January 30, 2018The Effect of the Level of Solute Concentration on Glycine maxPurposeIn Japan, soybeans (Glycine max) are commonly used in various traditional foods and cuisines and has a large significance in the Japanese food culture. Tofu, soy sauce, miso are a few examples of foods made from soybeans which are regularly used in Japanese food, which are regularly consumed by the Japanese people. On March 11th, 2011, a massive tsunami, following a magnitude 9 earthquake hit the coast of Tohoku. According to Ieyasu Tokumoto, thirteen thousand hectares of farmland was damaged by the tsunami, where the subsidence and high salinity of the groundwater originating from seawater made it difficult to remove salinity from the soil (2016). With having a Japanese cultural background where I consume soybeans on a daily basis, and having experienced the Tohoku earthquake, I decided to investigate how the salinity of the soil has an effect on the germination of Glycine max. Through this investigation, I will be able to explore the effect the tsunami in the Tohoku earthquake had on the soybean farms in Tohoku. Research QuestionHow does changing the solute concentration of NaCl (0.00M, 0.20M, 0.40M, 0.60M, 0.8M) (+0.05M) when germinating 10 Glycine max seeds affect the rate of CO2 production (ppm) (+50) produced over the course of five minutes (1 min, 2 min, 3 min, 4 min, 5 min) (+0.5 seconds)?Independent VariableThe amount of NaCl (0.00M, 0.20M, 0.40M, 0.60M, 0.8M) (+0.05M)Unit: MolesDependent VariableRate of CO2 Production (+0.5 ppm)Unit: ppm (parts per million over time)Controlled VariablesVariable Potential impact on resultsSpecific method for controlling variableNumber of Glycine max seedsThe different number of seeds in each bottle can affect the CO2 production, which could cause inaccuracies in the results. This is because The number of Glycine max seeds will be constant at 10 seeds per bottleTime (minutes) (+0.5 seconds)Changes in the different lengths of time when measuring the CO2 production can negatively affect my results, as the Glycine max will continue to germinate.The CO2 production will be measured through the course of 5 minutes and will be kept constant for each incrementTemperature (+0.05º)Changes in temperature can affect the rate of CO2 production as temperature is one of the factors that affect germination.The temperature will be kept constant by conducting the experiment in the same area of the room.Level of oxygen Changes in the level of oxygen present in the air can affect the rate of CO2 production, as oxygen is one of the factors that affect germination.The level of oxygen present in the air will be controlled as I will be collecting the data in the same room and in the same area of the room.Amount of water in each bottle (500ml (+0.5M))The amount of water I will be using to soak the Glycine max may affect the rate of CO2 production if different amounts of water are used for each trial. I will be using a beaker and standing from the same angle when measuring the amount of water.Uncontrolled VariablesVariablePotential impact on resultsBiological VariationAs the genes present for each Glycine max seed is different, the CO2 that the Glycine max produces during germination may differ from seed to seed. Therefore, this may cause uncertainties in my results.Background Information:According to Encyclopedia Britannica, “Germination, the sprouting of a seed, spore, or other reproductive body, usually after a period of dormancy” (n.d).  Dormancy is a time period where the seed metabolizes stored energy reserves and are unable to germinate. In some cases, specific temperatures and light intensities are required in order for the seeds to break past dormancy. In order for a seed to germinate, oxygen, water, a specific temperature, and a suitable soil pH level is required. Seeds will continuously deteriorate and eventually die unless they are in the process of germinating. This rate of deterioration is highly dependent on the temperature and amount of moisture (Roberts EH., n.d). Therefore, changes in the temperature and level of moisture will alter the rate of deterioration, and thus affect the rate or the chance of the seed germinating. cell respiration takes place to accumulate the high amount of energy used during the process of germination. During cell respiration, organic compounds are broken down in order to generate Adenosine Triphosphate, which is the energy used to carry out functions of life including germination. The process of cell respiration starts with glycolysis, where glucose molecules are converted into pyruvates by splitting glucose into two three-carbon molecules (Khan Academy, 2017). Then, the