Supplemental Materials for “Arabidopsis EDT1/HDG11 Confers
Drought Tolerance in Transgenic Rice without Yield Penalty” by Yu
et al.
Supplemental Tables
Supplemental Table 1. Biological process categories involved in photosynthesis,
oxidation-reduction are significantly enriched in DEGs between 2-56N and
ZH11N.
DEGs between 2-56N and ZH11N were classified based on Gene Ontology (GO)
through the Annotation Chart Tool of web tools DAVIA. All genes in the rice genome
were used as background for significance testing. The enriched functional annotation
terms were identified and listed according to their enrichment P-value. BP, biological
process; Count, number of genes; Benjamini, Benjamini-Hochberg adjusted P-value;
FDR, false discovery rate
Category
Term
Count
P-Value
Fold
Benjamini
FDR
Enrichment
GOTERM_BP_FAT
GO:0015979~photosynthesis
13
1.11E-08
9.2589848
2.59E-06
1.42E-05
GOTERM_BP_FAT
GO:0009853~photorespiration
4
5.95E-05
46.396671
6.94E-03
0.0764743
GOTERM_BP_FAT
GO:0055114~oxidation reduction
27
1.78E-04
2.1387734
1.38E-02
0.2279859
GOTERM_BP_FAT
GO:0016054~organic acid catabolic process
5
2.95E-04
15.035958
1.71E-02
0.3786341
GOTERM_BP_FAT
GO:0046395~carboxylic acid catabolic process
5
2.95E-04
15.035958
1.71E-02
0.3786341
GOTERM_BP_FAT
GO:0009072~aromatic amino acid family metabolic process
6
3.35E-04
9.743301
1.56E-02
0.4301892
GOTERM_BP_FAT
GO:0015977~carbon utilization by fixation of carbon dioxide
4
3.58E-04
27.064725
1.39E-02
0.4589755
GOTERM_BP_FAT
GO:0006558~L-phenylalanine metabolic process
4
1.26E-03
18.04315
4.12E-02
1.6055778
GOTERM_BP_FAT
GO:0043094~cellular metabolic compound salvage
4
1.48E-03
17.09351
4.24E-02
1.8873305
GOTERM_BP_FAT
GO:0009310~amine catabolic process
4
2.29E-03
14.762577
5.79E-02
2.9069701
GOTERM_BP_FAT
GO:0009063~cellular amino acid catabolic process
4
2.29E-03
14.762577
5.79E-02
2.9069701
GOTERM_BP_FAT
GO:0009698~phenylpropanoid metabolic process
5
3.36E-03
7.9602132
7.57E-02
4.2333395
GOTERM_BP_FAT
GO:0008610~lipid biosynthetic process
8
4.94E-03
3.7764732
1.00E-01
6.1701527
GOTERM_BP_FAT
GO:0016053~organic acid biosynthetic process
9
5.46E-03
3.3065501
1.01E-01
6.7960708
GOTERM_BP_FAT
GO:0046394~carboxylic acid biosynthetic process
9
5.46E-03
3.3065501
1.01E-01
6.7960708
GOTERM_BP_FAT
GO:0009074~aromatic amino acid family catabolic process
3
6.22E-03
24.358252
1.06E-01
7.7070513
GOTERM_BP_FAT
GO:0006559~L-phenylalanine catabolic process
3
6.22E-03
24.358252
1.06E-01
7.7070513
GOTERM_BP_FAT
GO:0019748~secondary metabolic process
6
6.22E-03
5.0746359
9.91E-02
7.712476
GOTERM_BP_FAT
GO:0006631~fatty acid metabolic process
6
7.08E-03
4.9208591
1.05E-01
8.7263588
GOTERM_BP_FAT
GO:0016051~carbohydrate biosynthetic process
6
8.34E-03
4.7297578
1.15E-01
10.205726
1/9
GOTERM_BP_FAT
GO:0005996~monosaccharide metabolic process
