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Introduction

All seed plants produce ovules, containing an egg cell, and pollen, containing sperm cells, and
when the sperm fertilizes the egg, a seed is created (Tschunko et. al 2007, p.51). A key part of the ovule
is the opening near the egg cell called the micropyle (Tschunko et. al 2007, p.51). Gymnosperms and
angiosperms are two categories of seed plants, and the seeds used in this experiment are from an
angiosperm. Fertilization and seed maturation occur in angiosperms by a pollen grain landing on the
pistil’s stigma and growing a long pollen tube through the style into the ovary, which then the sperm is
delivered to the egg through the micropyle opening (Tschunko et. al 2007, p.52). Now that fertilization
has occurred, the seed can begin to grow.

A seed is broken down into three parts: seed coat, embryo, and nutritive tissue. The seed coat is
material from the mother plant that acts as a protective covering and can structurally be modified to aid
in dispersal (Tschunko et. al, p.52). The embryo is the offspring that developed from fertilization and is
attached to fleshy, nutrient-rich structures called cotyledons (Graham, Graham, Wilcox 2007, p.51). The
nutritive tissue gives nutrients to the growing seedling for a limited time until it is able to do
photosynthesis (Tschunko et. al 2007, p.52). The nutritive tissue is also called endosperm in angiosperms
but is absent in mature seeds because it is absorbed into the cotyledons during seed development
(Tschunko et. al 2007, p.52).

Once a seed is mature, it is very dry so the first step of germination is imbibition, the uptake of
water (Tschunko et. al 2007, p.55). The seed swells and if viable, it activates enzymes to do their work
(Tschunko et. al 2007, p.55). Some enzymes digest starch into sugars and storage proteins into amino
acids which are then broken down even further during cellular respiration (Tschunko et. al 2007, p.55).
Energy is released to resume embryo growth while other enzymes and hydrated molecules go through a
cascade of biochemical processes that create a seedling and independent plant (Tschunko et. al 2007,
p.55). Some factors that affect the rate of germination are temperature, light exposure, acidity, and
salinity.

This experiment was done using the Lactuca sativa or known as its common name, Oakleaf
lettuce. It’s grown mostly in the United States and Spain in optimal growth conditions of 21-24 °C, pH
6.0-6.7, and ¼ to ⅜ inches underground (Křístková, Doležalová, Lebeda, Vinter, Novotná 2008, p.115).
It’s also deeply lobed and shaped like the leaves of an oak tree. These seeds were chosen because they
germinate quickly, have a thin tap root, are cheap and have a thin seed coat (Křístková, Doležalová,
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Lebeda, Vinter, Novotná 2008, p.115). Because environmental factors like salt runoff are common
causes of plasmolysis, this experiment was done to see if salinity would effect seed germination.
Knowing how much salt seeds can continue to grow in is important because it affects agriculture,
explains why plants die if exposed to a lot of salt, and helps the community know where to place certain
plants. The hypothesis for this experiment was if seeds are placed in varying salt concentrations, then
the higher concentrations would prevent seed germination.

Materials and Methods

9 petri dishes, 3 per salt solution, were gathered and labeled using a permanent marker with the
salt solution amount and replicate number. Paper towels were cut using scissors into circles and placed
into each petri dish. Graduated cylinders, beakers, balances, weighing paper and spatulas were used to
make salt solution solutions of 0%, 2%, and 5%. Then 16 oakleaf lettuce seeds were arranged in each
dish in rows of 4. Graduated pipets were used to place one drop of each salt solution on each seed,
depending on the designated salt solution for each replicate. Each dish was then sealed with parafilm M.
For 9 days, the parafilm was taken off to water seeds with distilled water then new parafilm was put on
to reseal each dish. Germination was checked daily and recorded using pictures to track the number of
seeds that successfully germinated.

Results

TABLE 1: SEED GERMINATION AVERAGES

Control Treatment #1 Treatment #2

Day 0% 2% NaCl 5% NaCl
0 0% 0% 0%
1 0% 0% 0%
2 0% 0% 0%
3 96% 0% 0%
4 98% 0% 0%
5 100% 0% 2%
6 100% 0% 2%
7 100% 15% 2%
8 100% 21% 4%
9 100% 33% 4%
Table 1 shows the averages for seed germination for all three replicates per salt solution.

