Investigating transpiration in plants using a potometer

Technical & Teaching Notes

Introduction and context

The rate of transpiration in a plant varies depending on a range of conditions, including light intensity, relative
humidity, wind speeds and temperature, as well as between plant species. Various manipulations to the plant can also
affect transpiration rate.

In this practical, students use a potometer to measure the rate of transpiration.

There are two main types of potometer used in schools – the mass potometer, which measures transpiration through
the loss of mass, and the moving-bubble potometer, which measures water uptake by the shoot. (Note that water
uptake in a cut stem is likely to be greater than that in a rooted plant. This is due to the direct access to xylem vessels
in a cut shoot, rather than resistance to this across the apoplast in a rooted plant.)

This design allows for the possibility of combining both measurement techniques, by placing the potometer on a
balance, and recording both the change in mass and the volume of water taken up.

Although many metabolic processes (e.g. respiration and photosynthesis) also change water content and mass of the
plant, their effects are usually ignored when conducting potometer experiments. This is because the volume of water
involved in metabolic processes is insignificant compared to the large volume constantly flowing through the plant in
the transpiration stream.

Safety Notes

We suggest that a technician sets up the potometers in advance. However, students may be able to do some of the
setting up themselves, perhaps from step 8 onwards.

 Some plants exude sap which can irritate some people‟s skin (eg Euphorbia spp.). Care should be taken in
the selection of plants, and technicians may prefer to use gloves when assembling the potometer.
 Students should be careful when moving the potometer from one situation to another.
 Caution should be taken when preparing holes in rubber bungs (stoppers) - guidance on appropriate methods
for cutting holes and inserting tubing can be found on the CLEAPSS website (search for „making a hole
through a stopper‟).
 Caution should be taken when pushing the graduated pipette, the cutting and the syringe into the bung. We
advise using plastic graduated pipettes to reduce chances of breakages/cuts (see supplier details).

Apparatus

 Small glass jar – glass baby food jars or 60ml specimen jars may be found to be suitable for this. Shorter jars
make it easier to assemble to whole unit under water to eliminate air bubbles.
 Rubber bung (to fit jar) with two 4 mm holes and one 2mm hole

3
1 cm plastic graduated pipette

3 3
10 cm or 20 cm plastic syringe
 Blunt cork borer / drill
 Paper towel
 Sink
 Woody plant cuttings, at least 15-20cm long with stem diameter of approx. 5mm, e.g. Sycamore
 Bucket of water for collection of cuttings
 Scissors or secateurs

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Potometer and water transport in plants – Teaching and Technical notes: p. 1
Other apparatus for extension activities

 Top-pan balance accurate to 0.1g

 Light source / fan / plastic bags
 Petroleum jelly
 Vase for cut flowers
 Suggestions of other species of plant for comparison
o Rosemary, Lavender
o Sycamore, Beech or woody shrubs found in school grounds or local green spaces
o Cut flowers (e.g. rose)

Information about constructing the potometer

The apparatus should be prepared by the technician in advance, to prevent accidents.

It is crucial to avoid air bubbles in the assembled potometer. This is the most common reason for potometer
failure.

1. Cut two 4mm holes and one 2mm hole in the bung (see
safety note).
3
2. Take a 1 cm graduated pipette and carefully push it into
one of the larger holes in the bung. (If the hole needs to
be enlarged, temporarily insert a no. 3 cork borer into the
hole until the graduated pipette is fitted.)

3. Insert the syringe nozzle into the small hole of the bung.

4. Fill a sink or large deep plastic container with water.

5. Take a plant cutting at least 15-20 cm long. Immediately plunge the cut end of the stem in the sink of water
and remove the bottom 2-3 cm of stem with sharp scissors. Cut the stem at an angle to make it easier to push
into the remaining bung. Leave the cutting in the sink. Try to avoid getting the leaves of the shoot wet when
assembling the potometer in the sink, as this significantly slows down subsequent transpiration.

6. To temporarily open the remaining bung hole further,

carefully put the no.3 cork borer through the hole,
starting from the narrow end of the bung. Immerse the
bung in the water.

7. Keeping the bung immersed, gently insert the cutting into

the cork borer far enough so that the cutting goes all the
way through the bung. Carefully remove the cork borer.
The stem of the cutting should now fit tightly in the hole.

8. Immerse the bung and syringe, and small jar into the
water, making sure both are full of water.

9. Place the bung in the neck of the small jar.

10. Remove the entire apparatus from the sink and remove
surface moisture with a paper towel.

11. Push syringe plunger down until the water level is at zero
in the graduated pipette. The potometer is now ready for
use.

