Table S1. Genetic constructs used in this study
Name
TA29::BARNASE::NOS
PTD::GUS::NOS
En35S::GUS::E9
EnACT11::GUS::E9
EnACT2::GUS::E9
Other
name
3PG
3SG
3A11G
3A2G
Binary vector backbone
Reference
pTTM8
modified pBI101
pMON10547
pMON10547
pMON10547
Li et al. (2007)
Sheppard et al. (2000)
Perlak et al. (1993)
Perlak et al. (1993)
Perlak et al. (1993)
Table S2. Primers used in Polymerase Chain Reaction (PCR)
Name
Sequence (5’ 3’)
ACT2.001
ACT2.002
ACT11.001
ACT11.002
NPTII-F
NPTII-R
TA29-pro-F02
TA29-term-R03
V35F5
GUS003
atggtacctaggcaactatttttatgtatg
aggatccagctgcaaacacacaaaaagag
taggtaccgctagcaaatgtcaaatggaatgcatc
gagaattctgtacatcctgtcaaaattgatataaa
cttcttgacgagttcttc
cgctgcctcgtcctg
tcctcacactaagtccatgtttgc
ggaaagtgaaattgaccgatcagag
aggactattctggcttcctcttac
ccagactgaatgcccacaggcc
Size
(bp)
1228
1405
340
656
880
Comment
Reference
Construction of
ACT2 promoter
Construction of
ACT11 promoter
Detection of
KanR gene
Detection of
BARNASE gene
Detection of
GUS gene
This study
This study
This study
This study
Skinner et
al. (2003)
Table S3. ANOVA table (one-way ANOVA) for the non-transgenic control trees that were
distributed between the four transgenic constructs in the reporter trial for 2001
Name
Construct
Error
Total
Degrees of freedom
3
6
9
F-value
2.700
p-value
0.139
Table S4. ANOVA tables (one-way ANOVA) for the non-transgenic control trees that were
distributed between the four transgenic constructs in the reporter trial for 2003
Name
Construct
Error
Total
Degrees of freedom
3
5
8
F-value
0.526
p-value
0.683
Table S5. ANOVA table (2-way ANOVA) with “Event” and “Block” as main effects for the
sterility trial model
Name
Event
Block
Error
Total
Degrees of freedom
18
2
46
66
F-value
52
8822522
Pr(>F)
<0.0001
<0.0001
Table S6. Dunnett’s test for the sterility trial
Linear Hypotheses
1 - control == 0
2 - control == 0
3 - control == 0
4 - control == 0
5 - control == 0
6 - control == 0
7 - control == 0
8 - control == 0
9 - control == 0
10 - control == 0
11 - control == 0
12 - control == 0
13 - control == 0
14 - control == 0
15 - control == 0
16 - control == 0
17 - control == 0
18 - control == 0
Estimate
-38.214
-36.771
-35.636
-29.152
-25.580
-24.984
-24.857
-22.705
-21.313
-20.932
-19.960
-19.490
-18.859
-17.734
-17.513
-17.216
-16.863
-12.241
Standard error
2.226
2.310
2.347
6.340
8.004
6.734
5.633
3.337
2.949
6.532
4.761
4.933
2.722
4.780
12.739
4.230
2.767
3.797
***: P<0.001, **: P<0.01, and *: P<0.05
z value
-17.167
-15.917
-15.182
-4.598
-3.196
-3.710
-4.412
-6.804
-7.227
-3.204
-4.192
-3.951
-6.929
-3.710
-1.375
-4.070
-6.094
-3.224
Pr(>|z|)
< 0.001
< 0.001
< 0.001
< 0.001
0.020
0.003
< 0.001
< 0.001
< 0.001
0.020
< 0.001
0.001
< 0.001
0.003
0.900
< 0.001
< 0.001
0.018
Significance
***
***
***
***
*
**
***
***
***
*
***
**
***
**
***
***
*
Table S7. ANOVA table (one-way ANOVA) with “Construct” as main event for the reporter
trial model for 2001
Name
