Table S1. Fold increase of gene expression levels and p-values of AY-WB candidate
effector (SAP) genes.
AY-WB
Gene1
pI03
22
32
33
73
127
145
146
148
169
189
203
212
224
2255
2295
236
237
245
259
263
269
275
280
294
295
329
339
340
3425
366
3675
3685
369
370
3765
387
402
433
480
529
562
568
588
624
Fold increase
ID2
in insect3, 4
SAP37
SAP21
SAP05
SAP06
SAP19
2.6
SAP27
SAP59
SAP26
SAP25
1.0
SAP61
SAP36
21.1
SAP55
SAP39
1.5
SAP54
SAP53
1.9
SAP62
1.7
SAP40
SAP41
SAP63
2.9
SAP43
SAP52
1.6
SAP64
11.8
SAP22
SAP44
SAP45
SAP51
3.7
SAP65
7.3
SAP49
SAP48
SAP47
SAP66
SAP56
SAP67
SAP68
SAP11
SAP09
1.8
SAP08
8.8
SAP30
SAP50
5.6
SAP69
1.8
SAP70
1.5
SAP34
2.3
SAP71
SAP35
SAP15
1.9
Fold increase
in plant3, 4
1.3
1.7
2.2
1.8
29.4
1.8
2.1
12.3
1.4
1.4
4.1
6.5
3.2
1.0
1.4
1.4
2.1
1.5
3.3
2.1
2.4
2.7
2.6
2.5
3.3
1.3
1.3
p-value4
0.148
0.209
0.007
0.247
0.005
0.01
0.094
0.01
0.869
0.007
0.006
0.093
0.25
0.028
0.01
0.008
0.007
0.013
0.003
0.014
0.003
0.002
0.833
0.099
0.23
0.003
0.003
0.004
0.081
0.007
0.009
0.01
0.01
0.014
0.01
0.082
0.003
0.011
0.005
0.012
0.003
0.003
0.002
0.014
0.008
640
645
667
1 Numbers
SAP13
SAP20
SAP72
2.5
10.9
1.2
0.011
0.013
0.003
refer to locus tags of GenBank accessions NC_0007716 (AYWB genes on
chromosome), and NC_007717 (pI03 on AY-WB plasmid).
2 Identities
(ID) correspond to secreted AY-WB proteins (SAPs), which are candidate
effectors, published in Bai et al., 2009 (Tables 1 and 2).
3 The
expression levels of the AY-WB genes shown in this table were normalized ith
those of AYWB_007 (dnaB), AYWB_064 (ftsY), AYWB_254 (gyrA), AYWB_434
(pykF) and AYWB_440 (pfkA).
4 Table
rows of genes up-regulated in plants versus insects are highlighted in dark
grey and those up-regulated in insects versus plants in light grey (p-value < 0.05).
Genes whose expression is not significantly different between the plant and insect
host are represented in white rows.
5
Due to identical sequences in the primer annealing sites, primers may not be gene
specific and may have amplified more than one target.
Table S2. Primers and other information for transgenic Arabidopsis lines that
express candidate AY-WB effectors.