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Potometer and water transport in plants – Teaching and Technical notes: p. 2
Suppliers

Suitable glass jars (sold either as 60ml powder bottles or 60ml wide necked bottles) are
available from school suppliers for around 70p each. Size 39 rubber stoppers fit this size
jar, e.g: http://www.timstar.co.uk/bo02880-extra-wide-mouth-bottles-clear-glass.html
http://www.timstar.co.uk/st14605-s-rubber-stoppers-solid.html

Alternatively, glass baby food or similar jars are suitable for this purpose. Rubber stoppers
(bungs) are available from standard school suppliers, and are listed according to smallest
diameter (bottom) measurement, costing around £1.50 ea.

1ml plastic graduated pipettes can be purchased from standard school suppliers, but are
available to purchase in small quantities from King Scientific www.kingscientific.co.uk (see
http://www.kingscientific.co.uk/lab-equipment/graduated-pipette-1ml ) where 10 pipettes are
available for ~£3.60 inc. postage. Sold as sterile disposable units these pipettes can be
reused multiple times for this purpose (the cotton wool plug should be removed before use).

Standard 10 or 20ml plastic syringes can be used, also available from King Scientific in small
quantities (10 x 10ml syringes for ~£2).

Teaching Notes

General practical tips

 Getting the potometer to work – the two main reasons for the potometer not working are leaks on the
equipment or blockages in the plant. Putting the plant cuttings in water as soon as possible after cutting,
trimming the plant cutting under water once it is in the potometer and assembling the potometer underwater
should minimise the chance of air bubbles blocking the xylem vessels of the plant. In order to minimise the
chance of leaks the plant cutting, graduated pipette and syringe need to fit tightly into the bung. Assembling
under water ensures all surfaces are wet which helps produce a good seal. Petroleum jelly could be added
around the bung edge and where the syringe, graduated pipette and plant cutting insert into the bung to try to
address leaks that can‟t be located.
 Time frame to collect data – it should be possible to see a change in the water level in about 10mins under
normal classroom conditions and students could do this every 10mins for 40mins to produce a reliable figure
for the transpiration rate of their plant specimen.
 Varying abiotic factors – it is likely to be difficult to obtain a wide variety of different values for an abiotic factor
but it should be possible to compare a “high” and a “low” value for some. It would be good practice to measure
the value of these two so that they can be recorded alongside the data (e.g. use a luxmeter to measure the
light intensity).

3 3
When the meniscus in the graduated pipette or tube drops to about 0.7cm , it may be moved back to 0.0 cm
by gentle pressure on the syringe plunger.

Students may need to be taught or reminded about

 Reading the water level from the appropriate part of the meniscus

Notes on plant material choice

 Woody species are needed in order to get the stem through the bung and create a good seal.
 Rosemary or lavender potted plants from a supermarket or garden centre should provide enough material for
several potometers and may promote discussion of adaptations to reduce water loss and which habitats they
are naturally found in.

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Potometer and water transport in plants – Teaching and Technical notes: p. 3
  Trees, shrubs, or hedgerow plants (without spines) that are often available in school grounds/local green
spaces and can also be used (e.g. Laurel (Prunus laurocerasus)).

Notes on student tasks

 As a minimum students should measure the rate of transpiration for their plant specimen in a pre-prepared
potometer. All the students need to do to start the investigation is to use the syringe to return the water level in
the graduated pipette to zero (or just take an initial reading so long as the water level is high enough).
 This can probably be done about every 10mins for about 40mins. This gives them a relatively reliable value
for transpiration for their specimen and the class data can be used to provide repeats (from different
specimens of the same species). An easy way to standardise data between repeats would be to report
transpiration rate per leaf (assuming they are all about the same size).
 What else could they do:
2 2
o Report transpiration rate per m or per cm (this may be an opportunity to get students using standard
form),
o Use the potometer on a balance and compare measurements by volume to measurements by mass.
3
Assuming 1cm of water has a mass of 1g do these different ways of measuring agree? How do the
percentage errors for each way of measuring compare?
o Students could compare transpiration rates in two or more species (maybe one with some obvious
adaptations to reduce water loss and one that doesn‟t).
o Students could manipulate the plant to investigate where the majority of water is lost from (e.g. use 4
different set ups – petroleum jelly on both upper and lower surfaces, just on the upper surface, just on
the lower surface, no petroleum jelly).
o Students could manipulate an abiotic factor (light intensity – using lamps, wind speed – using a fan,
temperature – using a hair dryer with a hot and cool setting, humidity – by putting the plant in a plastic
bag).
o Students could investigate the proportion of water lost by transpiration and evaporation in a vase of
cut flowers using a balance to measure water loss.

