Exercise 1

Seed Germination and

Seedling Growth
Introduction
A seed contains an embryonic plant in an inactive condition, and germination is
its resumption of growth. It is packed with enough stored energy to power the young
seed until it can capture its own energy, as a plant, from the sun (B. G. Bareja, 2011).
These important seed benefits would be impossible without the plant’s first life process,
which is seed germination. The simplest way to understand the germination of seeds is
to think of it as its transition from childhood into adulthood. The seeds grow or develop
into plants (Seed Germination, 2015).
The first change when seeds are placed to germinate is the imbibition of water.
The term imbibition means the absorption of water by the solid particles of an adsorbent
without forming a solution (S. Kumar, n.d.). The development starts as soon as water is
absorbed but in others germination does not take place until further requirements are
fulfilled. In order for a seed to germinate, it must be viable; the seed embryo must be
alive. The experiment assessed the effect of some external factors on germination and
growth, including: gas composition, temperature, hydrogen ion concentration (pH) and
osmotic concentration. These external factors must be favorable for chemical processes
to occur within the seed that allow germination to occur.
Even if all favorable environmental conditions needed for germination are
present, seed germination may still not occur. Several factors affect whether a seed will
germinate or not. Most seeds produced by mature plants pass through a period of
inactivity from germination. Seed dormancy has been defined as the incapacity of a
viable seed to germinate under favorable conditions (Bewley, 1997). Germination
process is an important step in the life of a seed plant because if it occurs during
unfavorable conditions the seedling is unlikely to survive.
The aim of this study is to have an overview of the relationship between the
physiology of the seed and its external environment as it goes through the initial state of
growth, to be able to assess the effect of some external factors on seed germination
and seedling growth. Using Vigna radiata (mungbean), it served as their model plant for
germination and growth. The mungbean seed is easily observable and has a short
germination time. This experiment is designed to introduce the seed germination along
with the growth of seedling and the effect of external factors that allows it to function. It
also provides them with some general background information of the seed germination.

Methodology

Plant used: Vigna radiata (mungbean)

Before preparing the materials needed, the students organized the seeds
depending on its shape or size and only a number of 10 seeds were grouped for each
external factor. All flasks and petri dishes are properly labeled.

Equation for germination percentage: GP = seeds germinated ÷ total seeds x 100

Equation for the mean length of the hypocotyl root axis: sum of the length of the
hypocotyl root ÷ no. of seeds germinated ÷ total seeds = mean length
• Effect of Gas Composition

The students subjected the seeds into different types of gas composition to
observe where seeds will germinate properly. To control the effect of gas composition in
the seed germination of mungbean, two Erlenmeyer flasks used that measures 250ml
and a rubber stopper-glass tubing. A piece of cotton was spread at the bottom of each
flask and poured 25ml of distilled water. Then, 10 same size seeds are placed in each
flask. The students then plugged the first flask with cotton after that the second flask
was plugged with a rubber tubing which is inserted into the rubber stoppers. The rubber
tubing was instantly closed with a pinch cock. Afterwards, the flasks stayed at a room
temperature for four days for which it was observed in the second and fourth day. The
students calculated the number of seeds that germinated and measured the length of
the hypocotyl-root axis. The germination percentage and average length of the
hypocotyl-root axis of the seedlings was then recorded.

• Effect of Temperature

The effect of temperature can be controlled by putting three petri dish, each dish
in different temperature: one in the room, other in the refrigerator and last in the oven.
The petri dishes were lined with filter paper which is moistened with 8 ml distilled water
and the 10 matching seeds in each petri dish were put. The students then recorded the
percent germination observed and measured the length of the hypocotyl-root axis of the
seeds in each of the 3 set-ups after second and fourth day.

• Effect of hydrogen ion concentration (pH)

The effect of pH was controlled when the group put water and different pH (pH 4,
pH 7, and pH 10) on four Petri dishes with filter paper. They moistened one petri dish
with 8ml distilled water, and the remaining three petri dishes was pipetted each with the
indicated 8ml treatment solution. Afterwards, they spread 10 similar seeds in each petri
dish and incubated at the room temperature for 4 days. They counted the number of
seeds that germinated and measured the average length of the hypocotyl-root axis of
the seedlings in each petri dish.

• Effect of varying Osmotic Concentration

The effect of osmotic concentration can be controlled by increasing the

concentration (0.5%, 1.0%, 1.5%) of NaCl in 4 Petri dishes. First, the students lined
each with filter paper then they moistened one Petri dish with 8ml distilled water and the
rest with 0.5%, 1%, 1.5% NaCl solutions. They have placed 10 seeds in each petri dish
and incubated all at room temperature. After that, they recorded the % germination,
observed and measured the length of the hypocotyl-root axis of the germinated seeds in
each after second and fourth day.