pyruvate molecules are arranged and combined through the Krebs cycle. During cell respiration, carbon dioxide is produced as a waste product. Therefore the rate of carbon dioxide production can determine the germination of seeds. As discussed, water is a key factor which is needed for a seed to germinate. Therefore, as NaCl is present in the water absorbed by the seed, the osmotic pressure created by sodium chloride prevents the water to pass the membrane and salt to enter the seed, therefore intoxicating it (Cocoponics, 2011). Furthermore, as there is a high solute concentration in the water surrounding the cell when germinating seeds, water leaves the seeds, where the seeds are unable to germinate as water is a requirement for germination.  Therefore, the salinity of the soil caused by the tsunami that hit the coast of Tohoku will toxicate the Glycine max seeds planted. HypothesisAs seen in the graph above, the rate of CO2 production of Glycine max will decrease over the course of five minutes as the solute concentration is increased. This is because the higher the NaCl concentration is in the water surrounding the Glycine max seeds, more water will diffuse out the Glycine max seeds, and slow the rate of CO2 production, which therefore shows the decrease in the rate of germination.Equipment and MaterialsItem QuantityGlycine max seeds50Plastic Bottles5Water500ml (+0.5M)Salt2M (+0.5M)CO2 Respirometer (+50) 1Data Logger1Timer1Image of Experimental SetupMethod:Add 100 ml of water into each of the 5 plastic bottles. Add 0.00M of NaCl into one bottle, 0.20M in another, 0.40M in another, 0.60M in another, 0.8 in another bottle. Label each bottle with the different solute concentrations (0.00M, 0.20M, 0.40M, 0.60M, 0.80M) (+0.5)Place 10 Glycine max in each bottleLeave Glycine max to soak for 24 hours Recording CO2 productionPlug in a CO2 respirometer attached to a data logger and stand one meter away for 1 minute to let the sensor stabilize and calibrate the probe. Record the rate of CO2 production of the initial atmospherePlug the CO2 respirometer into the bottle with 0.00M of salt and record the level of CO2 production (0 min).Set the timer to 5 minutes and record the CO2 production at 1 min, 2 min, 3 min, 4 min, and 5 min (+0.05 seconds) by stepping 1 meter away from the sensorRepeat steps 1 to 4, five timesRepeat step 5 with the four other bottles with different solute concentrations (0.20M, 0.40M, 0.60M, 0.8M)Ethical considerationsAlthough the soybeans will not be directly harmed, 50 soybeans will be used, which may lead to food waste.SafetyThere are no safety requirementsData Collection and Processing Raw Data 1:The amount of carbon dioxide (ppm) produced over the course of 5 minutes (+0.5 seconds) for 0.00M of NaClInitial atmosphere (0 min) (+0.5 seconds)1 min (+0.5 seconds)2 min (+0.5 seconds)3 min (+0.5 seconds)4 min (+0.5 seconds)5 min (+0.5 seconds)Change in CO2 Production (1 to 5 minutes) Trial 1 412 85010631311164218701458Trial 236088810231360170417871427Trial 334786012331302152017541407Trial 439992611761432160516971298Trial 536579812011466151418841519Standard Deviation81.13Raw Data 2:The amount of carbon dioxide (ppm) produced over the course of 5 minutes (+0.5 seconds) for 0.20M of NaClInitial atmosphere (0 min) (+0.5 seconds)1 min (+0.5 seconds)2 min (+0.5 seconds)3 min (+0.5 seconds)4 min (+0.5 seconds)5 min (+0.5 seconds)Change in CO2 Production (1 to 5 minutes) Trial 1 52896041411418096213462399423466Trial 261297231399318264204352400823396Trial 359495791416217954199872412223528Trial 461396341425418213214532395423341Trial 557391141383218572221412368423111Standard Deviation160.29Raw Data 3:The amount of carbon dioxide (ppm) produced over the course of 5 minutes (+0.05 seconds) for 0.40M of NaClInitial atmosphere (0 min) (+0.5 seconds)1 min (+0.5 seconds)2 min (+0.5 seconds)3 min (+0.5 seconds)4 min (+0.5 seconds)5 min (+0.5 seconds)Change in CO2 Production (1 to 5 minutes) Trial 1 6725016771010400128661506814396Trial 25965132780110220119461518714591Trial 36564997769710180127651554014884Trial 46325021780111452129221498814356Trial 56895124763411043125441523114542Standard Deviation208.91Raw Data 4:The amount of carbon dioxide (ppm)produced over the course of 5 minutes (+0.5 seconds) for 0.60M of NaClInitial atmosphere (0 min) (+0.5 seconds)1 min (+0.5 seconds)2 min (+0.5 seconds)3 min (+0.5 seconds)4 min (+0.5 seconds)5 min (+0.5 seconds)Change in CO2 Production (1 to 5 minutes) Trial 1 585486670769646121941446613881Trial 2623483271339732119951423313610Trial 3656491270239643121221431413658Trial 4599486571269731120131442413825Trial 5601492371119693123121441213811Standard Deviation116.54Raw Data 5:The amount of carbon dioxide (ppm) produced over the course of 5 minutes (+0.5 seconds) for 0.80M of NaClInitial atmosphere (0 min) (+0.5 seconds)1 min (+0.5 seconds)2 min (+0.5 seconds)3 min (+0.5 seconds)4 min (+0.5 seconds)5 min (+0.5 seconds)Change in CO2 Production (1 to 5 minutes) Trial 1 642464072369746122021449413852Trial 2631470171429699121111422113590Trial 3623463272519822119981432713704Trial 4612472371299701123121490114289Trial 5607459772449734124021434813741Standard Deviation270.38Calculating change in carbon dioxide production:Final – Initial= Change in carbon dioxide productionEx. 