LD50 Lab Report

Luke Johnson
Mrs. Norris
AP Environmental Science
11 October, 2014
Collaborators:
Hannah Whitt, James Hamil, Megan Redfern
Introduction:
It is suspected that salt (NaCl sodium chloride) applied to highways for deicing may affect the
growth of vegetation along the roadside and aquatic plants in nearby streams. Therefore, a dose-
response experiment must be conducted to determine how radish seeds will respond to various
concentrations of salt. Vegetation supports entire ecosystems by providing a source of food for animals.
Vegetation also prevents erosion along roads. If salt is affects aquatic and land vegetation, then
measures must be taken to prevent the use of salt to remove ice on roads.
Problem:
Do higher concentrations of salt solutions in water contribute to fewer germinating radish
seeds?
Hypothesis:
The higher concentration of salt solution in water, the fewer radish seeds will germinate. 100%
of seeds will germinate for the control group, 100% of seeds will germinate from the .75 g NaCl/L H2O,
90% of seeds will germinate from 1.5 g NaCl/L H2O, 80% will germinate from the 3 g NaCl/L H2O, 60%
will germinate from the 6 g NaCl/L H2O, 40% will germinate from the 12 g NaCl/L H2O. The seeds that
are able to germinate with higher salt solutions will experience depressed growth.
Parts of the Experiment:
Control Group: 10 mL of distilled water (Container 6)
Experimental Group: radish seeds that are exposed to various concentrations of salt
Independent Variable: concentration of salt solutions in g NaCl/L H20
Dependent Variable: Percentage of seeds that germinated and radicle growth
Controlled variable: Room temperature, environment, amount of water each seed
receives, amount of light each plant recieves
Materials: (Lab Protocol #1)
6 test tubes (size 20 X 150 mm)
Tape (for labeling test tubes)
Test tube rack
10 mL graduated cylinder or pipette
Distilled water
concentrated salt solution = 12.0 grams table salt (sodium chloride)/1 liter distilled
water
Materials: (Lab Protocol #2)
6 test tubes with serial dilutions of salt solutions (from part 1)
6-90 mm petri dishes with lids
3 pieces of unbleached paper towel
Pencil
Scissors
10 mL graduated cylinder or syringe
Tape and permanent marker
60 lettuce seeds (Radish seeds, 10/petri dish)
Tweezers
Plastic bag
Materials: (Lab Protocol #3)
Metric ruler
Calculator
2 sheets graph paper (75 square X 100 squares)
Methods: (Lab Protocol #1)
1. Set up 6 test tubes in a test tube rack and label the tubes with the following salt concentrations:
12.0 g/L, 6.0 g/L, 3.0 g/L, 1.5 g/L, 0.75 g/L and Control. See Table 1. Salt Solutions.
2. Add 10 mL of distilled water to test tubes #2-6.
3. Measure 20 mL of the concentrated solution (12.0 g/L) and pour into test tube #1.
4. Transfer 10 mL of salt solution from test tube # 1 to test tube #2.
5. Gently swirl test tube #2 to mix the salt solution.
6. Repeat steps 4 and 5 for test tubes #3-5 measuring 10 mL each time. DO NOT add any salt
solution to test tube #6.
7. Measure 10 mL of distilled water into test tube #6 to serve as the control. The control will
indicate whether or not your seeds are viable (capable of growing or developing). NOTE: The
total remaining solution in each test tube is 10 mL, except for test tube #5 will have 20 mL.
8. Unless you will be using the solutions right away, cover them tightly with plastic wrap to prevent
water loss through evaporation.
Methods: (Lab Protocol #2)
1. Obtain six petri dishes. Label each dish according to the concentration of salt solution to be
tested. See Table 1. Salt Solutions Concentrations.
2. Fold a half of sheet of paper towel or coffee filter into quarters. Cut it out so that it fits into the
bottom of the petri dish.
3. Measure 6 mL of salt solution and pour onto the paper towel in the appropriate petri dish. In
the control dish, add 5 mL of distilled water. The purpose of a control is to identify how well the
seeds will grow without any salt. NOTE: If using the same graduated cylinder, start with the
lowest salt solution (distilled water) to the highest salt solution so that the graduated cylinder is
not contaminated with a higher salt solution.
4. Add 10 lettuce seeds to each petri dish. Space the seeds out evenly on the paper towel so that
they do not touch each other or the sides of the dish.
5. Place the dishes in a plastic bag and seal it to retain moisture. Label your groups name on the
outside of the bag.
6. Incubate the seeds in a dark place at a constant temperature (preferably 24.5 degrees Celsius)
for 4-5 days.
7. Inspect radish seeds during incubation period. Record any observations. If the paper seems dry,
add a 1 or 2 more millimeters of the appropriate salt solution or distilled water (control).
Methods: (Lab Protocol #3)
1. Remove the lid of the control dish. Count the number of seeds that germinated (sprouted).
Calculate the percentage of seeds that germinated and record in Table 2. Radish Seed Results.
Note: If fewer than 80% of the seeds in the control sample germinated, this indicates a
problem with the experiment, e.g., bad seeds, poor incubation conditions, etc. The results
discarded or the test rerun.
2. To measure the length of the radicle (embryonic root), carefully remove the germinating radish
seed from the paper towel in one piece. The radicle may be growing into the layers of towel and
can break if you pull too hard.
3. Measure the length of the radicle for each of the germinating radish seeds to the nearest
millimeter (mm). Look carefully at each sprout to make sure you are measuring just the root, not
the shoot as well. Record data in Table 2. Radish Seed Results.
4. Repeat steps 1-3 for each petri dish.
5. For each treatment, calculate the mean (arithmetic average) radicle length for each salt
solution. Add the total radicle lengths for each salt solution and divide by the total number of
seeds that germinated. DO NOT INCLUDE data from seeds that did not germinate. Record data
in column labeled, Mean Radicle Length (mm) in Table 2. Radish Seed Results.
6. Make a line graph from the data collected to show a dose-response curve. The horizontal axis
should be for the independent variable, dose (concentration of salt solutions). The vertical axis
should be for the dependent variable, response (mean radicle length)/ Draw and label the axes.
In your group, you will need to come to some agreement about scales for these by looking at
your data. Remember to give your graph a title.
7. To help you answer the following question: Did the radicle length increase or decrease in
length as compared to the control? subtract the mean radicle length of ach treatment (T) from
the mean radicle length of the control (C). Record your answers in the column, Difference in
Radicle Length on Table 2. Difference in Radicle Length = C (control) T (treatment mean
radicle length)
8. Make a line graph to show the percentage of seeds that germinated for each salt solution.
Complete your lab report. See Student Handout 2: Laboratory Report Outline.