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Potometer and water transport in plants – Teaching and Technical notes: p. 4
Possible questions for students
1. Follow the protocol and answer the following questions

Name the pierce of equipment used to measure transpiration rate in plants.

Explain how a potometer works?
Why might a potometer fail to work? How would you know? What would you do about it?
How do you measure the position of water in the graduated pipette accurately?
3 3 3
How accurate is your reading of water level (to the nearest 1cm , 0.1cm , 0.01cm or something different)?
What is the percentage error in the values you get for water lost in a particular time period?
How could you modify the experiment to reduce this percentage error?
What could you do, to continue the experiment, if the water level was dropping close to the level of the bung and
therefore in danger of going out of sight?
How do you calculate the rate of water loss in your plant specimen?
Would you expect to see a difference in the rate of water loss between students who had plant specimens of different
sizes? What could you measure and what could you calculate to take this into account?
What factors do you need to keep constant in order to obtain reliable measurements from your plant specimen?

2. The data from your investigation

How do you convert readings of where the water level is to volume of water lost in a particular time period?
How do you work out the rate of water loss over a particular time period?
If you plotted time since the start of the experiment on the x-axis and water level of the y-axis what shows you the rate
of water loss? If this was a straight line what would this tell you?
Why might some data points (maybe in different repeats) deviate from the line of best fit?
How would you present data from an experiment that investigated the effect of an abiotic factor?
How would you present data from an experiment that investigated the effect of a manipulation of the plant compared
to a control (or of two or more different species)?

3. Explore the application of your findings

Scaling your data up

 What calculation could you do to estimate how much water would be lost from your plant specimen over a
whole day (24hrs)? What value do you get? What assumptions does this make?
 Your plant specimen is only part of a plant. What could you measure about your plant specimen, and of a
whole plant in order to estimate the rate of water loss of a whole plant? (given data or a whole plant – do this
– what value do you get?) What assumptions does this make?
 How could you estimate the water loss due to transpiration of a whole field (let‟s say 100m x 100m) of your
plant.

Watering flowers
 Assume your plant specimen has the same transpiration rate as a single rose (i.e. stem, leaves and flower, as
part of a bunch of flowers). How much would you have to top up the vase of water by every day if you had a
bunch of 12 red roses to account for the loss due to transpiration?
 How else is water lost from this vase of flowers?
 Using Fick‟s law justify a design of vase that would minimise water loss in this way.
 Design (and conduct if possible) an experiment to estimate the relative importance of water loss by both these
means.

Farming
 Using your thoughts for “scaling your data up” and “watering flowers” write a set of bullet points explaining
why transpiration rates and other factors relating to water loss are important for farmers.
 Design an investigation for a farmer to understand how water is lost from their field and suggest what they
could then do with this information.

Flood protection
 How does the transpiration rate of plants link to flood risk?
 Design an investigation into whether replacing a front lawn with gravel will increase flood risk.
 Research online (or use data provided by your teacher) to provide an argument, based on data, for the
maintenance (or planting) of woodland as a way of reducing flood risk.

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Potometer and water transport in plants – Teaching and Technical notes: p. 5
 4. Answer these questions about the biology of transpiration

Water and plants

 What is water for in plants?
 Where (exactly) is water gained and lost in plants (and how is this different in your investigation)?
 What route does water take through a plant?
 State and explain the theory of how water moves through plants.
 What is transpiration?
 Why is water transported by mass flow?
 How is the plant adapted to take in and transport water?
 Apart from transpiration what else reduces the number of water molecules in a plant?
 Apart from transpiration what else makes a plant gain or lose mass?

Factors affecting the rate of water loss

 What factors affect the rate of water loss in plants?
 How do different factors affect water loss in plants?
 What might happen to rate of water loss if we measured it in a whole plant rather than a cutting? Why?
 What might happen to the rate of water loss if the leaves are covered in petroleum jelly? Would it matter if
only one side was covered and would it matter which side? Why?
 What adaptations do plants have to reduce water loss? Why do these plants have these adaptations whereas
other plants don‟t?
 What is Fick‟s law?
 Use Fick‟s law to explain how different adaptations reduce water loss in plants.
 Use your knowledge of adaptations to reduce water loss and or Fick‟s law to explain the differences between
data collected using a potometer with different species or under different abiotic conditions.

