Lauren Epstein

Gunsher
APES 2nd
13 February 2015
Contributors: Bailey, Jared, and Devin
LD-50 Lab
In this lab you will be testing the effects of salt on the germination and growth of plants.
It is suspected that salt (NaCl - sodium chloride) applied to highways may affect the
growth of vegetation along the roadside and aquatic plants in nearby streams.
You will conduct a dose-response experiment to determine how seeds respond to various
concentrations of salt. You will make serial dilutions - a series of solutions, each of which
are half as concentrated (or two times more dilute) than the one from which it is made to
set up a wide range of concentrations of salt solutions to test on the seed growth.
Introduction
LD50 is the lethal dose that kills 50% of something and will be tested on radish
seeds during the lab. Dose-response curves will be to plot the results of the experiment.
It is a type of graph where the X-axis plots concentration of the substance, or in this
case, salt concentration and the Y-axis plots response, which could be almost anything,
but in this experiment its germination of the radish seeds. Another method to be used in
the experiment is Serial Dilution. Serial Dilution is a process by which the amount of salt
per mL is cut in half each time. This method is used to get accurate measurements of
salt quickly and to gather a wide range of concentrations.
The purpose of this lab is to measure the effects of salt on radish seeds to see how
much road salt could effect roadside vegetation. Salt is often used on highways during
the winter to melt ice off the roads. Ice usually forms when the temperature of water
reaches 32 degrees Fahrenheit or 0 degrees Celsius. When you add salt, that
temperature starts to drop. A 10-percent salt solution freezes at 20 F (-6 C) and a 20percent solution freezes at 2 F (-16 C) for example. Even though this helps cars to not
slide on the road, once the ice melts it turns into salt water as it mixes with the salt and
washes off the road into the ground. If the salt water is absorbed into the ground plants
will later take it up. If the salt concentration is high enough it could start killing off the
plants on the side of the road.
Problem
What is the LD50 of salt for radish seeds?
How does salt affect the initial growth of radish plants?
Hypothesis: Radish plants cannot survive in ocean water (35 g/L) therefore the more
salt that is added the more likely the seeds will die, or wont germinate. I believe the
LD50 will be 18 g/L because it is the closest to the concentration of ocean water but it
still isnt concentrated enough to kill 100% of the seeds.
Parts of the Experiment
Independent variable: Amount of Salt (18 g/L 0 g/L)

Dependent variable: How many seeds are germinated and the length of
the roots
Control: Pure water with no salt

Materials & methods

Using 20 ml of the original concentration of salt, perform a serial dilution to create
five different salt concentrations, plus a sixth with plain water.
Count out 60 seeds, 10 for each water sample
Put 10 seeds into a folded paper towel, space the seeds out evenly on the paper
towel so that they do not touch each other
Wet the paper towel/seeds and seal in a baggie, be sure to label the bags with the
salt concentration for that sample
After 4-5 days examine the seeds, counting the number of each that germinated
For the germinated seeds, measure the length of the radicle
Data Analysis:
According to our groups data, there would never be an LD50 based off of just the
percent germinated because all of our seeds germinated in all the concentrations, except
one in the plain water control group. This could have been that the seed rolled out of the
water or that it was a defective seed. Although there technically isnt a LD50 there would
be if the concentration were increased further because the radicle length was decreasing
as the salt increased. For our group the length decreased from 52.8 mm to 3.3 mm the
higher the salt concentration in the solution. These results partially support my
hypothesis because I stated that the higher the concentration the more likely the seeds
will not germinate or die, but I also stated that the LD50 would be 18 g/L which is not
true with this data table.
Although our groups data was nearly 100% germinated across the concentrations,
the other groups had different germination percents than ours. For example, Group 1
only had 20% germinated in the 18 g/L water which mostly increased as the salt
concentration became more diluted. This could be a more correctly done lab or the seeds
could have been defective. Even though the individual group results do not seem to be
very accurate, the averages for all of the groups seems to make more sense as the
average starts off at 74.3% for the highest concentration and steadily increasing to
93.3% for the control group. Therefore it appears to work better to have a wide range of
samples and seeds to get a better outcome of results.
Data
% salt
solutio
n

% seeds
germinat
ed

100%

100%

50%

100%

25%

100%

Radicle Length (mm)

Mean
Radic
le
Lengt
h

10

5m
m
20
mm
30

5m
m
15
mm
35

3m
m
30
mm
50

3m
m
15
mm
30

2m
m
8m
m
35

6m
m
5m
m
15

3m
m
5m
m

1m
m
4m
m

2m
m
15
mm

3m
m
30
mm

3.3
mm
14.7
mm
32.5

12.5%

100%

6.25%

100%

0%
(contr
ol)

90%

mm
34
mm
55
mm
47
mm

mm
30
mm
50
mm
90
mm

mm
50
mm
57
mm
40
mm

mm
35
mm
53
mm
30
mm

mm
25
mm
44
mm
50
mm

mm
35
mm
68
mm
45
mm

40
mm
43
mm

25
mm

25
mm

mm
33.2
mm
52.8
mm
50.3
mm

Class Data
Group 1 (names)
Jordan
Kendall
Melba
Sophia

Group 2 (names)
Amber
Samantha
Maggie
Logan L.

Group 3 (names)
Kylee
Christina
Logan
Julianne

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L
12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

% Germinated
20

Mean Radicle length

.45

90

2.7

80

3.8

90

3.1

80

2.5

90

3.9

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L
12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

% Germinated
100

Mean Radicle length

6

90

16.9

100

17.7

100

38.6

100

39.4

100

35.4

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L
12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

% Germinated
100

Mean Radicle length

17.4mm

100

9.7mm

50

2.2mm

70

1.5mm

100

17.9mm

30 omitted from the

results

1mm

Group 4 (names)
Andrew
Fatou
Julia
Alex

Group 5 (names)
Lauren
Livi
Miky
Youjin
Renee

Group 6 (names)
Matt
Lauren
Devin
Bailey
Jared

Group 7 (names)
Jerrel
Madison
JP

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L
12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

% Germinated
90%

Mean Radicle length

5.22mm

90%

2.78mm

90%

6.89

100%

4.1mm

100%

40.7mm

90%

9mm

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L
12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

% Germinated
50%

Mean Radicle length

0.2mm

100%

3.6mm

70%

4.3mm

90%

6.3mm

90%

6.1mm

90%

5.1mm

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L
12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

% Germinated
100%

Mean Radicle length

3.3mm

100%

14.7mm

100%

32.5mm

100%

33.2mm

100%

52.8mm

90%

50.3mm

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L

% Germinated
60%

Mean Radicle length

.6mm

100%

14mm

100%

26.1mm

Taylor

Averages

12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

100%

36.9mm

100%

47mm

100%

32.3mm

Salt Concentration
100%
18 g/L
50%
9 g/L
25%
4.5 g/L
12.5%
2.25 g/L
6.25%
1.25 g/L
0%
0 g/L control

% Germinated
74.3%

Mean Radicle length

4.73 mm

88.6%

9.2 mm

84.3%

13.36 mm

78.7%

17.67 mm

95.7%

29.49 mm

93.3%

19.57 mm

Using the % germinated data, construct a dose/response curve to

determine the LD50 of
salt for these seeds.
Conclusions (points to address in your write-up)
What was the LD50 for salt exposure to radish seeds?
How did the salt affect the growth?