Photos:


Data:
Group 6 Data
Treatments:
Concentration of
Salt Solution
(mg/L)
% Seeds
Germinated
1
mm
2
mm
3
mm
4
mm
5
mm
6
mm
7
mm
8
mm
9
mm
10
mm
T= Mean
Radicle
Length
(mm)
Difference
in Radicle
Length:
C- T
C = Control 100% 75 50 70 60 80 70 85 60 85 90 72.5 0
.75 100% 100 43 62 73 34 57 26 70 60 36 56.1 16.4
1.5 100% 95 62 116 70 80 50 75 100 60 85 79.3 -6.8
3.0 100% 45 30 95 40 75 30 45 30 35 60 48.5 24
6.0 100% 66 35 56 41 73 43 49 40 75 30 50.8 21.7
12.0 100% 16 15 20 5 20 0 0 0 0 0 7.6 64.9


Group Number
Mean Radicle Length (mm) 1 2 3 4 5 6 7 8
Control 81.5 78 89.7 121.8 35.3 72.5 41
19.5
12.0 34 26 29.5 35.0 49 7.6 42.5
.7
6.0 78.5 40 55 99.7 0 50.8 45
12
3.0 135.5 72 68.5 105.0 52.1 48.5 71
8.5
1.5 82.5 82 100.7 110.6 47.1 79.3 45.5
45.5
.75 97.8 100 118.6 83.34 50.7 56.1 22.5
15.5



Group 1 2 3 4 5 6 7
8
% Seeds Germinated

Control 100% 100% 100% 100% 100% 100% 100%
100%
12.0 80% 40% 40% 60% 70% 50% 90%
40%
6.0 100% 90% 80% 100% 0% 100% 100%
90%
3.0 100% 80% 80% 90% 80% 100% 100%
80%
1.5 90% 100% 100% 100% 70% 100% 90%
100%
.75 100% 100% 100% 90% 100% 100% 80%
90%
Difference in Radicle Length
(mm)
Length of Control 87.5 78

121.8 0

0
12.0 47.7 52 60.2 -86.8 -13.7 -64.9 -18.5
18.8
6.0 3 38 34.7 -22.1 35.3 -21.7 4.5
7.5
3.0 -5.4 6 21.2 -16.8 -16.8 -24 30
11
1.5 -1 -4 -11 -11.2 -11.8 6.8 4
26
.75 -16.5 -22 -28.9
-
38.47
-15.4 -16.4 1.5
4
















Mean Radical Length (mm)
(Figure 1)- Each line represents a separate group

% Seeds Germinated
(Figure 2)- Each line represents a different group

0
20
40
60
80
100
120
140
160
Control 12 6 3 1.5 0.75
1
2
3
4
5
6
7
8
0%
20%
40%
60%
80%
100%
120%
Control 12 6 3 1.5 0.75
1
2
3
4
5
6
7
8
Difference in Radical Length
(Figure 3)- Each line represents a different group

Conclusion:
The control group had 100% germination for 100% of the groups, meaning the lab was
performed correctly and most likely did not have bad seeds, or poor incubation conditions. Some groups
did experience heightened growth of radish seed roots as compared to the control in salt solutions such
as in groups three and five where the .75 g NaCl/L H20 had longer roots than the control. As the salt
solutions became increasingly concentrated with salt, the radicle length did decrease as compared to
the control a majority of the time. The only discrepancy may be the 0% of seeds germinate in figure 2 for
group 5s salt concentration of 6 g NaCl/L H2O because not a single other group had 0% germinated
even for higher concentrations of salt. It appears that lower concentrations of salt water do not greatly
-100
-50
0
50
100
150
Length of
Control
12 6 3 1.5 0.75
1
2
3
4
5
6
7
8
impact radish germination, but radicle growth is somewhat effected, though higher concentrations have
a very clear negative impact and should not be used to deice roads.
















Work Cited
"LD50 Lab Directions.pdf - Google Drive." LD50 Lab Directions.pdf - Google Drive. N.p., n.d. Web. 12 Oct.
2014.