Gene
SAP
AYWB_022
21
AYWB_032
05
AYWB_033
06
AYWB_073
19
Gene specific primer sequences to clone into pDONR
(5’ to 3’) 1
AAAAAGCAGGCTCCACCATGGAAAAAGCAAATAAT
CATATTG
AGAAAGCTGGGTGTTAATCTTCTATAATTTTAATTAA
TG
AAAAAGCAGGCTCCACCATGGCCCCTAATGAAGAG
TTTGTTGGC
AGAAAGCTGGGTGTTATTTTTGTTCATATTGCCAAA
A
AAAAAGCAGGCTCCACCATGTTTCCTGGAATAGATT
TAGATG
AGAAAGCTGGGTGTTAAATTCTTCTAGGACTGTTAG
AAAAAGCAGGCTCCACCATGAATAATAATTCATCAT
CGAATATAAAC
AGAAAGCTGGGTGTTAACGTCTTCTTCTATTGTTCG
T1 lines2
>10
>10
>10
>10
AYWB_127
27
AYWB_145
59
AYWB_146
26
AYWB_148
25
AYWB_152
60
AYWB_169
61
AYWB_189
36
AYWB_203
55
AYWB_212
39
AYWB_224
54
AYWB_225
53
AYWB_236
40
AYWB_237
41
AYWB_245
63
AYWB_258
42
G
AAAAAGCAGGCTCCACCATGCACAATGCAACAAAT
ATACAAAAAAAAAC
AGAAAGCTGGGTGTTAATCAGATAGAATATATTTTA
T
AAAAAGCAGGCTCCACCATGAAACAAGCAGCTCAT
AATTTG
AGAAAGCTGGGTGTTATTTAGTTAAATTATTAAC
AAAAAGCAGGCTCCACCATGAAAACTTATGATCAAA
ATCAACATAATTTA
AGAAAGCTGGGTG
TCAATATGTATCTTCGTCTTTATA
AAAAAGCAGGCTCCACCATGAAACAACAAGAACAA
TTTCAATC
AGAAAGCTGGGTGTTAATAATCTGTAATCTTTTTAA
C
AAAAAGCAGGCTCCACCATGCAAGAAAAAATAGATT
TAA
AGAAAGCTGGGTGTCATTTCAGAATTATTAA
AAAAAGCAGGCTCCACCATGGATGAAAAAAAATGT
CCAAAAG
AGAAAGCTGGGTGTTATGGCCAAAGTGCATTAACT
G
AAAAAGCAGGCTCCACCATGATTGGTTATTATTTTT
TTAAAAATC
AGAAAGCTGGGTGTTATTGGGTTTCTAAATTAGTTA
TTTG
AAAAAGCAGGCTCCACCATGGATGATTTAGTTAATG
AAATAAC
AGAAAGCTGGGTGTTATAAAATATTACAATTACAAC
C
AAAAAGCAGGCTCCACCATGTTAACTAATTACTTTT
CTAAAC
AGAAAGCTGGGTGTTATGCTAAATTAGAGATTTTAT
TTTC
AAAAAGCAGGCTCCACCATGGATAAAGATATTGCTA
GC
AGAAAGCTGGGTGTTAATTATTTTCATCATTTAAAG
AAAAAGCAGGCTCCACCATGGAAACAGGAGTCATC
AAACTCC
AGAAAGCTGGGTGTTAAGTAGTTACCACCTGGG
AAAAAGCAGGCTCCACCATGAATAATAATAACGTTG
GAACAAG
AGAAAGCTGGGTGTTAACGTCTTTGAGAAGAACG
AAAAAGCAGGCTCCACCATGGAAAACAATAATTTAT
TAGATTC
AGAAAGCTGGGTGTTAATAATTTATATTTAATTTTGT
AAAAAGCAGGCTCCACCATGATAGAAGATTGGCAT
AAATTAAAAAG
AGAAAGCTGGGTGTTATGTTTTTTCTATTCTATATTT
TA
AAAAAGCAGGCTCCACCATGATAAATAATTTAAATG
ATGAAAATTC
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
AYWB_259
43
AYWB_263
52
AYWB_269
64
AYWB_275
22
AYWB_280
44
AYWB_294
45
AYWB_295
51
AYWB_329
65
AYWB_339
49
AYWB_340
48
AYWB_342
47
AYWB_366
66
AYWB_367
56
