OCR A Biology

22.1

Practical cloning
1 Use a non-flowering stem – all plant resources available for growing new roots (1); make an oblique
cut in stem – maximises surface area available for rooting powder/new root development (1); use
hormone rooting powder – scientists unsure whether effect is the hormone directly or anti-fungal
action but seems to increase success rate (1); reduce leaves to two or four – minimises loss of water
by transpiration whilst maintaining photosynthesis (1); keep cutting well-watered – reduces water
stress (1); covering with plastic bag – keeps air humid and reduces water loss by transpiration.
2 Cuttings are genetically identical (1); so differences in rate of growth, time to flowering etc. down to
differences in conditions they are given (1).

Summary questions
1 Organ which contains stored food from photosynthesis e.g., potato (1); cloning – new bud/plants
may arise from the organ identical to original plant (1); allows plant to survive adverse conditions (1);
and produce a new shoot (1); using energy from food store (1).
2 One mark for each advantage or disadvantage, max 4. Must provide at least one advantage and
disadvantage. Cuttings – genetically identical to parent so likely to produce good crops (1); often
shorter time from planting to crop (1); reliable (1); don’t have to buy in (1); can use own plants (1).
Seeds – have genetic variation so more variability in quality of crop (1); but are more likely to
withstand disease of changes in circumstances (1); take time and right conditions to germinate and
grow to maturity (1); in some cases can collect seed and reuse for next planting but don’t always get
the same quality (1). Any other sensible suggestions.
3 Genetically identical because parts of the same plant (1); but eventually form will depend on
growing conditions – levels of light, water, temperature etc. (1); identical suggests appearance is the
same (1); cloned plants may not be identical because a mutation may take place in stem cells of
meristems (1); changing pattern of growth in the plant (1).

22.2

Yes, we have no bananas

1 All bananas are clones because they are sterile – so disease affected all of them (1).
2 Micropropagation allows disease-free Cavendish bananas to be produced in large numbers (1);
however, clones would still be all same strain so would be vulnerable to the same disease (1); but
micropropagation allows for mass production of genetically modified Cavendish plantlets immune to a
particular disease e.g., Black sigatoka (1). Banana explants could be used where parts of the plant
are transferred to a nutrient medium.
3 Any sensible points for example, from early humans taking plantlets that form at bottom of mutated
bananas (1); a technique used for centuries (1); culture methods remained same for a long time (1);
explanation of why asexual propagation needed (1); until introduction of micropropagation –
advantages it offers (1).

Summary questions
1 Variety of ways students might do this but answers must include: Tissue selection (1);  tissue
sterilisation (1);  tissue proliferation (callus formation) (1);  transfer of cells from callus to new
medium (1); for shoot and root development (plantlet formation) (1); plantlets potted into compost to
form small plants (1);  plants hardened off to be planted out and form mature plants (1).
2 Look for scientific explanation of whichever features chosen (6 marks max. 1 mark for each correct
feature, 2 marks for explanation of each feature). For example: Reliable way of increasing numbers of
endangered/rare plants. Will help towards maintaining biodiversity of ecosystems. Allows for rapid
production of large numbers of crops, very valuable for countries where there are food shortages.
3 a Advantages relatively easy (1); relatively cheap and readily available (1); history of use (1).
Disadvantages any disease in parent plant transferred with cutting etc. (1); limit to number of new

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 OCR A Biology

plants that can be formed so cannot keep up with demand if there is a major threat to crop (1); still
produces clones (1). Any other sensible suggestions. Max 6, one point for each argument. Student
must provide at least two arguments for and against.
b Advantages can produce disease-free plants (1); can produce plants engineered to be resistant to
disease (1); can produce almost limitless numbers of plants fast (1). Disadvantages relatively
expensive (1); needs some infrastructure (1); still produces clones (1). Any other sensible
suggestions. Max 6, one point for each argument. Must provide at least two arguments for and
against.

22.3

Cloning humans
1 DNA sequencing – if child is a clone of an adult they will have identical DNA apart from
mitochondrial DNA which will come from egg donor (1).
2 Cloning primates – even primitive primates is very difficult because spindle mechanism needed for
cell division is very close to nucleus in primate cells and so often destroyed or damaged when
nucleus is removed (1); and synchronisation between stage of embryo and state of maternal
reproductive organs has to be finely attuned (1); no DNA evidence of cloned humans (1); ethical
objections among many people, unlikely to get a team together to do the research etc (1). Any other
sensible points.

