Bubbling Cabomba pondweed Teaching Notes Introduction Like many other water plants, Cabomba has specialised tissue called aerenchyma that allows gases to diffuse inside the plant. Aerenchyma provide buoyancy and allows oxygen transport to the submerged roots that may otherwise be in hypoxic (low oxygen) conditions. If you cut through the aerenchyma, oxygen can escape. This can be made use of this in the classroom to show how the rate of photosynthesis is affected by light intensity or by the colour of light reaching the plant. Don’t worry if you have tried to do his with other pondweed in the past - Cabomba is very reliable and bubbles freely for hours. This document will explain how students can investigate the rate of photosynthesis by monitoring oxygen generation in Cabomba. It includes notes on how to do this by: 1) 2) 3) 4) Counting Cabomba bubbles in white light Counting Cabomba bubbles in different wavelengths (colours) of light Collecting Cabomba bubbles in a pipette Collecting Cabomba bubbles in a microsyringe Demo video We have produced a demo video showing how to get the most out of this protocol in the lab, available at https://www.youtube.com/watch?v=eIEJ0FfB-VI Safety and biosecurity Take care with use of water near electrical equipment. Ensure hands are dry when using lamps Keep all containers of water away from sockets and from areas where spills could drip into sockets. Boiling tube racks could be placed into trays to catch any spills and prevent water running onto electrical equipment. NB Cabomba is not native to the UK and should be disposed of in domestic rubbish and not put into ponds or waterways. Apparatus Per group to set up a piece of bubbling Cabomba Approx 7cm long piece of Cabomba Boiling tube Boiling tube rack (type that allows light to illuminate whole tube) Lamp (see ‘lighting’ in the technical notes) Meter ruler / measuring tape Stop clock Copyright: Science & Plants for Schools: www.saps.org.uk Bubbling Cabomba pondweed - teaching notes (revised 2012) 1 1% sodium hydrogencarbonate solution (using sodium hydrogen carbonate solution will ensure that CO2 does not become a rate limiting factor although these plants will bubble well for some time in water). To save time, the Cabomba could be cut into 5-7cm long sprigs before the lesson, ensuring the sprigs are returned to a large beaker/trough of water as soon as they have been cut. Cutting the stems under water will reduce air traps forming in the plant tissues. Each sprig should have only one cut end and the rest of the sprig should have several healthy ‘leaves’. Students will use these sprigs upside down (i.e., with the cut end of stem at the top and the ‘leaves’ at the bottom) in their boiling tube to allow oxygen bubbles to leave the cut tissues. Basic method To set up a piece of bubbling Cabomba: 1. 2. 3. 4. Half fill a boiling tube with 1% sodium hydrogen carbonate (or water) Place a piece of Cabomba into the boiling tube (upside down – cut end up) Gently push the Cabomba down, taking care not to snap the stem Cabomba has rubbery leaves that grip the side of the boiling tube so it does not need weighing down 5. Place the boiling tube in a rack 6. Position a lamp to shine onto the plant 7. Bubbles of gas should be seen emerging from the cut end after a short time (one minute is usually long enough) If bubbles do not appear after 2 minutes, the cut end has probably started to ‘heal’ up, so it will need to be snipped off again to freshen it up Snipping the cut end under water to refresh the sample Copyright: Science & Plants for Schools: www.saps.org.uk Bubbling Cabomba pondweed - teaching notes (revised 2012) 2 Investigation 1: counting gas bubbles at different light intensities Photosynthesis is a chemical reaction. Students should already be aware from other science lessons that several external factors can affect the rate of a reaction. These are known as rate limiting factors and most GCSE/14-16 students need to be able to identify and discuss these factors. The easiest rate limiting factor to investigate using bubbling Cabomba is light intensity. Method Students will require the apparatus listed on page 1 and should set up their Cabomba stems in boiling tubes of water or 1% sodium hydrogen carbonate (to ensure that CO2 does not become a rate limiting factor). Students move their Cabomba set distances away from the lamp and count the number of bubbles that emerge from the cut end of the plant. They should find that more bubbles are generated in a set time when the Cabomba is closer to the lamp. Small