AYWB_368
67
AYWB_369
68
AGAAAGCTGGGTGTTATCTAATTTTTTTTAATTTTAT
ATC
AAAAAGCAGGCTCCACCATGGAAACAAGACAAAAT
GAAAATAG
AGAAAGCTGGGTGTTAAATTATATTTTTAATAAAATC
AAAAAGCAGGCTCCACCATGGGAATTTTATGGCCG
AAAAATAC
AGAAAGCTGGGTGTCAGTTTTGATTTATTGCAACTT
C
AAAAAGCAGGCTCCACCATGAAAACGGACACCCAA
AAAGAACAAG
AGAAAGCTGGGTGTTAAAATGTTTTGGTTTCTTTAA
TAGTTCC
FOR:AAAAAGCAGGCTCCACCATGCCATTCACAACC
AACAATAACCAAAATACAC
M1R: GCTTTACCATGTGATTCTGCTTCTTTCC
M1F: GCTTTATGTAATGAAACCGAAGCAC
REV:AGAAAGCTGGGTGTTATTTATTTTCTTTTGTGT
TGTGTTTTTGG
AAAAAGCAGGCTCCACCATGGAAAAAAATAAACCC
CTTCAAAC
AGAAAGCTGGGTGTTATTTTAATTTATTTATTATATT
AAAAAGCAGGCTCCACCATGAATAATAATGAAGTTA
GAAATAT
AGAAAGCTGGGTGTTAATGTTTTAATTTTTTTATTAA
TTC
AAAAAGCAGGCTCCACCATGATTAATTATGAAAATG
AATCAATTTC
AGAAAGCTGGGTGTTATTTATTTAGATTGTTTTG
AAAAAGCAGGCTCCACCATGATCTGTAAACCATCA
GAAAAAATAC
AGAAAGCTGGGTGTTATTTTTTGGTTTCTTTGAG
AAAAAGCAGGCTCCACCATGATAGAAAATAAAAAAA
TATGCAATC
AGAAAGCTGGGTGTTAAAATATTTTTTTATACAATGA
ATTAA
AAAAAGCAGGCTCCACCATGTTCCCTGGTATAGATT
TAGACGC
AGAAAGCTGGGTGTTATAAATTAGTATCATTAAAAA
C
AAAAAGCAGGCTCCACCATGGAAACAGGAGTCATC
AAACTCC
AGAAAGCTGGGTGCTATCTGTTTCTTTCCCATTC
AAAAAGCAGGCTCCACCATGAATAACGATAATAACA
ATAATG
AGAAAGCTGGGTGTTATCGATTTAGGGGGTTATTT
GG
AAAAAGCAGGCTCCACCATGTTACCTGGAATAACTT
TAG
AGAAAGCTGGGTGTTATTGTATGATGTCTCGC
AAAAAGCAGGCTCCACCATGGGAAATAAAAATAGC
AGAAAGCTGGGTGTTAATCATTAATTATCACTC
AAAAAGCAGGCTCCACCATGCATAATGGTAATGCA
>10
>10
>10
>10
5
>10
>10
>10
>10
4
2
>10
>10
>10
>10
AYWB_370
AYWB_376
11
09
AYWB_387
08
AYWB_402
30
AYWB_433
50
AYWB_480
69
AYWB_529
70
AYWB_562
34
AYWB_588
35
AYWB_624
15
AYWB_640
13
AYWB_645
20
AYWB_667
72
pI03
37
pI04
01
pII03
02
AC
AGAAAGCTGGGTGTCAACGTCTTCTGTTATTAG
N.A.
AAAAAGCAGGCTCCACCATGTTAACTAATTACTTTT
CTAAAC
AGAAAGCTGGGTGTTATGATAAATTAGAGAT
AAAAAGCAGGCTCCACCATGGGTCAAGTTTATTCA
GAAAT
AGAAAGCTGGGTGTTATTTACAAGTTATTTTTTTTC
AAAAAGCAGGCTCCACCATGACTTGTAATAATAATT
CTGAAG
AGAAAGCTGGGTGTTATTTTTTCTTTTTATTAGG
AAAAAGCAGGCTCCACCATGTCCAAAAATTTTAATT
ATAATG
AGAAAGCTGGGTGTTAAATTTTGTAATAAAGTG
AAAAAGCAGGCTCCACCATGGATAACAACAATCAA
GTTTTAC
AGAAAGCTGGGTGTTAGAATTCGCTTTTGTAAGGTC
AAAAAGCAGGCTCCACCATGTATCATTTAGTTAAAG
ATTCAATAG
AGAAAGCTGGGTGTTATATTTTTTTTATTTGAGTAAT