7
9.29E-03
3.840265
1.21E-01
11.308276
GOTERM_BP_FAT
GO:0006575~cellular amino acid derivative metabolic process
6
9.75E-03
4.5529444
1.20E-01
11.832858
GOTERM_BP_FAT
GO:0006006~glucose metabolic process
6
1.22E-02
4.3111951
1.40E-01
14.551616
GOTERM_BP_FAT
GO:0045229~external encapsulating structure organization
6
1.65E-02
3.9931561
1.77E-01
19.241622
GOTERM_BP_FAT
GO:0007047~cell wall organization
6
1.65E-02
3.9931561
1.77E-01
19.241622
GOTERM_BP_FAT
GO:0006633~fatty acid biosynthetic process
5
1.99E-02
4.7761279
2.01E-01
22.759834
GOTERM_BP_FAT
GO:0043648~dicarboxylic acid metabolic process
4
2.40E-02
6.3681706
2.28E-01
26.860721
GOTERM_BP_FAT
GO:0019318~hexose metabolic process
6
2.51E-02
3.5820959
2.28E-01
27.856675
GOTERM_BP_FAT
GO:0009250~glucan biosynthetic process
4
3.21E-02
5.6978368
2.72E-01
34.224535
GOTERM_BP_FAT
GO:0019438~aromatic compound biosynthetic process
4
3.50E-02
5.5046898
2.84E-01
36.754441
GOTERM_BP_FAT
GO:0033692~cellular polysaccharide biosynthetic process
4
3.81E-02
5.3242082
2.95E-01
39.304496
GOTERM_BP_FAT
GO:0044042~glucan metabolic process
5
4.03E-02
3.8299139
3.00E-01
41.055124
GOTERM_BP_FAT
GO:0000271~polysaccharide biosynthetic process
4
4.30E-02
5.0746359
3.07E-01
43.14483
GOTERM_BP_FAT
GO:0006091~generation of precursor metabolites and energy
8
4.65E-02
2.4057533
3.19E-01
45.806066
GOTERM_BP_FAT
GO:0005976~polysaccharide metabolic process
6
4.77E-02
3.0071917
3.17E-01
46.635347
GOTERM_BP_FAT
GO:0006457~protein folding
6
4.87E-02
2.9887426
3.14E-01
47.387284
GOTERM_BP_FAT
GO:0019439~aromatic compound catabolic process
3
5.77E-02
7.6119539
3.52E-01
53.393894
GOTERM_BP_FAT
GO:0019253~reductive pentose-phosphate cycle
2
5.95E-02
32.47767
3.53E-01
54.569573
GOTERM_BP_FAT
GO:0019685~photosynthesis, dark reaction
2
5.95E-02
32.47767
3.53E-01
54.569573
GOTERM_BP_FAT
GO:0016052~carbohydrate catabolic process
6
6.00E-02
2.8159829
3.47E-01
54.855346
GOTERM_BP_FAT
GO:0006096~glycolysis
4
6.34E-02
4.330356
3.54E-01
56.894247
GOTERM_BP_FAT
GO:0005978~glycogen biosynthetic process
2
7.10E-02
27.064725
3.80E-01
61.2036
GOTERM_BP_FAT
GO:0005977~glycogen metabolic process
2
7.10E-02
27.064725
3.80E-01
61.2036
GOTERM_BP_FAT
GO:0006112~energy reserve metabolic process
2
7.10E-02
27.064725
3.80E-01
61.2036
GOTERM_BP_FAT
GO:0018130~heterocycle biosynthetic process
4
7.17E-02
4.1110975
3.75E-01
61.579095
GOTERM_BP_FAT
GO:0008299~isoprenoid biosynthetic process
3
7.45E-02
6.5833115
3.79E-01
63.023686
GOTERM_BP_FAT
GO:0006073~cellular glucan metabolic process
4
8.05E-02
3.9129723
3.96E-01
66.013128
GOTERM_BP_FAT
GO:0044264~cellular polysaccharide metabolic process
4
8.98E-02
3.7330655
4.23E-01
70.159834
Supplemental Table 2. Clusters of photosynthesis-related genes have higher
enrichment score in DEGs between 2-56N and ZH11N.
