Effect Of Different Concentrations Of Vitamin C

Effect Of Different Concentrations Of Vitamin CThis project was designed to wonder the effect of different assimilations of vitamin C upshot on root spr bring outing of Brassica parachinensis or Choy Sum. Seeds treated with vitamin C solutions of various soaking ups were utilize and sprouting parting was calculated. Seeds were soaked in vitamin C solutions of increasing densitys for 12 hours, dried and then sown in germinating trays. The sheds were urineed with the same add up of vitamin C solution of individual s starting time-wittednesss. After 36 hours, the number of germinated microbes was counted and the germination parcel was calculated and recorded. Results showed that the germination lot increases as low engrossment of vitamin C solution was utilise. It was shown that 1% of vitamin C solution gave the highest germination percentage. A statistical synopsis use Pearson product-moment correlation coefficient showed a statistically monumental negative linea r relationship amidst concentration of vitamin C and germination percentage since the calculated r value was greater than critical value at 5% significant level.IntroductionIn array to increase productivity, it is important to improve overall growth and performance of agricultural crops. Germination rate in particular is crucial because an increase in germination rate is usually followed by an improvement of overall sowling performance thus enhancing overall plant growth and productivity.(P atomic number 18ra and Cantliffe, 1991).There are many accompanimentors affecting seed germination such as temperature, humidity, presence of light and seed dormancy. Recently, there is also evidence to support a link amid certain biochemical characteristics and seed postcode.(Randhir and Shetty, 2003) Fast oxidation of fatty acids and proteins can reduce viability, heftiness and germination percentage in seeds.11Vitamin C or ascorbic acid is one of the almost widely taken dietary supple ment.5 It is an antioxidant which is an essential nutrient for living organisms as it protects the body against oxidative focal point. Vitamin C is made naturally in plants precisely the amount depends on the variety of plant and external factors on which the plants grow.1Ascorbic acid is an important metabolite involved in many cellular processes, including cell division (De Gara et al., 2003). Ascorbate has been shown to play tenfold roles in plant growth, such as in cell division, cell wall expansion, and other developmental processes. It can act as a coenzyme for legion(predicate) metabolic enzymes and has recently been shown to be a potent antioxidant.12Figure 1 The oxidation of ascorbic acid to dehydroascorbic acid 6(410 words)Vitamin C interacts with oxidants in an oxidation reception where ascorbic acid is oxidised to dehydroascorbic acid through an ascorbyl radical intermediate.8 The oxidized forms of ascorbate are relatively un oxidizable, and do non cause cellular damage. They can be reborn back to ascorbate by cellular enzymes.4 Application of ascorbic acid exogenously may influence various processes in plants including seed germination, ion uptake and transport, and membrane permeability.12Structure of vitamin C or L-ascorbic acidor Dehydroascorbic acidL-ascorbate 4 (nominal oxidized formof Vitamin C) 4Previous reports show approximately conflicting evidence about the effect of vitamin C towards seed germination. While some researchers say that vitamin C can promote seed vigour as in pea (Pisum Sativum) seeds, 12 some put in that its presence will inhibit seed germination as in soybean seed.13Defined in the qualitative term, seed vigour is the sum of those properties of theseedwhich determine the potential level ofactivity and performance of theseed or seedlot during germination and seedling emergence.12Seeds of genus Brassica are used in this essay because the seeds are small and have a short life cycle. Therefore, experiment can be make within few days and the seeds can be used in large number with less space needed.Hence, this experiment was aimed to explore the benefits of vitamin C which is an antioxidant which may help to increase plant resistance that leads to grow crops.(647 words)Experimental HypothesisThe lower the concentration of vitamin C solution, the high the germination percentage of Brassica parachinensisNull HypothesisThere is no correlation between different concentrations of vitamin C solutions and the germination percentage of Brassica parachinensis(685 words)PlanningTrialsTrial experiment was conducted to determine the best range of concentrations of vitamin C solution to be used and the reasonable methods and procedures so that good allow fors are obtained.Methods InvolvedPreparing vitamin C solution of different concentrationsSince the source of vitamin C used is the vitamin C tablet, the best method elect is the serial dilation method. During trial, concentrations established are 100 %, 75%, 50%, 25%, 10% and 0%. The basic stock solution of 100% was alert by dissolving 100mg of vitamin C tablet in 100ml distilled water in a 300ml beaker. Other concentrations were made by serial dilution method. To prepare 50% concentration of vitamin C solution, same amount of 100% solution and distilled water were added together. Equal amount of 100% and 50% concentrations were mixed up to make 75% concentration and so on.Sowing the seedsThe seeds were primed in 6 different Petri dishes for 12 hours according to its respective concentration. Another 6 Petri dishes were labelled. Layers of cotton of the same thickness were put into Petri dishes and weaken with the same volume of vitamin C solution of different concentrations. Soaked seeds with similar size were chosen and took out of Petri dish using a p communication channel of forceps. They are then dried using constitution towels and were sown in labelled Petri dishes.