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Lily Pads & Sunlight

By Bilal Hussain

Rationale: My interest in science has led me to study A Level Biology and during my studies I have been given the opportunity to investigate an Abiotic-biological relationship using scientific methods. A visit to Epping Forest was organized by the school specifically for this reason, and being of Bangladeshi heritage I was compelled to investigate the water lily, not only because I am fascinated by this flowers tendency to open in the day and close at night, but also because it is the national flower of Bangladesh. During tour of the ponds, I decided to investigate and compare he growth of water lilies between two different ponds and after a trial study recording the conditions of the two ponds, I realized there were too many variables between two ponds like the temperature of the water and the volume of water in the ponds and also the minerals in the water which may also affect the growth to make a fair comparison, so I needed an alternative to my initial plan. In one of the ponds ingeniously named Lily Pond where the abundance and diversity of lily pads was significant, I noticed that in one side of the pond the lily pads appeared to be slightly larger. My initial assumption was that the difference in age was the reason behind this, but I was told otherwise by the well informed staff at the site. After further deductions I spotted a difference between the microclimates of the same pond, it was amount of sunlight received throughout the day. One side of the pond was constantly in the shade because of a tree next to the pond; while the other side was further away from the tree, thus free from the shade and open to direct light throughout the day. Being well-versed in the concept of plant photosynthesis, it seemed legitimate to assume that the lack of light was behind the under development of the plants, as light is one of the main components needed for plant photosynthesis to occur, but I am obliged to confirm my theory. Aim: I have decided to study the relationship between the lily pad size and the light received. My experiment will nearly mimic an experiment performed in the journal Effects of Light and Nutrients on Leaf Size, CO2 Exchange, and Anatomy in Wild Strawberry[6] but my experiment will be restricted to light intensities only and performed on pond plants. Research: The water lily plant which I am focusing on is referred to as Nymphaea Odorata in the Epping Forest plant identification book; it is a stunning flowering plant that blossoms fragrant white flowers and boasts large round floating leaves. They appear in dense patches often in fresh water ponds and small lakes. In moderate quantities the fragrant water lily can also benefit the lake by providing shelter and habitat for fish and invertebrates and shade to cool the water. [1] This quote may lead you to believe that the water lily is a magnificent flower, but it has also been proven to be a nuisance, as large populations can have adverse effects to the aquatic ecosystem. In a journal I an experiment, similar to the one I intend to do, was performed on wild strawberries, the dependent variable he observed was the thickness of the leaves. As well as having the resources to measure CO2 exchange of the plants at certain conditions. He found Leaf size in these experiments was apparently determined by nutrient and carbon supply but I want to know if the light level will have an effect on the leaf. Being a plant the Water Lily is a Phototroph, which in the online biological dictionary states it is An organism, typically a plant, obtaining energy from sunlight as its source of energy to convert inorganic materials into organic materials for use in cellular functions [2] The biological dictionary describes a prototroph using photosynthesis to make food, the overall and most commonly known chemical equation is: 6CO2 + 6H20 C6H12O6 + 6O2 The significance of the products of photosynthesis is immense, and deserves much more credit than any one individual can give. The main purpose to the plant it to produce glucose, as oxygen is a by-product. The word equation for photosynthesis is: Carbon dioxide + Water Glucose + Oxygen

But this equation does not do justice to the complexity and magnificence of the process of photosynthesis. The process begins with the compilation of the components needed for the process. Water is absorbed by the roots by osmosis and transported via the xylem. Carbon dioxide is taken from the air by the leaves and well as light is absorbed by the leaf. The leaf is the most important organ of the plant for respiration majority of the processes of photosynthesis takes place here. But it doesnt happen like a chemical reaction that the equation may suggest, it happens in a series of steps, and has many intermediates which I will discuss. Eventually the products mentioned in the simple equation are produced. The whole process can be split into 2 parts, the reactions which require light are called light dependant reactions, and the reactions that do not require light are called light independent reactions. In the light dependant stage, light is used to split water by photolysis into hydrogen atoms, oxygen and excited electrons which in turn replaces the electrons which have moved to a lower energy state in photosystem I and II; this electron carrier chain is used to force hydrogen across a gradient, and the gradient is used to run ATP-Synthase to generate ATP by combining ADP and Phosphate groups. The production of ATP here is referred to as cyclic photophosphorylation. In another related process called non-cyclic photophosphorylation, NADP+ eventually accepts the electrons at the end of the chain and becomes NADPH. A byproduct of non-cyclic photophosphorylation is oxygen, which goes back to the atmosphere. In the light independent stage, otherwise known as the Calvin Cycle, the products from the light dependent stage and CO2 from the atmosphere are used in this process. Ribulose-1, 5bisphosphate, a 5 carbon molecule reacts with a CO2 molecule to make an unstable intermediate which eventually disassembles to make 2 molecules Glyceraldehyde 3phosphate, which is a 3 carbon molecule. The Calvin Cycle ultimately makes carbohydrate molecules to make glucose, sucrose or fructose. These products are then used as an energy source for growth, repair and reproduction. As mentioned earlier, all the reactions mentioned take place in the leaf. In a palisade cell the chloroplast an organelle. If we take a closer look at this particular organelle we will find the sub units of the organelle. These are starch molecules, granum and the stromal lamellae and the stroma.

