Supporting Information Legends
Fig.S1. Phenotypes of the sad1-1 mutant. (a) Mature fifth leaves of wild-type and sad1-1 plants. (b)
Heights of wild-type and sad1-1 seedlings. (c) Lamina joints of leaves of wild-type and sad1-1 plants.
(d–g) Cross sections of the fourth leaves of wild-type (d,e) and sad1-1 (f,g) plants cut at 50% (d,f) and
5% (e,g) distance from the base of the leaf. (h–i) Close-up views of plastochron 3 in wild-type (h,i)
and sad1-1 (j,k) plants. The arrow indicates an area of many small cells in the wild-type plant. (l,m)
Longitudinal sections of shoot apices in two-week-old wild-type (l) and sad1-1 (m) plants. Arrows
indicate the third and fourth leaf base nodes.
Fig.S2. Phenotypes of transgenic plants. (a) Complimented phenotype of sad1-1 by the cDNA of the
C1 gene. m3, m4,m8,m9 are segregated siblings containing the introduced SAD1 cDNA in sad1-1
mutant background. H3 and H6 are segregated siblngs with the introduced SAD1 cDNA in sad1-1/wt
genotype background. (b) Phenotypes of the C1 and C2 RNAi lines.
Fig. S3. Amino acid alignments of SAD1 orthologs. SAD1 (Os08g0422600), SAD1LIKE
(Os01g0259900), At,Arabidopsis thaliana (AT5G64680), Gm, Glycine max (XP_003529342,
LOC100790461), Lj, Lotus japonica (chr6.LjT22E11.60.r2.m), Bd,Brachypodium distachyon
(Bradi2g09420), Zm, Zea mays (GRMZM2G132944), Vv, Vitis vinifera (VIT_01s0150g00220), Pt,
Populus trichocarpa (POPTR_0001s12900), Sb, Sorghum bicolor (Sb03g009990), Sl, Solanum
lycopersicum (Solyc01g008760.2), Ps, Picea sitchensis (WS04716_G08), Pp, Physcomitrella patens
(XP_001757296), Es, Ectocarpus siliculosus (CBJ26277), Cv, Chlorella variabilis (XP_005843761), Stp,
Strongylocentrotus purpuratus (XP_789115), Nv, Nematostella vectensis (XP_001640164), Pm,
Petromyzon marinus. (AAN64297), Hs, Homo sapiens (NP_036231), Mm, Mus musculus (BAD02839),
Bt, Bos Taurus (XP_005900384), Rn,Rattus norvegicu (NP_001102886), Xt,Xenopus tropicalis
(XP_002932650), Mz, Maylandia zebra (XP_004569655), Yl, Yarrowia lipolytica (XP_502300),
Dh,Debaryomyces hansenii (XP_457489), Ca,Candida albicans (XP_723578), Sc,Saccharomyces
cerevisiae (NP_012387), Kl,Kluyveromyces lactis (XP_451266), Eg,Eremothecium gossypii
(NP_983645), Ast, Aspergillus terreus (XP_001214870), Sp, Schizosaccharomyces pombe
(NP_595723), An,Aspergillus niger (EHA24120), Nf,Neosartorya fischeri (XP_001265787),
Mg,Meyerozyma guilliermondii (EDK37676).
Fig. S4. Amino acid alignments of RPA49 orthologs. Os, Oryza sativa (Os11g0615100),
At,Arabidopsis thaliana (AT3G13940), Gm, Glycine max (XP_003546428), Lj,Lotus japonicus
(AFK41205), Pt, Populus trichocarpa (XP_002339506), Vv, Vitis vinifera (XP_002278228), Sl, Solanum
lycopersicum (XP_004240985), Bd,Brachypodium distachyon (XP_003577359), Zm, Zea mays
(AFW60354), Sb, Sorghum bicolor (XP_002449857), Ps, Picea sitchensis (ABR17135), Pp,
Physcomitrella patens (XP_001777438), Cv, Chlorella variabilis (XP_005845401),
Stp,Strongylocentrotus purpuratus (XP_793654), Nv,Nematostella vectensis (XP_001625428),
Hs,Homo sapiens (NP_071935), Mm,Mus musculus (NP_001272729), Bt,Bos taurus (NP_001029580),
Rn,Rattus norvegicu (NP_001101408), Xt,Xenopus tropicalis (NP_001120306), Mz, Maylandia zebra
(XP_004564331), Yl, Yarrowia lipolytica (XP_503575), Dh,Debaryomyces hansenii (XP_459948),
Ca,Candida albicans (XP_719593), Sc,Saccharomyces cerevisiae (NP_014151), Kl,Kluyveromyces lactis
(XP_452329), Eg, Eremothecium gossypii (XP_003644181), Sp, Schizosaccharomyces pombe
(NP_594382), Mg,Meyerozyma guilliermondii (EDK41550).
Fig. S5 Phylogenetic tree of RPA49 orthologs in different species. Gene names were showed in
legend of Fig. S4.
Fig. S6. Interaction of SAD1 with components in Mediator. SAD1 interacts with the MED4 subunit of
Mediator in the yeast two-hybrid assay. The SAD1 N terminal half is more important than the C
terminal half for interactions with MED4 and the MED4 C terminal half. The N terminal of SAD1
contains the structural domain for heterodimer formation and shows higher level of conservation
than the C terminal region.
Fig. S7. Roles of Mediator in Pol I, Pol II and Pol III function in plants. (a) Yeast two-hybrid assay of
OsMED4 in the bait vector and OsTFIIF1, OsTFIIF2, or OsRPC53 in the prey vector. Co-transformed
yeast cells were assayed for growth on selective media [–Leu (L), –Trp (T), –His (H), and –Ade (A)].
MED4 interacts with OsTFIIF2 and OsRPC53. Empty vectors were used as negative controls. (b)
Model of OsMED4 interaction with RNA Pol I, Pol II and III.
Fig. S8. Phenotypes of Arabidopsis atmed4 mutants. (a) Genomic organization of the atmed4-1 and
atmed4-2 T-DNA insertion alleles. (b) Siliques from wild-type, AtMED4/atmed4-1 and
AtMED4/atmed4-2 lines. Asterisks indicate aborted embryos.
Fig. S9. Levels of plant hormones in sad1-1. Endogenous plant hormones in shoots (a) and roots (b)
of wild type and sad1-1 mutant. GA,gibberellin; IAA,indole-3-acetic acid; IAAsp,indole-3acetylaspartic acid; ABA, abscisic acid; JA,jasmonic acid; SA,salicylic acid; N.D., not detected.
Supplemental Table 1. Primers used in the experiments