STUDENT WRITE - UPS

o To investigate the behaviour of Daphnia pulex in

response to different light intensities and wavelengths

o An investigation to discover whether xylem ends in leaves

are always closely associated with stomata
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 Sr1
B iology I nvestigation Student’s report of an AS-level investigation in Biology
www.curriculumpress.co.uk (Assessor’s comments are inserted in the report in bracketed italics)
AIM: To investigate the behaviour of Daphnia pulex in response to different light intensities and
wavelengths.
Null hypotheses: 1. Daphnia shows no response to different light intensities.
2. Daphnia shows no response to different light wavelengths.
Introduction
Daphnia pulex is a crustacean that is very common in freshwater ponds. It is commonly known as a ‘water flea’. It grows
to a length of about 1 mm and shows a characteristic ‘dancing’ swimming movement. The animals jerk up and down in the
water, in an upright position, with the head either tilted forward or backward. Low power microscopical examination of
the animal reveals five pairs of locomotory limbs, a pumping heart, digestive canal and a prominent compound eye, which
appears as a large dark body in the head. The second antennae are very large and are the main organs of propulsion.

Drawing to show appearance of Daphnia under low power of the light microscope.

Second antenna

Carapace
Compound eye

Excretory organ

First antenna Heart

Limbs, used for swimming Digestive tract

Brood pouch
Setae on limbs, used for
filtering food
Developing embryos

Reduced abdomen

• The possession of an eye suggests that Daphnia is sensitive to light and may orientate itself to light. It may show
taxes or kineses in response to light.

• A kinesis is a random movement in which the rate of movement is related to the intensity of the stimulus, but not
to its direction. An orthokinesis involves changes in speed of movement and a klinokinesis involves changes in the
rate of turning. The lower the rate of turning the more likely it is that the animal will leave an unfavourable area.

• A taxis is a movement in response to the direction of a stimulus. Movements towards a stimulus are called ‘positive’.
Movements away from a stimulus are called ‘negative’.

• Thus it can be expected that if Daphnia is placed in a gradient of white light intensity, from dim to bright light, by
kinetic and taxic movements the Daphnia will find and congregate in its most favoured light intensity. Similarly, if
Daphnia is placed in a spectrum of light wavelengths, it should also find and congregate in its favourite light colour
by kinetic and taxic movements.

• The relative light intensity illuminating the Daphnia can be measured as 1/d2 where d is the distance of Daphnia
from the light source. (A reasonably comprehensive introduction)
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B iology I nvestigation Student’s report of an AS-level investigation in Biology
Apparatus
• Long, narrow aquarium tank • Pieces of cardboard, to act as light shields
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• 2 dm3 beaker • Metre rule

• Culture of Daphnia pulex • Pipette for transferring Daphnia. (The end opening
• Lamps containing 60W white light bulbs was wide enough to allow the Daphnia through).
• Prism • Stop watch

Method
The investigation was carried out at room temperature which was 18°C. A thermometer was kept in the water
containing the Daphnia and regular checks were made in case the water was heated by the light bulbs.

450 cm3 of tap water was placed in a 2 dm3 beaker and one Daphnia transferred from the culture into the tap water,
using the pipette. The Daphnia was left for 10 minutes to acclimatise to its new surroundings. A 60W lamp was placed
either side of the beaker at a distance of 2 metres away. The rest of the room was in darkness and cardboard shields
were used to direct the light onto the beaker without illumination from unwanted reflections. This is shown in the
diagrams below:

Side view of experimental set up

Top of cardboard cut away to

Position of observer’s eye
allow observation

Back side of cardboard shield

60W lamp Daphnia in beaker of water
(tunnel shaped)

