table s1. tissues and developmental stages throughout the ...3 calli 3 calli screening stage 4 calli...
TRANSCRIPT
![Page 1: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/1.jpg)
Sample No Abbreviation Tissues or organ Developmental stages
1 Calli 1 Calli 15 DAS (days after subculture)
2 Calli 2 Calli 5 DAR (days after regeneration)
3 Calli 3 Calli Screening stage
4 Calli 4 Calli 15 DAI T2 (days after induction)
5 Calli 5 Calli 15 DAI T3 (days after induction)
6 Seed imbibition Seed 72 h after imbibition
7 Seed germination Embryo bud and radicle 3 days after germination
8 Plumule/Dark Plumule 48 hours after emergence, Dark
9 Plumule/Light Plumule 48 hours after emergence, Light
10 Radicle/Dark Radicle 48 hours after emergence, Dark
11 Radicle/Light Radicle 48 hours after emergence, Light
12 Seedlings Seedling Three-leaf stage
13 Young shoot Shoot Seedlings with 2 tillers
14 Young root Root Seedlings with 2 tillers
15 Mature leaf Leaf Stage 3 (secondary branch primordium differentiation stage)
16 Old leaf Leaf 4– young panicle
17 Mature sheath Sheath Stage 3
18 Old sheath Sheath 4– young panicle
19 Young flag leaf Flag leaf 5 DBH (5 days before heading)
20 Old flag leaf Flag leaf 14 DAH (14 days after heading)
21 Panicle, stage 3 Young Panicle Stage 3
22 Panicle, stage 4 Young Panicle Stage 4 (pistil/stamen primordium differentiation stage)
23 Panicle, stage 5 Young Panicle Stage 5 (pollen-mother cell formation stage)
24 Young panicle Panicle 4– young panicle
25 Old panicle Panicle Heading stage
26 Young stem Stem 5 DBH
27 Old stem Stem Heading stage
28 Hull Hull 1 DBF (1 day before flowering)
29 Spikelet Spikelet 3 DAP (3 days after pollination)
30 Stamen Stamen 1 DBF
31 Endosperm 1 Endosperm 7 DAP (7 days after pollination)
32 Endosperm 2 Endosperm 14 DAP (14 days after pollination)
33 Endosperm 3 Endosperm 21 DAP (21 days after pollination)
Table S1. Tissues and developmental stages throughout the life cycles of two rice varieties.
![Page 2: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/2.jpg)
Table S2. Gene primers used for PCR amplification.
Genes Purpose Forward primer (5’-3’) Reverse primer (5’-3’)
OsSUS3 Gene clone TCCCCCGGGATGGGGGAAACTACTGGAG TCCCCCGGGCATTGCCCATTCTTTTCAC
OsSUS3 Q-PCR GTGAGACACGCCGCTACCTGGAGA CCCTGAAATCAACTAAACCCT
OsCESA1 Q-PCR GGCATGGTGGCGGGTATAT AGGCGACGGCTTTCTGAGT
OsCESA3 Q-PCR GTGTAGATAGAAACACGAAAGGTG CAATGGGCACCCACACACGT
OsCESA4 Q-PCR CCATCGTCTACCCGTTCACC TCCAGCCCAGCAATCATCTT
OsCESA7 Q-PCR TTAACGGGGTTCAAGATGC ATGGTGTTGGTGTAGGCGA
OsCESA8 Q-PCR TGGTCGGTGTTGTTGCTGG ATGGTCGGAGTGCGGTTTT
OsCESA9 Q-PCR TACAAGAACGGCAACCTCA AAGAACAAACTCGCAAACG
OsIRX9 Q-PCR ACAAGGAGATGATGATTTTAGCA AGGCGGAGGTCGTAGATGTCG
OsIRX14 Q-PCR GATGGATGGTGTGATTGTG TTGAACTGGAAGAGGCATA
OsGH9B1 Q-PCR ACTTCCTCTACAAAACCCCAAC ATAATCCACTCGCTGAACCATA