Diagram 1: Chloroplast

Here is the diagram of the thylakoid membrane where PSI and PSII can be perceived as proteins. The light dependent reactions take place in the thylakoid membrane which is displayed in diagram 2. And the light independent reactions take place in the stroma.

Diagram 2: Thylakoid Membrane

Trials: The facilities existing in Epping Forest Field Centre gave me access to multiple ponds, all in a variety of conditions. This allowed me to conduct a pilot study to become more familiar with the ponds in Epping Forest. This also allowed me to acquaint myself with scientific methods of investigation. Before I had decided to focus on the Lily Pond, I operated trials to discover the conditions of all the ponds. In my first pilot study I used a sweep net on the surface and in the water of each pond approximately 20 times and recorded all the organisms I had caught categorizing them into subdivisions of the pond. . I also conducted observations of the conditions of the water like PH and temperature as well as light intensity. My results of all three ponds were as follows: Lily Pond is a medium sized pond as it is 120m2 and practically lily pad shaped. The dimensions of it are shown in the satellite picture named diagram 4. On one side of the pond there is a tree, which gives half of the pond constant shade. There are many plants present in this pond such as the Yellow Flag Iris and Nuttalls Pond Weed, but the most populous and dominant plant is the White Water Lily. Out of all ponds the lily pond was indeed the most diverse. There were a huge range of invertebrates and they were very common to the point of uncountable number of water mites almost every time the sweep net was used. Table 2 Organisms Pond Skater Water boatmen Mayfly Nymph Water Mite Table Hornet3 Temp Lux Midges 10C Larvae 60,910 Newt lux Fish Above 0 0 0 Surfac e 8 0 0 0 1 2 6 0 6 1 1 Lily Pond Open water Below Vegetatio n 0 1 5 >100 0 4 1 1 Sedimen t 0 2 3 >50 0 1 0 0

0 0 Lily Pond0 1 PH 0 Neutral 0 0 (7) 0 0 0

During my condition trials, I recorded the temperature and Lux 5 times in the shaded and 5 times in the non-shaded in 1 hour intervals and then found an average of the whole pond by adding them up and diving them by 10, I also used a PH test strip to measure the PH of the water by dipping the test strip into the water for about 5 seconds and then manually using a chart to work out the PH. The temperatures of all the ponds were taken by submerging an analogue thermometer into the water and reading the temperature.

Diagram 3: Google Maps, Satellite view of Paul's Nursery Rd, High Beech, Loughton IG10 4AF [3]

Table Oak Plain Pond 5 Tem Lux PH p 19 153,100 Neutral C lux (7) Table 4 Organisms Dragonfly Stonefly Newt 2 1 0 Above

Oak Plain Pond is a large pond stretching an area of 470m2 and is surrounded by open grassland, allowing the pond unobstructed daylight during the day. There are 2 patches of water lilies in this pond, but it mainly consists of Nuttalls Pond Weed and Floating Sweet-grasses. Oak Plain Pond Below Vegetation Sediment 0 0 0 0 1 0

Table 7 Tem p 9C

Frog Pond Lux 1,090 lux PH Neutral (7)

Frog pond is a relatively small pond being only 110 m2 and is completely packed with vegetation like Bur-reed, Yellow Flag Iris and Nuttalls Pond Weed. This pond constantly in the shade as it is surrounded by tall trees. When the sweep net was used the daphnia seemed to be exclusive to the pond, but after more sweeps near the middle of the pond other invertebrates were caught.