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Potometer and water transport in plants – Teaching and Technical notes: p. 6
Getting learning value from the practical

It is important to consider the purpose of practical work in Biology lessons and the following information provides
suggested areas where this practical can be used to help students develop their practical, mathematical and subject-
knowledge-based skills and understanding. The latter part of this section shows how this practical (or an extension of
it) can also be used to meet some of the A-level specification requirements for the Use of apparatus and techniques
as well as the Common practical assessment criteria (CPAC).

PRACTICAL SKILL DEVELOPMENT

You should be able to:
 take accurate readings from a graduated pipette to identify the height of the water in the pipette
 accurately measure change in volume of water in the pipette over a set time period
 record data in a suitable table
 identify whether the potometer is working effectively and making adjustments or replacing as necessary
 identify and control (or measure) factors that should be constant between repeats
 Suggest and conduct an appropriate number of repeats in order to get a reliable value for transpiration rate in the
specimen
 Suggest (and collate from the class) an appropriate number of repeats in order to get a reliable value for
transpiration rate under specific conditions in one species

EXTENSION TO PRACTICAL SKILL DEVELOPMENT

You should be able to:
 vary one abiotic factor in a standardised way in order for measurements to be taken at multiple values of this
abiotic factor
 suggest possible plant manipulation experiments to explore specific questions relating to transpiration in plants
 describe a suitable control experiment for your suggested plant manipulation experiment
 design (and conduct) an experiment to compare transpiration rates in two (or more) different species.
 Suggest a way to estimate the leaf surface area of the specimen
 Compare the pros and cons of measuring water loss by change in volume or change in mass

MATHS SKILLS DEVELOPMENT

You should be able to:
 perform calculations to identify the change in volume of water (water loss) over a set time period
 perform calculations to identify the rate of water loss over a set time period
 Calculate an average rate of water loss over a set time period
 Calculate the percentage error in the measurements taken

EXTENSION TO MATHS SKILL DEVELOPMENT

You should be able to:
 plot a line graph, scatter graph or bar chart to show the effect of an abiotic factor on transpiration rate
 plot a bar chart to compare water loss in different species or an experimental group with a control group
 Calculate the surface area of a leaf
 Calculate the transpiration rate per cm of leaf surface area
3

 Estimate the surface area of a tree (or other plant)

 Estimate total water loss of a tree over a whole day (or other plant)
 Identify the assumptions made when estimating the total water loss of a plant per day (or other plant)

ASSOCIATED SUBJECT KNOWLEDGE DEVELOPMENT

You should be able to:
1. Describe in outline the route water takes through a plant
2. Draw and annotate a diagram of a root cross-section showing the routes water can take from the soil into
xylem vessels
3. Describe the two routes water takes from the soil into xylem vessels
4. Describe the structure and location of the Casparian strip and explain its importance
5. Define transpiration

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Potometer and water transport in plants – Teaching and Technical notes: p. 7
6. Describe the route water takes from xylem vessels through a leaf
7. List 3 environmental factors that affect transpiration rate in plants and, for each, explain how they influence
transpiration rate
8. State the name of a piece of equipment used to measure transpiration rate in plants and describe how it is
used
9. List 3 adaptations some plant species have to reduce transpiration from leaves and, for each, use Fick‟s law
to explain how they reduce transpiration
10. Describe the cohesion-tension theory of water transport in plants
11. Define “Water Potential”, state the symbol used and the units, and use it to explain the movement of water
through plant tissue

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Potometer and water transport in plants – Teaching and Technical notes: p. 8
RECORDING EVIDENCE OF STUDENTS‟ WORK

Students can use the worksheet provided to produce evidence of their work in this practical that can contribute to their
practical skills accreditation.

MEETING ASPECTS OF THE USE OF APPARATUS AND TECHNIQUES

The full practical can allow students to meet various components of Appendix 5c of the GCE AS and A level subject
content for biology, chemistry, physics and psychology (DfE, April, 2014) in bold (with clarification where needed):

1. Use appropriate apparatus to record a range of quantitative measurements (to include mass, time,
volume, temperature, length and pH).
2. Use appropriate instrumentation to record quantitative measurements, such as a colorimeter or
potometer.
3. Use laboratory glassware apparatus for a variety of experimental techniques to include serial dilutions.
4. Use of light microscope at high power and low power, including use of a graticule.
5. Produce scientific drawing from observation with annotations.
6. Use qualitative reagents to identify biological molecules.
7. Separate biological compounds using thin layer/paper chromatography or electrophoresis.
8. Safely and ethically use organisms to measure:
a. Plant or animal responses
b. Physiological functions
9. Use microbiological aseptic techniques, including the use of agar plates and broth
10. Safely use instruments for dissection of an animal organ, or plant organ.
11. Use sampling techniques in fieldwork.
12. Use ICT such as computer modelling, or data logger to collect data, or use software to process data.
(tabulating results of water loos over time, doing calculations, presenting results graphically, statistical
analysis of results)