Summary questions
1 Natural twinning early embryo splits (1); and two fetuses go on to develop (1); from the two halves
of divided embryo (1). Artificial twinning split in early embryo is produced manually (1); number of
identical embryos may be replaced in surrogate mothers (1); to produce a number of identical high
quality animals (1).
2 Observing births and recoding twin births when animals appear the same (1); genetic testing of any
twin cattle of the same gender (1).
3a Both processes involve removing eggs from an animal (1); both involve surrogate parents (1); both
potentially produce a number of genetically identical organisms (1). Any other sensible suggestion (3
max).
b In twinning either gametes meet outside the body (1); and early embryo develops before being split
(1); or early embryos flushed from the mother (1); egg cell contributes all maternal DNA (1); embryos
produced from gametes (1); embryos genetically related to two parents (1).
4 Outline of process: Egg cells retrieved are enucleated so only contribute mitochondrial DNA (1);
nucleus comes from an adult (somatic) cell of the animal to be cloned, not from gametes (1); an
electric shock is needed to stimulate development (1); no male gamete involved (1); embryos
genetically related only to adult cell nucleus donor (1). Any other sensible points. Look for evidence of
research and understanding by students. Look for evaluation of the evidence – indications of time
involved and cost balanced against potential benefits etc. Max 6, max 2 for decision and max 4 for
description of process.

22.4

Extra microorganisms
1 Reliable source of chymosin not dependent on numbers of animals slaughtered for meat (1); pure
enzyme – no contamination with animal products therefore less allergy risk (1); vegetarians can eat
cheese without ethical problems (1). Any other sensible points.

Summary questions
1 Points can include: no welfare issues (1); big range of microorganisms available to suit different
reactions (1); can genetically manipulate microorganisms to carry out required synthesis reactions

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 OCR A Biology

e.g., human proteins (1); microorganisms have a very short life-cycle and rapid growth rate so can
produce large manufacturing volumes in short time space (1); nutrient requirements of
microorganisms often relatively cheap (1); can be genetically manipulated some microorganisms can
be modified to utilise materials which would otherwise be wasted (1); microorganisms produce own
enzymes so catalyse the reactions (1); processes using microorganisms use relatively low
temperatures and pressures (1). Any other sensible points, max 6.
2 Baking: Mixed with sugar and water (1); respires aerobically (1); carbon dioxide produced used to
make bread rise (1); yeast killed by heating as bread cooks (1); takes a couple of hours (1). Brewing:
mixed with malted barley and hot water (1); respiration (fermentation ) continues for days in anaerobic
conditions (1); ethanol produced as waste product (1); yeast eventually inhibited (not killed) by rising
pH (1); build-up of ethanol and lack of oxygen (1) (max 6, up to 3 marks for each process).
3a To destroy bacteria that would make it go bad rapidly (1); or cause diseases such as TB (1)
b The fat droplets are spread evenly through milk so cream doesn’t separate out (1); and creates a
uniform product (1).
c Cheese whole milk used (1); bacteria used to separate the curds from the whey (1); changing
texture, and bacteria ripen or mature the cheese in controlled slow reactions at low temperatures to
change taste (1); out-compete bacteria that would make the cheese go bad (1); takes weeks, months
or years (1); can last for years (1). Yoghurt skimmed milk powder added to milk to enrich it (1);
°
specific bacteria added and incubated at 45 C for 4–5 hours to produce extracellular polymers that
give the texture to yoghurt (1); lasts 2–3 weeks in a fridge (1) (1 mark per difference, 2 marks max).

22.5

Plants and bioremediation

1 No naturally occurring microorganisms that can accumulate heavy metals (1); interfere with
biological pathways, so genetic modification unable to find a way to use them as yet (1); plants have
cell walls, lignified areas, xylem etc that are not metabolically active or dead, where heavy metal ions
can be accumulated without doing any damage (1). Any other sensible point.