From the results drawn from my trials, it was obvious that a comparison of the ponds could not be made, as there were too many variables to consider. So I have decided to do a trial of my main experiment to gain a better understanding of this investigation, and then evaluate my current plan to ensure all the variable are taken into account. In lily pond where my trial will take place, I will select lily pads and measure the radius with a ruler; I then used the radius putting it into the formula to work out the area of a circle using the radius, because lily pads are roughly all circular. I then used a lux meter to take a Table 8 Pond 1 (Lily Pond) measurement of the light intensity at the location of the leaf, Lily Radi Area Lux to measure how much light was being received by the plant. Pads us 1 2 3 4 5 6 7 8 9 10 11.2 5 13.5 13.7 5 13 9.5 14.2 5 13.5 9 9 10.5 397. 6 572. 6 594 530. 9 283. 5 638 572. 6 254. 5 254. 5 346. 4 609 0 887 0 637 0 582 0 437 0 752 0 506 0 666 0 316 0 321 0 Here are my results:
700 600 500 400 300 200 100 0 0 2000 4000 6000 8000 10000
L h In sity ig t ten

Gra 1 ph

Area of L P s ily ad

Lily P ond

When looking at the results put on a scatter graph my data seems to be relatively conclusive as there is visible correlation, the light intensity increases the size of the lily pads measured increased. For example at around 3000 lux the areas of the lily pads were approximately 300cm2 and at 9000 lux the areas were around 590 cm2. I have also completed a similar trial to this, but on Oak Plain pond, to be used as a control. Oak plain ponds lily pads are consistently in the open so there are absolutely no restrictions of light, from this fact I expect that if the results were plotted on the same graph all the results will be at the top right hand corner. To do a true comparison I will plot both Graph 2 results in the same graph.
Areaof Lily Pa ds

Table 9 Lily Pads 1 2 3 4 5 6 7 8 9 10 is:

Pond 2 (Oak Plain Pond) Radi Lux Area us 14.5 660. 1920 2 0 12.7 510. 1740 5 4 0 14.2 637. 1450 5 6 0 13.7 593. 1530 5 7 0 15 706. 1300 5 0 12 452. 1580 16 0 16 803. 1230 84 0 13.5 572. 1540 3 0 13.5 572. 1210 3 0 12.5 156. 1430 3 0

1000 800 600 400 200 0 0 5000 10000 15000 20000 25000 Lig Intens ht ity

Lily Pond Oa Plain k

There seems to be indications of correlations but to be certain, I will need to use a statistical method to calculate the product-moment correlation coefficient. Although this is a relatively long mathematical calculation especially since I will be doing it twice, it is appropriate for my experimental design and will give me results which mathematically indicate the correlation, if any present. To work out R I need to find the sum of both X and Y and both sums of their squares. The formula for the product-moment correlation coefficient

r=

S xy S xx S yy
S xx = x
2

Where:

( x) 2
( y ) 2
n
n

S yy = y 2

S xy = xy

( x )( y )
n .

Table

Y (Lux)

X2

Y2

XY

10 660.2 510.4 637.6 593.7 706.5 452.16 803.84 572.3 572.3 156.3 Table 11 5665.3

(Area) 19200 17400 14500 15300 13000 15800 12300 15400 12100 14300 X (Area) Y (Lux) 149300 X2 3484619.5 81 435864.04 260508.16 406533.76 352479.69 499142.25 204448.6656 646158.7456 327527.29 327527.29 24429.69 Y2 22737300 00 368640000 302760000 210250000 234090000 169000000 249640000 151290000 237160000 146410000 204490000 XY 840748 10 12675840 8880960 9245200 9083610 9184500 7144128 9887232 8813420 6924830 2235090

I have worked out: Sxx= 275057.172 Syy= 44681000 Sxy= -508119 So for Oak Plain R= -0.145 Table 12 397.6 572.6 594 530.9 283.5 638 572.6 254.5 254.5 346.4 Table 13 4444.6 X (Area) Y (Lux) 6090 8870 6370 5820 4370 7520 5060 6660 3160 3210 X (Area) Y (Lux) 57130 X2 218546 7.8 X2 158085.76 327870.76 352836 281854.81 80372.25 407044 327870.76 64770.25 64770.25 119992.96 Y2 35611050 0 Y2 37088100 78676900 40576900 33872400 19096900 56550400 25603600 44355600 9985600 10304100 XY 269191 09 XY 2421384 5078962 3783780 3089838 1238895 4797760 2897356 1694970 804220 1111944

And for these results I have worked out: Sxx= 210021 Syy= 29726810 Sxy= 1527109.2 And for Lily Pond R= 0.611 These results show that there is a regular positive correlation of the lily pads in Lily pond. Correlation is portrayed by this method as a number between -1 and 1. The numbers -1 means perfect negative correlation, 0 being no correlation and 1 meaning perfect positive correlation.

