Effect Of Different Concentrations Of Vitamin C Solution Biology Essay

This undertaking was designed to look into the consequence of different concentrations of vitamin C solution on seed sprouting of Brassica parachinensis or Choy Sum. Seeds treated with vitamin C solutions of assorted concentrations were used and germination per centum was calculated. Seeds were soaked in vitamin C solutions of increasing concentrations for 12 hours, dried and so seed in shooting trays. The seeds were watered with the same sum of vitamin C solution of several concentrations. After 36 hours, the figure of germinated seeds was counted and the sprouting per centum was calculated and recorded. Consequences showed that the sprouting per centum additions as low concentration of vitamin C solution was used. It was shown that 1 % of vitamin C solution gave the highest sprouting per centum. A statistical analysis utilizing Pearson product-moment correlativity coefficient showed a statistically important negative linear relationship between concentration of vitamin C and sprouting per centum since the deliberate R value was greater than critical value at 5 % important degree.

Introduction

In order to increase productiveness, it is of import to better overall growing and public presentation of agricultural harvests. Germination rate in peculiar is important because an addition in sprouting rate is normally followed by an betterment of overall seedling public presentation therefore heightening overall works growing and productiveness. ( Parera and Cantliffe, 1991 ) .

There are many factors impacting seed sprouting such as temperature, humidness, presence of visible radiation and seed quiescence. Recently, there is besides grounds to back up a nexus between certain biochemical features and seed energy. ( Randhir and Shetty, 2003 ) Fast oxidization of fatty acids and proteins can cut down viability, energy and sprouting per centum in seeds.11

Vitamin C or ascorbic acid is one of the most widely taken dietetic supplement.5 It is an antioxidant which is an indispensable food for populating beings as it protects the organic structure against oxidative emphasis. Vitamin C is made of course in workss but the sum depends on the assortment of works and external factors on which the workss grow.1

Ascorbic acid is an of import metabolite involved in many cellular procedures, including cell division ( De Gara et al. , 2003 ) . Ascorbate has been shown to play multiple functions in works growing, such as in cell division, cell wall enlargement, and other developmental procedures. It can move as a coenzyme for legion metabolic enzymes and has late been shown to be a powerful antioxidant.12

Figure 1: The oxidization of ascorbic acid to dehydroascorbic acid 6

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Vitamin C interacts with oxidizers in an oxidization reaction where ascorbic acid is oxidised to dehydroascorbic acid through an ascorbyl extremist intermediate.8 The oxidised signifiers of ascorbate are comparatively unreactive, and do non do cellular harm. They can be converted back to ascorbate by cellular enzymes.4 Application of ascorbic acid exogenously may act upon assorted procedures in workss including seed sprouting, ion consumption and conveyance, and membrane permeability.12

Structure of vitamin C or L-ascorbic acid or Dehydroascorbic acid

L-ascorbate 4 ( nominal oxidized signifier of Vitamin C ) 4

Previous studies show some conflicting grounds about the consequence of vitamin C towards seed sprouting. While some research workers say that vitamin C can advance seed energy as in pea ( Pisum Sativum ) seeds, 12 some province that its presence will suppress seed sprouting as in soya bean seed.13

Defined in the qualitative term, seed energy is the amount of those belongingss of the seed which determine the possible degree of activity and public presentation of the seed or seed batch during sprouting and seedling emergence.12

Seeds of genus Brassica are used in this experiment because the seeds are little and have a short life rhythm. Therefore, experiment can be done within few yearss and the seeds can be used in big figure with less infinite needed.

Therefore, this experiment was aimed to research the benefits of vitamin C which is an antioxidant which may assist to increase works opposition that leads to heighten harvests.

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Experimental Hypothesis

The lower the concentration of vitamin C solution, the higher the sprouting per centum of Brassica parachinensis

Null Hypothesis

There is no correlativity between different concentrations of vitamin C solutions and the sprouting per centum of Brassica parachinensis

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Planing

Tests

Trial experiment was conducted to find the best scope of concentrations of vitamin C solution to be used and the sensible methods and processs so that good consequences are obtained.

Methods Involved

Fixing vitamin C solution of different concentrations

Since the beginning of vitamin C used is the vitamin C tablet, the best method chosen is the consecutive dilation method. During test, concentrations tested are 100 % , 75 % , 50 % , 25 % , 10 % and 0 % . The basic stock solution of 100 % was prepared by fade outing 100mg of vitamin C tablet in 100ml distilled H2O in a 300ml beaker. Other concentrations were made by consecutive dilution method. To fix 50 % concentration of vitamin C solution, same sum of 100 % solution and distilled H2O were added together. Equal sum of 100 % and 50 % concentrations were assorted up to do 75 % concentration and so on.

