relative availability of each ion.
Investigating plant mineral deficiencies
Procedure

1. Using a measuring cylinder, fill a beaker with the ‘all nutrients present’ solution.
2. Collect seedlings of same species and age
3. Gently push the plantlet roots through the hole so it is in the solution below. Connect one seedling as shown in the above diagram.
4. Wrap the beakers in black paper or aluminium foil to exclude light from the solutions and to prevent growth of algae., place the beakers under a
light bank for five days
5. Keep the temperature, humidity and the concentrations of common ions constant in each beaker.
6. Measure the height initially and after five days to allow comparison of the treatments.
7. Repeat the experiment with one mineral deficient two times and calculate the mean increase in height.
8. Repeat the entire experiment with other solutions.

• Brophyllum plantlets are suggested for this investigation. This is because they are not relying on stored nutrients.
• Test tubes or boiling tubes can be used but smaller tubes work better.
• The tubes should be wrapped in aluminium foil to exclude light from the solutions and to prevent growth of algae.

Using plant starch and fibres

Food for Thought:
• People have always exploited plants to provide material for building, clothing, medicines, food and drinks, dyes and for fuel.
• Plants are central to the human diet, they provide macro and micronutrients also contain fibres which helps the working of the gut.
• Some plants are grown as food staples – these are basic energy-supplying foods in the diet, containing many amyloplasts (store starch). Many seeds contain very
rich stores of starch. These seeds, therefore, provide plenty of carbohydrates, some proteins and oils as well as small amounts of valuable
micronutrients
• We use other seeds, such as sunflowers, linseed, oil-seed rape and many nuts, for the oils they contain. Pulses (beans, peas, lentils) provide much of the protein requirement
for people who eat little or no meat.
• Fleshy, succulent fruits are important as sources of sugar and vitamins.
Plant fibres:
• Plant fibres have been used for years to make rope, paper and cloth.
• these fibres are very long sclerenchyma cells and xylem tissue which are very tough.
• Cellulose is not easily broken down by enzymes or chemicals. However, the matrix of pectates and other compounds around the fibres (including lignin) can
usually be dissolved or removed.
• Plant fibres have great tensile strength; they cannot be easily broken by pulling. This, along with their flexibility, makes them very useful.
• Usually occur in bundles of fibres which are much stronger than the individual cells. More comfortable to wear as they are more absorbent
• The cellulose fibres make the wood very resistant to compression so it is excellent for weight-bearing in buildings.-Wood also retains some of the flexibility of the matrix and
doesn’t tend to crack.
• Wood is also a good insulator, homes built from wood need less heating in the winter than a brick house.
• It is a sustainable resource and Carbon neutral - taking in carbon as it grows and releasing it when it is burnt.

Synthetic fibres
They are made from chemicals derived from a non-sustainable resource which gets increasingly expensive and is rapidly being used up. However, they cannot soak up
body fluids, such as sweat and are non-biodegradable.

Plastics:

• Plastics are synthetic polymers

• These are produced from oil-based chemicals and made up of repeating small units called monomers
• They are used to make a wide range of products however, now, modern materials are being developed from natural products as the environmental problems caused by
plastics are becoming increasingly obvious

• Most plastics are polymers made from petrochemicals originating from oil which is a non-renewable source.
• These are non-biodegradable which has led to plastic pollution on a grand scale.
• Some plastics may be melted down and recycled, but many cannot.

Biological Polymers (Bioplastics): are a form of plastics derived from renewable biomass sources, such as vegetable fats and oils, corn starch, pea
starch or microorganisms. Common plastics, such as fossil-fuel plastics, are derived from petroleum- these plastics rely more on scarce fossil fuels
and produce more greenhouse gas when incinerated. Some, but not all, bioplastics are designed to biodegrade. Biodegradable bioplastics can
break down in either anaerobic or aerobic environments, depending on how they are manufactured. Some common applications of bioplastics are
packaging materials, dining utensils, food packaging, and insulation.