Construct
Error
Total
Degrees of freedom
4
36
40
F-value
0.212
p-value
0.930
Table S8. ANOVA table (one-way ANOVA) with “Construct” as main event for the reporter
trial model for 2003
Name
Construct
Error
Total
Degrees of freedom
4
36
40
F-value
0.263
p-value
0.900
Table S9. Dunnett’s test table for the reporter trial data from 2001
Linear Hypotheses
3A11G - control == 0
3A2G - control == 0
3PG - control == 0
3SG - control == 0
Estimate
-3.583
-12.603
-41.193
0.556
Standard error
127.260
124.383
127.035
124.768
z value
-0.028
-0.101
-0.324
0.004
Pr(>|z|)
1
0.999
0.935
1
Table S10. Dunnett’s test table for the reporter trial data from 2003
Linear Hypotheses
3A11G - control == 0
3A2G - control == 0
3PG - control == 0
3SG - control == 0
Estimate
-1586.1
-3041.2
-4206.2
-593.9
Standard error
8020.9
8075.5
8054.7
8391.2
z value
-0.198
-0.377
-0.522
-0.071
Pr(>|z|)
0.993
0.937
0.848
1
Table S11. ANOVA tables (one-way ANOVA) with “Event” as main effect for each construct in
the reporter trial for 2001
3SG construct
Name
Degrees of freedom
9
Event
9
Error
18
Total
3PG construct
Name
Degrees of freedom
9
Event
11
Error
20
Total
3A2G construct
Name
Degrees of freedom
9
Event
10
Error
19
Total
3A11G construct
Name
Degrees of freedom
9
Event
10
Error
19
Total
F-value
4.146
p-value
0.023
F-value
7.495
p-value
0.001
F-value
17.356
p-value
1e-04
F-value
4.026
p-value
0.020
Table S12. ANOVA tables (one-way ANOVA) with “Event” as main effect for each construct in
the reporter trial for 2003
3SG construct
Name
Degrees of freedom
9
Event
8
Error
17
Total
3PG construct
Name
Degrees of freedom
9
Event
10
Error
19
Total
3A2G construct
Name
Degrees of freedom
9
Event
9
Error
18
Total
3A11G construct
Name
Degrees of freedom
9
Event
10
Error
19
Total
F-value
15.816
p-value
3e-04
F-value
3.954
p-value
0.022
F-value
77.816
p-value
<0.0001
F-value
3.464
p-value
0.033
Table S13. Dunnett’s test for the sterility trial data comparing catkin mean length of control to
that of transgenic events for catkin collection from March 10, 2009
Linear Hypotheses
Event 7 - control == 0
Event 9 - control == 0
Event 12 - control == 0
Event 14 - control == 0
Estimate
-3.709
-3.662
-2.783
-4.047
Standard error
0.308
0.291
0.362
0.281
z value
-12.059
-12.573
-7.682
-14.387
Pr(>|z|)
< 2e-16
< 2e-16
1.58e-14
< 2e-16
Table S14. Dunnett’s test for the sterility trial data comparing catkin mean length of control to
that of transgenic events for catkin collection from March 17, 2009
Linear Hypotheses
Estimate
Event 7 - control == 0
Event 9 - control == 0
Event 12 - control == 0
Event 14 - control == 0
Event 17 - control == 0
-2.341
-2.822
-3.079
-4.103
-5.257
Standard
error
1.049
0.459
0.854
0.618
0.295
z value
Pr(>|z|)
-2.231
-6.152
-3.605
-6.636
-17.805
0.114
< 0.001
0.002
< 0.001
< 0.001
Table S15. Catkin angle per event
Event
7
9
12
14
17
Average
St. error
Angle
72.83
90.89
77.83
130.26
83.28
91.02
10.26