Sowing the seeds

The seeds were primed in 6 different Petri dishes for 12 hours harmonizing to its several concentration. Another 6 Petri dishes were labelled. Layers of cotton of the same thickness were put into Petri dishes and moisten with the same volume of vitamin C solution of different concentrations. Soaked seeds with similar size were chosen and took out of Petri dish utilizing a brace of forceps. They are so dried utilizing paper towels and were sown in labeled Petri dishes.

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In the first test, 25 seeds were soaked in different concentrations of vitamin C solution while another 25 seeds were non soaked before being sown. This is to find whether it is necessary to soak seeds or non before seeding.

36 hours after seeding seeds

Concentration/ %

Number of seeds germinated

Soaked seeds

0

15

10

16

50

1

Table 1 shows the figure of besotted and unsoaked seeds after 36 hours

In the 2nd test, each group of 25 seeds were soaked in different concentrations before being sown. This is to find the best scope of concentrations of vitamin C for chief experiment.

36 hours after seeding seeds

Concentration/ %

Number of seed germinated

Trial 1

Trial 2

Mean

0

15

16

16

10

18

16

17

25

5

5

5

50

1

3

2

75

2

1

2

100

1

0

1

Table 2 shows sprouting per centum of seeds soaked in different concentrations

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Consequences

After 36 hours of sowing, the figure of seeds germinated was counted and sprouting per centum was calculated. Germination is claimed to hold occurred when the radicle was visibly extended from the seed, stick outing through a break in the seed coat. The consequence was tabulated.

Based on the consequences obtained after tests were carried out, there were a few things that could be done to better consequence. First, more seeds should be used so that the consequence will be more important. For the existent experiment, 50 seeds were used alternatively of 25 seeds.

Seeds were primed for 12 hours because priming is of import as seed coat contains some chemicals that inhibit sprouting. Seed priming is a common agronomic technique shown to do an addition seed energy, every bit good as synchronize and accelerate sprouting, better emphasis opposition, and heighten overall works growing and productivity.12 However, some of the seeds start to shoot and breaks on seed coat could be observed if the seeds were primed for excessively long.

Besides, the scope of concentration to be used in existent experiment was determined to be between 0 % to 10 % concentrations because both showed high per centum of seed sprouting with lone little difference in value.

Seeds soaked in 100 % concentration showed the lowest sprouting per centum. The sprouting of seeds was about wholly inhibited in this solution. Fractures on the seed coats were observed but radicles were non extended significantly.

Besides, after transporting out test experiment, I realized that it was better to seed seeds in shooting tray instead than in Petri dishes as Petri dish is round-shaped which made it hard for me to number the figure of seeds germinated.

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Main experiment

Variables

Manipulated: Concentration of vitamin C solution

Different concentrations of vitamin C are prepared utilizing the same consecutive dilution technique as in tests. But this clip utilizing smaller scope of concentrations of 10 % , 7.5 % , 5 % , 2.5 % , 1 % and 0 % ( distilled H2O ) . But for smaller concentration, micropipette is used alternatively of mensurating cylinder to do up the entire volume of 100ml.

Reacting: Germination per centum ( % )

Germination per centum is calculated utilizing the undermentioned equation ;

Germination is said to happen when radicle emerges ( & A ; gt ; 2mm ) after 36 hours.

Changeless: mass of vitamin C ( 100mg/tablet ) , volume of solution used for soaking seeds ( 20ml/Petri dish ) and volume of solution used to wash cotton ( 60ml/tray ) and environmental factors ( light strength, temperature, and humidness ) , period of soakage, type and figure of seeds. All trays were placed under normal white fluorescent lamp and on a research lab tabular array at 28EsC.

Apparatus and stuffs

Apparatus:

300ml beakers, Petri dishes, mensurating cylinders, shooting trays, stirring rod, micropipette, and stamp and howitzer.

Materials:

Distilled H2O, 100mg vitamin C tablet, cotton, choy sum seeds, and paper towels.

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Safety safeguards

Baseball gloves were worn when managing vitamin C tablet to forestall tablet from taint.

Micropipette was used with attention. The tip of micropipette was ever checked to guarantee that there is no air bubbles indoors it as this may cut down the existent volume required for each concentration.

Glass wares like beakers and mensurating cylinders were handled gently since the setup may interrupt easy. This may take to hurts.

Soaked seeds were transferred gently utilizing forceps as they are really fragile.

Data aggregations of chief survey

Number of seeds used in every concentration: 50

Soaking for 12 hours

Observations after soaking seeds for 36 hours

Concentration ( % )

Number of seeds germinated

Average figure of seeds germinated

Replicate 1

Replicate 2

Replicate 3

0.0

45

44

43

44

1.0

48

48

47

48

2.5

42

39

38

40

5.0

41

42

43

42

7.5

35

35

34

35

10.0

34

33

33

33

Table 2 shows the sprouting per centum for seeds treated with different concentrations

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Sample computation ( taken values from 0 % concentration )

Germination per centum:

=44/50 x 100

=88 %

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Statistical analysis

For this experiment, Pearson product-moment correlativity coefficient ( PMCC ) was used to mensurate the strength of additive dependance between the two variables ; the different concentrations and the sprouting per centum. The values obtained are usually distributed.