There is a weak negative correlation of the lily pads in Oak plain, but that is insignificant as this will be the last time I experiment at this pond. This trial was initiated to be evaluated to find flaws and also to use those flaws to modify and to improve the main experiment. Hypothesis: My primary hypothesis is that in the side of the pond which is not constantly under the shade of the tree the plants access to light will not be limited and thus will have no restrictions in growth allowing the lily pads to grow to their maximum potential. On the other hand the water lily plants in the shade will be inhibited and therefore have smaller leaves. The null hypothesis (H0) is that there will be no significant difference between the plants on the side with shade and the plants on the side without. Main Plan & Equipment: I intend to measure the area of each leaf and then plot the figures against the light intensities I also measure, on a graph, thus allowing me to see a clear depiction if there is a significant difference between the plants on the side with shade and the plants on the side without. As lily pads are almost circles I will measure the diameter and half it to get the radius and use the formula Arena = r2 to find an approximate area. And then use a lux meter and record the light intensity that particular leaf receives. All the equipment I have and will use, are all in the following table: Table 1 Name 30m Tape Measure Equipment List Diagram Use Used to measure the length and width of Lily Pond.

Sweep Net

Used to catch aquatic animals and invertebrate. Used to accumulate the organisms caught by the sweep net

Trays and Pots

Lux meter

Used to measure light intensity

Thermometer

Used to measure temperature

PH Test Strips

Used to measure PH of the water Used to measure the diameter of the lily pads

Ruler

Waders

Used to protects and keep legs dry during going into the ponds

Clip Board

Used to lean on to draw around lily pads

Graph paper

Used to measure the area of the lily pads

First Aid Kit

Always carried around

Safety and risks: To properly measure the area of the lily pads, I was required to go into the water; the fear of drowning was always a concern as I do not know how to swim but I was told the water was fairly shallow. There were many safety issues I had to keep in mind, such as the slippery bottom of the ponds, due to the loose sediment at the bottom was a safety concern as well as many branches from trees and large rocks below the water. I used a metre stick to check for anything before stepping further into the pond. The ponds were at first glance free from rubbish, but there was no guarantee that visitors of the forest did not throw sharp objects into the water. To prevent any harm to myself I was required to wear very thick and heavy waders which protected my legs and also kept me dry. All students were required to tape any healing or open wounds secure during pond trials. It was easy to get lost as Epping Forest is a fairly large forest and also it is hard to make out direction especially when it is the first time visiting this location. To avoid any unnecessary concern all students had to sign out by writing when they were leaving and also when they expect to be back. The forest is a fairly dangerous place as there are lots of threats such as animals, some thorny plants, and even people as well as rubbish found on the ground, there was even a road nearby. Although there was no need as it was common sense we were told by the forest people to not talk to strangers and not to eat anything from the forest. We were also told not to stray from the group and especially not to cross a main road without being careful. All groups were required to carry first aid kits at all time. Whilst in the water the frightening sizes of the dragonflies were intimidating, but the staff of the forest guaranteed they were of no harm. Modification & Variables: One of the main criticisms of the plan, I have noticed reviewing my trial, is that there was absolutely no structured or random system used to pick the 10 lily pads used in this trial; I had picked a lily pad and then measured the radius with a ruler, then recorded the light intensity. This kind of unintentional subconscious bias may have falsely led to my conclusive data. There are two significant ways to overcome this kind of prejudice. I could either use a systematic sampling method or a random sampling method, but the best technique to use in this situation is a random sampling technique. This will be achieved by splitting the pond into sections, one side which is constantly in the shade and the other open, and using those sections I will record the light intensity every 45 minutes from 8:00 to 16:45 to get a clear picture of the difference in light intensity between the 2 sections. Then I will sub divide those sections into smaller sections to split up the water lily population, and from each of the three sections I will take 5 lily pads chosen by random. The lily pads will be chosen by using a numbered compass (as shown in diagram 2) at the centre of the sub-section and then using a