Bioplastics have three large potential benefits

1. They are a sustainable resource – can be grown easily to supply the needs of the bioplastics industry.
2. They are biodegradable as they are based on biological molecules that can be broken down.
3. They are carbon neutral

The Facts - Plastics made from petrochemicals have extremely useful properties that aren’t easy to achieve in bioplastics. Bioplastics are much more expensive than oil-
based plastics. Tensile strength of oil based plastic is greater than bioplastic. There is also tension for the use of crops for food, biofuels or bioplastics.

Plant based medicines:

Over centuries of experimentation, people have found that chemicals produced by plants are also of great benefit in helping the human body fight discomfort or
disease.
The foxglove was known for centuries to have medicinal qualities and in particular was used to treat a condition known as dropsy. However, it was
not until a country doctor called William Withering published A Treatise on the Foxglove in 1775 that it became an accepted form of medicine.

Developing new drugs

The work of William Withering:
A patient came to him complaining of a serious heart condition. He had no effective treatment, so the patient went to see a local ‘wise woman’,
who used herbs to cure a number of conditions. He bought the recipe, seeing how it helped the patient. He discovered that foxgloves were the active ingredient
Withering had heard of the foxglove’s curative properties for dropsy, but his attention was focused when he met Mrs. Hutton, a ‘wise woman’ who
was showing signs of the disease. She assured him that she would be all right after she’d had a cup of her special tea. Imagine Withering’s surprise
when he visited her again and found that she had recovered.
Talking to Mrs Hutton, Withering found out that foxglove was amongst the twenty or so herbs used in the potion. He suspected that it was
something in the foxglove which was the active ingredient. Later Mrs. Hutton sold her recipe to Withering who began to investigate the plant
further. One of his first patients was a brewer suffering from swollen limbs and an irregular heartbeat. After a few doses of Withering’s ‘digitalis
soup’ he became healthy and his pulse became ‘more full and regular’. Unfortunately his next patient, an old woman, nearly died from the
treatment so Withering gave up his investigations.

Sequence of steps that Withering took when testing his drug

• Extracted the active ingredient from the drug
• He tested the drug on patients with symptoms of the disease.
• Recorded any side effects.
• Used standard procedure to discover the effective dosage; slowly increased dose until patients experienced diarrhoea and vomiting and
then reduced slowly.
• Recorded all results and published results.

Drug testing today – These days, medicines brought onto the market have undergone several years of research and development. The new medicine has to be:
1. Effective 4. Safe, non-toxic and without any unacceptable side-effects
2. Easily taken and removed from your body 5. Capable of being made in a large scale
3. Stable -able to be stored for some time and used under normal conditions 6. capable of making a profit

When scientists think they have a compound that might make a useful medicine, they patent it. This patent gives the inventor to be the only one to make and sell
their invention for the next 20 years. Potential substances are analysed and the active ingredient (the drug compound which may bring about a cure) is
identified and copied so that it can be manufactured synthetically. Slight variations of the chemical structure are made just in case they might have
a better effect.
Today a potential new drug must pass a series of tests if it is to be developed into a new product. A series of trials of the compound is required.
There are five stages in all.

Pre – clinical testing (non-human trial)– Animal studies are laboratory studies or isolated cells and tissue cultures assess toxicity, safety and side
effects.
 Before a drug can be tried on people, it needs a way of getting into them. At this stage, the potential drug will be tested on animals to find out how it works in a
whole organism.
 This will also show if the drug gets taken into cells, if it is changed chemically in the body and if it is excreted safely.
 Mammals are used for drug testing as they are as similar as possible to humans. The most widely used mammals are rats and mice. Some tests have to be carried on
both rodents and non-rodents. These tests are expensive and time consuming and centre of much ethical debate.
 Wherever possible, animals are replaced by tissue cultures and computer models. These tests are also refined to cause minimum distress. However, at this moment, the
information from computer modeling and from tests on cell or tissue cultures is not sufficient enough to test drugs safely on people without animal testing, so, the law
states that animal testing must be carried out at this stage.
 Many people have ethical objections against this but the use of rodents is perceived as much less emotive.
 If the animal testing is successful, the first human trials follow. Before trying the drug on people, you have to apply for a clinical trial authorization
with the “Medicines and Healthcare products Regulatory Agency (MHRA)”.
 They take decisions about the testing and licensing of new medicines.

