Supplemental figures
Supplemental tables
Table SI:
Phenotypic variables of non-symbiotic MtSucS1-antisense and corresponding
control lines cultured under non-mycorrhizal, Pi-deprived or fully supplemented
growth conditions
A, Final harvest vegetative and reproductive characters of 64 days old non-symbiotic MtSucS1antisense and control lines (equivalent to 50 dpi-old mycorrhizal lines)
non-mycorrhizal, Pi-deprived
c
FWplant
[g]
5.65
FWroot
[g]
3.77
DWshoot
[g]
0.36
RSR
2.06
RWR
0.67
(±0.55)
(±0.36)
(±0.04)
(±0.12)
(±0.01)
as12
4.78
3.24
0.31
1.79
0.69
(±0.37)
(±0.23)
(±0.03)
(±0.11)
(±0.01)
yield
3.50
k
0.48
(±0.74)
0.38
(±0.61)
2.50
4.70
3.25
0.29
2.26
0.69
(±0.31)
(±0.21)
(±0.02)
(±0.07)
(±0.01
0.32
(±0.33)
FWroot
[g]
1.64
DWshoot
[g]
0.69
RSR
0.61
RWR
0.38
k
c
FWplant
[g]
4.35
yield
7.75
FWseed
[mg]
4.08
seedlot
vigor [%]
79.4
(±0.29)
(±0.12)
(±0.03)
(±0.04)
(±0.02)
(±0.25)
(±0.07)
(±3.8)
as12
5.35
1.86
0.74
0.57
0.36
3.74
59.9
(±0.65)
(±0.14)
(±0.06)
(±0.09)
(±0.04)
1.96
8.00
(±0.41)
(±0.05)
(±10.9)
1.13
7.75
4.12
70.7
(±0.25)
(±0.17)
(±13.4)
as19
2.74
fully supplemented
as19
6.27
2.79
0.73
0.80
0.44
(±0.78)
(±0.37)
(±0.08)
(±0.02)
(±0.01)
1.83
B, Photochemical yield of non-mycorrhizal MtSucS1-antisense and control lines at time point 28 dpi
Φ non-mycorrhizal, Pi-deprived
c
as12
as19
n
40
20
20
oldest LP
561.6 (±16.2)
525.9 (±22.9)
512.6 (±35.1)
youngest LP
694.9 (±2.9)
686.4 (±6.5)
686.3 (±4.1)
Φ fully supplemented
avg. Φ
628 (±8)
606 (±13)
600 (±19)
n
20
10
10
oldest LP
708.0 (±6.4)
734.7a (±3.0)
704.8 (±7.4)
avg. Φ
718 (±6)
739a (±2)
716 (±5)
youngest LP
727.8 (±5.2)
743.2a (±2.8)
728.1 (±5.2)
C, Nutritional status of the shoot tissues of non-symbiotic MtSucS1-antisense and controls plants
non-mycorrhizal, Pi-deprived
c
as12
as19
n
10
5
5
N [%]
2.20 (±0.08)
2.10 (±0.11)
1.96a (±0.05)
C [%]
42.28 (±0.18)
41.94 (±0.25)
42.44 (±0.08)
fully supplemented
C/N
19.36 (±0.81)
20.46 (±1.11)
21.76a (±0.55)
n
10
5
5
N [%]
2.35 (±0.10)
2.66 (±0.10)
2.12 (±0.18)
C [%]
42.35 (±0.36
42.53 (±0.11)
42.82 (±0.16)
C/N
18.28 (±0.75)
15.99 (±0.80)
20.58 (±1.67)
D, Attributes of root system morphology as total root length, surface area, volume, and root tip number
of 64 days old non-symbiotic, fully supplemented MtSucS1-antisense and control lines
n
c
10
TRL
[cm]
982.9
RSa
[cm2]
184.7
aRD
[mm]
0.60
RV
[cm3]
2.79
RT
1465
sRL
346.3
sRL
[% c]
100
sRT
519
sRT
[% c]
100
(±72.6)
(±16.3)
(±0.09)
842.9
145.3
0.55
(±0.31)
(±93)
(±72.6)
(±21)
(±28)
(±5)
2.00a
1230a
275.8
80
372a
(±8.1)
(±0.06)
72a
(±0.13)
(±196)
(±36.9)
(±11)
(±48)
(±9)
as12
5
(±42.1)
as19
5
1167.1
228.2
0.62
3.57
1302
375.3
114
403
82
(±122.9)
(±27.7)
(±0.10)
(±0.50)
(±88)
(±77.6)
(±22)
(±52)
(±12)
Data are means (±SE).Data are obtained from A and B, n> 10 non-mycorrhizal, Pi-deprived or fully
supplemented plants per transgenic line; C and D, ten control and five MtSucS1-as line root systems
and shoot samples, respectively. Significantly different means are indicated by a (P< 0.05) and
b (P< 0.01). Abbreviations: FW, fresh weight; DW, dry weight; RSR, root to shoot ratio based in FWs;
RWR, root weight ratio as DW root per DW plant; k, allometric coefficient; yield, generative yield; TRL, total
root length; RSa, root surface area; aRD, average root diameter; RV, root volume; RT, root tip
number; sRL and sRT, DW shoot-normalized specific root length and root tip number; Φ, photochemical