ATC
AAAAAGCAGGCTCCACCATGAAATTAAGCCAACCC
CTTCC
AGAAAGCTGGGTGTTATATATTTAATTTATCAGATTC
AAAAAGCAGGCTCCACCATGGATAACTCCTATAAC
CAAAAAACCC
AGAAAGCTGGGTGTTAATTAGAAGAAGAAGAAGCA
GA
AAAAAGCAGGCTCCACCATGAAAAATGCCAAAACT
GTTTCAAAAG
AGAAAGCTGGGTGTTATTGTTTTAATTCTTTTTCTAT
AAAAAGCAGGCTCCACCATGACGCCTTATAATGTTC
CTTTAAAAAC
AGAAAGCTGGGTGTTATAACCTACTAATAGGTTTTT
TG
AAAAAGCAGGCTCCACCATGATAAATAAAGATATTG
CTTC
AGAAAGCTGGGTGTTATTTTTTTATAAAGGAAACAT
TATC
AAAAAGCAGGCTCCACCATGTTAAGAAATAATCATG
AACAAGAATTA
AGAAAGCTGGGTGTTATGATTCTGCTTTTTGTTGAG
CTTCG
AAAAAGCAGGCTCCACCATGATTAATAAACCTCAAC
CTAAAACTG
AGAAAGCTGGGTGTTAATTAAAATGCTTTACTTTTTT
TTC
AAAAAGCAGGCTCCACCATGCAATCTCAATATCAAA
CTCAATATATC
AGAAAGCTGGGTGTTAAGCAATTGAAACTAATTTAC
TTG
AAAAAGCAGGCTCCACCATGATTAATAAACCTCAAC
>10
6
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
pIII02
73
pIV03
76
1With
CTAAAACTG
AGAAAGCTGGGTGTTATTCATTAGAATGATAAGTGT
C
AAAAAGCAGGCTCCACCATGAAATCAGAAAAAGATT >10
TAAAAAAAG
AGAAAGCTGGGTGTTATTCCCAATCAAAAAAAGTTT
C
AAAAAGCAGGCTCCACCATGATTAATAAACCTCAAC >10
CTAAAACTG
AGAAAGCTGGGTGTTATTCATTAGAATGATAAGTGT
C
the exception of AYWB_275 (SAP22) all phytoplasma effector genes were
amplified by one-step PCRs using specific primers with attB1 and attB2 adapter
sequences at the ends. AYWB_275 (SAP22) was generated by generating two PCR
fragments using primer pairs For&M1R and M1F&Rev and a subsequent second
round of PCR in which the two PCR products were combined and fused into one
PCR product by an overlap PCR procedure using attB1 and 2 adapter primers (see
Materials and Methods).
2 Number
3
of T1 lines obtained and analyzed for phenotypes.
N. A. = not applicable; the SAP11 gene was synthesized and not amplified.
Table S3. Primers used for RT-PCR to examine SAP54 gene expression in stable
transgenic Arabidopsis lines.
Gene ID
Primer name
Primer sequence (5’ to 3’)
AYWB_224 (SAP54)
SAP54For
GGGGACAAGTTTGTACAAAAAAGC
SAP54For
TTTGATAATTGTTGTATTTCGTTTTC
Arabidopsis actin
ActinFor
GCACCCTGTTCTTCTTACCG
(GenBank accession
ActinRev
AACCCTCGTAGATTGGCACA
number: AT3G18780)
Table S4. Primers used in expression analysis of AY-WB genes.