Experimental set up from above

Top of cardboard cut away to

allow observation 60W lamp

60W lamp Daphnia in beaker of water

The Daphnia was observed for six 1 minute periods and the number of times it changed its direction of movement per
minute was recorded. A time of 1 minute was allowed to elapse between each 1 minute recording period. Each direction
change was recorded by a pencil dash on paper, thus - ///// ///// ///// /. Direction changes caused by the Daphnia
hitting the side of the beaker were not recorded. (It might be hard to see whether the turn was due to hitting the
glass or not)

This procedure was repeated with the lamps moved successively closer to the beaker, at 1.75m, 1.5m, 1.25m, 1.0m,
0.75m, 0.5m and 0.25m. (Possible heating effects?)
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B iology I nvestigation Student’s report of an AS-level investigation in Biology
The aquarium tank was then filled with tap water, the culture of Daphnia added and allowed to equilibrate for 10
minutes with the aquarium evenly lit by the normal laboratory lighting. Using the 60W lamps at different distances
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from the tank and pieces of cardboard to act as light shields, a gradient of light intensity was established along the
length of the aquarium. At the start of the investigation the laboratory lights were switched off so that the room was
dark and the 60W lamps switched on. This is shown in the diagram below:

Aquarium

60W lamps

Cardboard shields
Observer

Group of Daphnia

2 metres

The complete apparatus was covered over the top with a cardboard shield to prevent extraneous light reaching the tank.

The water in the aquarium was agitated gently to disperse the Daphnia throughout the gradient of light intensity. The
Daphnia were observed at 10 minute intervals for half an hour to see if they were tending to accumulate in an area of
particular light intensity.

The aquarium was then illuminated with a spectrum of light achieved by passing the white light from a 60W bulb
through a prism. The lamp and prism distances from the aquarium were adjusted so that the aquarium was lit at the
optimum light intensity found in the previous investigation (1.3 lux) and the visible spectrum covered almost the whole
length of the tank. (The ends of the tank would be illuminated by infra-red and ultra-violet wavelengths to which the
Daphnia may be sensitive). Pieces of cardboard were used as light shields to prevent the tank from being illuminated
by extraneous white light. This is shown in the diagram below:

Cardboard light shield

Violet

Group of Daphnia Prism

White light from

60W bulb

Observer

Aquarium

Red Cardboard light shield

Spectrum of light

The complete apparatus was covered over the top with a cardboard shield to prevent extraneous light reaching the tank.

The Daphnia were observed at 10 minute intervals for half an hour to see if they were tending to accumulate around
any particular light wavelength. (Would all wavelengths have the same light intensity? Use of a light meter?)
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B iology I nvestigation Student’s report of an AS-level investigation in Biology
Results
Table 1. Klinokinetic responses of Daphnia to different light intensities
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Test Distance of light bulb from Daphnia / metres

0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00
1 11 13 13 15 18 28 28 29

Number of turns
2 11 11 12 16 19 30 29 28

per minute
3 12 11 13 15 19 30 28 27
4 10 12 11 17 20 29 30 28
5 11 11 12 16 18 28 30 29
6 12 12 11 17 20 28 29 28
mean 11.17 11.67 12.00 16.00 19.00 28.83 29.00 28.17
Relative light 16.0 4.00 1.78 1.00 0.64 0.44 0.33 0.25
intensity/ 1/d2 (76.4 lux) (19.1 lux) (8.49 lux) (4.77 lux) (3.05 lux) (2.12 lux) (1.30 lux) (1.19 lux)

Table 2. Taxic response of Daphnia to light intensity

Time/mins 0 10 15 20 30
Comment about Daphnia evenly Few Daphnia Most Daphnia in Daphnia now Nearly all Daphnia
distribution distributed remaining at ends area illuminated crowded into area congregated in
throughout tank. of tank, tending by lamps between illuminated by area illuminated
to move towards 1.25m and 2.25m lamps between by lamp at 1.75m
centre. distance. 1.5m and 2.0m distance (1.30 lux)
distance.

Table 3. Taxic response of Daphnia to light wavelength.