yield of photosystem II, LP, leaf pair; C/N, carbon to nitrogen ratio; n, number of replicates; SE,
standard error; % percentage normalized to controls set to 100%
Table SII:
c
as12
as19
Leaf developmental attributes of mycorrhizal MtSucS1-antisense and control lines
n
35
20
20
RLi
[leaf primordia/dpi]
0.94 (0.06)
0.74a (0.07)
0.73b (0.06)
RLa
[leaves/dpi]
0.78 (0.06)
0.74 (0.07)
0.60b (0.05)
plastochron [d] phyllochron [d]
1.06 (0.06)
1.29 (0.08)
1.35b (0.09)
1.35 (0.09)
b
1.36 (0.08)
1.68b (0.12)
Data are means (SE) calculated for the time interval between 0 and 39 dpi. Significantly different
means are indicated by a (P< 0.05) and b (P< 0.01). Abbreviations: n, number of replicates; RLi, rate of
leaf initiation; RLa, rate of leaf appearance; dpi, day post inoculation; d, day; SE, standard error
Table SIII: Primer sequences used in real time RT-PCR experiments on total RNA derived from
mycorrhizal root tissue
ID
putative annotation
left primer
right primer
SucS
TC100410
TC95820
TC94447
TC98648
TC99016
Sucrose synthase MtSucS1
Sucrose synthase MtSucS2
Sucrose synthase MtSucS3
Sucrose synthase MtSucS2/MtSucS4
Sucrose synthase MtSucS5
cttttggaatttctcagactt
aaacaaggacttgatttcact
atatgagcatctacttcccat
aagatactctttctgctcacc
gtagtctctggcatcaatgta
gacatcatcttgagcaaagta
tcaggataacattgtaactcg
tctaagtctttggattccttc
gttcttctttgaagctgagat
gaagaatcctccaacaattac
AM marker
TC96500
TC106954
TC94453
TC100720
TC111031
TC95776
TC95651
TC95400
TC95018
TC95549
TC107796
TC98216
Blue copper-binding protein MtBcp1
Serine carboxypeptidase MtScp1
Inorganic P transporter MtPT4
Glutathione-S-transferase MtGst1
Aquaporin MtNip4
Annexin 2 MtAnn2
1-deoxy-d-xylulose-5-P synthase MtDxs2
H+-ATPase MtHa1
Germin-like prot. Glp1
H(+)/hexose cotransporter MtSt1
Caffeic acid O-methyltransferase COMT
Chalcone reductase CHR
catctttgcatgagagactta
ggtttctctttatgcttgttt
tgagttgttggttcttgttag
atcattgctaacatgtcaaag
gtcaagcttttgctttagaat
gcaaaagttaccatgtgatta
aatttcattcattctcacctt
aggttctaacatctattggga
caaaatttgaagaaaggtttt
attgaagagatggacagagtt
tattgtgagaaatgacgagag
ataggaatgggaacttcagta
aatgaatgggagagttacaat
tcagctagtcttagctcctct
tataaaagagcgaagaggttt
ataccagactcaagaacaggt
tttgtagttgtttgcagctat
attgattcatgaagaatgtca
gatcttccattcatctttttc
gacaacttatgtgaaccaatg
atttaagatgaagaaagcacc
aataatcccgtaattttgaag
cttttcacttaaaggagcaat
agatcataatctgcatcagtg
Inv
BQ148180
TC107108
TC96236
TC103706
TC98116
TC99238
BG455025
AJ497350
TC106499
TC106886
BQ148153
TC99506
BQ144400
Cell wall invertase I
Cell wall invertase
Vacuolar acid invertase
Cell wall invertase II
Cell wall invertase II
Alkaline/ neutral invertase
Alkaline/ neutral invertase
Neutral invertase
Neutral invertase
Alkaline/ neutral Invertase
Vacuolar acid invertase PsI-1
Acid invertase
Alkaline/neutral invertase-like
gttgaagtgaaagggattact
aaatctacgtatgaacccagt
ttgtgaagcttgataagaaga
accattgagttacaaaggact
gtttgattgatcacttctgtg
tctagtctggttagcaatgtg
tggattattttactacgagca
gaggattctgatcttgtgaat
atttgacatgtttccttctct
cagatgaaaccagtgattaaa
gattttatcccatgaagaaac
gtgttaaaggctagtttggat
ctgtgcagaagaacagattta
gaatcacgtgtttatttgaag
cacatgagaaccaaattctta
aggagtgtactcagaaagagc
taatccattcccttagaaaag
ccactcctactttctgcttat
agatggaataaaatcacgaat
taatatgaaatgagagagcga
taagacagtttcatcacaacc
acttgttgacagcatcagtag
acatgcaattaatgaaacaag
gacgtacgtttgaatctgtaa
gcaaattagttccagtcttct
tgcataacgaaagtgagtaat
1-FEH
TC96754
TC99051
TC100756
TC100757
TC109927
Fructan 1-exohydrolase 1-FEH prec.
