FACULTY OF AGRICULTURE

UPM CAMPUS SERDANG

BACHELOR OF AGRICULTURAL SCIENCE

AGR 3301 CROP PHYSIOLOGY
TITLE: LAB REPORT SEED VIABILITY

LECTURER: DR. MUHAMMAD NAZMIN BIN

YAAPAR

LAB GROUP :2

NO NAME MATRIX NUMBER

1 Nur Asyran Bin Alimin 210792
2 Aiman Danish Koh Bin Affandi Koh 210783
3 Jeevanish Mohanasuntheram 21070
4 Nur Alyaa Najwa Binti Amran 210617
5 Nurul Fasihah Binti Abu Ubaidah 210750
ABSTRACT
This experiment examined the seed quality and viability by conducting two
types of test which is Tetrazolium test by using a dilute (0.1%) solution of 2,3,5-
triphenyltetrazolium chloride and other conventional method which is using sand
media and paper towel media. A total of 10 seeds for 2 replications are obtained to
undergo TZ test which is maize seeds and bean seeds. The conventional method was
conducted using 10 seeds for 2 replications of maize and beans seed as well. As for
the TZ test, we discovered that the seeds turn red after 1 hour in an incubated oven for
35°C at parts such as scutellum, coleoehizae, radicle and plumule. As for the
conventional method, the amount of germinated seeds are counted after a week. The
germination percentage is required to determine the viability of a population seeds.
The equation to calculate germination percentage is GP = seeds germinated/total
seeds x 100.

INTRODUCTION
A seed is officially considered to have germinated when the young root, called
the radicle, emerges from the seed coat. To germinate, a seed requires three things
which is water, oxygen and a suitable temperature. Water uptake, also called
imbibition is the first stage of seed germination. During this process the dry seed,
which typically has a water content of less than 10% absorb water and swells. This
process serves to hydrate the dry components of the seed and activate the metabolic
machinery necessary for germination. A seed that has no germinated because it is
lacking one or more of the necessary requirements for germination is termed
quiescent. The seeds are simply “resting”, waiting for the appropriate conditions for
germination. The purpose of laboratory testing of seed germination is to assess seed
quality or viability and to predict performance of the seed and seedling in the field.
Germination percentage is a good measurement of see viability.

OBJECTIVES

 To determine the seed viability of maize and bean seed using TZ method

 To compare TZ method with other conventional germination methods

METHODOLOGY

Exercise 1:Tetrazolium test (Chemical test which gives a quick estimate of

seed viability (24-48 hrs.)
i) Maize
1. Obtain 10 seed for 2 replications that were pre-soaked overnight
2. Cut the grains in half (longitudinal) through the embryo
3. Randomly, place 10 seeds (half seed) into a petri dish respectively
4. Pour tetrazolium chloride into petri dish respectively. Make sure all seeds are
covered with tetrazolium chloride
5. Closed the petri dish and allow it incubate in oven (35°C) for about 1 hour or
until seed changes to red. When working with tetrazolium, wear gloves and use a
forceps to handle the grains
6. After taking out the seeds from the oven, wash the seeds with water. Make sure
the TZ does not spill on the bench as color will stain the table
7. Determine the viability percentage by observing parts that are supposed to
become red in color (Diagram is provided). Parts that should be red in color if the
seed is viable are scutellum, coleorhizae, radical and plumule
ii) Bean
1. Same as maize, determine the viability percentage for 2 replications of 10 bean
seeds that pre-soaked overnight
2. Peel off the testa and only half of the beans, which remained the plumule and
radical will be soaked in tetrazolium chloride.
Follow protocol on maize above

Exercise 2: Germination test (test determines the percentage of normal

seedlings that develop under ideal conditions. The percentage of hard or
dormant seed is also determined and reported)
a) Sand media (maize and bean)
1. Obtain 10 seeds for 2 replications for each grain
2. Put clean, sterile sand into trays without hole
3. Water the sand until it is moist. Do not use excess water
4. Make holes at about the same depth as the size of the seeds for each replicate of
the seeds. Ideally, the distance between the holes should be at least three to five
times the seed diameter
5. Fill seeds from each replicate into the holes and cover with sand
6. Water the sand again to cover the seeds, but do not make it too wet
7. Prepare a label with the species, date of sowing and replicate of the test and
place in each tray
8. Count for the germination percentage after 5 -6 days
b) Paper towel media (maize and bean)
1. Obtain 10 seeds for 2 replications for each grain
2. Place two paper towels in the bottom of a tray, one on top of the other
3. Wet the tower thoroughly and tilt the tray up on one end so that excess water
runs off the tray.
4. Place seeds in between the moist paper towels.
5. Roll the paper loosely
6. Place the rolled paper towel in the tray
7. Cover the tray with cling film to prevent the towels from drying out.
8. Place the tray in an upright position in a location of diffuse, not direct light, such
as a north window
9. The location should be warm enough for good plant growth
10. After 5 to 7 days, unroll the paper carefully to avoid tearing the paper or
damaging the roots of the young seedlings. Count the number of germinated seeds
with intact tap roots and shoots.
11. Do not count moldy seed or diseased seedlings.
RESULT :

Conventional methods
GP TZ-TEST SAND MEDIA PAPER TOWEL MEDIA
MAIZE BEAN MAIZE BEAN MAIZE BEAN
REPLICATION 40% 60% 20% 90% 100% 100%
1
REPLICATION 40% 40% - - 100% 80%
2
AVERAGE 40% 50% 20% 90% 100% 90%