Time/mins 0 10 15 20 30
Comment Daphnia evenly Few Daphnia Daphnia tending Nearly all the Nearly all the
about distributed remaining at red to collect towards Daphnia present Daphnia present
distribution throughout tank. illuminated area. blue illumination. in the blue-violet in the blue-violet
illumination. illumination.

Analysis of data
• The values of light intensity expressed in lux were calculated 30
and inserted in table 1. The formula for doing this was: 28
Mean number of turns per minute

Lux = Wattage of bulb 26

2
4 πd 24
where d was the distance of the lamp in metres from the Daphnia. 22
• The mean values for the number of turns per minute of Daphnia
20
in different light intensities were calculated and inserted in
table 1 above. (Why not measure directly with a light meter?) 18
• A graph was plotted of the mean number of turns of Daphnia 16
per minute against the light intensity expressed as ‘distance of 14
lamp from Daphnia/metres’. 12
• The Daphnia showed the greatest rate of turning (klinokinesis) in
10
the lower light intensities particularly when the lamp was at a
distance of 1.75 metres (giving a relative light intensity of 0.33
and an absolute light intensity of 1.30 lux). The rate of turning was
0 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00
similar when the illumination was varied between 0.25 and 0.44 lux. Distance of lamp from beaker/metres
• Daphnia showed the lowest rate of turning when the lamp was
0.75 metres away (giving a relative light intensity of 1.78 and an absolute light intensity of 8.49 lux). Increasing
the light intensity beyond this had little effect although the rate of turning decreased slightly.
• These results suggest that Daphnia prefers to be in low light intensities and tries to escape from high light
intensities by swimming in a straight line rather than frequently changing direction.
• The phototaxic response of Daphnia to a light intensity gradient resulted in the animals congregating in the relative
low light intensity where the lamp was 1.75 metres away from the tank, (a relative light intensity of 0.33 and an absolute
light intensity of 1.30 lux). It took 30 minutes before the majority of the Daphnia were in this area in the tank.
• The phototaxic response of Daphnia to light wavelength resulted in the animals congregating in the region of the
tank illuminated by the blue-violet portion (300 – 400 nm) of the spectrum. The response to light wavelength was
completed more quickly (20 minutes) than the response to light intensity (30 minutes).
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B iology I nvestigation Student’s report of an AS-level investigation in Biology
Conclusions
The proposed null hypotheses were not true. Daphnia did show responses to light intensity and to light wavelength.
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Daphnia showed phototaxism by moving towards low light intensities and towards light of shorter wavelength (blue-
violet). The congregation of Daphnia in low light intensities and in blue-violet light was achieved by klinokinetic
movements (which caused a slower rate of turning in disliked conditions, resulting in faster movement out of those
conditions).

Discussion
The movement of Daphnia towards its preferred light intensity and colour is a phototaxis and is enabled by klinokinetic
movements. When suffering ‘discomfort’ due to the ambient light intensity the rate of turning of Daphnia decreases
which means that the Daphnia stands a greater chance of leaving that light intensity. When in a comfortable light
intensity the Daphnia turns more frequently and so is more likely to remain in that intensity. This type of behaviour is
inherent, simple and automatic, and must have evolved because it has survival value for the animal. A possible advantage
is that the concentration of Daphnia within a particular region increases the chances of finding a mate and successfully
reproducing. Another advantage is that with many Daphnia within a small area, greater water turbulence is caused by
the jerky dancing movements and this could improve the dispersion of available food so that all animals receive a supply.
It would also tend to disperse carbon dioxide, excreted via the gills, into the water and perhaps improve oxygen
diffusion gradients to the gills. It may be that a higher density of plankton, particularly phytoplankton species such as
the photosensitive Euglena, concentrate in the light intensities preferred by the Daphnia. These organisms are used
by Daphnia as food and are collected by filter feeding. The concentration of Daphnia together can also increase
survival chances against predators such as sticklebacks and other fish. A Daphnia swimming alone is more likely to be
‘focussed on’ and eaten by a predator, but the predator is likely to be confused by the presence of many Daphnia moving
haphazardly and jerkily.