Gene ID_(forward
or reverse
primer)
AYWB022_fwd
AYWB022_rev
AYWB032_fwd
AYWB032_rev
AYWB033_fwd
AYWB033_rev
AYWB073_fwd
AYWB073_rev
AYWB127_fwd
AYWB127_rev
AYWB145_fwd
AYWB145_rev
AYWB146_fwd
AYWB146_rev
AYWB148_fwd
AYWB148_rev
AYWB169_fwd
AYWB169_rev
AYWB189_fwd
AYWB189_rev
AYWB203_fwd
AYWB203_rev
AWYB212_fwd
AWYB212_rev
AYWB224_fwd
AYWB224_rev
AYWB225_fwd
AYWB225_rev
AYWB229_fwd
AYWB229_rev
AYWB236_fwd
AYWB236_rev
AYWB237_fwd
AYWB237_rev
AYWB245_fwd
AYWB245_rev
AYWB259_fwd
AYWB259_rev
AYWB263_fwd
AYWB263_rev
AYWB269_fwd
AYWB269_rev
AYWB275_fwd
AYWB275_rev
AYWB280_fwd
AYWB280_rev
AYWB294_fwd
Primer sequence (5’ to 3’)
TCAAGTAATGGCGATGGAAAAAG
TAATGTATTTTTTCTACAATTTGTTTTTGATCTC
AGCAATGGCTGCCCCTAAT
TTAACTATTCTCATGTCGCCAACAA
ATGGTTGCAATTTCAAACACAATAG
CGTCGAGAGTTGATTGTTTGTTG
CGTTGTATGCTTCTGAATCACAAA
TTGCGCTTTTTTTCGATCAATAT
AAATAGACTCGGATTCATCATCTGAAT
CTCTCATCGAATGTAGCGAAACA
CCTGTTAACTGGCTTTCTCCTAAAA
ACTGAATTATACAAATTATGAGCTGCTTG
CTTGGTCACGTTCGGCAATAT
GATTCATTATCAACCCGATGGAA
AAATTGAAAAAGAAGTTGATGCATTAAC
TTAGTATTCGGCAAAAAAATAACGTTT
TGGACACACTTCGCGATTATATTT
TGTACATCTTCCCAATCTTCAACAAC
TTTAATTCAGAAGAAATTTGAGGGTCT
AAAACTGCTTTAGAAAACCAAAAAACA
TCGTTGCCAGAATCTATTTGACTT
GGGACATGCGTTAGTAGCTTATGAT
TACGATGATACTAAACGCCCACAA
TTGGATGTTAAGTTTTCCTAAGCTGTT
AATGTATTTGTATTCGCTTTTTTAGGATT
GCAATATCTTTATCCATCCCCATTACT
ATATAATTGGCTGTCGGCTTTTG
GACGGTTTCGACAGTGCATCT
GCACTGTCGAAACCGTCTAAAGT
CGATGGAATAAAGATGGCGATACT
TGCGATGAATAATAATAACGTTGGA
CTAAAGAATTATAAACATGCTCGCTTGT
AGATTCAAGAGCATCAGAGGAACA
ACGGACAAACTCCAAATCTTATTGT
TTTAATGACAACAGAACAAACAGATAGAGT
AGAGTGGCTTCTTGTTCAAAAATAACT
TATGCTTCGGATGTTGATAATATGG
GAAAAGAAGGCACAAGAAACAAATT
GATTTTTGTTTAAGCCTGCAACAA
TGATTTCTGCTTTAAATTCCAATGAC
TCGATTGTTTCATTAGCTTTTGTGT
TGTCCGTTTTAGCTTGAGGTTTATT
AACCCATTTGACAAATGGTACGA
AACCATCAAACTCATCTACTCGCAT
AATCATCAAGTAATGGCAATGGAA
TGGTTTCAATGTTATAAATCGTTTGAA
ATGAAGTTAGAAATATTAATGATTTATCGATTG
AYWB294_rev
AYWB295_fwd
AYWB295_rev
AYWB329_fwd
AYWB329_rev
AYWB339_fwd
AYWB339_rev
AYWB340_fwd
AYWB340_rev
AYWB342_fwd
AYWB342_rev
AYWB366_fwd
AYWB366_rev
AYWB367_fwd
AYWB367_rev
AYWB368_fwd
AYWB368_rev
AYWB369_fwd
AYWB369_rev
AYWB370_fwd
AYWB370_rev
AYWB376_fwd
AYWB376_rev
AYWB387_fwd
AYWB387_rev
AYWB402_fwd
AYWB402_rev
AYWB433_fwd
AYWB433_rev
AYWB480_fwd
AYWB480_rev
AYWB529_fwd
AYWB529_rev
AYWB562_fwd
AYWB562_rev
AYWB568_fwd
AYWB568_rev
AYWB588_fwd
AYWB588_rev
AYWB624_fwd
AYWB624_rev
AYWB640_fwd
AYWB640_rev
AYWB645_fwd
AYWB645_rev
AYWB667_fwd
AYWB667_rev
AYWBpI03_fwd
AYWBpI03_rev
TCATTGTATTTAGATTAAGTCGATCATATTGT
CATTAATTGTATTCATCTTGTATGAAATTGA
TCCGTTATGGCGATTAATTATGAAA
ACCTCAATATGATGGTTGGATAGCT
TCTTGTCCATTAAATGAAGGGAAAC