14494 – 642= 13852Change in carbon dioxide production = 13852 ppmCalculating standard deviation on Excel:Ex. Formula=STDEV(H22:H26)Processed Data Processed Data 1:Average amount of carbon dioxide (ppm) produced over the course of 5 minutes (+0.05 seconds) for 0.00M, 0.20M, 0.40M, 0.60M, 0.8M (+0.5) of NaCl0 min (+0.5 seconds)1 min(+0.5 seconds)2 min (+0.5 seconds)3 min (+0.5 seconds)4 min (+0.5 seconds)5 min (+0.5 seconds)Change in CO2 Production (1 to 5 minutes) Standard Deviation0.00M (+0.5)3778641139137415971798142181.130.20M (+0.5)58495311407118220210722395223368160.290.40M (+0.5)6895124763411043125441523114542208.910.60M (+0.5)613488070949689121271437013757116.540.80M (+0.5)623465972009740122051445813835270.38Calculating average amount of CO2 produced: Ex.412 + 360 + 347 + 399 + 365=18831883/5= 376.6Data Presentation: Line GraphAnalysisAnalysing the data presented in Figure 1, all increments show a constant and upward trend in the average rate of CO2 production, indicating that the Glycine max soaked in different moles of NaCl all germinated, however at a different rate depending on the concentration of NaCl. This can be seen in the trendline shown for each increment, where data for 0.00M, 0.80M, and 0.6M of NaCl establish a stronger correlation to the trendline compared to the data for 0.20M and 0.40M of NaCl, although all trendlines are somewhat supported. Furthermore, as there are no significant anomalies, this demonstrates the reliability and the accuracy of my method. The error bars present also indicate the accuracy of my results. As seen in Figure 1, the low standard deviations and the small error bars, indicate that the uncertainty of the data is low. This shows that the method was reliable and well conducted, leading to accurate and reliable results. On the other hand, the minimal error seen in the error bars were due to a number of uncontrolled variables. First of all, biological variation in the Glycine max could have affected my results. As no two Glycine max are identical, the amount of CO2 production could have differed in each seed. Furthermore, the uncertainty when collecting data from the CO2 respirometer each minute, and when timing each trial could have affected the data collected, and thus, caused the uncertainty and low standard deviation in my data.ConclusionsThe graph above (Figure 1) depicts the changes in the average carbon dioxide (ppm) production in Glycine max over the course of five minutes. Analysing the data presented in the graph, all Glycine max seeds germinated over the course of five minutes, no matter how many moles of NaCl was added to the concentration the seeds were soaked in. However, analyzing each increment further in depth, when no moles of NaCl was added to the water when germinating the beans, the data shows that there was only a slight increase in the rate of CO2 production over five minutes. Furthermore, when 0.40, 0.60, and 0.80 moles of NaCl were added to the water when germinating Glycine max for the different trials, the data shows that they all increased in the average change in the rate of CO2 production over five minutes approximately at the same rate. Not only was the rate of CO2 production higher compared to when no NaCl was added, the average amount of CO2 produced increased. Finally, as 0.20 moles of NaCl was added to the water when germinating Glycine max, the average rate of CO2 production over 5 minutes increased at a higher rate compared to the other increments. In addition, the average amount of CO2 produced was higher compared to the other increments, indicating that the Glycine max was germinating at a higher rate than other increments. However, a decrease in the average rate of CO2 production and the decrease actual amount of CO2 produced can be seen in Figure 1 from when more than 0.20 moles of NaCl was added to the water when germinating the seeds. This illustrates the negative effect the NaCl had on the Glycine max during the process of germination. This is because the NaCl present in the water that the Glycine max has absorbed, hindered the process of germination, causing