DEGs between 2-56N and ZH11N were analyze by the Functional Annotation
Clustering Tool through the web tools DAVIA. The clusters are ordered by group
enrichment score. BP, biological process; Count, number of genes; Benjamini,
Benjamini-Hochberg adjusted P-value; FDR, false discovery rate.
Category
Term
Count
P-Value
Fold
Benjamini
FDR
Enrichment
Annotation Cluster 1. Enrichment Score: 3.500417088054158
GOTERM_BP_FAT
GO:0009853~photorespiration
4
5.95E-05
46.39667
0.006909
0.076422
GOTERM_BP_FAT
GO:0015977~carbon utilization by fixation of carbon
4
3.58E-04
27.06472
0.0138
0.458663
2/9
dioxide
GOTERM_BP_FAT
GO:0043094~cellular metabolic compound salvage
4
0.001481
17.09351
0.042248
1.886056
Annotation Cluster 2. Enrichment Score: 2.5880866748204343
GOTERM_BP_FAT
GO:0046395~carboxylic acid catabolic process
5
2.95E-04
15.03596
0.017042
0.378376
GOTERM_BP_FAT
GO:0016054~organic acid catabolic process
5
2.95E-04
15.03596
0.017042
0.378376
GOTERM_BP_FAT
GO:0009072~aromatic amino acid family metabolic
6
3.35E-04
9.743301
0.015506
0.429897
process
GOTERM_BP_FAT
GO:0006558~L-phenylalanine metabolic process
4
0.001258
18.04315
0.041043
1.604492
GOTERM_BP_FAT
GO:0009310~amine catabolic process
4
0.002292
14.76258
0.057678
2.905018
GOTERM_BP_FAT
GO:0009063~cellular amino acid catabolic process
4
0.002292
14.76258
0.057678
2.905018
GOTERM_BP_FAT
GO:0009698~phenylpropanoid metabolic process
5
0.003359
7.960213
0.075403
4.230516
GOTERM_BP_FAT
GO:0006559~L-phenylalanine catabolic process
3
0.006219
24.35825
0.105791
7.702005
GOTERM_BP_FAT
GO:0009074~aromatic amino acid family catabolic
3
0.006219
24.35825
0.105791
7.702005
process
GOTERM_BP_FAT
GO:0019748~secondary metabolic process
6
0.006224
5.074636
0.098689
7.707427
GOTERM_BP_FAT
GO:0006575~cellular amino acid derivative metabolic
6
0.009748
4.552944
0.119095
11.82529
3
0.057656
7.611954
0.351049
53.36963
process
GOTERM_BP_FAT
GO:0019439~aromatic compound catabolic process
Annotation Cluster 3. Enrichment Score: 2.136664422663831
GOTERM_BP_FAT
GO:0008610~lipid biosynthetic process
8
0.004942
3.776473
0.099617
6.166079
GOTERM_BP_FAT
GO:0046394~carboxylic acid biosynthetic process
9
0.00546
3.30655
0.100844
6.791599
GOTERM_BP_FAT
GO:0016053~organic acid biosynthetic process
9
0.00546
3.30655
0.100844
6.791599
GOTERM_BP_FAT
GO:0006631~fatty acid metabolic process
6
0.007077
4.920859
0.104458
8.720677
GOTERM_BP_FAT
GO:0006633~fatty acid biosynthetic process
5
0.019888
4.776128
0.199794
22.74623
Annotation Cluster 4. Enrichment Score: 1.472260217080035
GOTERM_BP_FAT
GO:0008610~lipid biosynthetic process
8
0.004942
3.776473
0.099617
6.166079
GOTERM_BP_FAT
GO:0008299~isoprenoid biosynthetic process
3
0.07447
6.583311
0.394003
62.9986
GOTERM_BP_FAT
GO:0006720~isoprenoid metabolic process
3
0.104077
5.412945
0.481381
75.63173
Annotation Cluster 5. Enrichment Score: 1.4565624806002893
GOTERM_BP_FAT
