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Supporting Information Figs S1–S4 and Tables S1–S5
Fig. S1 Anther development of WT and efd plants. Anther sections of WT and efd are shown. (a,
b) WT (a) and efd (b) anthers at stage 7, callose wall generated in both WT and efd anthers. (c, d)
WT (c) and efd (d) anthers at stage 8, microspore cytoplasm in efd anther looks more condensed.
(e, f) wt (e) and efd (d) anthers at stage 9, arrow in F indicates empty efd microspore. (g, h) WT (g)
and efd (h) anthers at stage 10. Pollen grains already look irregular in shape at this stage. (i, j) WT
(i) and efd (j) anthers at stage 11. The arrow in (j) indicates arrested uni-nuclei microspore at stage
11. (k, l) WT (k) and efd (l) anthers at stage 12, showing tapetum remanet and few pollen grains in
efd anther. Msp, microspore; T, tapetum; Tds, tetrads; PG, pollen grain; RM, remanet. Bars, 10
μm.
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Fig. S2 SEM observation of dehiscent anthers and mature pollen grains of WT and efd. A, B
are anthers of WT (A) and efd mutant (B) plants. A, elliptic wild-type mature pollens; B, mature
efd pollen grains which are irregular in shape. Arrow in B indicates membrane-like structure that
wraps around efd pollen grains. C and D are mature pollen grains of WT (C) and efd mutant (D)
plants. C, sculpture structure of WT pollen exine. D, abnormal exine structures of efd pollen grains.
Arrow in C indicates normal sculpture exine of WT mature pollen；Arrow in D indicates coniform
protrusions on the surface of efd pollen grains. Bars: (A, B) 50 μm; (C, D) 5 μm.
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Fig. S3 Characterization of EFD protein. (a–c) The protein conformation of a typical DNA MT
revealed by X-raystressanalysis (a) and EFD predicted by Swiss pdb-viewer (b), and then merged
(c). In B, blue and green indicate α-helix and β-sheet, respectively. (d, e). Sub-cellular location of
EFD in onion epidermal cell. EFP is location in nucleus. (f, g) Sub-cellular location of GFP driven
by CAMV35s promoter. Bars: (d–g) 50 μm.
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Fig. S4 Phylogenetic analysis of EFD. Sequences of DNA methyltransferases were obtained
from
NCBI
database
and
then
analyzed.
MET1
(P34881),
CMET5
(AF007807),
DanioDnmt1(AF097875), MusDnmt1 (P13864) and ZMET1 (AF0063403) are members of
MET1/Dnmt1 family; AtDnmt2 (AED93453) and MusDnmt2 (AF045889) are members of Dnmt2
family; DRM1 (B62154), DRM2 (AED92056), DanioDnmt3 (AF135438), MusDnmt3A
(AF068625), MusDnmt3B (AF068628) and ZMET3 (A1736568) are members of Dnmt3 family;
CMT1 (AF039372), CMT2 (AL021711), CMT3 (AEE34973) and ZMET2 (AF243043) are
members of CMT family.
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Table S1 Results of reciprocal cross between WT and efd
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♀
♂
Number
efd
WT
38
WT
efd
6/30
11.5normal vs 37.2abnormal ovules
WT
efd
14/30
Siliques elongate, but no or few normal ovule
WT
efd
10/30
No developmental ovules
Description
49.6 normal vs 1.8 abnormal ovules per silique average
Table S2 Statistical data of efd anther dehiscence
Number
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Description
59/180
Anthers that didn’t dehisce.
88/180
Anthers that dehisced but no pollen could be observed.
33/180
Anthers dehisced with only a few pollen grains.
Table S3 Genes with greatest down-/up- expression in efd buds
Gene ID
Description
Change fold
AT4G08670
putative lipid transfer protein
–22.3273
AT4G35010
β-D-galactosidase
–13.732
AT2G07560
putative plasma membrane protein, ATPase
–12.220
AT5G14380
AGP6
–13.169
AT3G01270
putative pectate lyase
–13.143
At1g12010
putative amino-cyclopropane-carboxylic acid oxidase
+23.610
AT5G23010
2-isopropylmalate synthase-like
+4.926
AT2G18550
putative homeodomain transcription factor
+4.822
AT2G05540
putative glycine-rich protein
+4.160
AT1G61800
glucose 6-phosphate/phosphate-translocator precursor
+3.920
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Table S4 Altered exine formation related genes in efd buds
Locus
Description
Log2 fold change in efd
AT2G13680
synthesis of callose, CALS5
-3.51234
At5G40260
MtN3/saliva family protein, RPG1
-5.84866
AT1G01280
sporopollenin synthesis, CYP703A2/DEX2
-11.4447
Table S5 Altered sporopollenin secretion and translocation related genes in efd
Locus
Description
Log2 fold change in efd
AT3G55100
ABC transporter
-2.15336
AT2G29940
putative ABC transporter
-2.4811
AT4G08670
putative lipid transfer protein
-22.3273
AT3G18280
lipid transfer protein
2.00977
AT1G18280
lipid transfer protein
-7.68879
AT2G25890
Glycine-rich protein
-6.05866
AT5G07510
Glycine-rich protein (GRP14)
-2.23752
AT5G07520
Glycine-rich protein (GRP18)
-2.01245
AT5G07530
Glycine-rich protein (GRP17)
-2.03534
AT5G07540
Glycine-rich protein (GRP16)
-2.24775
At2G07560
Oleosin-like protein (GRP20)
-13.2201
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