Transcript Arabadopsis - PlantingScience.org

Dru Cowan
Spencer Sanson
Frederick Smith

The purpose of this eight week pilot project is
to test the anthocyanin phenotype, erecta
phenotype, and the trichome counts in
Arabadopsis plants. The ultimate goal is to
determine whether each of these traits is
continuous or discrete.
•Erecta
phenotype: continuous
•Anthocyanin phenotype: discrete
•Trichome number phenotype: continuous

Preparing the seeds

Cold shock treatment
 Rinse seeds with bleach
 Rinse seeds five times with sterilized water


Place seeds on wet paper towel in sections on Petri
dish
Petri Dishes wrapped in saran wrap and placed in
fridge for three days

Light Box


Dishes of seeds placed in light box
Dishes rotated every other day
 Distributes heat and light more evenly

Preparation of second set of seeds



Cold shock treatment
Place five seeds in one Jiffy © plug
The two plugs for each line are placed in a 2x2
cardboard pot

Trichome number Assay


Take 5 of the first true leaves from each line
Place leaf under dissecting scope and count the
number of trichomes on each leaf

Anthocyanin Assay

Preparation of sucrose solution
 Put 100 grams of sucrose per 100 mL of water in dilute
fertilizer
 Place in microwave for three minutes
 Let cool at room temperature
 Place .25 mL of solution in each microfuge tube

Anthocyanin Assay (cont.)



Slice mature leaf at petiole
Place cut end of leaf in sucrose solution
Place in light box for two days at 22° C
Trichome Count
80
75.2
70
Trichome Number
60
55.2
50
40
35.8
33.6
30.2
30
20
17.4
15
21.8
19.8
10
0
70
177
72
64
62
Line Number
66
82
78
65
Average Trichome Number
80
70
Trichome Number
60
50
40
30
Yucca
Botan-x
20
Genus Geniuses
10
RAEJ
0
53
25
43
55
23
54
59
155
47
70
177
72
64
62
66
82
78
65
134
138
141
142
146
147
149
150
152
178
85
89
92
94
101
120
127
Line Number
132
161
183
ColumbiaLandsberg
Frequency
4.5
4
3.5
3
2.5
Frequency
2
Poly. (Frequency)
1.5
1
0.5
0
0
20
40
60
80
•Not
able to test erecta phenotype
•Anthocyanin assay deemed failure
•Trichome number assay results were
inconsistent
•Trichome number—continuous trait?







Uneven heat distribution in light box
Too much heat killed plants
Overbleached seeds
Susceptibility to disease
Hard to differentiate first true leaves in
trichome number assay
Alteration of trichome count
Leaves submerged in sucrose solution in
anthocyanin assay
Failures: Discrete or continuous?
Procedure still under review
Pilot study: successful
•Dr.
Marshall Sundberg
•Dr. Allison Landry
•Fellow classmates