The correlativity coefficient, R scopes from -1 to 1. A value of 1 implies that a additive equation describes the relationship between X and Y absolutely for which Y increases as Ten additions. A value -1 implies correlativity in which Y decreases as Ten additions. A value of 0 implies that there is no additive correlativity between the variables.

ten

0.0

1.0

2.5

5.0

7.5

10.0

?x=26

Y

88

96

80

84

70

66

?y=484

x2

0.0

1.00

6.25

25.0

56.25

100.0

?x2=188.5

y2

7744

9216

6400

7056

4900

4356

?y2=39672

XY

0.0

96

200

420

525

660

?xy=1901

Table 3 shows the computations done in ciphering the values of R

Sx = x2 –

= 188.5 – ( 26 ) 2/6

= 75.833

Sy = y2 –

= 39672 – ( 484 ) 2/6

= 629.33

Sxy = xy –

= 1901 – [ ( 26 ) ( 484 ) ] /6

= -196.33 ( 1933 words )

To happen correlativity coefficient, R

R =

R = -196.33/ ( 75.8331/2 x 629.331/2 )

R = -0.899 ( negative mark indicates negative correlativity between variables )

Degree of freedom = N -2

n = # of braces of informations

= 6 – 2 = 4

Degree of significance for two-tailed trial

.10

.05

.02

1

.988

.997

.9995

2

.900

.950

.980

3

.805

.878

.934

4

.729

.811

.882

5

.669

.754

.833

6

.621

.707

.789

Table 4 shows the critical values for PMCC

The value of R obtained was compared with the critical value from Table 4 at 5 % significance degree.

R = 0.899 & A ; gt ; 0.811 ( critical value ) at 5 % significance degree

An analysis utilizing PMCC demonstrated a statistically important negative linear relationship between concentration of vitamin C and sprouting per centum since calculated R value was greater than the critical value at 5 % significance degree.

Therefore, void hypothesis can be rejected.

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Datas analysis

Based on the informations collected in Table 2, it was shown that low concentration of vitamin C did advance sprouting of choy amount to a certain extent. The statistical analysis utilizing PMCC farther verified the correlativity. The tabular array besides revealed that merely low concentration of vitamin C was needed to rush sprouting since under 10 % concentration, seeds germinated at a lower rate ( 66 % ) .

Graph 1 illustrates clearly the general tendency and relationship between the variables, the lower the concentration of vitamin C, the lower the sprouting rate of choy amount. The highest sprouting 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 additive graph demoing that the values obtained were due to certain mistakes that lead to some anomalous consequences. At 5 % concentration of vitamin C, we can see that the sprouting rate is somewhat higher than that of 2.5 % but it is non supposed to be so.

Some of mistakes that may happen during experiment are

Blending mistake when fixing solutions of different concentrations by consecutive dilution.

The thickness of cotton bed in shooting tray may non be changeless throughout the tray so vitamin C solution with thinner cotton bed may acquire evaporated faster therefore soaking up of solution is less for the seed in that tray

Some of seeds may hold been ab initio damaged so sprouting may non go on

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The sprouting per centum was compared with the consequence from other published academic diaries. The sprouting per centum against twenty-four hours graph below shows the consequences from diary titled Effect of vitamin C and folic acid on seed energy response and phenolic-linked antioxidant activity from beginning 12. It shows that the optimal concentration of vitamin C for pea seed to shoot is 500micromol/dm3.

Using appropriate computations, it can be calculated that 10 % concentration is about equal to 500micromol/dm3.9 Thus, the sprouting per centum of pea seed in 10 % concentration is highest compared to other concentrations. However, the consequence I obtained for highest sprouting per centum for Brassica parachinensis was different. This can be due to some mistakes and restrictions occurred during experiment or the fact that they are of different types of seed which is further discussed in restriction and alteration portion.

In this experiment, the invariable or controlled variable is the usage of distilled H2O ( 0 % concentration ) . Since the consequences showed that the difference between sprouting rate between 0 % and 1 % concentration was really little ( merely approximately 8 % ) , this indicated that the consequence of vitamin C on seed energy may be excessively small. Even though the consequence is little, it can non be denied that this small aid from vitamin C can give a immense impact on agricultural sector as it is able to advance seed sprouting therefore bettering mass production of workss instead than irrigating seeds with H2O merely.