AATTCCGCTATCAAAAAAAACAAATAA
TGGCGTATCAAAGAACCTCCTAAT
TTGATGTTTTCTTGCGTCTAAATCTATAC
TTAATACTAAATGTTTGCATGCTTTCC
TGGGTCTTTAGGGAGTTTGATGA
TGGCAGCTTTTATTGGGTTTATC
GGTTGTTGCCCGATAATTCTTAAT
ATATCAGAAACCGAGCGATATATTTAGA
TCTTTCTTCTAATCGGATCTTTTGTG
AATGTTTGTATGCTTTACCTGGAATAAC
AAGGCATATTCTTCATCATTGGAAA
GCGATGGGAAATAAAAATAGCAA
TTGTAAAATGCGCGCTTGAGT
TGCAACACCAAATAACGGTCAT
TTTAAATTTTTCGTTTCCTCTTCGG
TTAGAAAACCCTGAATTAATGGAAATATT
AGTTGCTTCGTGTTCGATCGT
ACCAATTCCCATTCCAAACTTAAA
TCAAATAATTTTTCCGAAGAATCTGTT
AATTTGGTTTATGCTGGTCAAGTTTA
CTTTTTTAGGATTATTTTTTATCACCAACA
CTTCAGAATTATTATTACAAGTTGCCATTAC
GTTTATTTTGATGGAGGCACAAGA
TTGAGCGCCATTACCTAAAGTTT
GAGGAGCAGGAATAACTTGTTTGAC
TTAACTTTTTTGCGTGTTTCTGAAA
AAAAAGAATTCAGCCAAACAGCTATAG
TATCATGGATACAACGAAAAAAACGAT
AAAAGAAAAACCAAATCCCACTCTTA
TGTTATGCAATAATTGGTCTTGAAAAGT
GAATTGAACAAAGAAAAGCGGTTT
TTATTGCCGTTGGTATTTTGTTATCT
ATTCTTGCAACAAACACAAAGCTAA
TGCCACAAGATAATTTTCAACGTT
CATCCTCCTTGCCTTTTTGATTTAT
TGCAGTTAGATGAAGCAAATACCAA
GCGACGCCTTATAATGTTCCTTTA
TTTTCTTTAATATGGTTGTTTTGAATTTG
TGGTTCTTTTCCAACTACAGAAGGT
ATGATTGTGGATGGCTTTGTTGT
TTGTTTCTGGGCAAGGTTCTAATT
AGTTCCCCTTGATGGATTTAAAGA
TGGAATTGTTGGTTATATTGCAAAA
TTGAATTATATCACCTGTGTCTTTGCT
Figure S1. AY-WB infection stunts leaf growth growth and leads to anthocyanin
accumulation in Arabidopsis. Shown are rosette leaves sampled from 10-week old
infected (above) or healthy (below) Arabidopsis plants (6-weeks post inoculation).
Plants were grown in short day photoperiod. Scale bar, 1 cm.
Figure S2. AY-WB-infected plants produce narrower and paler leaves than healthy
plants. Shown are 6-week old healthy (left) and infected (right) plants (3-weeks postinoculation). Top panel represents close-up images of whole plants below. Plants
were grown in short day photoperiod. Scale bar, 1 cm.
Figure S3. Rosettes of AY-WB-infected Arabidopsis plants have a reduced diameter.
A, Image of healthy (left) and infected (right) plants grown in long day (LD)
photoperiod (4-weeks post-inoculation). B, Image of healthy (left) and infected (right)
plants grown in short day (SD) photoperiod (6-weeks post-inoculation). C, Rosette
diameter is slightly reduced in infected plants grown in LD. D, Rosette diameter is
severely reduced in infected grown in SD. Error bars represent standard error. N ≥
10. Scale bar, 1 cm.
Figure S4. Plants infected with AY-WB have a decreased height. A, Healthy (left)
and infected (right) 9-week old plants (5-week post-inoculation). Scale bar, 1 cm. B,
Quantification of height differences between healthy and infected plants. Error bars
represent standard error. Plants were grown in long day photoperiod.