Results

• Effect of Gas Composition

• Ordinary Air (control)
Day Day 2 Day 4
 GP: 20% GP: 90%
Mean Length: 0.2cm Mean Length: 2.34cm

• Carbon Dioxide
Day 1 Day 2 Day 4

GP: 0% GP: 0%
Mean Length: 0cm Mean Length: 0cm

Graph 1: Bar Graph for the Mean length of the hypocotyl-root axis in the Effect of
Gas Composition

• Effect of Temperature

• Room Temperature (30-32 ˚C)

Day 1 Day 2 Day 4

 GP: 60% GP: 100%
Mean Length: 0.82cm Mean Length: 1.87cm

• Low Temperature (refrigerator: 10˚C below)

Day 1 Day 2 Day 4

GP: 0% GP: 0%
Mean Length: 0cm Mean Length: 0cm

• High Temperature (oven: 70˚C)

Day 1 Day 2 Day 4

GP: 0% GP: 0%
Mean Length: 0cm Mean Length: 0cm
Graph 2: Bar Graph for the Mean length of the hypocotyl-root axis in the Effect of
Temperature

• Effect of pH

• Distilled water (control)

Day 1 Day 2 Day 4

GP: 50% GP: 80%

Mean Length: 0.56cm Mean Length: 1.71cm

• pH 4
Day 1 Day 2 Day 4

GP: 0% GP: 20%

Mean Length: 0cm Mean Length: 0.55cm

• pH 7
Day 1 Day 2 Day 4
 GP: 0% GP: 0%
Mean Length: 0cm Mean Length: 0cm

•pH 10
Day 1 Day 2 Day 4

GP: 0% GP: 0%
Mean Length: 0cm Mean Length: 0cm

Graph 3: Bar Graph for the Mean length of the hypocotyl-root axis in the Effect of
Hydrogen Ion Concentration (pH)
• Effect of varying Osmotic Concentration

• Distilled water (control)

Day 1 Day 2 Day 4

GP: 70% GP: 100%

Mean Length: 0.34cm Mean Length: 1.79cm

• 0.5% NaCl

Day 1 Day 2 Day 4

GP: 70% GP: 90%

Mean Length: 0.34cm Mean Length: 0.88cm
• 1.0% Nacl
Day 1 Day 2 Day 4

GP: 0% GP: 80%

Mean Length: 0cm Mean Length: 0.48cm

• 1.5% NaCl
Day 1 Day 2 Day 4
 GP: 0% GP: 70%
Mean Length: 0cm Mean Length: 0.30cm

Graph 4: Bar Graph for the Mean length of the hypocotyl-root axis in the Effect of
Osmotic Concentration

Discussion of Results

Effect of Gas Composition

Air is essential for plants to germinate. The students subjected the seeds into
different types of environment to observe where seeds will germinate properly. (S.
Samiksha, n.d.) Failure to control this factor will have an incomplete observation. It is
required by the germinating seed for metabolism. It is also used as a part of aerobic
respiration until it manages to grow green leaves of its own. The effects of oxygen on
seed germination are complex and not always very well understood. Most seeds require
oxygen to germinate, but this requirement largely depends on the species and the
dormancy state of the seeds. It is the embryo that requires oxygen, but the covering
structures may reduce oxygen supply to it and thus determine seed sensitivity to this
gas. Moreover, partial or total deprivation of oxygen, which may occur in the natural
habitat of plants, may exert peculiar effects that are by no means negligible. (J. Kigel,
1995)
The ordinary air (control) is the most favorable for any plants as the result shows
that the control has the number of most germinated seeds. It is obtained in 90%
germination in an ordinary air since it is composed of mostly oxygen gas whereas most
seeds require an adequate supply of oxygen during germination. Oxygen is required for
respiration to oxidize starches, fats, and other food reserves, and its utilization is
proportional to the amount of metabolic activity. And seeds undergo aerobic respiration
in which oxygen is required to produce 36 ATP. It needs oxygen and produce carbon
dioxide (CO2). This carbon dioxide needs to be able to move away from the seed. If the
soil or media is not well aerated due to overwatering or compaction, the CO2 will not
dissipate and seeds can suffocate (S. T. DuPont, n.d.) In Carbon Dioxide, seeds do not
germinate in pure carbon dioxide gas. No germination occurred because there is no
ATP produced and seeds undergo in aerobic respiration in which oxygen is required to
produce ATP enabling them to germinate.