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Evaluation

Seeds are exposed to assorted biotic and abiotic emphasiss during sprouting. Oxidative emphasis for illustration, caused by the production of high degree of reactive O intermediates such as O2, O2- , H2O2, and HO- can take to the devastation of cells.4 Presence of low concentration of exogenic vitamin C stimulates seed vigour as it minimises oxidative emphasis by scavenging these reactive O species.

Many oxidizers contain an odd negatron so they are extremely reactive. These free groups are damaging because they can interact with other molecules ensuing in a whole concatenation of free extremist reactions. Ascorbate can end these chained extremist reactions by functioning as a stable ( electron + proton ) giver in interactions with free radicals.4

However, high concentration of vitamin C solution inhibits seed sprouting because as solution becomes more concentrated, more H2O from cells diffuse out doing cellular membrane leakage.3

Besides that, being a good negatron giver, extra ascorbate in the presence of free metal ions can non merely advance, but besides initiate free extremist reactions, therefore doing it a potentially unsafe pro-oxidative compound in certain metabolic contexts.8 We hypothesize that whether vitamin C has a net pro oxidizer or antioxidant consequence depends on the concentration of vitamin C solution.8

With relation to bettering seed energies through stimulation of phenolic synthesis, it has been suggested that exogenously applied phenolic antioxidants may be able to excite endogenous phenolic content in workss ( Randhir and Shetty, 2003 and Randhir et al. , 2002 ) .

Due to its built-in antioxidant potency, vitamin C seems to be a suited growing regulator in increasing seed energy and phenolic evocation. In this survey, we have hypothesized that the intervention of workss with vitamin C as exogenic seed interventions may ensue in an addition in both biochemical and traditional agronomic indexs of seed vigour.11

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Restrictions and alterations

One of the restrictions in this experiment is the altering encompassing temperature. The seeds were left in research lab room where the temperature may fluctuate over clip. This may impact the germinating clip of the seeds and the figure of germinated seed after 36 hours. Besides temperature, humidness and light strength were besides uncontrolled. Since all seeds were exposed to the same environmental factors, the consequence is considered to be dependable.

Besides that, some seeds may hold been ab initio damaged before soaking or seeding. Therefore, vitamin C solution may non be the existent cause of suppression of seed sprouting. The seeds are chosen merely by physical steps such as similarity in size, form and coloring material and seed coat is non broken.

However, restrictions can be overcome by utilizing a big figure of seeds and the experiment is repeated for a few times to guarantee that consequence obtained is dependable and adequate to back up decision. This probe tests merely on the effectivity of vitamin C solution on seed sprouting without exposing seeds to other biotic emphasis such as salt or abiotic emphasis such as exposure to sunshine.

Alterations could be made by presenting a biotic or abiotic emphasis to the seed and detect if vitamin C still stimulates seed sprouting or otherwise. Different types of seed may hold different consequence with vitamin C solution. So, utilizing seeds from one type of works may non be able to bespeak how vitamin C really acts on seed. As vitamin C may heighten seed energies in certain species, it could make otherwise to other species. Hence, the same experiment should be done to seeds from other species to see how they are affected by vitamin C solution.

Decision

The lower the concentration of vitamin C, the higher the sprouting rate of Brassica parachinensis seeds. This is due to the antioxidant belongingss of vitamin C which acts against the oxidative emphasis. The optimal vitamin C concentration for seed sprouting is 1 % .

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Beginning Evaluation

I have referred to assorted beginnings in carry oning my research.

Beginnings 1, 2, 3 and 4 are from Wikipedia which has been the largest and most popular general mention work on the cyberspace. Although many have uncertainties over its dependability as it can be edited by anyone with entree to the site but it is cited and linked to many other literatures and academic researches around the Earth. Beginning 5 is a website written by wellness experts on wellness and medical specialty. Beginning 6 is a website that provides information written by university bookmans with mention to other academic researches. Beginning 7 is dependable as it is written by Dr. Bryan D. McKersie of University of Guelph and is posted on the cyberspace in 1996. Beginning 8 is a dependable beginning as it is an article by The American Society for Nutritional Sciences and was published in The Journal of Nutrition web site in 2004. Beginning 9 has been a referral site to many people around the universe. The information seems dependable as it is written by experts.

Beginning 10 is a published book therefore the information is dependable.

Beginning 11 is a diary from ScienceDirect which I have accessed from my campus library. ScienceDirect is a taking full-text scientific database offering journal articles and book chapters from more than 2,500 peer-reviewed diaries and more than 11,000 books. There are presently more than 9.5 million articles/chapters, a content base that is turning at a rate of about 0.5 million add-ons per twelvemonth. It is operated by the publishing house Elsevier. Source 12 is a diary advised by an international Editorial Board and is abstracted or indexed by assorted research databases like the EBSCO and Elsevier. Source 13 is an article from the Journal of Biological Chemistry.