Clinical trials- In the development of new drugs, three-phased testing will only take place on those drugs that pass the pre-clinical testing stage.
a. Phase 1 –
• A small group of volunteers are told about the drug and given range of doses to determine safe dose
• The trial confirms whether or not the compound is being absorbed, distributed, metabolized and excreted by the body in the way predicted
by the laboratory tests
• The effects of different doses are monitored and also to check the safety of drug.
• Scientists continue to look for longer-term effects in previous animal testing.
• If the drug is successful, it moves onto phase 2.
• Here an independent body of scientists decides whether work can progress to phase 2.

b. Phase 2 –
• Trialinwhicha new drug is given to a small group of volunteer patients affected by the condition, the drug is designed to treat between 100-300 patients.
• Doctors see how the new medicine affects the disease in real patient. Volunteers are monitored closely to find out more about the ideal dose, the effectiveness
of the drug, safety on patients and any side-effects
• If the results are promising, Phase (iii) trials are set up.
• Success at this stage means the compound has a good chance of becoming a useful medicine

c. Phase 3 –
• The testing is to find out if drug is effective; a large group of patients (1000 – 3000 people) are selected and divided randomly into two groups.
• One is given the compound being investigated.
• The second is given an inactive dummy compound known as a placebo, this removes bias
• It is important that neither the patients nor the doctors know who is having the compound under investigation and who is having the placebo
or standard treatment. This is known as a double blind randomized controlled trial
• Number of patients involved is large, so there is a better chance of any unexpected adverse side-effects to show up (The steps also look for any
adverse reactions in the patients).
• The way is now open to license the compound as a drug after which it can be marked.

After licensing –
Trials continue to collect data in the effectiveness and safety of a new drug after the drug has been licensed.
Note:
Phase 2 and 3 trials are normally carried out as double-blind trials (neither patient nor doctor know whether the drug given is the new medicine, a control
medicine or a placebo).-
Placebos are used because patients often appear to respond to a treatment they believe will do them good, this is known as the placebo effect
It is difficult to achieve a complete set of results in clinical trials because many patients stop taking the medicine for various reasons or do not take it regularly.-The number of
patients enrolling in a trial is not necessarily the number of patients who complete it
In some trials, the new drug or drug combination is so successful that the trial is halted early.-Thebenefits of a medicine must always outweigh the risks
Questions:
1. Explain why the review team in the phase 1 trials needs to be independent.
It is difficult for those involved in the production tool the new compound to be completely objective about it.
2. Explain why it is sensible to test the drug on healthy volunteers in phase 1 before testing it on patients in phase 2.
If the drug is unsafe then less likely to cause serious harm as volunteers are well due to the effectiveness of body defense system
3. If phase 2 is a success, why is phase 3 needed?
It is more rigorous Larger sample size hence more reliable Double blind Use of statistics

4. Is it ethical to give some patients a placebo when it is known that the compound is likely to have a beneficial effect?
Yes, because it is not known for certain if the new compound is effective; patients freely consent to participate in trials.
5. Sometimes patients on a placebo will show an improvement in their condition. Suggest why this may be so?
The influence of mind over body; the patient may have been getting better anyway due either to chance or their own immune response
6. Suggest why a placebo is used as part of phase 3 of a drug trial protocol.
To compare the response of drug with placebo and the difference will the actual effect of the drug.
To get rid of non-clinical response.
List the similarities and differences between modern protocols and withering’s digitalis soup
Differences

Modern protocols Withering’s digitalis soup

• Animal testing No animal testing
• Tested on healthy people No healthy people were involved
• Double blind trial No Double blind trial
• Placebo used No placebo
• Controlled by regulatory authorities Not controlled
 • Large sample size Small sample
Similarities
• Both isolated a possible drug
• Both initially tested on a small group of patients and then a larger group of patients.