Effect of Temperature
Temperature greatly affects the germination; it causes different enzymes to
activate. We will subject the seeds into different temperature to observe what suitable
temperature for them to germinate is. This affects growth rate as well as metabolism of
the seed. Quite obviously different seeds require different optimum temperatures (E. H.
Roberts, 1988). Temperature affects the rate at which water is imbibed as well as the
rate of metabolic processes such as the translocation of nutrients and hormones, cell
division and elongation, and other physiological and biochemical processes. According
to Hartmann et al. (n.d.), temperature is the single most important factor in the
regulation of the timing of germination, because of its role in dormancy control and/or
release, or climate adaptation. A suitable temperature is necessary for the germination
of seeds and their further growth. For every seed minimal, maximal and optimal
temperatures exist and can be conveniently worked out (C.S.V., 2006). The minimal
and maximal temperature varies for plant to plant. Seeds not usually germinate below
5˚C and above 45˚C and the best temperature often lies between 25-30˚C.
Each individual species has a base and ceiling temperature that represents the
extremes at which germination can occur. In the case of mungbeans, the base and
ceiling temperatures are 7.5 0C and 40 0C respectively. Temperature affects the
percentage of seeds that germinate. Seeds kept in higher temperatures are more likely
to deteriorate and not germinate. Same in low temperature, the extreme temperature of
the refrigerator and oven inhibited the germination of the seeds. If there is a presence of
extreme temperature, enzymes will be denatured and be ineffective, thereby no seed
germination could take place. Thus, only the room temperature, an optimal temp,
obtained a 100% of germination among other temperatures.

Effect of pH
Hydrogen ion concentration is an important factor in seed germination.
Germination will not proceed if the environment is too basic or too acidic. The influence
of pH value on cumulative seed germination indicated no germination below pH 3 or
above 12. Different seeds displayed characteristic pH tolerance limits and requirements.
However, when the effects of acidity are eliminated, the inhibitory effect which is due to
a toxic substance can therefore be considered (H. Lieth, 1993).
The hydrogen ion concentration (pH) also affects the seed germination. The data
indicated that either the lower or higher the pH concentration, lower the chance to
germinate. Plant cannot grow without nutrients. As the result, the mungbeans that were
placed in pH 4 didn’t grow on the second day but germinate up to 20% and has a mean
length of 0.55cm. According to Cheung, Wong and Yen (1998), “When the plants are
placed in acidic environment, there is not enough nutrients for the plant because
hydrogen ions remove the other positively charged ions from the environments where
the plants are in.” In pH 7 and pH 10, no seeds germinated because higher pH level will
hinder germination and growth. As for the control group (distilled water), 100% of the
seeds germinated. This is because germination happens at pH that is most favorable
and distilled water does not have any impurities so the germination is high. On the
second day the seed germinated up to 50% with the length of 0.56cm and as expected
the percent of germination increases up to 80% with the mean length of 0.56cm and as
expected the percent of germination increase up to 80% with the mean length of
1.71cm. At high or low pH, protein is being denatured resulting to low germination. The
acidic pH level aid slow process of germination as well as the alkaline environment.

Effect of Varying Osmotic Concentration

Seed germination is also affected by varying osmotic concentration. The solution
may able to inhibit the seed germination and to suppress the growth of mungbeans
when the osmotic concentration of a solution is low. The analysis of the above results
indicated significant decrease in germination percentage of the mungbean as the
salinity of the NaCl increases. Data showed that the mungbeans from 1.5% NaCl has
the least germination percentage of 70% with 0.30 cm length of the hypocotyl-root axis
on day 4. According to the study of Aref, El-Juhany, & Elkhalifa (n.d.), “the inhibition of
seed germination under NaCl concentrations is explained as a result of lowering
osmotic porential”. The passage of NaCl ions along the normal diffusion gradient od
seeds resulted in both change in ionic composition of the cell and decrease in water
potential (Crawford, 1978). This is the reason why high concentrations of salt reduces
the rate of seed germination, thereby also the germination percentage. In other words,
its salinity prevents the seed from absorbing water or moisture which is needed for them
to sprout. Followed by the germination of seed on 1.5% NaCl, the 1.0% NaCl having
80% germination and 0.48 cm mean length has fewer seeds germinated than with 0.5%
NaCl due to the same explanation about varied NaCl concentration. On the other hand,
the mungbean from the distilled water raised from 70% to 100% with 1.74 cm on day 4
indicating that it has the most seeds that germinated. Thus, when the concentration of a
plant extract is high, the suppression of growth may be due to the osmotic effect. This
controlled variable resulted in such because the distilled water does not contain any
concentration of sodium chloride that could make the seeds difficult to conserve
moisture properly, thus making them susceptible to dry up. This entails that the seeds
germinated better in distilled water than any other concentrated solutions.