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Diagram 2: compass, edited replacing angles with numbers [4]

random number generator and using those numbers point out random lily pads on the compass. Another criticism of the trial is that my method of measuring leaf area by the circle formula. The formula is used for circle and although lily pads are circular, the method leaves room for inaccuracies. The most accurate way of measuring the leaves I can think of is by using graph paper (each square will be 1cm2) and counting the number of squares a leaf covers. Although this is a lengthy and monotonous process the areas found will be significantly more accurate than the older method. I will use a clip board to lean on and also to steady the graph paper to draw around the leaf. Majority of the variables will be under control during this assessment; I have been ensured by the staff at the centre, the PH of the water will remain the same, and well as the temperature of the water may vary throughout the day but will not vary from one side of the pond to the other. But I will do further trials to ensure it is as they claim. I was also told the age of all the plants are all the same, but there is no way at my level of knowledge to be certain they are. Although the water lily is the dominant plant in this pond, another factor to take into considerations which may affect the growth of the plants is the competition with other plants. To ensure this does not affect the experiment I will ensure my subdivisions are not near the growth of the other plants. The effectiveness of this alteration will be discussed in the evaluation. Main Implementation: I will begin my main experiment by first confirming I have all the equipment needed, I have made a checklist consisting of all the apparatus I have used and will use. The next step of preparation is to put on the waders I have been set, as well having a spare set of clothes to change into after the experiment. And lastly I will check myself for any open wounds which may need to be taped up before going into the water. At exactly 0800 hours I will record the light intensity at each side of the pond. I will use the x10 setting of the lux meter for both sections as the intensity is relatively low at this time of day. I will note the light intensities of both areas; after 45 minutes have passed I will repeat the measurements once again and will continue doing this every 45 minutes for the rest of my time in the forest centre. During the time between the light measurements I will draw around the chosen lily pads. The lily pads have been pre chosen for each sub section by the random number generator which will allow me to choose 5 lily pads from each sub sections of each side of the pond. I will use my clip board to lean of while drawing around the lily pads of the graph paper. I did not anticipate the graph paper getting wet during the experiment, so some of the graph papers did get wet during the illustrations. After collecting all my results, I will allow my sketches to dry, and then begin to count the squares. This is an extremely long process, as some lily pads had an area of up to 500cm2. Just to be sure I measured the temperature of the water on both shade and non-shade several times. Table Light Intensity Throughout the Results: Table 14

Lily Pad 1 2 3 4 5 6 7

Area of Lily Pad (cm2) NonShade shade


144 154 168 221 236 251 259 113 164 390 444 456 468 482

15 Time (HH:MM) 8:00 8:45 9:15 10:00 10:45 11:15 12:00 12:45 13:15 14:00 11 14:45 15:15 16:00 16:45

Day (Lux) Shade 1070 1180 1280 1280 2550 2560 2030 2150 2500 8700 15000 10500 9900 10300 Non-Shade 3800 4900 5600 6000 15000 15800 25800 36900 37000 52800 69900 45000 50600 40000

8 9 10 11 12 13 14 15

260 265 276 317 355 369 372 380

486 491 503 508 512 515 518 540

To properly express my results I will plot the area on a bar graph. The Purple represent the Non-Shaded section, and the blue represents the Shaded.

G raph 4 600 500


Area (cm 2)

400 300 200 100 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15


S alles to Larg t m t es

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I will also plot my light measurements on a line graph, the squares represent the Non-Shaded section, and the diamonds represents the Shaded.