Table 1 shows average seed germination %

Seedling Vigor
Seeding Vigor Paper Towel Media
Maize Bean
Replication 1 Replication 2 Replication 1 Replication 2
Height mean 5.48 4.1 5.18 2.05
(cm)
Fresh Weight 0.1146 0.1252 0.2152 0.259
mean (g)
Table 2 shows the average height and fresh weight of paper towel method
Seeding Vigor Sand Media
Maize Bean
Height mean (cm) 19.25 23.75
Fresh Weight mean 0.4535 0.909
(g)

Table 2 shows the average height and fresh weight of sand media method

Discussion
The viability of a seed accession is a measurement of how many seeds are viable
under ideal conditions and capable of growing into self-replicating plants. It's
important to know that the seeds we used and bought will grow into plants when
planted in the field, as well as to forecast seed and seedling performance. As a result,
they must be highly viable at the outset and during storage. Within the parameters of
the environment, the viability of the seed at the time of storage has an impact on the
accession's storage life. The most accurate viability test is the germination test. The
TZ-test was the germination test that we utilised in the practical lesson, along with
other traditional germination test methods.
The tetrazolium test is a quick chemical test that can be conducted in a short
period of time with minimal equipment. In this test, a seed is incubated in a diluted
0.1% solution of 2,3,5-triphenyltetrazolium chloride. From data table 1 above shows
the percentage of germination TZ-test of maize and bean seeds reading of capable
germination test from our result. For the maize seed the germination percentage for
both replicate 1 and replicate 2 are same which is 40% and the average is 40% while
for the bean seeds germination percentage for replicate 1 is 60% and 40% for replicate
2 and the average is 50%. The average for both maize and bean seeds germination test
indicate that they are most non-germinable seeds.

Generally, our result for TZ-test from data table 1 were determined by these figures below
which it can identify the criteria of maize and bean seed germination.
 Criteria of TZ-test for Maize and Bean Germination

Criteria of TZ-test Result and Description

Maize Germination

Figure 1 Figure 2 Figure 3

Figure 1 and Figure 2 above show the criteria for Figure 4 shows the maize seed is non-
interpreting Tetrazolium test result on maize seed germinable because the scutellum entirely
germination test. unstained.

Bean Germination

Figure 4 Figure 5 Figure 6

Figure 4 and figure 5 above show the criteria for Figure 6 shows the bean seed is non-
interpreting Tetrazolium test result on bean seed germinable because the upper end of
germination test. cotyledon is unstained.
From the table and figures above show the criteria of TZ-test for maize and bean
germination included the result and description for the examples of non-germinable of
maize and bean seeds that we mostly we got from this experiment. As we know that
the red areas indicate stained and considered as living tissues and the white areas
represent of unstained or unwanted dead tissue inside the seed.

The conventional germination test method is a method used to observe and

calculate the germination percentage of maize seed and beans seeds. There are 2 types
of media that we use which are sand and paper towel. For the paper towel media,
replication 1 uses 1 layer of paper towel and replication 2 uses 2 layers of paper towel
media. The bean seeds and maize seeds will be planted into these 2 types of media
and left for a week. After a week, we observed how many seeds are germinate and
non-germinate.
Data table 1 shows the percentage of germination of maize seeds and bean
seeds. It shows that replication 1 and 2 for maize seeds in sand media is 20%. For the
bean seed in sand media, the germination percentage is higher than maize seed in sand
media which is 90% and for the maize seeds in paper towel media, the germination
percentage in replication 1 and 2 is 100% while replications 1 bean seeds germination
percentage is 100% & which is higher than replications 2 which is 80%. Next, data
Graph 1 shows that for both sand media and paper towel media, the highest average
germination percentage was maize seeds than bean seeds. The maize seeds were
100% average germination percentage for paper towel method and bean seeds in sand
media is 90% and in paper towel media is also 90%. From the both data also we can
see that maize seeds more germinate in paper towel media compared to sand media.
While the bean seeds capable to germinate from both media.
 120%
GERMINATION PERCENTAGE OF BEAN AND MAIZE SEEDS
1
100%
0.9 0.9

80%

60%
0.5
0.4
40%

0.2
20%

0%
.
MAIZE BEAN

TZ TEST SAND MEDIA PAPER TOWEL

Graph 1: Average germination percentage

From the result, we can compare between the chemical and conventional treatment.
We can observe that sand media method and paper towel method is more effective in
identifying germination of seed than chemical treatment (TZ). For overall, data table 1
shows the TZ-method has a lower germination
percentage compared to other conventional test methods for both seeds which are
maize seeds and beans seed except sand media method for maize seeds. Maybe there
some error in preparing the experiment, which maybe we accidentally applied too
much water which cause excessive water due to no holes for drainage. Data graph 1
also shows the average germination percentage of TZ-method is lower than other
conventional methods for both seeds which are maize seeds and beans seeds.

Rhetorical question:
Why both of the seeds in paper towel method germinates smaller size compared to
sand media method? Perhaps both of the seeds germinate bigger in sand media is due
to the sand are more suitable compared to paper towel, the plant will receives enough
oxygen and perfect temperature. While another reason which the plants are smaller in
paper towel method is due to the plants compete with each other for food and not
enough aeration.
Does the seeds have any changes in different measure of humidity?
The maize seeds do not get mouldy in the paper towel media, but the beans seeds do.
Because of the high humidity level in the paper towel media, the beans seeds became
mouldy. The fact that maize seeds contain Thiram is the reason why they do not
mould. Thiram's job is to protect the seed from fungal infection.