The responses of Daphnia to light must be due to automatic reflex action. The compound eye is the photoreceptor
which receives the stimulus and transduces it to nerve impulses. The limb muscles are the effectors and work to cause
the jerky turning movements when stimulated to do so by the nerve impulses received via the reflex arcs. Because
Daphnia turns less frequently in unfavourable light and more frequently in favourable light, inhibitory synapses must
be involved in unfavourable light, and excitatory synapses involved in favourable light. (Good discussion but no
indication of sources of information.)

Evaluation
In retrospect it was realised that the light intensity for each colour illuminating the aquarium may not have been the
same. This could have been checked using a light meter or a camera exposure meter. It may have been better to use
coloured celluloid in the light path rather than the prism. To ensure the same light intensity for every colour lamp
distances could have been adjusted, or extra celluloid pieces for particular colours inserted to increase the depth of
celluloid through which the light had to pass. (Problem recognised in retrospect and possible solution suggested).

Light bulbs could have produced a heating effect which may have influenced the distribution of Daphnia. It may have
been better to use fluorescent light tubes which do not produce much heat. The thermometer only checked the water
temperature in one part of the tank – other parts may have been at a slightly higher or lower temperature.

Errors in counting the number of turns made by the single Daphnia in the beaker could have been caused by confusion
as to whether turns made at the sides of the beaker were truly klinokinetic or caused because the Daphnia hit the
glass. Also some turns may have been missed due to blinking by the observer. However, these types of error were
probably standard in every count performed. The methods of assessing Daphnia’s preferences for particular light
intensities and wavelengths were valuable, but not suited to generating quantitative data, apart from suggesting that
the response to light wavelength was faster than the response to light intensity.

When plotting the graph, figures were rounded to the nearest number possible to plot. For example, 28.83 was rounded
to 30.0.

Bibliography
Barnes, R.D. (1974) Invertebrate Zoology. W B Saunders. ISBN 0-7216-1562-7.
Brum, G., McKane, L. and Karp, G. (1994) Biology: Exploring Life, 2nd ed. John Wiley & Sons Inc. ISBN 0-471-59806-2.
Kent, M. (2001) Advanced Biology. Oxford University Press. ISBN 0-19-914195-9.
Speed and Smith (1975) Investigations in Behaviour and Elementary Neurobiology. Macmillan Education Ltd. (no ISBN).
Internet sites: search for ‘taxes and kineses in Daphnia’.
(Some or all of these references should have ben referred to in the discussion.)
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B iology I nvestigation Student’s report of an AS-level investigation in Biology
Assessor’s Report
(Performance levels available, 0, 1, 2, and 3). Students should consult the specifications of the syllabus they are following for
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the Awarding Body specific requirements for assessment criteria and performance levels.

• Planning:
Although protocol notes of planning were not included, from the written introduction and method it appeared that the
investigation had been carefully considered. The methods of changing the independent variables relevant to the problem
under consideration could be followed and implemented with little further clarification of detail. Consideration was given to
controlling some of the other variables, such as temperature, which might have influenced the results. The method of
assessing Daphnia’s klinokinetic responses to changing light intensities generated useful quantitative data, but
quantitative data was not generated in the other parts of the investigation. Because of this, and because not all variables
were controlled, performance level 2 was awarded rather than performance level 3.

• Implementing:
The experiments were conducted safely, in a well organised way, with reasonable skill and, for the klinokinesis
investigation generated appropriate quantitative results of reasonable accuracy. The results for finding the preferred light
intensity and wavelength only generated qualitative results although they were valuable and seemed conclusive. All results
were displayed clearly in tables in organised fashion. The assessor felt that a performance level grading of between 2
and 3 was appropriate and awarded 3.