GO:0007047~cell wall organization
6
0.016486
3.993156
0.176066
19.22986
GOTERM_BP_FAT
GO:0045229~external encapsulating structure
6
0.016486
3.993156
0.176066
19.22986
organization
GOTERM_BP_FAT
GO:0044042~glucan metabolic process
5
0.040282
3.829914
0.298694
41.03388
GOTERM_BP_FAT
GO:0030243~cellulose metabolic process
3
0.13627
4.595897
0.539646
84.77171
Annotation Cluster 6. Enrichment Score: 1.3541124578965305
GOTERM_BP_FAT
GO:0016051~carbohydrate biosynthetic process
6
0.008339
4.729758
0.114799
10.19914
GOTERM_BP_FAT
GO:0009250~glucan biosynthetic process
4
0.032061
5.697837
0.271204
34.20575
GOTERM_BP_FAT
GO:0033692~cellular polysaccharide biosynthetic
4
0.038094
5.324208
0.29394
39.28383
process
3/9
GOTERM_BP_FAT
GO:0044042~glucan metabolic process
5
0.040282
3.829914
0.298694
41.03388
GOTERM_BP_FAT
GO:0000271~polysaccharide biosynthetic process
4
0.042973
5.074636
0.306153
43.12294
GOTERM_BP_FAT
GO:0005976~polysaccharide metabolic process
6
0.047678
3.007192
0.315738
46.6125
GOTERM_BP_FAT
GO:0006073~cellular glucan metabolic process
4
0.08052
3.912972
0.410593
65.98812
GOTERM_BP_FAT
GO:0044264~cellular polysaccharide metabolic process
4
0.089779
3.733066
0.4383
70.13522
GOTERM_BP_FAT
GO:0034637~cellular carbohydrate biosynthetic process
4
0.106994
3.455071
0.482717
76.63144
Annotation Cluster 7. Enrichment Score: 1.2318929845707975
GOTERM_BP_FAT
GO:0005996~monosaccharide metabolic process
7
0.009291
3.840265
0.120094
11.30102
GOTERM_BP_FAT
GO:0006006~glucose metabolic process
6
0.012158
4.311195
0.1393
14.54246
GOTERM_BP_FAT
GO:0019318~hexose metabolic process
6
0.025079
3.582096
0.226862
27.84062
GOTERM_BP_FAT
GO:0006091~generation of precursor metabolites and
8
0.046535
2.405753
0.318092
45.78343
energy
GOTERM_BP_FAT
GO:0016052~carbohydrate catabolic process
6
0.059989
2.815983
0.353895
54.83087
GOTERM_BP_FAT
GO:0006096~glycolysis
4
0.063362
4.330356
0.361467
56.86951
GOTERM_BP_FAT
GO:0006007~glucose catabolic process
4
0.106994
3.455071
0.482717
76.63144
GOTERM_BP_FAT
GO:0046365~monosaccharide catabolic process
4
0.106994
3.455071
0.482717
76.63144
GOTERM_BP_FAT
GO:0019320~hexose catabolic process
4
0.106994
3.455071
0.482717
76.63144
GOTERM_BP_FAT
GO:0046164~alcohol catabolic process
4
0.122693
3.247767
0.516242
81.39306
GOTERM_BP_FAT
GO:0044275~cellular carbohydrate catabolic process
4
0.125388
3.215611
0.516138
82.11422
GOTERM_BP_FAT
GO:0015980~energy derivation by oxidation of organic
3
0.174672
3.92875
0.621823
91.51005
compounds
Annotation Cluster 8. Enrichment Score: 0.7214338916359302
GOTERM_BP_FAT
GO:0018130~heterocycle biosynthetic process
4
0.071707
4.111097
0.39064
61.55403
GOTERM_BP_FAT
GO:0051186~cofactor metabolic process
4
0.299889
2.068641
0.81012
98.97465
GOTERM_BP_FAT
GO:0051188~cofactor biosynthetic process
3
0.318549
2.591303
0.826609
99.27532
Annotation Cluster 9. Enrichment Score: 0.6394489919209264