intervals of 2-5cm work best when using 40W/60W desk lamps. Sample data set Lamp Distance from plant (cm) Number of bubbles per minute Test 1 Test 2 Average 0 2 4 6 8 10 105 95 88 74 66 51 112 91 80 70 56 45 This continuous data can be plotted on a line graph with a line/curve of best fit 108.5 93.0 84.0 72.0 61.0 48.0 Discussing the data It frequently occurs that the second reading in each repeat is lower than the first. Possible reasons that can be discussed The plant was still adjusting to the lower light intensity during the first reading The plant was still adjusting to the lower temperature caused by moving away from a filament lamp The cut end of the plant may be healing as time progresses There could be another rate limiting factor as time progresses Evaluating the investigation Students who have been well trained in fair testing can be very vocal about the shortcomings of this method, which can lead to effective ‘How Science Works’ discussions. Was it really possible to get the lamp at 0cm? How could you really make sure the plant had adjusted to the new light intensity each time? Were there some variables we could not control even though we should have? Did temperature play a large part in this investigation? If the lamp had produced no heat, how might this have changed the shape of the graph? Would energy saving bulbs reduce the heating effect? Were all the bubbles the same size? Why is it more sensible to compare the shapes of the graphs in the class rather than to compare the numbers of bubbles? Copyright: Science & Plants for Schools: www.saps.org.uk Bubbling Cabomba pondweed - teaching notes (revised 2012) 3 Investigation 2: counting oxygen bubbles at different wavelengths of light Photosynthesis is a chemical reaction. Students should already be aware from other science lessons that several external factors can affect the rate of a reaction. These are known as rate limiting factors and most GCSE/14-16 students need to be able to identify and discuss these factors. An interesting factor to investigate using bubbling Cabomba is how wavelength (colour) of light affects the rate of photosynthesis. Method Students will require the apparatus listed on page 1 and: Scissors Sticky tape Acetate film in clear / red / blue / green 1% sodium hydrogen carbonate (or water) Students set up their Cabomba stems in boiling tubes of 1% sodium hydrogen carbonate. Students then place their Cabomba a set distance (e.g. 5cm) away from the lamp. Starting with clear acetate, students wrap each of the different coloured acetates in turn around the boiling tube, and secure with sticky tape. With the sticky tape facing away from the lamp, they should then record the number of bubbles produced by the Cabomba in a set time (e.g. a minute) in each of the different light conditions. Students should find that green acetate causes photosynthesis to stop as this light is reflected by chlorophyll. Sample data Filter (acetate) colour Clear / none Red Blue Green Colour of light getting into the boiling tube All colours Red Blue Green Number of bubbles per minute Test 1 Test 2 Average 80 42 50 10 84 48 56 8 82 46 54 9 Discussing the data It may occur that the second reading in each repeat is higher than the first. Possible reasons: o The plant was warming up which increased photosynthesis It is usually the case that there is still some photosynthesis in green light Possible reasons: o The plant has not fully adjusted yet o Small amounts of red or blue light are getting through the acetate o The plant contains a photosynthetic pigment that can absorb green light (this is true of some algae, although these tend to be red algae) The difference between photosynthesis in red and blue light can also be discussed o Is the difference significant? How could this be tested further? o Can we link this potential difference to the environment the plant lives in? Copyright: Science & Plants for Schools: www.saps.org.uk Bubbling Cabomba pondweed - teaching notes (revised 2012) 4 Investigation 3: measuring the volume of oxygen with a pipette Photosynthesis is a chemical reaction. Students should be aware from previous science lessons that several external factors can affect the rate of a reaction. These are known as rate limiting factors and most GCSE/14-16 students need to be able to identify and discuss these factors. The easiest rate limiting factors to investigate using bubbling Cabomba is light intensity. Two methods are outlined that will allow students to determine the volume of oxygen produced in different light