(909 words)In the first trial, 25 seeds were soaked in different concentrations of vitamin C solution while another 25 seeds were not soaked before being sown. This is to determine whether it is necessary to soak seeds or not before sowing.36 hours later on sowing seedsConcentration/%Number of seeds germinatedSoaked seeds0151016501 duck 1 shows the number of soaked and unsoaked seeds after 36 hoursIn the second trial, each group of 25 seeds were soaked in different concentrations before being sown. This is to determine the best range of concentrations of vitamin C for main(prenominal) experiment.36 hours after sowing seedsConcentration/%Number of seed germinatedTrial 1Trial 2Mean015161610181617255555013275212100101Table 2 shows germination percentage of seeds soaked in different concentrations(1075 words)ResultsAfter 36 hours of sowing, the number of seeds germinated was counted and germination percentage was calculated. Germination is claimed to have occurred when the radicle was visibly extended from the seed, protruding through a fracture in the seed coat. The result was tabulated.Based on the results obtained after trials were carried out, there were a few things that could be done to improve result. Firstly, more seeds should be used so that the result will be more significant. For the square experiment, 50 seeds were used instead of 25 seeds.Seeds were primed for 12 hours because priming is important as seed coat contains some chemicals that inhibit germination. Seed priming is a common agronomic technique shown to cause an increaseseed vigour,as well as synchronize and accelerategermination, improve sample resistance, and put up overallplantgrowth and productivity.12 However, some of the seeds start to germinate and fractures on seed coat could be observed if the seeds were primed for too long.Besides, the range of concentration to be used in real experiment was determined to be between 0% to 10% concentrations because both showed high percentage of seed germination with entirely slight difference i n value.Seeds soaked in 100% concentration showed the lowest germination percentage. The germination of seeds was almost totally inhibited in this solution. Fractures on the seed coats were observed but radicles were not extended significantly.Besides, after carrying out trial experiment, I realized that it was better to sow seeds in germinating tray quite an than in Petri dishes as Petri dish is round-shaped which made it difficult for me to count the number of seeds germinated.(1351 words)Main experimentVariablesManipulated Concentration of vitamin C solutionDifferent concentrations of vitamin C are prepared using the same serial dilution technique as in trials. But this time using smaller range of concentrations of 10%, 7.5%, 5%, 2.5%, 1% and 0% (distilled water). But for smaller concentration, micropipette is used instead of measuring cylinder to make up the total volume of 100ml.Responding Germination percentage (%)Germination percentage is calculated using the following equat ionGermination is said to occur when radicle emerges (2mm) after 36 hours.Constant mass of vitamin C (100mg/tablet), volume of solution used for soaking seeds (20ml/Petri dish) and volume of solution used to moisten cotton (60ml/tray) and environmental factors (light intensity, temperature, and humidity), period of soaking, type and number of seeds. All trays were placed under normal unclouded fluorescent lamp and on a laboratory table at 28C.Apparatus and materialsApparatus300ml beakers, Petri dishes, measuring cylinders, germinating trays, stirring rod, micropipette, and pestle and mortar.MaterialsDistilled water, 100mg vitamin C tablet, cotton, choy sum seeds, and paper towels.(1535 words)Safety precautionsGloves were worn when handling vitamin C tablet to prevent tablet from contamination.Micropipette was used with care. The tip of micropipette was always checked to ensure that there is no air bubbles inside it as this may reduce the actual volume required for each concentratio n.Glass wares like beakers and measuring cylinders were handled gently since the apparatus may shimmy easily. This may lead to injuries.Soaked seeds were transferred gently using forceps as they are very fragile.Data collections of main studyNumber of seeds used in both concentration 50Soaking for 12 hoursObservations after soaking seeds for 36 hoursConcentration (%)Number of seeds germinatedMean number of seeds germinatedReplicate 1Replicate 2Replicate 30.0454443441.0484847482.5423938405.0414243427.53535343510.034333333Table 2 shows the germination percentage for seeds treated with different concentrations(1712 