Graph 3
L ht Intensity (L ) ig UX

80000 60000 40000 20000 0


8: 00 9: 15 10 :4 5 12 :0 0 13 :1 5 14 :4 5 16 :0 0

T e (HH:MM) im

Analysis of results: My results appear to be irrefutable to my hypothesis, that there are larger lily pads in the nonshaded area of the pond. The results of the light intensity show that at any time of the day the non-shaded area of the pond had significantly higher light intensity than the shaded. As expected the largest lily pad was found in the non-shaded area of the pond which was an incredible 540cm2, this particular leaf consumed lot of my time. The smallest lily pad was surprisingly also found in the non-shaded area which was 133cm2. This could possibly be an anomaly; if this were an anomaly it could have occurred if the lily pad was originally from the shaded and somehow became detached and floated to the non-shaded side. As unlikely as it sounds, that may be the case. Another observation I have noted is that when the sun rose and set, the surrounding forest trees did not allow light to hit the shaded side directly, this observation may be used in support for the integrity of my experiment. As an added numerical insurance I will work out averages for my results. For this particular experiment the statistical method to find the product-moment correlation coefficient cannot be used, nor can other methods like the Mann-Whitney U test or Spearmans rank. The most effective method will be to find the mean. Table Non-shaded Mean 14 Lux 29221.43 Percentage difference Table Non-shaded Mean 15 Area 439.3333 Percentage difference Shaded Mean Lux 5071.429 17.35% Shaded Mean Area 268.4667 61.1%

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From working out the mean a significant difference between the two sides of the pond can be seen, and a conclusion can be drawn. But if we work out a percentage, to intensify the criticism of our results, we see that the percentage difference between the variable dont seem to be proportional. This could be due to the light intensity recording being taken on one day, and also the lack of samples of lily pads. Conclusion: Assuming there were no other factors involved, we can deduce that light intensity will increase the size of lily pads, indicating a significant factor regarding the development of the Nymphaea Odorata. If we broaden this conclusion we can assume that all water lily plants will have this correlation, but it will uncertainly not go beyond this particular species as not all types of plant are affected by light. Some plants thrive in the shade and cannot function to its best in direct sunlight, from the top of my head I recall a rubber plant should not be placed in direct sunlight. The main reason is because particular plants are adapted to absorb low light intensities, because of the limitations of the pigments not all intensities can be absorbed by plants. Another factor to take into consideration is the limitations of aquatic light which is mentioned in the book, Light and Plant Responses written by T.H Attridge. A section in the book describes how the penetration of light is dependent on many factors, such as solar elevation and water purity, it can also be affected by cloud cover and wind speeds. Other factors include algal absorption. The factors mentioned could greatly affect the results of my experiment, as I did not even consider that some of the submerged lily pads may have had completely different light exposures to the non-submerged lily pads. Evaluation: Unfortunately I could only perform my experiment once, and also I have a limited number of lily pad samples. If I were to expand my sample size or even record the areas of all the lily pads in the pond, and even go to the extent of performing the same experiment in a different pond. This will include finding a suitable pond which is half in the shade and not. This could also lead to preparing a fully controlled experiment, where water volume, temperature, PH and even the contents of the water will be under control. The dependant variable will be the light intensity, and using many different intensity levels and not just 2. To be fair the light will be turned off to mimic day and night as well as temperatures will be decreased when lights are turned off. Even if I went to that extreme it would maybe be used to find if there truly is a correlation but experiment like those cannot relate to real life as everything will be artificial. But not only will the resources needed have to be extensive; it will be expensive and time consuming. It can be disagreeable that the growth of the plants can be measured by its leaf size. If cactuses were brought into the question measuring the area of leaves would seem absurd. Unfortunately it is the best I could do with my limited time and resources. To properly measure plant growth either the biomass or dry mass will need to be recorded. But that may require me to take the plant out of the ground, dehydrate it and weigh its dry mass or even crush it to obtain its volume. The procedure to obtain precise results all consist of the destruction of the plant, this can be avoided if a common and an unwanted weed was used, but that will change the experiment entirely. When using my statistical formula book, I noticed a statement Two variables may be strongly correlated without a cause-and-effect relationship existing between them [5] The statement highlights difference between causation and association should be carefully considered in any evaluation. In water lily plants it is unlike most water plants as only the leaves and flowers remain above water, majority of this plant is the leaf. In hindsight I noticed another factor I did not take into consideration, which was the thickness of the leaf. Another unforeseen factor is that the number of leaves on both side. A better and possible measurement would be to work out the total surface area of all the leaves and to find an average. The size and number of the flowers could have also been measured as I would not have exerted any more effort, than that which was already engaged. All these shortcomings have led to revoke my conclusion.

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Bibliography [1] http://www.ecy.wa.gov/programs/wq/plants/weeds/lily.html [2] http://www.biology-online.org/dictionary/Phototroph [3] http://maps.google.co.uk [4] http://austintaylorphotography.com: permission given to use if origin of picture was mentioned [5] [DOC] Scatter Graphs and Correlation www.schoolworkout.co.uk/documents/s1/PMCC.doc

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