Figure S5. AY-WB infected Arabidopsis plants produce an increased number of
axillary stems (witches’ broom phenotype) at an early stage of infection. Stem
number was measured at 22 days (A), 26 days (B), and 35 days (C) post-inoculation.
Error bars represent standard error. Data in A and B are statistically significant, as
indicated by an asterisk; Students t-test, p-value <0.05.
Figure S6. AY-WB infected plants produce leaf-like flowers. A, AY-WB infected
plant, boxes encompass large leaf-like flowers that produce shoots in lieu of carpels.
B, Flowers from infected plant have stellate (branched) trichomes on green petals
(pe), sepals (se), and carpels (ca), and produce truncated virescent stamens (st) that
do not produce pollen. Scale bar, 1 mm.
Figure S7. AY-WB genes encoding effector proteins are expressed in infected Arabidopsis.
Figure S8. SAP54 expression in Arabidopsis transgenic lines. Data show RT-PCR
results from total RNAs isolated from Arabidopsis lines as indicated. RT-PCR of
Arabidopsis Actin 2 (AT3G18780) was used as control.
Figure S9. Expression of SAP54 induces leaf-like stamens. A, Flowers of
35S::SAP54 transgenic lines frequently produce an increased number of stamens
that undergo conversion to leaf-like organs. In this image, a plant has produced
seven stamens (as numbered) whereas wild-type Arabidopsis plants yield six
stamens. B, Stamens undergo transformation, as evidenced by production of
trichomes (indicated by arrowheads), virescence, and outgrowth of leaf-like
structures (C). C, Image of leaf-stamen hybrid. The anther is indicated by an asterisk
and arrowheads delineate the stalk of the stamen. Plants were grown in short day
photoperiod and flower structures of 11 week-old plants were photographed. Scale
bar, 1mm.
Figure S10. Carpels of 35S::SAP54 Arabidopsis line 1 are bulbous and possess
trichomes (tr). Carpels collected from 11 week-old plants were dissected and
processed for cryo scanning electron microscopy. Figure at right is enlargement of
leaf-like structure (lls) inside the carpel in figure at left. Plants were grown in short
day photoperiod. Scale bar, 200 μm. Abbreviations: lls, leaf-like structure; ov, ovule;
tr, trichome.
SAP54
Maryland AY
Spiraea stunt
Onion Yellows
1
1
1
1
MFKIKNNLLLLLNVF-VFAFLGLFLINNNQQVMGMDKDIASTSNNNPNITNYSIEENIIN
MFKIKNNLLLLLNAFIVFTLLGLFLITNNQQVMGMNKDIASASNNNQNITNYSIEENIIN
MFKIKNNLLLLLNAFIVFTLLGLFLITNNQQVMGMNKDIASASNNNQNITNYSIEENIIN
MFKIKNNLLLLLNSFIVFAFLGLFLIANNQQVMGMNKDIASASNNNQNITNYSIEENIIN
************* * **::****** ********:*****:**** *************
SAP54
Maryland AY
Spiraea stunt
Onion Yellows
LKYKIRENAVKKINIENEIQQLSNNEPRKNTLLTLKKNLENLINNQKEQLKTYQILLKTL
LKYKIRENAVKKINTESEIQQLSNNDPKKNTLLALKQNLENLINNQKKQLKTYQTLLKTL
LKYKIRENAVKKINTESETQQLSNNDPKKNTLLALKQNLENLINNQKKQLKTYQTLLKTL
LKYKIRKNAVKKINTEREIQQLSNNDPNKNTLLALKQNLENLIHNQKEQLKTYQKLLKTL
******:******* * * ******:*.*****:**:******:***:****** *****
SAP54
Maryland AY
Spiraea stunt
Onion Yellows
NDENN
NDENN
NDENN
NDENN
*****
124
125
125
125
Figure S11. Alignment of an AY-WB phytoplasma effector
that induces
morphological changes in flowers. SAP54 homologues are encoded in the genomes
of Maryland Aster Yellows phytoplasma (Maryland AY; ABH11652.1), Spiraea stunt
phytoplasma (ABU55747.1), and OY-M (NP_950301.1). The signal peptide of SAP54
is underlined and conserved amino acids are indicated by an asterisk.