words)Sample calculation (taken values from 0% concentration)Germination percentage=44/50 x 100=88%(1749 words)Statistical analysisFor this experiment, Pearson product-moment correlation coefficient (PMCC) was used to measure the strength of linear dependence between the two variables the different concentrations and the germination percentage. The values obtained are norm ally distributed.The correlation coefficient, r ranges from -1 to 1. A value of 1 implies that a linear equation describes the relationship between X and Y perfectly for which Y increases as X increases. A value -1 implies correlation in which Y decreases as X increases. A value of 0 implies that there is no linear correlation between the variables.x0.01.02.55.07.510.0x=26y889680847066y=484x20.01.006.2525.056.25100.0x2=188.5y2774492166400705649004356y2=39672xy0.096200420525660xy=1901Table 3 shows the calculations done in calculating the values of rSx = x2 = 188.5 (26)2/6= 75.833Sy = y2 = 39672 (484)2/6= 629.33Sxy = xy = 1901 (26) (484)/6= -196.33 (1933 words)To find correlation coefficient, rr =r = -196.33/ (75.8331/2 x 629.331/2)r = -0.899 (negative sign indicates negative correlation between variables)Degree of freedom = n -2n = of pairs of data= 6 2 = 4Level of significance for two-tailed test.10.05.021.988.997.99952.900.950.9803.805.878.9344.729.811.8825.669.754.8336.621.70 7.789Table 4 shows the critical values for PMCCThe value of r obtained was compared with the critical value from Table 4 at 5% significance level.r = 0.899 0.811 (critical value) at 5% significance levelAn analysis using PMCC demonstrated a statistically significant negative linear relationship between concentration of vitamin C and germination percentage since calculated r value was greater than the critical value at 5% significance level.Therefore, null hypothesis can be rejected.(2093 words)Data analysisBased on the data collected in Table 2, it was shown that low concentration of vitamin C did promote germination of choy sum to a certain extent. The statistical analysis using PMCC further verified the correlation. The table also revealed that only low concentration of vitamin C was needed to hasten germination since under 10% concentration, seeds germinated at a lower rate (66%).Graph 1 illustrates understandably the widely distributed trend and relationship between the variab les, the lower the concentration of vitamin C, the lower the germination rate of choy sum. The highest germination rate (96%) was recorded by seeds sown in 1% vitamin C concentration which was the lowest concentration used.However, it can be seen that the line is fluctuating downwards and there is a linear graph present that the values obtained were due to certain errors that lead to some anomalous results. At 5% concentration of vitamin C, we can see that the germination rate is slightly higher than that of 2.5% but it is not supposed to be so.Some of errors that may occur during experiment areMixing error when preparing solutions of different concentrations by serial dilution.The thickness of cotton layer in germinating tray may not be constant throughout the tray so vitamin C solution with thinner cotton layer may get evaporated faster thus absorption of solution is less for the seed in that traySome of seeds may have been initially damaged so germination may not happen(2342 wor ds)The germination percentage was compared with the result from other published academic ledgers. The germination percentage against day graph below shows the results from journal titled Effect of vitamin C and folic acid on seed vigour response and phenolic-linked antioxidant activity from source 12. It shows that the optimum concentration of vitamin C for pea seed to germinate is 500micromol/dm3.Using appropriate calculations, it can be calculated that 10% concentration is approximately equal to 500micromol/dm3.9 Thus, the germination percentage of pea seed in 10% concentration is highest compared to other concentrations. However, the result I obtained for highest germination percentage for Brassica parachinensis was different. This can be due to some errors and limitations occurred during experiment or the fact that they are of different types of seed which is further discussed in limitation and modification part.In this experiment, the constant or controlled variable is the use of distilled water (0% concentration). Since the results showed that the difference between germination rate between 0% and 1% concentration was very small (only about 8%), this indicated that the effect of vitamin C on seed vigour may be too little. Even though the effect is little, it cannot be denied that this little help from vitamin C can give a huge carry on on agricultural sector as it is able to promote seed germination therefore improving mass production of plants rather than watering seeds with water only.