Antibacterial chemicals in plants

Plants are susceptible to infection by bacteria and fungi; they do everything to repel such attacks. Several plants are known to, or thought to,
destroy or inhibit the growth of certain bacteria. A plant with this property is known as antibacterial.
Preparing agar plates - Pouring agar plates

Safety - Aseptic techniques should be used throughout to avoid contamination. (updraught of a flame)

Procedure
▪ Collect a bottle or test tube containing 15 cm3 of sterile nutrient agar.
▪ Melt the agar by placing the bottle or tube in a hot water bath (agar melts at 97°C). If the bottle has a screw cap it should be loosened to allow
air to escape.
▪ Once all the agar has melted remove the bottle. You will need to use a cloth to do this. Allow the agar to cool to about 50°C, a temperature at
which you can handle the bottle. The agar will start to solidify at about 42°C. Take care not to let it cool too much or it will set as you pour it into
the Petri dish.
▪ Pipette 1 cm3 of bacterial broth into a sterile Petri dish using an aseptic technique. The lid of the Petri dish should only be lifted enough to allow
entry of the pipette. See Figure 2 below.
▪ Pour the 15 cm3 of molten agar into the Petri dish and replace the lid. Gently push the plate back and forth, N-S, NE-SW and NW-SE to mix the
bacteria with the agar and allow the agar to set.
▪ Please note: It is essential that the plates are used for the investigation an hour or so after the agar has set, otherwise once the bacteria have
started to grow they will be unaffected by the antimicrobial agent.

Inoculation of bacteria
Safety- Methylated spirits is toxic and highly flammable and because of the latter hazard should not be used while naked flames are in use – which
happens in the preparation and pouring of agar plates.
Use aseptic techniques. Do not open Petri dishes containing growing microorganisms.
Only bin used Petri dishes after they have been autoclaved.
Procedure
▪ Agar plates seeded with suitable bacteria need to be prepared. This may have been done for you in advance; if not, follow the instructions on
the sheet pouring agar plate.
▪ Obtain a plant extract by crushing 3 g of plant material with 10 cm3 of industrial methylated spirit and shake it from time to time for 10
minutes. The advantage of using methylated spirits instead of water is that it kills any bacteria that might otherwise contaminate the extract.
▪ Pipette 0.1 cm3 of extract onto a sterile discs cut from new filter paper using a hole punch can be used.
▪ Let the paper discs dry for 10 minutes on open sterile Petri dishes.
▪ Repeat steps 1 to 4 for other plants, making separate test discs for each extract.
▪ Use sterile forceps to place the test discs onto the bacterial plate together with the suitable control per plate. Three test discs and a control can
be placed on a single Petri dish. Ensure that you can distinguish between the different discs by marking the underside of the Petri dish.
▪ Close the Petri dish and tape it as shown in Figure below. Do not tape all round the dish because this can lead to

▪ The growth of anaerobic bacteria, some of which may be harmful.

Figure 1 A convenient way of taping a Petri dish without allowing anaerobic conditions to develop.
▪ Incubate the plates for 24 hours at 25 °C.
▪ Observe the plates without opening them. Bacterial growth on an agar plate looks cloudy. Make any appropriate measurements that will
enable you to compare the antibacterial properties of the different plant extracts.
▪ The simplest measurement would be to use a ruler and measure the diameter of the cleared area. It is straightforward to compare the results
of different treatments if the clear areas are perfect circles. If the diameter varies, one possibility is to measure at the widest point. For a more
precise measurement the area of the clear zone would have to be determined.
▪ Do not throw the plates in the bin. (The unopened Petri dishes will need to be autoclaved before disposal in the dustbin.)
▪ Wash your hands thoroughly with soap and water after completing the practical.