OsGH9B3 Q-PCR TCCGATCTCCTCTACGACTGA CCCCGTTCCTGCCATTT
OsGH9B16 Q-PCR CGGAACCACCAAGTAGCCAT GAATACACGACAAAGTCAACGAAC
OsXAT2 Q-PCR GGCGAAGAGGATAATAAGAAC TCACGGATGGTATAATTGGA
OsXAT3 Q-PCR AAGCAAGAAGGTATGAATAATGT AGTGGCAGTTGGATGAAT
OsGSL1 Q-PCR TGAGGACCTGCCACGATT CACGCTGATTGCGAACAT
OsGSL3 Q-PCR TGGCAAGCGACCACATAG AGACCTTAGCACGGACTG
OsGSL5 Q-PCR GTGGTGTCCCTGCTATGA GTTGTTTGCTATTCTCCC
OsSG1 Q-PCR GGCGTATGGGACAAAGGA TTCAGAGGCGAAGGATGG
OsGNS5 Q-PCR TTGCGGCCATTCCTACAGT TGGTGAGGGCGATGCTTG
OsUBQ1 Q-PCR CAGTAAGTCCTCAGCCAT AGACCAGACAACCATAGC
![Page 3: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/3.jpg)
Transgenic
line
Biomass production (g/plant)
2013 2014 2015
ZH11 ZH11 31.81±0.96 27.95±1.37 24.73±1.29
Vector EV 31.63±1.18 26.58±1.93 23.92±0.63
SUS3-OE
1 35.27±1.39** +11%† 34.49±1.55** +30% 27.10±1.11** +13%
2 33.89±1.01* +7% 32.43±1.56** +22% 26.73±0.88** +12%
3 35.77±1.12** +13% 33.78±1.73** +27% 26.87±1.06** +12%
4 36.55±2.13** +15% 35.57±1.77** +34% 28.62±1.30** +20%
Table S3. Biomass production in three-year field experiments.
* and **, Indicated significant difference between transgenic lines and EV control by t-test as P < 0.05 and 0.01 (n=10).†Percentage of increased or decreased level between transgenic line and EV by subtraction of two values divided by EV.
![Page 4: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/4.jpg)
Transgenic
line
Total grain yield (g/plant)
2013 2014 2015
ZH11 ZH11 34.01±1.69 27.82±1.14 23.58±0.90
Vector EV 33.56±2.24 28.44±2.21 23.42±1.25
SUS3-OE
1 37.56±2.83* +12%† 33.91±0.77** +19% 25.43±0.86* +9%
2 36.26±1.47* +8% 33.90±3.65** +19% 25.22±0.33* +8%
3 36.68±2.98 +9% 31.07±1.27 +9% 25.53±0.87* +9%
4 35.72±1.80* +6% 31.39±3.03* +10% 27.68±1.60** +18%
Table S4. Total grain yield in three-year field experiments.
* and **, Indicated significant difference between transgenic lines and EV control by t-test as P < 0.05 and 0.01 (n=10).†Percentage of increased or decreased level between transgenic line and EV by subtraction of two values divided by EV.
![Page 5: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/5.jpg)
Transgenic
line
Leaves Stems
Cellulose Hemicelluloses Lignin Cellulose Hemicelluloses Lignin
ZH11 ZH11 19.02±0.19 16.21±0.12 12.55±0.16 22.35±0.13 16.34±0.10 12.85±0.17
Vector EV 18.98±0.44 16.16±0.37 12.61±0.19 22.42±0.44 16.36±0.10 12.79±0.95
SUS3-
OE
1 23.92±0.73** +26%@ 17.57±0.38** +9% 12.86±0.20 25.02±0.28** +12% 18.10±0.28** +11% 12.38±0.83
2 21.67±0.29** +14% 17.06±0.31** +6% 12.08±0.19 24.41±0.17** +9% 17.41±0.18** +6% 12.11±0.43
3 21.50±0.44** +13% 17.28±0.32** +7% 12.70±0.27 24.43±0.28** +9% 18.22±0.28** +11% 13.76±0.44
4 22.88±0.38* +21% 18.26±0.44** +13% 12.02±0.19 25.74±0.23* +15% 17.69±0.23** +8% 13.67 ±0.37
Table S5. Three major wall polymer contents (% dry matter) in leaves and mature stems of transgenic rice lines.