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Potometer and water transport in plants – Teaching and Technical notes: p. 9
MEETING ASPECTS OF THE COMMON PRACTICAL ASSESSMENT CRITERIA (CPAC)

Competency Practical Mastery Evidence from this practical / or

opportunity to train students in this
skill using this practical
Follows Correctly follows instructions to carry out the Observational evidence from the teacher.
written experimental techniques or procedures. Positive outcome for the student‟s
procedures photometer set up.
Being able to answer questions about the
procedure.
Applies Correctly uses appropriate instrumentation, Use of potometer and tools to manipulate
investigative apparatus and materials (including ICT) to carry abiotic factors or the plant in some way
approaches out investigative activities, experimental (and data/statistical package).
and methods techniques and procedures with minimal
when using assistance or prompting.
instruments Caries out techniques or procedures methodically, Follow procedure – identify whether there
and in sequence and in combination, identifying are any issues with the set up and adjust
equipment practical issues and making adjustments when accordingly (or just set up again).
necessary.
Identifies and controls significant quantitative If manipulating abiotic factors or the plant
variables where applicable, and plans approaches in some way there should be an emphasis
to take account of variables that cannot readily be on only changing one thing at a time and a
controlled. comparison using a suitable control.
Selects appropriate equipment and measurement Compare the measurement of volume of
strategies in order to ensure suitably accurate water lost by the potometer and the
results. measurement of mass of water lost using
a balance.
Safely uses a Identifies hazards and assesses risks associated Possible opportunities if the potometer is
range of with these hazards when carrying out set up by the student.
practical experimental techniques and procedures in the
equipment lab or field.
and materials
Uses appropriate safety equipment and Possible opportunities if the potometer is
approaches to minimise risks with minimal set up by the student.
prompting.
Identifies safety issues and makes adjustments
when necessary.
Makes and Makes accurate observations relevant to the Accurate readings taken from the
records experimental or investigative procedure. potometer.
observations
Obtains accurate, precise and sufficient data for Readings from the potometer taken over
experimental and investigative procedures and time, recorded and used appropriately.
record this methodically using appropriate units
and conventions.
Researches, Uses appropriate software and/or tools to process Graph drawing packages. Statistical
references data, carry out research and report findings. analysis packages. Presentation
and reports packages. Report writing packages (e.g.
Word). Internet search engines.
Sources of information are cited demonstrating Referencing in final report or presentation.
that research has taken place, supporting
planning and conclusions.

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Potometer and water transport in plants – Teaching and Technical notes: p. 10
Background information

The learning outcomes document for this practical summarises the key background information relating to the
transport of water through a plant.

Sample data

Photinia plant stem in potometer

set up.

The table below shows some data collected on a reasonably cool day, with the potometer (in the photograph above)
under normal classroom lighting condition

3
Time since start of Location of water Water loss / cm Average rate of water
experiment / min surface in graduated loss since start of the
3 3 -1
pipette / cm experiment / cm min
0 0.37 0.00 0.0000
15 0.41 0.04 0.0027
45 0.49 0.12 0.0027
75 0.58 0.21 0.0028

Further Investigations

As mentioned within this document there is scope to develop this practical in three directions:
 Exploring the link between different abiotic factors and transpiration rate
 Manipulations of plants with comparison to a control
 Comparisons between different species of plant

Reference

Burgess J.A. (1983) „An improved potometer‟, School Science Review , 64 (229) 699-701

The animation on the SAPS website (http://www.saps.org.uk/secondary/teaching-resources/1274) is directly relevant

to the study of this topic showing transport of water and sugar as well as growth of a stem.

Acknowledgements

Protocol and support materials re-developed by Dr Chris Graham, Long Road Sixth Form College, Cambridge and
Gail Webdell, Faculty of Education, University of Cambridge. Thanks to members of the SAPS Teachers and
Technicians Advisory Group for their input into a revision of the resource, and the science club at Didcot Girls School
for trialling the activity.
Science & Plants for Schools: www.saps.org.uk
Potometer and water transport in plants – Teaching and Technical notes: p. 11