Fructan 1-exohydrolase 1-FEH
Fructan 1-exohydrolase 1-FEH
Fructan 1-exohydrolase 1-FEH
Fructan 1-exohydrolase 1-FEH prec.
tttctttgcaacagatgatac
attggctatagagaaagatgc
cataaattggactcatttgaa
aacaatatgcatcaagaaatg
actttagagacccttcaactg
taaccctttccatagtcaaat
tatggatgtccaatgaaagt
actcacctacttcaactccat
tccatctccaaaactctctat
ttatcttcaaaacatgcctaa
TC130469
MtTef1a
actgtgcagtagtacttggtg
aagctaggaggtattgacaag
AY193825
H+-ATPase GmHA5
Isocitrate lyase GmICL
PEP carboxykinase GmPEPCK
Phosphat transporter GmosPT
Translation elongation factor GmTEF1
Internal transcribed spacer 28SrRNA Gm1ITS12
Glyceraldehyd 3-P dehydrogenase GmGAPDH
3-phosphoglycerate kinase Gm3-PGK
H+-ATPase GmPMA1
Glycogen synthase GmGlyS
Malate synthase GmMS
Fatty acid elongase GmMAELO
G. mosseae BEG12 large subunit 28S rRNA
atttcttcatgtcctcatttt
aaaactcgaaatacgaaagtc
gttgttacgccaaatgtatta
ggcaaaattttagttggtact
ttgacaaacgtacaatagagaa
gaaatgcgataagtagtgtga
tataaaggtcaaattgaagca
attgccttataaaaaggtgag
tgtcacaaaggaatagtgaag
ttgttaaagtatgagtcaccg
gatatgtcttataagtctgttccc
caattcgtaattgatttgttc
gatctctcggtgtactttctc
taatttggcagtaagttcaaa
ttgaacaaagaccacaattt
tttggatgatctgatgaaata
acccacgacaataaagagtat
gcagcaataattaaaatagca
actacatgagatggtacgtca
attcacaccacatacaaacat
ctgggccttttataatttct
aaatcaacccaatcttgtact
cgtataaattcctccaacttt
atatgagcagtttgtggtaaa
actactcaaaatccaacatcc
gtcattacgtcaacatcctta
fungal
genes
DQ074452
X84232
AF311438
AY149918
AY541918
Table SIV: Ratios of polar metabolite steady-state levels in MtSucS1-antisense and control
lines at 50 dpi
L-Asparagine (L-Asn)
L-Homoserine (L-HSer)
Pyroglutamate
5-Methylthioadenosine
a-Ketoaminobutyrate
Raffinose
Maleate
Xylose
4-Aminobutyrate (GABA)
Mannitol
2-Phosphoglycerate (2-PG)
L-Histidine (L-His)
Ornithine-Citrulline-Arginine
Spermidine
Spermine
L-Isoleucine (L-Ile)
Glucose 6-P
Myo-Inositol-P
Glycerol 3-P
L-Tyrosine (L-Tyr)
D/L-Diaminopimelate
Shikimate
3-Phosphoglycerate (3-PG)
DHAP
L-Tryptophan (L-Try)
Glycerol 2-P
Fructose 6-P
Trehalose
L-Threonine (L-Thr)
α-Ketoglutarate
2-Methylcitrate
L-Proline (L-Pro)
L-Valine (L-Val)
2-Isopropylmalate
2-Methylisocitrate
L-Leucine (L-Leu)
S-Methylcysteine
β-Alanine (β-Ala)
Glycine (Gly)
L-Serine (L-Ser)
Pinitol
Ribulose 5-P
Fructose
L-Arginine (L-Arg)
O-Acetyl-L-Serine
Glucose
Urea
L-Alanine (L-Ala)
Erythrose 4-P
L-Norleucine
2-Aminoadipate