an osmotic pressure preventing water to pass and salt to enter through the membrane of the Glycine max, thus, intoxicating it (Cocoponics, 2011). Referring back to my hypothesis, the data does not support my hypothesis or my background research in any way. Overall, observing the data collected through this experiment, it can be concluded that the level of NaCl when germinating Glycine max does not affect the rate in CO2 production up until a point (0.20M), where a significant level of solute in the water when germinating the beans will slow the rate and the amount of CO2 produced over the course of five minutes.Evaluations & ImprovementsThere are various limitations to the method of this experiment. First of all, since the first set of data (0M of NaCl) and the other four set of data (0.2M, 0.4M, 0.6M, 0.8M) was collected on different days, this could have affected the amount of CO2 produced, thus, causing the fluctuations in my data for the initial amount of CO2 production for every trial. The factors such as the changes in temperature in the classroom could have affected the amount of CO2 produced in the initial time as the temperature is one of the factors that affect germination. Therefore, this affected the accuracy of the data collected, as in some trials, data fluctuated or differed depending on the day I collected the data. Secondly, biological variation in the Glycine max could have affected the precision of my data collected, which can be seen, as varying amounts of CO2 was produced for the five trials. The biological variation is present due to slight genetic differences in each Glycine max seed used in this experiment. Furthermore, as the data was collected by recording the amount of CO2 produced displayed on the GLX each minute, this affected the accuracy of my data collected. First, as the number displayed on the GLX was constantly changing due to the slight changes in the CO2 production, it was a struggle to record the amount of CO2 produced exactly after each minute passed. Second, as it was necessary to approach the GLX connected to the CO2 sensor in order to collect data, this could have affected the accuracy of my data, as slight movement and change in temperature can affect the number displayed on the sensor due to the high sensitivity. On the other hand, improvements can be made to the method in order to limit inaccuracies in the data collected from this experiment. First of all, the data should be collected on the same day, where the room temperature can be moderated at a consistent temperature, which can limit inaccuracies in the data. Furthermore, although factors such as random biological variation cannot be maintained for each trial and increment, the accuracy of the data can be improved by carrying out more trials for each increment (1 min, 2 min, 3 min,  4 min, 5 min), as the probability of accurate data collected from measuring the amount of CO2 produced will increase. ExtensionThrough this experiment, I was able to explore the effect of the change in NaCl concentration on the amount of carbon dioxide produced over five minutes. To further extend this experiment, the exploration of other factors such as pH level, light, and temperature would be ideal, as through those explorations, I will be able to explore the process of germination in greater depth. Furthermore, I would like to explore whether if other factors such as radiation from the destruction of the nuclear power plant in Fukushima caused by the March 11, 2011 Tohoku earthquake could have an affect on the germination of Glycine max seeds in order to further explore the effect this earthquake had on plants in Tohoku.References:Cellular respiration | Biology | Science. (n.d.). Retrieved January 30, 2018, from https://www.khanacademy.org/science/biology/cellular-respiration-and-fermentationHow salt affects Seed Germination. (2015, July 29). Retrieved January 30, 2018, from http://www.cocoponics.co/germination-2/salt-affects-seed-germinationRoberts, E. H. (n.d.). Temperature and seed germination. Retrieved January 29, 2018, from https://www.ncbi.nlm.nih.gov/pubmed/3077854Soybeans. (n.d.). Retrieved November 07, 2017, from http://www.tokyofoundation.org/en/topics/japanese-traditional-foods/vol.-3-soybeansThe Editors of Encyclopædia Britannica. (2016, October 18). Germination. Retrieved January 28, 2018, from https://www.britannica.com/science/germinationTokumoto, I., Chiba, K., Mizoguchi, M., & Miyamoto, H. (2016). Investigation of rootzone salinity with field monitoring system at tsunami affected rice fields in Miyagi, Japan. SOIL Discussions, 1-28. doi:10.5194/soil-2016-12

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