GRAPH 1: SEED GERMINATION FOR REPLICATES

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Graph 1
shows the comparison of the average seed germination percentage per salt solution over the course of
nine days.
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Figure 1.1 Day 5 of Control Group (0%) Figure 1.2 Day 5 of 2% Solution

Figure 1.3 Day 5 of 5% Solution

Figure 1.4 Day 9 of Control Group (0%)

Figure 1.5 Day 9 of 2% Solution Figure 1.6 Day 9 of 5% Solution

The first seeds to germinate were the ones placed in the 0% salt solution, the control group.
According to Table 1, 96% of the control seeds germinated on Day 3 and by Day 5, all of them
germinated. Unexpectedly, the seeds placed in the 2% solution took the longest to germinate because it
wasn’t until Day 7 that the first 15% germinated. Also, these had a steady germination rate as seen in
Graph 1, as opposed to the control group that spiked and then plateaued in germination. The 5%
solution germinated the least, as expected, but germinated two days earlier than the 2%. Only 2 seeds
from replicate 1 of the 5% grew in Figure 1.6, giving the average percentage of 4% in Table 1.

Discussion

For mature seeds to germinate they must uptake water, imbibition. Salt however can create an
osmotic potential that prevents water uptake or have toxic effects on germinating seeds due to the Na +
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and Cl- ions (Kaya et. al 2005). When the seeds were placed in the 2% and 5% salt solutions, the outside
of the cell solution became hypertonic which causes water to move out of the cell to dilute the high
concentration. This causes the cell to shrink and more water leaves than being taken in, resulting in
delayed germination (Kaya et. al 2005). Watering the seeds daily provided water for the seeds to
potentially take in, but the higher salt concentrations made the imbibition process more difficult (Kaya
et. al 2005). Seeds in the 5% solution had the most osmotic potential and therefore resulted in the data
shown in Graph 1 and Figure 1.6 to have 4% germination and only two seeds from one of the replicates
to successfully germinate.

Another factor of seed germination that was inhibited was the activation and productivity of
enzymes (Nasri et. al 2017). Enzymes in viable seeds digest storage proteins into amino acids and starch
into sugars through cellular respiration, then go through a cascade of biochemical processes to develop
the embryo but the presence of salt decreased this entire process (Tschunko et. al 2007, p.55). The
binding of an enzyme to its substrate is very specific at the active site and anything that can bind to the
active site that may be foreign is considered an inhibitor (Robinson 2015). The Na + and Cl- ions were
foreign to the seeds and acted as competitive inhibitors which resulted in the lower seed germination
rates for the 2% and 5% solutions in Table 1 (Kaya et. al 2005).

Modifications done to the materials and methods were using 3 petri dishes per salt solution,
totaling 9 petri dishes used and the salt solutions used were 0%, 2%, and 5%. Sixteen seeds were
arranged in rows of four in the petri dishes and only 1 drop of solution was put on the seeds. Other
changes that could be made to see the amount of salt needed to affect germination is to use several salt
solutions, possibly 0.5%, 1%, 3%, and 4%. Also, the salt solution seeds could be watered twice a day
while the control seeds are watered once to see if the more chances the salty seeds have to uptake
water would effect germination. Larger petri dishes could be used so the seedling could freely grow
without the lid squishing the top stems. Planting the seeds in soil could be done to see if the soil would
absorb some of the salt and allow for more imbibition and better germination.
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Bibliography

Kaya, Mehmet Demir, et al. “Seed Treatments to Overcome Salt and Drought Stress during Germination
in Sunflower (Helianthus Annuus L.).” European Journal of Agronomy, vol. 24, no. 4, 2006, pp.
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Křístková, E., Doležalová, I., Lebeda, A., Vinter, V., & Novotná, A. (2008). Description of morphological
characters of lettuce (Lactuca sativa L.) genetic resources. Horticultural Science, 35(No. 3), 113–
129. doi: 10.17221/4/2008-hortsci

Nasri, Nawel, et al. “Effect of Salinity on Germination, Seedling Growth and Acid Phosphatase Activity in
Lettuce.” American Journal of Plant Sciences, vol. 06, no. 01, 2015, pp. 57–63.,
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Robinson, P. K. “Enzymes: Principles and Biotechnological Applications.” Essays In Biochemistry, vol. 59,
2015, pp. 1–41., doi:10.1042/bse0590001.

Tschunko, A., Graham, Graham, & Wilcox. (2007). Plant biology: laboratory manual. Upper Saddle River,
NJ: Pearson.