Study Questions

1. What is the purpose of displacing air with CO2 gas?

Displacing air with CO2 will help in eliminating oxygen thereby reducing oxygen
pressure. Lack of oxygen retards aerobic respiration but it then subjected to anaerobic
respiration due to the absence of O2. Due to the displacement of air with Carbon
Dioxide, the effect of gas composition on seed germination may be evaluated. This gas
seems useful to plants only insofar as they can break it down, and seeds, at the earliest
stage of their development, do not seem to carry out this decomposition appreciably.
However, as germinating seeds produce too much carbon dioxide gas for us to deprive
them of it entirely, it is impossible to decide if its complete absence is harmful or helpful
to them.

2. Why is germination usually low in soils that contain high amount of partially
decomposed organic matter?
 Germination is the process by which a plant grows from a seed while
decomposition is a biological process that includes the physical breakdown and
biochemical transformation of complex organic molecules intro simple molecules.
Successive decomposition of dead material and modified organic matter results in the
formation of a more complex organic matter called humus. Humus affects soil
properties. As it slowly decomposes, it colors the soil darker, increases soil aggregation
and aggregate stability, increases the ability to attract and retain nutrients, and
contributes nitrogen, phosphorous, and other nutrients. In that case, it is needed for
germination to take place in an environment which contains high decomposed organic
matter in order to get enough nutrients essential to grow plants.

3. Does light affect seed germination? What are photodormant seeds?

In order for the seed to germinate, favorable environmental conditions are

needed for them to grow. In some seeds, light does not have an effect on them. They
do not require light for germination to occur since most of their nutritional needs are
already contained in the seed itself. However, there are also some seeds that do require
light to germinate. These are the seeds that depend on light wherein if sufficient light is
absorbed, seedlings will begin to sprout.

Photodormant seeds refers to the seeds that wither require light or dark
conditions to germinate. It is said to be a form on endogenous nondeep physiological
dormancy. Examples of such are lettuce seeds and tobacco seeds. Dark-germinating
seeds include corn and mungbean seeds.

Conlcusion

The germination of a seed and seedling growth varies greatly with respect to
some external factors or environmental conditions that may either enabling their growth
or inhibiting them. From the gathered data, the effects of gas composition, temperature,
hydrogen ion concentration (pH) and osmotic concentration on seed germination were
determined through the analysis of germination percentage and the mean length of the
hypocotyl-root axis. In the effect of gas composition, seeds would germinate through on
ordinary air since it is composed mainly of oxygen, carbon dioxide, nitrogen and any
other elements essential for the seed to grow. This means that seeds will germinate in
an oxidized environment. But in carbon dioxide, imbibition of growth would definitely
occur. In the effect of temperature, mungbean grows best on room temperature rather
than on low or high temperature since the said type of seed is root capable to germinate
on extreme temperature or environment. seeds will only germinate on its optimum
environment. Furthermore, in the effect of pH, optimum hydrogen ion concentration (pH)
enables the seed to germinate. Seeds will germinate on environment where there is
right pH. However, if introduced in more acidic or basic environment, nutrients are being
denatured by the change in pH causing for deterioration of enzymes responsible for
growth. Lastly, in the effect of osmotic concentration, high concentration of salt results
on a decrease in the percentage germination since it reduces moisture for the seeds to
germinate. Seeds will germinate in an environment where water is deficit; seeds will
germinate if water is present, water is a prerequisite for triggering germination.
Generally, to be able for the seeds to germinate and grow properly we should consider
the requirements for oxygen, temperature, moisture and other factors.

References

Ben G. Bareja. Crops Review (2011, November) The Reproductive Organ of the
Angiosperms Plant Seed. Retrieved from http://www.cropsreview.com/plant-
seed.html

Byhju’s. (2015, December 24) Seed Germination. Retrieved from

http://byjus.com/biology/seed-germination/

Helmuth Leith and Ahmed A. Al Masoom. (1993). Towards the rational use of high
salinity tolerant plants. Vol. 1 p. 430

Jonathan, D.i (n.d.) Seed Plants. Retrieved from http://www.ck12.org/biology/Seed-

Plants/lesson/Seed-Plants-BIO/
Srinibas Kumar. Biology Discussion (n.d.) Imbibition: Conditions, Characteristics,
Factors, Role and Significance. Retrieved from
http://www.biologydiscussion.com/plants/imbibition

Leónie Bentsinka and Maarten Koornneef. The Arabidopsis Book (2009) Seed
Dormancy and Germination Retrieved from https://www.ncbi.nlm.nih.gov

Samiksha S. Your Article Library (n.d.) Major Factors Necessary for Seed Germination
in Plants (Internal and External Factors). Retrieved from
http://www.yourarticlelibrary.com/biology/plants/

S. Tianna DuPont. Penn State Extension. (n.d.) Seed and Seedling. Retrieved from
http://extension.psu.edu
Suyash Kothari. Competition Science Vision (CVS) (2006, November) Germination and
Dormancy of Seeds. pp. 1224-1225