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An Investigation into the effect on Phaseolus vulgaris (var.

Improved Golden

Wax) seeds in different solutions

Dr. Melville

ED 4205 YA

Marnie Merchant

Ashley Johnston

10 Pages

Introduction
For this experiment, we will be investigating the effect on the
germination of Phaseolus vulgaris (var. Improved Golden Wax) seeds of
different solutions. As a guiding scientific question, we want to see how the
mass percentage increases in seeds of Phaseolus vulgaris (var. Improved
Golden Wax), based on the liquid they are soaking in after 168 hours.
Germination is a complex metabolic process that involves many steps
in the initial stages of a plants life cycle.1 The overall process oxidizes the
lipids and carbohydrates within the seed and breaks down storage proteins
so that it can release the energy and amino acids that are needed for further
plant development.1 The three main stages involved are pre-germination,
germination and post-germination. 1 For the purposes of this research we are
just focusing on the pre-germination stage which describes the absorption of
water uptake by the bean seeds, as well as the different liquids that may
have an effect on the results. 1
Germination begins with imbibition, which is the uptake of water by the
dry seed. This process is usually completed when the radicle, which is part of
the embryo, pushes through the structure that is surrounding it.1 During
imbibition, the amount of water uptake by the dry seeds is quite large, which
greatly increases the seed size before the initiation of active metabolism
begins.2 The many other events occurring include protein hydration,
subcellular structural changes, respiration, macromolecular syntheses, and
cell elongation. Overall, these processes together result in transforming a
dehydrated embryo to an embryo with vigorous metabolism in growth.3
Some external factors that need to be considered in the germination
process are favourable temperature, light, oxygen, and water. Temperature
can affect the germination percentage as well as the germination rate.4 Most
plants germinate best at a room temperature of 21C. Light is not essential
in the early stages of germination, but it does play a main role later on in the
cycle. Depending on the crop, light can either stimulate or inhibit seed
growth.4 Oxygen is necessary for plants to grow as it is involved in cellular
respiration. If there is a lack of oxygen, there may be a disturbance in the
germination process.4 The most important factor being water, is essential to
the germination process. Since dry seeds will not germinate, the water is
needed to trigger the process of seed germination.4
The independent variables used in this experiment are the four
different solutions. The solutions were changed by adding salt, sugar, and
vinegar to the tap water. The dependent variable used in this experiment
was the mass of Phaseolus vulgaris (var. Improved Golden Wax) seeds. The
mass is expected to change based on the different solutions. The control set-
up used in this experiment was the tap water solution. This was used as a
baseline to compare the mass increase of the other solutions.
Since this study is focusing on the mass percentage increase of the
seeds based on their absorption of the liquid, the effects of salt, sugar, and
vinegar will be observed. There have been previous studies completed on
the effects of salinity in germination. The effect of salinity can either create
osmotic potential which will prevent the uptake of water or by the toxic
effects of ions on the embryo viability.1 Since salt absorbs and retains water
so strongly, it can cause a stress which may cause significant reductions in
the rate of final germination percentage which can lead to reduction of
yields.1 Since the mass percentage increase of the seeds is the object of
concern, the effect from the salt is not as detrimental as if the seeds were
going through the full germination process. The effect of sugar water can
also play a role in osmosis. The sugar water will lower the osmotic potential
because there will be less water available to the seed. The acidity of water
can also have an effect on the germination process.
Focusing on just the water uptake of the seeds and its absorption, for
this experiment, it is predicted that the tap water solution will absorb the
most water after the 168 hours. This is due to the fact that the salt, sugar
and vinegar concentrations cannot interfere and absorb any of the water
itself. Based on previous research, it is predicted that the salt water will have
the least mass increase because of the potential for osmosis to occur.
Hypothesis

If a Phaseolus vulgaris (var. Improved Golden Wax) seed is placed in


tap water, then the mass percentage will increase more than in other liquids.

Materials

4 Phaseolus vulgaris (var. Improved Golden Wax).


250 mL tap water
1 Cup (14.79 g) of Redpath granulated table salt (Sodium chloride-
NaCl) in 250 mL of water at 21C.
1 Cup (14.79 g) of Redpath special fine granulated sugar (sucrose-
C12H22O11) in 250 mL of water at 21C.
14.79 mL Heinz distilled white vinegar (CH3COOH) in 250 mL of water
21C.
1 JZ Scale 115
4 - 500 mL Mason Jars
1 - 250 mL Beaker
Measuring Spoons
Thermometer
Box
Stirring Rod