GOTERM_BP_FAT
GO:0019725~cellular homeostasis
4
0.168186
2.799799
0.614598
90.61179
GOTERM_BP_FAT
GO:0042592~homeostatic process
4
0.186376
2.662104
0.632563
92.93324
GOTERM_BP_FAT
GO:0045454~cell redox homeostasis
3
0.385012
2.234702
0.88677
99.8061
Annotation Cluster 10. Enrichment Score: 0.5440801519096107
GOTERM_BP_FAT
GO:0030001~metal ion transport
5
0.177
2.255394
0.619285
91.81255
GOTERM_BP_FAT
GO:0006811~ion transport
7
0.286086
1.557149
0.798594
98.68235
GOTERM_BP_FAT
GO:0006812~cation transport
5
0.460565
1.424459
0.926819
99.96401
Annotation Cluster 11. Enrichment Score: 0.34479520462167906
GOTERM_BP_FAT
GO:0044271~nitrogen compound biosynthetic process
6
0.429754
1.383992
0.915356
99.92652
GOTERM_BP_FAT
GO:0008652~cellular amino acid biosynthetic process
3
0.436169
2.013079
0.915617
99.93646
GOTERM_BP_FAT
GO:0009309~amine biosynthetic process
3
0.492878
1.804315
0.9407
99.98372
Annotation Cluster 12. Enrichment Score: 2.17243430528008E-6
4/9
GOTERM_BP_FAT
GO:0006468~protein amino acid phosphorylation
3
0.999994
0.217097
1
100
GOTERM_BP_FAT
GO:0006796~phosphate metabolic process
4
0.999994
0.249828
1
100
GOTERM_BP_FAT
GO:0006793~phosphorus metabolic process
4
0.999994
0.249828
1
100
GOTERM_BP_FAT
GO:0016310~phosphorylation
3
0.999998
0.20081
1
100
Supplemental Table 3. Genes related to photorespiration, carbon fixation,
regulation of transcription and oxidation-reduction are significantly enriched in
DEGs between 2-56D and ZH11D.
DEGs between 2-56D and ZH11D were classified based on Gene Ontology (GO)
through the Annotation Chart Tool of web tools DAVIA. All genes in the rice genome
were used as background for significance testing. The enriched functional annotation
terms were identified and listed according to their enrichment P-value. BP, biological
process; Count, number of genes; Benjamini, Benjamini-Hochberg adjusted P-value;
FDR, false discovery rate.
Category
Term
Count
P-Value
Fold
Benjamini
FDR
Enrichment
GOTERM_BP_FAT
GO:0009853~photorespiration
4
8.16E-04
19.269585
0.2731947
1.128125
GOTERM_BP_FAT
GO:0006536~glutamate metabolic process
4
1.87E-03
14.987455
0.3069366
2.5734138
GOTERM_BP_FAT
GO:0015977~carbon utilization by fixation of carbon dioxide
4
4.59E-03
11.240591
0.4513242
6.2018197
GOTERM_BP_FAT
GO:0045449~regulation of transcription
49
5.72E-03
1.4558299
0.4292769
7.6665234
GOTERM_BP_FAT
GO:0009415~response to water
4
7.28E-03
9.6347926
0.4351508
9.6558017
GOTERM_BP_FAT
GO:0006470~protein amino acid dephosphorylation
7
1.59E-02
3.4210496
0.647395
19.93844
GOTERM_BP_FAT
GO:0043094~cellular metabolic compound salvage
4
1.74E-02
7.0993209
0.6244537
21.631556
GOTERM_BP_FAT
GO:0006006~glucose metabolic process
9
1.85E-02
2.685805
0.5976642
22.815449
GOTERM_BP_FAT
GO:0019318~hexose metabolic process
10
1.92E-02
2.4795422
0.5685496
23.584593
GOTERM_BP_FAT
GO:0016311~dephosphorylation
7
2.05E-02