intensities. Investigation 3 uses disposable plastic pipettes and Investigation 4 uses microsyringes. This investigation allows students to measure the volume of gas produced during photosynthesis more accurately than by counting bubbles. In order to produce quantities of oxygen that can be measured, plants may have to be left for longer time periods e.g. an hour. Students may therefore have to set up several sprigs of Cabomba at different distances from the lamp Students will require all of the apparatus listed on page 1 and should set up their Cabomba stems in boiling tubes containing 1% sodium hydrogen carbonate (or water). They will also require: Scissors Blu-tackTM Small beakers or weighing boats Disposable plastic pipettes (3cm 3) (2) Students fill a plastic pipettes with 1% sodium hydrogen carbonate solution so that there is still air in the bulb of the pipette. With the tip pointing up so that the solution pours backwards into the bulb, they then dry the tip end of the pipette using a paper towel, and then seal the end using blu-tack. With the tip pointing down students then snip off the bulb of the pipette, catching the spilled liquid in a small beaker or weighing boat. The snipped pipette can be topped up with the spilled liquid and then inverted over a cut Cabomba stem in a boiling tube of 1% sodium hydrogen carbonate solution. If placed near a lamp, oxygen generated by the plant will build up in the pipette below the blu-tack seal and can be measured using the volumetric markings on the pipette. Students could set up several of these at different illumination levels to compare the volumes of oxygen produced. This could be organised as a class practical where results are collated to produce a class set. Tip of pipette sealed with Blu-tack. The pipette bulb has been cut off and the cut end placed over the cut Cabomba stem Suggested discussion points Plotting a graph of distance from lamp against volume of oxygen generated Plotting a graph of light intensity (1/(D2)) against volume of oxygen produced Fairness if collating class data (could control variables be controlled?) Fairness of the test if different sprigs had to be used for each distance How to test that the gas is oxygen Copyright: Science & Plants for Schools: www.saps.org.uk Bubbling Cabomba pondweed - teaching notes (revised 2012) 5 Investigation 4: measuring the volume of gas with a microsyringe Photosynthesis is a chemical reaction. Students should be aware from previous science lessons that several external factors can affect the rate of a reaction. These are known as rate limiting factors and most GCSE/14-16 students need to be able to identify and discuss these factors. The easiest rate limiting factor to investigate using bubbling Cabomba is light intensity. This investigation allows students to measure the volume of gas produced during photosynthesis more accurately than by counting bubbles. In order to produce quantities of oxygen that can be measured, plants may have to be left for longer time periods e.g. an hour. Students may therefore have to set up several sprigs of Cabomba at different distances from the lamp. Students will require the apparatus listed on page 1 together with: Blu-tackTM Microsyringes (2) and tips (1) Clothes peg (1) Beaker (1) Students seal off the end of a dry microsyringe with blu-tack. They then use another microsyringe fitted with a tip to fill the first microsyringe with 1% sodium hydrogen carbonate solution. This is then inverted over a cut Cabomba stem in a boiling tube of 1% sodium hydrogen carbonate solution. If placed near a lamp, gas generated by the plant will now build up in the microsyringe below the blu-tack seal and can be measured using the volumetric markings on the microsyringe. Students could set up several of these at different illumination levels to compare the volumes of gas produced. If required, the filled microsyringe can be balanced over a beaker using a peg while the Cabomba is set up if required. Filled microsyringe sealed with blu-tack and placed over the cut Cabomba stem to collect oxygen. Suggested discussion points Plotting a graph of distance from lamp against volume of oxygen generated Plotting a graph of light intensity (1/(D2)) against volume of oxygen produced Fairness if collating class data (could control variables be controlled?) Fairness of the test if different sprigs had to be used for each distance How to test that the gas is oxygen Copyright: Science & Plants for Schools: www.saps.org.uk Bubbling Cabomba pondweed - teaching notes (revised 2012) 6