Method

In this experiment, four different liquids are looked at to see the mass
percentage increase in Phaseolus vulgaris seeds after soaking in the liquids
(salt water, sugar water, tap water and vinegar water) for 168 hours. Three
seeds were used in each liquid to get an average mass percentage increase
for the seeds.
To begin the experiment, weigh out three beans using a scale ( 0.02 g)
for one mason jar and label the jar. Repeat this procedure three more times.
Label the mason jars (salt water, sugar water, tap water and vinegar water,
each with three beans per jar. Next, using a beaker, measure out 250 mL of
tap water at 21C and place in a mason jar. Do this for all four mason jars.
Using a measuring spoon, measure 14.79 g of Redpath granulated table salt
and mix into the mason jar labelled salt water. Using a stir stick, stir the
contents for approximately two minutes, or until the salt appears fully
dissolved. Rinse the stir stick off with tap water.
Next, repeat the above procedure using granulated sugar. Using the
measuring spoons, measure 14.79 g of Redpath granulated sugar and mix
into the mason jar labelled sugar water. Using a stir stick, stir the contents
for approximately two minutes, or until the sugar appears fully dissolved.
Rinse the stir stick off with tap water.
Repeat the above procedure using Heinz Vinegar. Using the measuring
spoons, measure 14.79 mL of Heinz distilled vinegar and mix into the mason
jar labelled vinegar water. Using a stir stick, stir the contents for
approximately two minutes, or until the vinegar appears fully mixed. Rinse
the stir stick off with tap water.
Finally, when all four mason jars are labelled and filled with their liquids,
place the relative three beans into each of the mason jars. Place all four
mason jars in a dark box in a room temperature of 21C. The box will remain
in the dark for 168 hours. Once the 168 hours are up, using the scale, weigh
each individual bean seed and record its mass in a table. Using the table,
calculate the mass percentage increase using the formula found in the
appendix. This percentage will show how much water the bean seed has
absorbed during the 168 hours.

Results

Table 1: Mass of Bean prior to absorption and after absorption.


Ingredient Bean Mass Prior to Bean Mass After Absorption Mass
s Absorption (168 Hours) Percenta
ge
Increase
Tap Water Bean #1: 0.33g Bean #1: 0.79g 56%
21C Bean #2: 0.34g Bean #2: 0.78g
Bean #3: 0.35g Bean #3: 0.76g
Average bean Mass: Average bean mass: 0.78g
0.34g

Salt Bean #1: 0.26g Bean #1: 0.29g 52%


Water Bean #2: 0.26g Bean #2: 0.64g
Bean #3: 0.26g Bean #3: 0.68g
Average bean Mass: Average bean mass: 0.54g
0.26g

Sugar Bean #1: 0.41g Bean #1: 1.04g 59%


Water Bean #2: 0.42g Bean #2: 0.95g
Bean #3: 0.43g Bean #3: 1.09g (0.38 +
Average bean Mass: 0.71bean split in half)
0.42g Average bean mass: 1.03g

Vinegar Bean #1: 0.33g Bean #1: 0.72g 54%


Water Bean #2: 0.34g Bean #2: 0.77g
Bean #3: 0.35g Bean #3: 0.72g
Average bean Mass: Average bean mass: 0.74g
0.34g