Putrescine
Pyruvate
2-Hydroxyglutarate
MtSucS1as19
-6.00a
-5.15b
-4.21
-3.05
-2.66a
-2.60
-2.51
-2.48a
-2.33
-2.31a
-2.20a
-2.05a
-2.00
-1.94
-1.93
-1.92a
-1.84a
-1.70a
-1.65a
-1.64
-1.64
-1.62
-1.58
-1.51
-1.49
-1.48
-1.45
-1.43b
-1.43a
-1.38
-1.37
-1.36
-1.36
-1.36
-1.35
-1.35a
-1.35
-1.32
-1.31
-1.30a
-1.27
-1.26
-1.24
-1.23
-1.23
-1.20
-1.19
-1.19
-1.19
-1.19
-1.16
-1.16
-1.16
-1.15
MtSucS1as12
-3.65a
-2.90b
-1.92
-1.04
-2.06
-1.80
-2.00
-1.20
-1.78
-2.30a
-1.24
1.04
-1.21
-1.85
-1.38
-1.01
-1.38
-1.22
-1.14
-1.17
1.57
-1.00
-5.53
-1.31
-1.03
-2.11
-1.17
-1.72b
-1.30b
-1.17
-1.08
-2.50
-1.23
-5.09
-1.40
1.04
-1.25
-1.14
-1.18
-1.09
-1.13
-1.59
1.08
-1.07
-1.29
-1.08
-1.24
-1.12
1.29
-1.07
1.06
-1.23
-1.09
-1.12
α-Ketocaproate
Glycerate
Salicylic acid
L-Methionine (L-Met)
Succinate
Panthotenic acid
L-Aspartate (L-Asp)
Glucoronic acid
Cellobiose
L-Lysine (L-Lys)
Gluconate 1,5-lactone
Phosphoenolpyruvate (Pep)
Ribose
Thymine
L-Homocysteine (L-HCys)
Fumarate
Ribose 5-P
Gluconate 6-P
N-Acetyl-Glutamate
Gluconate
L-Cysteine (L-Cys)
L-Phenylalanine (L-Phe)
Uracil
Galactose
Melibiose
O-Acetyl-L-Homoserine
myo-Inositol
Fructose 1,6-P
Citrate
Adenine
Malate
Rhamnose
Arabinose
L,L-Cystathionine
L-Glutamine (L-Gln)
L-Glutamate (L-Glu)
Sucrose
Isocitrate
cis-Aconitate
Adenosine
Cytosine
L-Cystathionine
S-Adenosyl-Homocysteine
Maltose
-1.14
-1.12
-1.12
-1.11
-1.11
-1.08
-1.08
-1.07
-1.06
-1.04
-1.03
-1.03
-1.02
1.00
1.01
1.02
1.04
1.05
1.05
1.05
1.07
1.08
1.08
1.10
1.11
1.13
1.14
1.14
1.17
1.19
1.22
1.24
1.30
1.31
1.33
1.35
1.40
1.42
1.42
1.50
1.62
1.68a
1.79
2.26
1.01
-1.03
-1.38
1.07
-1.09
-1.21
-1.10
-1.12
-5.00
-1.05
1.13
-1.09
1.16
1.02
1.40
-1.00
-1.25
1.22
1.08
1.07
1.12
1.26
1.02
1.26
1.07
1.14
1.19
-1.10
1.08
1.55
1.25
2.10
-1.13
1.30
1.38
1.31
1.46
1.39
1.28
-1.64
1.35
1.45a
1.13
2.60
Metabolites are sorted via x-fold changes with respect to as19. Relative values for lines as19 and as12
denote mean metabolite pool size ratios between MtSucS1 antisense and control lines. Significantly
different means are indicated by a (P< 0.05) and b (P< 0.01). Metabolite ratios exceeding the criteria of
significance of both a twofold threshold and P< 0.05 are shown in italic boldface. Ratios achieving only
one criterion are marked in bold. Abbreviations: dpi, day post inoculation; P, phosphate