• Analysing, drawing conclusions, synthesising and evaluating:

The mean values were calculated correctly and an appropriate line graph plotted. All conventions for doing this were
correctly followed and no errors were apparent. The data allowed the candidate to draw appropriate conclusions about the
Daphnia’s behavioural responses to changes in illumination. The null hypotheses could clearly be rejected without the
need for statistical testing (which is generally a requirement for A2 investigations but not for AS investigations). The
candidate gave an excellent evaluation of the usefulness of the photo-responses of Daphnia to the animal and also made
a reasonable evaluation of the limitations of the method, recognised some of the shortcomings, and suggested possible
improvements. Performance level 3 was awarded.

• Retrieval of information and communication:

The candidate found relevant information from a good range of sources, although they did not specify sources in the
discussion, and used the information to good purpose in the introduction and evaluation. The report was well written,
logically developed, used good English and appropriate scientific terminology. Performance level 3 was awarded.
 Sr7
B iology I nvestigation Student’s report of an A2-level investigation in Biology
www.curriculumpress.co.uk (Assessor’s comments are inserted in the report in bracketed italics)
AIM: An investigation to discover whether xylem ends in leaves are always closely associated with stomata.
Null hypothesis: Xylem endings in leaves are not always closely associated with stomata.

Introduction
Water, supplied to the leaves via the xylem vessels, is lost through the stomata (as water vapour) by transpiration.
Transpiration is important to the plant because it draws water up to the leaves from the soil (also draws up salts and
has a cooling effect). It would probably make transpiration more efficient if the bundle ends containing the xylem
vessels were near to the stomata so that water (vapour) could escape directly. (No reference to disadvantages of
transpiration - dehydration/wilting - to reduce the chances of this it would be better if xylem ends were not close to
stomata).

In this investigation leaves will be decolorised in alcohol so that the green colour of chlorophyll does not obscure the
xylem vessels, stained in ammoniacal fuchsin to demonstrate the lignin in the bundle ends and then observed in surface
view under the low power of the microscope. The relationship of the bundle ends to the stomata should be visible.
(Which leaf surface? - Under surface in a dorsi-ventral leaf. What microscope power - x40 or x400? Transmitted or
reflected light or both?). Micrometry will be used to measure the distances of bundle ends from the nearest stoma.
(What distance between xylem ends and stomata will be considered to be ‘closely associated’?)

Apparatus and reagents

• Dicotyledonous leaves of lettuce. (Why chosen - thin, • Calibrated eyepiece micrometer. (How was it
not much waxy cuticle so easily decolorised?) calibrated and in what units?)
• Absolute alcohol (ethanol). • Glycerine (a temporary mountant). (Glycerine 50 cm3,
• Ammoniacal Fuchsin (5.0g basic fuchsin added to 100 water 50cm3, crystal of thymol).
cm3 of 0.88 specific gravity ammonium hydrate and • Small beakers.
filtered. This reagent should be straw coloured). It • Forceps for handling leaves.
stains lignified tissue red but leaves other tissues • Bunsen burner
clear and colourless. • Large beaker.
• Distilled water. • Tripod.
• Microscope. • Gauze.
• Slides and coverslips. • Boiling tube.

Method
The chlorophyll was removed from the leaves by immersing them in boiling water for 1 minute to kill them and then
immersing them in boiling alcohol for a few minutes, using the bunsen burner, tripod, gauze and large beaker, as a
waterbath, in which to heat the boiling tube containing the alcohol and leaves. Providing the leaves were not boiled too
violently the histology of the leaf was not destroyed. (Safety hazard - was the bunsen burner switched off before the
alcohol tube was put in the boiling water?).

The leaves were then washed in cold water and stained by immersing them in ammoniacal fuchsin in a small beaker for
10 minutes. The leaves were then washed in small beakers in three changes of cold water to remove excess stain. (How
long were the leaves left in each change of cold water?)