Discussion

In this experiment, the main purpose of this investigation was to look


at the effect on Phaseolus vulgaris (var. Improved Golden Wax) seeds in
different solutions. Specifically, we wanted to see how the mass percentage
increases in seeds of Phaseolus vulgaris (var. Improved Golden Wax), based
on the liquid they are soaking in after 168 hours. The four different liquids
studied were tap water, salt water, sugar water and vinegar water. The
average bean mass prior to absorption in tap water was 0.34 g and the
average mass after absorption was 0.78 g. This resulted in a 56% mass
increase. The average bean mass prior to absorption in salt water was 0.26 g
and the average mass after absorption was 0.54 g. This resulted in a 52%
mass increase. The average bean mass prior to absorption in sugar water
was 0.42 g and the average mass after absorption was 1.03 g. This resulted
in a 59% mass increase. The average bean mass prior to absorption in
vinegar water was 0.34 g and the average mass after absorption was 0.74 g.
This resulted in a 54% mass increase. Over all, our hypothesis was not fully
supported by our results, since we originally assumed that the beans in tap
water would increase the most, but instead, the beans placed in sugar water
increased the most.
We originally assumed that our seeds placed in just tap water would
absorb the most liquid because this would be its more natural form of water
intake. However, the sugar water seeds mass increased the most. We believe
this is because sugar is used as a natural fertilizer in hydroponic plants. We
believe that the seeds placed in salt water had the lowest mass because the
salinity put stress on the seeds which made it difficult for them to absorb the
water concentration necessary for the enzyme production. Since salt lowers
the osmotic potential of water located in soil, making water less available to
plants and potentially shrinking the cell membrane, there is scientific
reasoning that explains why the seeds placed in salt water increased the
least. In addition, Janousek and Folger research proves that nearly all plants
germination stage is negatively impacted when salinity concentrations are
high. In their study, the seeds they placed in fresh water grew stronger than
the plants that were placed in an area with a high concentration of salt.
Another important aspect in our observations was that only one of the seeds
in the salt water held close to the same mass. We believe that since the
other two seeds increased slightly more than the one seed, that an error
occurred. This could be simply due to a corrupt seed from the packaging.
This would be an error from the manufacturer of the seeds. Gardeners
commonly see this occur: for example, when they plant a hundred seeds
they often expect a few seeds not to grow. According to Azooz and Parvaiz,
certain seeds outperform others, due to their stress tolerance. There are
several factors that affect the stress of the soil, such as salinity and acidity of
water and/or soil concentrations (Azooz Parvaiz).
As predicted, the beans in the vinegar water also had a difficult time
absorbing water. This is most likely due to the acidity of the water which has
harmful effects on beans. All seeds require a specific pH level in order to
succeed full potential. The optimal pH range for most plants is between 5.5
and 7.0; however, many plants have adapted to thrive at pH values outside
this range (Soil Acidity). Moreover, we believe that the vinegar changed
the pH levels enough that the seed was unable to germinate to its full
potential.
Therefore, since seeds naturally require water in the imbibition phase
of germination, it makes sense that all of the seeds should increase in mass.
The seeds placed in sugar water most likely increased the most since sucrose
can be used as an enzyme to enhance plant growth.
Possible sources of error could have occurred in the set up of the
experiment involving human error. Such error could include inaccurate
measuring of the liquids or loss of products during transfer. Other sources of
error could include uncontrollable temperature changes in the household the
mason jars were kept in. Even with the house thermostat being set at 21C,
there could always be a slight change throughout the day. Lastly, it is likely
that some beans were defective and unable to perform the process of
germination due to the manufacturing.
If this experiment were to be carried out again in the future, there
should be some modifications to the procedure. Firstly, determining a better
ratio of sugar, salt and vinegar to water would be recommended. In our
experiment, the salt and sugar solutions broke down after sitting for two
days; meaning that the solid (sugar and salt) sat at the bottom of the Mason
jars with the seeds. Perhaps decreasing the amount of each solute might
allow for the solutions to stay mixed for the required time length. It is
possible that once the solution became undissolved and solutes laid
concentrated at the bottom, that there is a higher stress level on the seeds
during the imbibition stage of germination, thus leading to less water uptake.
Conclusion

In conclusion, the results of this experiment suggest that the


Phaseolus vulgaris (var. Improved Golden Wax) seeds placed in sugar water
had the largest mass increase of 59% compared to the suspected mass
increase of 56% in regular tap water. An average mass between three seeds
was taken to determine the mass increase. The overall results showed the
sugar water increasing the most (59%), followed by the tap water (56%),
vinegar water (54%) and lastly salt water (52%).
Further research should be done on the effects of sugar and salinity on
plant germination since our environment and food security may be impacted
due to an increase in population, urbanization, climate change and
industrialization. Research has proven that Crops experience different
climate stresses during development. The magnitude of damage will depend
on the phenological stage of the crop and the stress duration. Climate
change could intensify some or all of these stresses, thus negatively
impacting agriculture (Ramirez et al.). It is important to find ways that help
plants adapt to changing/stressful environments. Moreover, we believe this is
a matter of concern for plant biologists trying to reach the goal of food
security.

Appendix

Sample Mass Percentage Increase Calculation:

Tap Water:

Mass of seed after absorption mass of seed prior


absorption x 100
mass of seed after absorption
0.78 g 0.34 g
= 0.78 g
x 100

= 56 %

References

1. Berhanu, A. T.; Berhane, G. African Journal of Plant Science 2014, 8 (5),


225231.

2. Bewley, J. D. American Society of Plant Physiologists 1997, 9, 1055


1066.

3. Bewley, J. D.; Black, M. Seeds: physiology of development and


germination; Plenum Press: New York, 1985.

4. Factors Affecting Germination of Seeds


http://www.tutorvista.com/biology/factors-affecting-germination-of-
seeds (accessed Jan 20, 2017).
Azooz, M. M., and Parvaiz Ahmad. Plant-Environment Interaction : Responses
And Approaches To Mitigate Stress. Hoboken: Wiley-Blackwell, 2016.
Discovery eBooks. Web. 10 Feb. 2017

Janousek, C. N; and Folger, C. L. Inter-specific variation in salinity effects on


germination in Pacific Northwest tidal wetland plants. Elsevier B.V.
November 2013

Ramirez-Cabral, N. Lalit, K., and Subhashni, T. Crop Niche Modeling Projects


Major Shifts In Common Bean Growing Areas. Agricultural And Forest
Meteorology 218-219.(2016): 102-113. ScienceDirect. Web. 10 Feb. 2017.

Soil Acidity. 2017 Soil Quality Pty Ltd. Retrieved from


http://soilquality.org.au/factsheets/soil-acidity . Web January 15 2017.

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