3.2335948
0.5543218
24.974161
GOTERM_BP_FAT
GO:0009063~cellular amino acid catabolic process
4
2.59E-02
6.1312317
0.6064072
30.557943
GOTERM_BP_FAT
GO:0009310~amine catabolic process
4
2.59E-02
6.1312317
0.6064072
30.557943
GOTERM_BP_FAT
GO:0009064~glutamine family amino acid metabolic process
5
2.72E-02
4.3233044
0.5926834
31.832814
GOTERM_BP_FAT
GO:0005996~monosaccharide metabolic process
10
3.10E-02
2.2784983
0.6126302
35.489928
GOTERM_BP_FAT
GO:0016052~carbohydrate catabolic process
11
3.22E-02
2.1441591
0.5986468
36.518152
GOTERM_BP_FAT
GO:0006575~cellular amino acid derivative metabolic process
8
3.87E-02
2.5212541
0.6427746
42.246885
GOTERM_BP_FAT
GO:0030001~metal ion transport
11
4.05E-02
2.0607751
0.6359405
43.718936
GOTERM_BP_FAT
GO:0016054~organic acid catabolic process
4
4.42E-02
4.9958184
0.6462683
46.641255
GOTERM_BP_FAT
GO:0046395~carboxylic acid catabolic process
4
4.42E-02
4.9958184
0.6462683
46.641255
GOTERM_BP_FAT
GO:0006457~protein folding
10
5.21E-02
2.0688205
0.6871847
52.467278
GOTERM_BP_FAT
GO:0055114~oxidation reduction
40
5.62E-02
1.3159717
0.6958133
55.245
GOTERM_BP_FAT
GO:0019320~hexose catabolic process
7
5.91E-02
2.511196
0.6963517
57.153269
5/9
GOTERM_BP_FAT
GO:0006007~glucose catabolic process
7
5.91E-02
2.511196
0.6963517
57.153269
GOTERM_BP_FAT
GO:0046365~monosaccharide catabolic process
7
5.91E-02
2.511196
0.6963517
57.153269
GOTERM_BP_FAT
GO:0009072~aromatic amino acid family metabolic process
5
5.94E-02
3.3721774
0.6801642
57.307149
GOTERM_BP_FAT
GO:0019748~secondary metabolic process
7
6.42E-02
2.4588794
0.6925842
60.253534
GOTERM_BP_FAT
GO:0006355~regulation of transcription, DNA-dependent
27
6.48E-02
1.409424
0.6798829
60.603002
GOTERM_BP_FAT
GO:0051252~regulation of RNA metabolic process
27
6.77E-02
1.4029089
0.6808932
62.268587
GOTERM_BP_FAT
GO:0046164~alcohol catabolic process
7
7.51E-02
2.3605242
0.7050557
66.219193
GOTERM_BP_FAT
GO:0010876~lipid localization
6
7.69E-02
2.6276707
0.6996425
67.105877
GOTERM_BP_FAT
GO:0006869~lipid transport
6
7.69E-02
2.6276707
0.6996425
67.105877
GOTERM_BP_FAT
GO:0044275~cellular carbohydrate catabolic process
7
7.80E-02
2.3371527
0.6913719
67.650256
GOTERM_BP_FAT
GO:0016051~carbohydrate biosynthetic process
7
8.39E-02
2.2917711
0.705893
70.427958
GOTERM_BP_FAT
GO:0009069~serine family amino acid metabolic process
4
8.91E-02
3.7468638
0.7159121
72.681073
GOTERM_BP_FAT
GO:0000160~two-component signal transduction system
4
8.91E-02
3.7468638
0.7159121
72.681073
(phosphorelay)
GOTERM_BP_FAT
GO:0006098~pentose-phosphate shunt
3
9.75E-02
5.6202957
0.7374673
75.985232
GOTERM_BP_FAT
GO:0006739~NADP metabolic process
3
9.75E-02
5.6202957
0.7374673
75.985232
Supplemental Table 4. Primer used for Q-PCR
Name
rice Actin1 LP
rice Actin1 RP
SNAC1 FP
SNAC1 RP
AREB2 FP
AREB2 RP
SAPK4 FP
SAPK4 RP
OsHsfA3 FP
OsHsfA3 RP
OsNCED3 FP
OsNCED3 RP
OsP5CS FP
OsP5CS RP
SNAC2 FP