(2577 words)EvaluationSeeds are exposed to various biotic and abiotic stresses during germination. Oxidative stress for example, caused by the production of high level of reactive oxygen intermediates such as O2, O2-, H2O2, and HO- can lead to the destruction of cells.4 Presence of low concentration of exogenous vitamin C stimulates seed vigour as it minimises oxidative stress by scavenging these reactive oxygen species.Many oxidants contain an unpaired electron so they are highly reactive. These free radicals are damaging because they can interact with other molecules resulting in a whole chain of free radical reactions. Ascorbate can terminate these chained radical reactions by serving as a stable (electron + proton) conferrer in interactions with free radicals.4However, high concentration of vitamin C solution inhibits seed germination because as solution becomes more concentrated, more water from cells diffuse out causing cellular membrane leakage.3Besides that, being a good electron donor, excess ascorbate in the presence of free metal ions can not only promote, but also initiate free radical reactions, thus making it a potentially dangerous pro-oxidative compound in certain metabolic contexts.8 We hypothesize that whether vitamin C has a net pro oxidant or antioxidant effect depends on the concentration of vitamin C solution.8With relation to improvingseed vigourthrough stimulation ofphenolicsynthesis, it has been suggested that exogenously appliedphenolic antioxidants may be able to stimulate endogenousphenoliccontent in plants(Randhir and Shetty, 2003andRandhir et al., 2002).Due to its inbornantioxidantpotential, vitamin C seems to be a suitable growth regulator in increasingseed vigour and phenolicelicitation. In this study, we have hypothesized that the treatment of plants withvitamin Cas exogenousseed treatments may result in an increase in both biochemical and traditional agronomic indicators ofseed vigour.11(2873 words)Limitations and modificationsOne of the limitations in this experiment is the changing surrounding temperature. The seeds were left in laboratory room where the temperature may fluctuate over time. This may affect the germinating time of the seeds and the number of germinated seed after 36 hours. Besides temperature, humidity and light intensity were also uncontrolled. Since all seeds were exposed to the same environmental factors, the result is considered to be reliable.Besides that, some seeds may have been initially damaged before soaking or sowing. Therefore, vitamin C solution may not be the actual cause of prohibition of seed germination. The seeds are chosen only by physical measures such as similarity in size, shape and colour and seed coat is not broken.However, limitations can be overcome by using a large number of seeds and the experiment is repeated for a few times to ensure that result obtained is reliable and enough to support conclusion. This investigation tests only on the effectiveness of vitamin C solution on seed germination without exposing seeds to other biotic stress such as salinity or abiotic stress such as exposure to sunlight.Modifications could be made by introducing a biotic or abiotic stress to the seed and observe if vitamin C still stimulates seed germination or otherwise. Different types of seed may have different result with vitamin C solution. So, using seeds from one type of plant may not be able to indicate how vitamin C actually acts on seed. As vitamin C may enhance seed vigour in certain species, it could do otherwise to other species. Hence, the same experiment should be done to seeds from other species to see how they are impact by vitamin C solution.ConclusionThe lower the concentration of vitamin C, the higher the germination rate of Brassica parachinensis seeds. This is due to the antioxidant properties of vitamin C which acts against the oxidative stress. The optimum vitamin C concentration for seed germination is 1%.(3200 words)Source EvaluationI have referred to various sources in conducting my research.Sources 1, 2, 3 and 4 are from Wikipedia which has been the largest and most popular generalreference workon theinternet. Although many have doubts over its reliability as it can be edited by anyone with access to the site but it is cited and linked to many other literatures and academic researches around the globe. Source 5 is a website written by health experts on health and medicine. Source 6 is a web site that provides information written by university scholars with reference to other academic researches. Source 7 is reliable as it is written byDr. Bryan D. McKersie ofUniversityofGuelph and is posted on the internet in 1996. Source 8 is a reliable source as it is an article by The American Society for Nutritional Sciences and was published in The Journal of Nutrition website in 2004. Source 9 has been a referral site to many people around the world. The information seems reliable as it is written by experts.Source 10 is a published book thus the information is reliable.Source 11 is a journal from ScienceDirect which I have accessed from my campus library. ScienceDirect is a leash full-text scientific database offering journal articles and book chapters from more than 2,500 peer-reviewed journals and more than 11,000 books. There are currently more than 9.5 million articles/chapters, a content base that is suppuration at a rate of almost 0.5 million additions per year. It is op erated by the publisher Elsevier. Source 12 is a journal advised by an international column Board and is abstracted or indexed by various research databases like the EBSCO and Elsevier. Source 13 is an article from the Journal of Biological Chemistry.