* and **, Indicated significant difference between transgenic lines and EV control by t-test as P < 0.05 and 0.01 (n=3);@ Percentage of increased or decreased level between transgenic line and EV by subtraction of two values divided by EV.
![Page 6: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/6.jpg)
Transgenic
lineDP
Monosaccharides of hemicelluloses (% total, mol/mol)
Rha Fuc Ara Xyl Man Glu Gal
ZH11 ZH11 1207±11 0.14 0.04 8.42 88.16 0.07 1.00 2.17
Vector EV 1223±8 0.25 0.03 8.11 87.78 0.08 1.15 2.60
SUS3-OE
1 1401±10** +15%@ 0.27 0.04 10.03 85.59 0.10 0.69 3.28
2 1323±10** +8% 0.20 0.03 9.11 87.06 0.08 0.62 2.91
3 1350±11** +10% 0.16 0.06 9.14 87.32 0.09 0.81 2.43
4 1411±12** +15% 0.25 0.04 11.39 84.33 0.11 0.71 3.18
Table S6. Wall polymer features in mature stems of transgenic rice lines.
* and **, Indicated significant difference between transgenic lines and EV control by t-test as P < 0.05 and 0.01 (n=3);@ Percentage of increased or decreased level between transgenic line and EV by subtraction of two values divided by EV.
Rha, Rhamnose; Fuc, Fucose; Ara, Arabinose; Xyl, Xylose; Man, Mannose; Glu, Glucose; Gal, Galactose.
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Transgenic
line
1% NaOH 1% H2SO4
Total sugar yield
(% dry matter)
Hexoses yield
(% dry matter)
Bioethanol yield
(% dry matter)
Sugar-ethanol
conversion rate (%)
Total sugar yield
(% dry matter)
Hexoses yield
(% dry matter)
Bioethanol yield
(% dry matter)
Sugar-ethanol
conversion rate (%)
ZH11 ZH11 24.45±0.21 16.27±0.31 10.65±0.11 65.46±0.63 10.74±0.16 9.07±0.18 5.35±0.06 58.99±0.81
Vector EV 24.47±0.37 16.29±0.24 10.52±0.26 64.58±0.78 10.98±0.29 9.32±0.29 5.41±0.26 58.05±0.89
SUS3-
OE
1 28.25±0.29** 19.44±0.23** 14.61±0.13** 75.15±0.73** 13.57±0.20** 11.85±0.24** 8.53±0.18** 71.98±0.74**
2 27.61±0.23** 18.90±0.21** 12.77±0.25** 67.57±0.45** 12.64±0.39** 10.86±0.18** 7.15±0.09** 65.84±0.76**
3 27.79±0.32** 19.03±0.14** 13.48±0.26** 70.87±0.68** 13.65±0.19** 11.68±0.20** 7.44±0.11** 63.70±0.68**
4 30.08±0.38** 20.22±0.28** 15.88±0.29** 78.54±0.73** 14.34±0.28** 12.45±0.30** 8.89±0.18** 71.41±0.73**
Table S7. Biomass enzymatic saccharification and ethanol productivity in in mature stems.
* and **, Indicated significant difference between transgenic lines and EV control by t-test as P < 0.05 and 0.01 (n=3).