Summary questions
1 Fungi produce penicillin naturally (1); bacteria genetically engineered/modified to produce human
insulin (1).
2 Use of microorganisms or plants to break down pollutants and contaminants in soil or water (1);
often carried out on site (1); because area of contamination may be very large so not practical to
remove contaminated material/too expensive to remove contaminated material (1); organisms
involved in bioremediation grown and break down contaminants in situ (1); living organisms so they
grow and spread (1); may be harvested and contaminants retrieved (1). Any other sensible points.
3 Gene for making human insulin cut out of human genome (1); using restriction endonucleases (1);
 inserted into a bacterial plasmid using DNA ligase (1);  plasmid containing human insulin gene
inserted into bacteria through transformation e.g., electroporation (1);  bacteria grown in a fermenter
(1);  mixture removed and penicillin extracted in process called downstream processing (1).

22.6

Investigating factors which affect the growth of microorganisms – serial dilutions and bacterial
counting
3
a Allowing for the fact that only 0.1 cm is plated you can think of the effective dilutions as follows (if
3 –6 –7 –8 –5 –6 –7
you continue to work in cm ): A = 10 B = 10 C = 10 . Also credit A = 10 B = 10 C = 10 as
these are the absolute dilutions of samples.
–3 8
b A number of bacteria cm = 500 × 10 000 000 = 500 000 000 or 5.00 ×10
–3 8
B number of bacteria cm = 52 × 100 000 000 = 520 000 000 or 5.2 × 10
–3 8
C number of bacteria cm = 4 × 1 000 000 000 = 400 000 000 or 4 × 10
c Credit students choosing to take the mean of the results for A, B and C:
–3
(A + B + C) /3 = 4 733 333 333 or 4.73 × 108 bacteria cm in the original sample. However, also

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 OCR A Biology

credit students who rely on the lowest dilution which gives countable results as the most accurate
guide to the true number of bacteria in the original sample. This means, since the plates are not
8 8
shown, that either plate A (giving an answer of 5.0 ×10 ) or plate B (5.2 × 10 ) could be the best
–3
answer for the true mean number of bacteria cm . Plate A if the colonies are small enough that few
or none will have merged (causing undercounting) or plate B if merging of colonies in plate A gives a
risk of undercounting. Additional credit should be given to students who make a distinction between
the number of bacteria as stated in the question and the number of viable bacteria or colony forming
units.

Summary questions
1 Both provide nutrients, suitable pH, moisture etc (1); both need to be maintained at optimum
temperature for growth (1); both must be kept sterile until inoculated with microorganisms (1); both
can be shaken at intervals to aerate it (1); agar plates remain closed once made up (1); broth is mixed
with known volumes of culture medium (1); agar plates inoculated using sterile wire loop and culture
medium (1); numbers in broth counted using turbidity, serial dilutions, and microscope graticules (1);
numbers on agar calculated using colony counting (1) (6 max, must be at least two similarities and
differences).
2 In large closed culture nothing gets in or out (1); initially, growth can be at theoretical rate as no
factors are limiting (1); as culture continues, numbers increase, food and oxygen are used up and
waste products build up often affecting pH (1); microorganisms run out of food or oxygen, are
inactivated by pH changes affecting enzymes or poisoned by waste products (1); so whilst theoretical
growth curve is exponential, real growth curve reaches a peak, plateaus, and declines (1).
3 a Vinegar is ethanoic acid therefore has a low pH (1); and inhibits bacterial growth (1)
b As temperatures fall bacteria growth slows but does not stop (1); so in fridge bacteria grow slowly
and eventually destroy food (1).
c Reactions in bacteria and fungi that act as decomposers affected by temperature (1); in August
temperatures relatively high so decomposition reactions relatively fast. In December, the
temperatures are much cooler so slower reactions in decomposers and rotting slower (1).
4 Could be done in a number of ways (max 6) e.g., set up series of cultures identical in every way
EXCEPT for factor you are investigating e.g., temperature, presence of a nutrient or antibiotic etc 
culture for several days  for each culture carry out a serial dilution  plate each dilution, making
streaks on agar plate with sterile inoculating loop and label carefully to show which original culture it
has come from and what dilution factor is repeat for all original cultures  culture agar plates for 5
°
days at 20 C for each factor find plate with number of bacterial colonies that can be counted. Work
out original number of bacteria using dilution factor  compare results across different cultures to see
effect on bacterial growth of factor being investigated.