The leaves were then mounted under coverslips on microscope slides using glycerol as the temporary mountant. (Were
the leaves cut up into pieces small enough to accommodate under the coverslip and was the underside of the leaves
uppermost - to show the stomata?)

The leaves were then examined under the low power of the microscope and a low power map made to show the
distribution of stomata and xylem endings. (What area of the leaf was selected for mapping and why? Was it randomly
selected and if so did you use random numbers to obtain microscope stage co-ordinates (if a mechanical stage was
used). Was only one area of the leaf sampled?)

Measurements were then taken, using the eyepiece micrometer, of the distances between the xylem endings and the
nearby stomata. (How many xylem endings and stomata were measured in this way and were they all in the same leaf
area?).
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B iology I nvestigation Student’s report of an A2-level investigation in Biology
Results
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Tissue map drawing of underside of leaf to show relative positions of stomata and bundle ends.

Position of stomata
on lower epidermis

Branch of main vein

(shown by red
stained lignin in
xylem vessel walls)

Xylem endings

(What magnification was this and what were the dimensions of the leaf area shown? Was this drawn from a view
near the midrib or near the edge of the leaf?)

Table 1. Distances of stomata from xylem ends

Xylem ending Distances of nearest ten stomata/µm

1 3 3 4 6 7 9 10 12 12 13
2 3 4 5 7 8 10 11 12 14 15
3 2 3 3 5 9 11 12 13 13 14
4 1 2 2 4 5 7 8 10 11 12
5 2 2 3 3 6 8 9 12 12 14
6 3 3 5 6 8 9 9 12 14 15
7 1 2 3 4 6 8 10 11 12 14
8 2 2 5 6 8 10 11 11 12 12
9 3 3 4 6 7 9 10 10 13 14
10 4 5 7 7 9 10 11 11 12 15

(Were all these measurements made from the same leaf area? Were measurements rounded to the nearest whole
number?)
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B iology I nvestigation Student’s report of an A2-level investigation in Biology
Analysis of data
From microscopical observation and the tissue map there seems to be no clear association in the positions of xylem
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endings and stomata. Stomata do not appear to be specially grouped around xylem endings but appear to be distributed
evenly in the lower epidermis. (Was this the case over the whole leaf area?)

The mathematical data was sorted to show the number of stomata at each distance from the nearest stomata. This is
shown in Table 2.

Table 2. Number of stomata at particular distances from nearest xylem ending.

Distance from xylem 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
ending/µm
Number of stomata 2 8 12 6 6 6 6 6 7 8 8 12 4 6 3

The following histogram illustrates this data:

12 (Assessor’s comment - The
11 x-axis seems to have
10 category labels rather than
a scale and both axes need
9 labelling.)
8
7
6
5
4
3
2
1
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
To test whether there was any association between xylem bundle endings and stomatal position a Chi-squared (χ2) test
was used. If the null hypothesis was true then an equal number of stomata should be found at each distance measured
from the bundle ends. The contingency table was worked out on this basis. (Should the values for E have been rounded
up to 6.7?)

Table 3. χ2 contingency table.

Distance from xylem 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
ending/µm
Number of stomata 2 8 12 6 6 6 6 6 7 8 8 12 4 6 3
(Observed)
Number of stomata 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7
(Expected)
(This is not a contingency table, but goodness of fit, although this has not affected the calculations in this case.)

Table 4. Processed data for calculating χ2.

(O - E)2 22.09 1.69 28.09 0.49 0.49 0.49 0.49 0.49 0.09 1.69 1.69 28.09 7.29 0.49 13.69
2
(O - E) 3.297 0.252 4.193 0.073 0.073 0.073 0.073 0.073 0.013 0.252 0.252 4.193 1.088 0.073 2.043
E

(O - E)2
χ2 = χ2 = 16.63
E
Number of degrees of freedom = (2 - 1) X (15 - 1) = 14

At a 5% significance level and 14 degrees of freedom the critical value from the χ2 table is 23.685.