![Page 8: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/8.jpg)
Transgenic
line
Strain IV173 Strain ZH173
2014 Wuhan 2015 Wuhan 2015 Hanchuan 2015 Wuhan
Lesion length (cm) Lesion length (cm) Lesion length (cm) Lesion length (cm)
ZH11 ZH11 8.18±1.07 16.57±0.72 17.63±2.76 6.54±0.83
Vector EV 7.34±1.22 16.58±0.73 16.8±0.85 6.77±0.79
SUS3-OE
1 4.26±1.06** -48%† 14.53±1.76** -12% 13.50±0.88** -23% 5.05±0.23** -25%
2 6.12±0.96** -25% 15.07±1.06* -9% 13.56±0.79** -23% 5.63±0.49** -17%
3 3.39±2.08** -59% 15.73±0.39** -5% 14.70±0.75** -17% 4.66±0.37** -31%
4 5.10±1.41** -38% 15.35±0.93** -7% 12.98±0.43** -26% 4.57±0.41** -32%
* and ** Indicated significant difference between transgenic line and EV by Student’s t-test at P < 0.05 and 0.01 (n=30).
† Percentage of decreased level between transgenic line and EV by subtraction of two values divided EV.
Table S8. Lesion lengths of transgenic leaves infected with bacterial blight for 20 days.
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Number of lesions per leaf Lesion length (cm)
Strain RB22 R01-1 MT31-3 RB22 R01-1 MT31-3
EV 21±2 30±3 19±2 0.35±0.04 0.27±0.02 0.16±0.02
SUS3-1 15±1** -29%@ 19±1** -37% 9±2** -53% 0.29±0.02** -17% 0.24±0.03** -11% 0.12±0.03** -25%
SUS3-4 15±1** -29% 17±2** -43% 11±2** -42% 0.28±0.03** -20% 0.22±0.03** -19% 0.11±0.02** -31%
* and ** Indicated significant difference between transgenic line and EV by Student’s t-test at P < 0.05 and 0.01 (n=30).@ Percentage of decreased level between transgenic line and EV by subtraction of two values divided by EV.
Table S9. Lesion number and length of transgenic leaves infected with blast strains.
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Table S10. Feeding behaviors of BPH recorded by electronic penetration graph (EPG)#.
EPG waveform
Nonprobing Pathway Phloem Ingestion Xylem Ingestion
EV 23.15±7.54 99.41±7.28 205.97±14.94 56.67±4.33
SUS3-1 57.32±8.98** 2.47& 159.98±12.92** 1.61 59.54±6.16** 0.29 104.36±11.37** 1.84
SUS3-4 48.45±7.73* 2.09 204.28±1.99** 2.05 45.85±6.32 ** 0.22 67.80±5.12** 1.20
* and ** Indicated significant difference between transgenic line and EV by Student’s t-test at P < 0.05 and P < 0.01 (n=20);& Ratio of values between transgenic plants and EV.# Total duration (min) of each EPG waveform type on different host plants over 6-h recording period.
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Z1 Z3 Z5 Z7 Z9 Z11 Z13 Z15 Z17 Z19 Z21 Z23 Z25 Z27 Z29 Z31 Z33
0
20000
40000
60000
80000
100000 OsSUS1 OsSUS2 OsSUS3
OsSUS4 OsSUS5 OsSUS6
OsS
US
s e
xp
res
sio
n
Fig. S1
M1 M3 M5 M7 M9 M11M13M15M17M19M21M23M25M27M29M31M33
0
20000
40000
60000
80000
100000 OsSUS1 OsSUS2 OsSUS3