22.7
Summary questions
1 Continuous processes run continuously once fermentation is started (1); sterile nutrient medium
added continuously once culture is growing exponentially (1); culture broth continually removed so
product can be processed and culture volume remains the same (1). Batch process everything added
at beginning in fixed volume of medium (1); nutrients used up and microorganisms, products, and
waste products build up (1); may be stationary phase when secondary metabolites formed, process
stopped, products extracted, reactor cleaned, and new process begun (1).
2 Any three from (max 2 per point): Temperature if temperature too low microorganisms will not grow
quickly enough, too high and enzymes will start to denature and microorganisms are inhibited or
destroyed. Bioreactors often have a heating and/or a cooling system linked to temperature sensors
and a negative feedback system to maintain optimum conditions. Nutrients if microorganisms use up
food supply they will start to die off so need a mechanism to keep food supplied, nutrient medium can
be added in controlled amounts to broth when probes or sample tests indicate that levels are

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decreasing to be mixed in using stirrers/paddles as will not spread through fast enough by diffusion
alone. Oxygen if microorganisms use up oxygen they will start to die off so need a mechanism to
keep nutrient medium oxygenated, oxygen is bubbled through broth when probes or sample tests
indicate that levels are dropping to be mixed in using stirrers/paddles as will not spread through fast
enough by diffusion alone. pH if waste products of microorganisms e.g., carbon dioxide build up then
pH of mixture will decrease. Change in pH can affect enzyme action and stop growth, buffers are
added to mixture and stirred in or alkaline solution added to maintain optimum pH.
3a Penicillin (1)
b Reaction continues after the maximum cell mass has been reached (1) – when food products would
be harvested (1); secondary metabolite is labelled (1).
c Graph should show process stopping once maximum cell mass is reached (1); could show removal
of cell mass (1); and addition of more nutrient medium and repeat of the exponential growth stage (1).

22.8

Immobilised enzymes in medicine

1 Advantage enzyme specificity means can get a very reliable response to one specific molecule in
the blood or urine (1). Any other sensible suggestion. Possible disadvantage enzymes usually work
best at a very specific pH and pH of urine can vary considerably which might limit use of biosensors
for chemicals in urine. Any other sensible suggestion.
2 Immobilising enzymes makes them less exposed to environment changes (1); buffers to maintain
steady pH could be included in immobilising structure (1); enzymes could be encapsulated in
membranes which only allow text molecules to pass through. Any other sensible point linking
immobilisation to protection from pH or other changes.

Summary questions
1 a Enzymes attached to an inert support system (1); over which the substrate passes and is
converted to product (1).
b More efficient (1); more specific; can optimise conditions for specific enzyme (1); less downstream
processing (1).
c Can be reused (1); easily separated from reactants and products (1); more reliable as control over
process (1); greater temperature tolerance(1).
2 Surface immobilisation – absorption onto inorganic carriers (1); covalent or ionic bonding onto
inorganic carriers (1); entrapment in a matrix (1); entrapment in membrane bound microcapsules (1).
3a Enzymes are accessible to substrates (1); allow continuous production by a continuous flow of
medium over the enzyme (1); conditions can be very tightly controlled over the enzyme beds (1);
changes in pH and temperature have less effect (1); any other sensible suggestions (3 max).
b Immobilising an enzyme may affect its ability to catalyse a reaction (1); diffusion of substrate to and
from active site of enzyme can be inhibited (1); by immobilising matrix or capsule and so slow reaction
(1); in surface immobilisation enzymes may be lost from matrix relatively easily (1). Any other sensible
points (4 max).
4 Look for evidence of research and of student having identified a suitable reaction catalysed by
immobilised enzymes – either one mentioned in the text or something different. Whichever reaction is
chosen the student should: give the reaction chosen (1); give the enzyme involved (1); explain how
the enzyme is immobilised (1 or 2 – one for simple statement of method, 2 for more explanation);
discuss the importance of the process (1); give the advantages of using immobilised enzymes over
other ways of catalysing the process (1 or 2 one for simple statement, 2 for more reasons or deeper
discussion) (6 max).

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