This means that the null hypothesis must be accepted because the statistical value calculated is less than the critical
value from the table. Thus there is no apparent association between the positions of xylem endings and stomata.
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B iology I nvestigation Student’s report of an A2-level investigation in Biology
Conclusions
There is no apparent association between the positions of xylem endings and stomata in the leaves of lettuce plants.
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Discussion
It was initially thought that bundle ends would be closely associated with stomata because this would increase the
efficiency of transpiration and so increase the efficiency of water and salt uptake and transport. Alternatively bundle
ends might be distant from stomata so that transpiration would not be too efficient, resulting in wilting, dehydration
and possible death of the plant.

The reality is (in lettuce leaves) that the stomata and xylem ends seem to be independently distributed. The water
from the xylem leaks over the surfaces of the mesophyll cells and is absorbed into the cytoplasm and vacuoles due to
the osmotic gradient. (Water potential gradients should be referred to rather than osmotic gradients.) Water
evaporates from the wet mesophyll surfaces into the intercellular air spaces and then diffuses along the concentration
gradient out of the leaf via the stomata. To make a supply of water equally available to all the mesophyll cells, the
xylem ends should be evenly distributed through the leaf and this would appear to be so from the data obtained. (but
apparently only one area of the leaf was assessed and it was not made clear which area this was.)

(Would it also be of advantage to the plant if the stomata were evenly distributed and could their distribution be
surmised from the data? No reference made to sources of information.)

Evaluation
Only one part of the leaf was sampled and it may not have been representative of the whole leaf. Further areas should
have been sampled, including areas near the midrib and near the leaf margins. It was not certain that the leaf was fully
grown - the distribution of stomata and xylem ends may alter due to growth changes.

Leaves of other species were not investigated, but should be.

(Was the decolorisation effective and did it damage the leaf histology in any way that interfered with the
observations? Was the staining technique adequate? How hard was it to see the stomata against the background of
the other leaf cells? Would it have been useful to check the overall stomatal distribution over the leaf surface -
perhaps by using the ‘nail varnish technique’? How accurate was the technique of micrometry?)

Bibliography
Ashby (1934). School Science Review. vol.XV. Murray.
Esau (1960). Anatomy of Seed Plants. John Wiley & Sons Inc. No ISBN.

Assessor’s Report
(Performance levels available, 0, 1, 2, and 3). Students should consult the specifications they are following for the Awarding
Body specific requirements for assessment criteria and performance levels.
• Planning:
Protocol notes of planning were not available but the consideration giving to planning could be inferred from the written
introduction and method. From the various omissions of detail indicated in the text above it was considered that planning
left something to be desired. The title provided an indication of the problem to be investigated but the introduction, although
relevant as far as it went, was superficial. The description of the method omitted various practical details. Thus these skills
were only awarded performance level 1. The method of measuring the data provided adequate data and was awarded
performance level 2.
• Implementation:
The experiment was carried out adequately but a safety hazard was not mentioned (although it was probably taken
account of). However, the leaf was not sampled over a range of areas. Because of these deficiencies performance level
1 was awarded.
• Analysing evidence and drawing conclusions:
The candidate made a reasonable drawing showing the distribution of xylem ends and stomata in one area of the leaf,
and commented on the distribution the drawing appeared to show. The candidate did a chi-squared test, which was
pertinent to the problem, and correctly deduced that the null hypothesis should be accepted. The candidate attempted to
relate the findings to the biological processes going on in the leaf but omitted to include all possible details and points
(indicated by the assessor in the text above). Performance level 2 was awarded.
• Evaluating errors and procedures:
This was superficial and did not recognise all the sources of error or limitations of the method. Some of these have been
indicated by the assessor in the text above. Performance level 1 was awarded.