OsSUS4 OsSUS5 OsSUS6
OsS
US
s e
xp
ress
ion
(a) (b)
![Page 12: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/12.jpg)
Fig. S2
0
20
40
200
300
400 **
**
**
**
OsS
US
3
exp
ressio
n
ZH11 EV SUS3-OE
1 2 3 4
0
1
2
3
4
5
**
**
*
**
**
SUS1 SUS2 SUS3 SUS4 SUS5 SUS6
Gen
e e
xp
ressio
n
(24h
/0h
)
Mock
IV173
0
1
2
3
4
5
6
7
**
****
****
**
SUS1 SUS2 SUS3 SUS4 SUS5 SUS6
Gen
e e
xp
ressio
n
(24h
/0h
)
Mock
RB22
0
1
2
3
4
*
* **
*
**
SUS1 SUS2 SUS3 SUS4 SUS5 SUS6
Gen
e e
xp
ressio
n
(6h
/0h
)
Mock
BPH
(b)(a)
(c) (d)
![Page 13: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/13.jpg)
Fig. S3
(a) (b) (c)
0
50
100
150
200
250
300
**
**
****
**
**
EV
OE-1
OE-4
CESA1 CESA3 CESA8
CesA9
Rela
tive e
xp
ressio
n
0
30
60
90
120
150
180
** ****
**
**
**
EV
OE-1
OE-4
CESA4 CESA7 CESA9
CesA9 R
ela
tive e
xp
ressio
n
0.0
0.2
0.4
0.6
2
4
6
8
**
**
****
EV
OE-1
OE-4
IRX9 IRX14
Rela
tive e
xp
ressio
n
(d) (e)
0
5
10
15
20
25
**
**
**
**
EV
OE-1
OE-4
XAT2 XAT3
Re
lati
ve e
xp
res
sio
n
0.0
0.5
1.0
200
300
400
500
**
**
****
**
**
EV
OE-1
OE-4
GH9B1 GH9B3 GH9B16
Re
lati
ve
ex
pre
ss
ion
![Page 14: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/14.jpg)
0
20
40
60
80
100
716466725859
10%
23%14%24%
****
****
1% H2SO
4 pretreatment
Su
ga
r-e
tha
no
l
co
nv
ers
ion
rate
(%
)
1 2 3 4ZH11 EV SUS3-OE
0
20
40
60
80
100
797168756565
10%5%
22%16%**
******
1% NaOH pretreatment
Su
gar-
eth
an
ol
co
nvers
ion
rate
(%
)
1 2 3 4
ZH11 EV SUS3-OE
0
20
40
60
80
100
484745484140
15%18%
9%17%
********
1% H2SO
4 pretreatment
He
xo
se
s r
ele
as
ed
(% c
ellu
lose
)
1 2 3 4ZH11 EV SUS3-OE
0
20
40
60
80
100
7978787872
9%9% 9%8%
********
1% NaOH pretreatment
He
xo
se
s r
ele
as
ed
(% c
ellu
los
e)
1 2 3 4
ZH11 EV SUS3-OE
72
(a)
(b)
Fig. S4
![Page 15: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/15.jpg)
(a)
0
1
2
3
4
5
**
Ge
ne
ex
pre
ss
ion
(24
h/0
h)
EV
OE-1
OE-4
GSL1 GSL3 GSL5 SG1 GNS5
**
**
**** **
**
IV173 infection (b)
0
1
2
3
4
5
6
7
****
**
RB22 infection****
Gen
e e
xp
res
sio
n
(24
h/0
h)
EV
OE-1
OE-4
GSL1 GSL3 GSL5 SG1 GNS5
*
*
** *0
1
2
4
8
12
16
20
**
BPH attack
****
**
Gen
e e
xp
res
sio
n
(6h
/0h
)
EV
OE-1
OE-4
GSL1 GSL3 GSL5 SG1 GNS5
**
**
*
(c)
Fig. S5
![Page 16: Table S1. Tissues and developmental stages throughout the ...3 Calli 3 Calli Screening stage 4 Calli 4 Calli 15 DAI T2 (days after induction) 5 Calli 5 Calli 15 DAI T3 (days after](https://reader034.vdocument.in/reader034/viewer/2022042920/5f64c2731235a26f6c703d92/html5/thumbnails/16.jpg)
Probe Amplifier
Converter
R
Ground
-+
Nonprobing
Pathway
Xylem ingestionPhloem ingestion
28 %
7 %
59 %
6 %
38 %
28 %
19 %
15 %
49 %
22 %
15 %
14 %
EV SUS3-OE-1 SUS3-OE-4 (a) (b)
Fig. S6