SNAC2 RP
OsNAC5 FP
OsNAC5 RP
OsLEA3-1 LP
OsLEA3-1 RP
Sequence (5’ to 3’)
CGCAGTCCAAGAGGGGTATC
TCCTGGTCATAGTCCAGGGC
GGGTGCTGTGTCGGCTGTA
CGACTGCGACGTAACCATGT
GCTGAACGATGAACTCCAGA
CTTTGCTGTCGGTCCAACTT
ATGTGATTTTGGCTACTCCAAG
ACCATACATCTGCAAGCTTTCC
TGCTGCCAGAGAACATAGGACTT
GTGTGTCAAATCCCTGAACCAA
ACGTGATCAAGAAGCCGTACCT
GCTGGTCGAGCGGGATCT
AGCCTGTTCAGGAGTAGGTGAA
ACCTCCAAAATCTTGCTCTCAG
CCAGCCCAAGATCAGCGAGT
TCATGTACTGGGGCAAGCCAAGCCA
CAGCAGCTGATGGTATTGTC
AGAGACCTGTTTGGCACGAA
CCTTCGTCATCATCCTC
CGGGGTCAAATTCAGT
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Supplemental Figures
Supplemental Figure 1. Generation of transgenic plants overexpressing
AtEDT1/HDG11. To express efficiently the AtEDT1/HDG11 in rice, we cloned the
AtEDT1/HDG11 cDNA into the expression binary vector pCB2006 (Lei et al., 2007),
in which the AtEDT1/HDG11 cDNA is under the control of the rice Actin1 promoter
(A). Transgenic rice lines were generated by Agrobacterium-mediated transformation
using the japonica variety Zhonghua 11 (ZH11) as the recipient. More than 140 T0
independent transgenic plants were generated. Positive T2 homozygous transformants
were prescreened for drought resistance in the greenhouse and more than 80%
transgenic line showed improved drought tolerance during seedling stage. Several
drought tolerance lines were selected for further analysis. RT-PCR analysis indicated
that AtEDT1/HDG11was expressed in all of the seven lines, whereas no signals were
detected in ZH11 non-transgenic plant (B). Southern blot analysis indicated that most
of the selected lines had 1–2 insertion copies (C).
A. Schematic diagram of the T-DNA region of expression vector
pCB2006-AtEDT1/HDG11. LB left border, RB right border, 35S polyA
Cauliflower mosaic virus (CaMV) 35S polyA , bar phosphinotricin
acetyltransferase gene, 35S promoter CaMV 35S promoter, NOS polyA nopaline
synthetase gene polyA.
B. AtEDT1/HDG11 expression analysis of the putative transgenic lines using RT-PCR.
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C. Southern blot analysis of the transgenic lines using bar as probe.
Supplemental Figure 2. Reduced leaf stomatal density and enlarged stomatal size
of the flag leaf at flowering stage.
Comparisons of stomatal density (A) and stomatal dimension (B) of the flag leaf at
flowering stage. Three flag leaves were sampled for each plant and ten plants were
sampled for both the ZH11 control and the transgenic plants. Values are mean ± SD
(*** P < 0.001).
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Supplemental Figure 3. Quantitative real-time PCR validation of the results of
DGE tag profiling.
The transcript levels of 9 stress-related genes were determined by quantitative
RT-PCR as described in Experimental procedures. Values are mean ± SD of three
independent experiments (* P < 0.05, ** P < 0.01).
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