NappHS. rrna. transcript abundance. NappHS relative con W+W 0.8. nicotine [µg mg -1 FM]
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1 (A) W+OS 3 min 6 min con L S L S RNA loding control NppHS rrna (B) (C) 8 1 k NppHS reltive trnscript undnce *** *** *** *** 3 k line 1 line (D) nicotine [µg mg -1 FM] con W+W Supplementl Figure S1: (A) Northern lot nlysis of NppHS trnscripts in wild-type () plnts. Rosette-stge leves (position +) were wounded with pttern wheel (w), nd µl of 1:1 (v/v) diluted M. sext orl secretions (OS) ws pplied to the resulting puncture wounds. Treted (L) nd systemic (S) leves from OS-elicited plnts t position were hrvested fter 3 min nd 6 min, s well s unwounded (con) leves t position + from five plnts. (B) Southern lot nlysis of, line 1 nd line plnts. Genomic DNA (5µg) ws digested with BmH1, lotted onto nylon memrne, nd hyridized with PCR frgment of the hptii gene, selection mrker on the trnsformtion vector presc. (C) Reltive NppHS trnscript undnce in locl (+) nd systemic () leves of W+OS treted Arizon () nd pphs-silenced plnts ( line 1 nd ), (*** = P <.1, ANOVA). (D) Men ± SE lef (+) nicotine content of 5 control (white rs), line 1 nd line plnts. Nicotine ccumultion ws dditionlly determined in rosette-stge leves tht were wounded, followed y the ppliction of µl wter (3 in row, every 3 min), nd hrvested four dys fter the tretment (lck rs).
2 (A) 8 8 stlk length [cm] 6 4 * * 6 4 line 1 line line 1 line dys fter elongtion (B) 1 1 flowers/ plnt (C) 1 oveground iomss [g] Supplementl Figure S: NppHS-silenced plnts do not differ from wild-type () plnts in flower production or stlk elongtion. (A) Men ± SE growth rtes of 1 to 15 (lck squres), nd the two lines of pphs-silenced plnts in two ccessions: line 1 (empty squres) nd line (gry squres) of (left pnel) nd (right pnel) ccession. Asterisks represent significnt differences from vlues of ech ccession (s determined y ANOVA followed y Bonferronicorrected post-hoc test). First flowering is indicted y rrows in oth pnels (: lck rrows, line 1: white rrows nd line : gry rrows). Numer of flowers (B) nd ove-ground iomss (C) in (lck rs) nd pphs-silenced plnts: line 1 (white rs); line (gry rs) of the (left pnel) nd the (right pnel) ccession. Brs represent mens ± SE of 9 to 1 replictes. Different letters indicte significnt differences mong the genotypes.
3 line 1 Supplementl Figure S3: Viility of pollen from nd flowers. Pollen were stined with fluorescine dicette nd visulized using fluorescence microscope.
4 flowers with elongted pistil [percentge per plnt] *** *** *** *** Supplementl Figure S4: Percentge of flowers with elongted pistils. Flowers with norml pistil growth in (lck oxes), line 1 (white oxes) nd line (gry oxes) of (left pnel) nd (right pnel) were counted nd divided y the totl numer of flowers per plnt (n = 8 to1 plnts/ per genotype).
5 uds [5-1mm] (A) ABA [ng g -1 FM] uds [3-4mm] (B) 16 (C) flowers Supplementl Figure S5: Men ± SE scisic cid (ABA) content of 4 to 5 smll (A, 5-1mm), elongted (B, 3-4mm) uds nd flowers of (lck rs) nd line 1 (white rs) nd line (gry rs) (left pnel) nd (right pnel) ccessions. Different letters indicte significnt differences mong plnts nd their respective -silenced lines (ANOVA followed y Bonferroni-corrected post-hoc test).
6 IAA [ng g -1 FM] , Supplementl Figure S6: Indole-3-cetic cid (IAA) concentrtion in nd uds. Men ± SE IAA content (n = 5) in uds of (left pnel) nd (right pnel) (lck rs) nd their respective lines 1(white rs) nd lines (gry rs). Different letters indicte significnt differences s determined y ANOVA (Bonferroni-corrected post hoc tests, P <.5).
7 (A) self-pollintion 5 5 Ethylene [nl/g/h] 4 3 1, 4 3 1, (B) 5 hnd-pollintion 5 Ethylene [nl/g/h] line 1 line 1 Supplementl Figure S7: Ethylene emissions re reduced in self-pollinted ut not in hnd-pollinted flowers. Ethylene emissions were mesured y stop-flow mesurements of single flowers. Men vlues of nd (lck rs), line 1 (white rs) nd line (gry rs) flowers re shown, error rs represent SE (n = 4-6), nd different letters indicte significnt differences s determined y ANOVA (Bonferroni-corrected post hoc tests, P <.5).
8 NppHS reltive trnscript undnce s- ud l- ud flo s- ud l- ud flo Supplementl Figure S8: Reltive trnscript undnce of NppHS in smll uds (s-ud), uds (lud) nd flowers (flo) of (gry rs) nd (lck rs) plnts. Different letters indicte significnt differences (ANOVA followed y Bonferroni-corrected post-hoc test, n = 5).
9 linolenic cid LOX HPOT 5 1 AOS llene oxide OPDA JA reltive trnscript undnce Ile JAR4 4 4 JA-Ile JAR coroll pistil stmen coroll pistil stmen Supplementl Figure S9: Trnscript ccumultion of genes involved in jsmonte synthesis nd in the conjungtion step of JA with Ile in the oxylipin pthwy. Left pnel shows simplified model of the oxylipin pthwy: A 13-lipoxygense (LOX3) oxygentes linolenic cid to 13-HPOT (13- hydroperoxide linolenic cid) which is further converted to llene oxide nd OPDA (1-oxophytodienoic cid) y AOS (llene oxide synthse). OPDA is converted to JA,nd the conjugtion of JA to Ile is medited y JAR4 (jsmonte resistnt 4) nd JAR6 (jsmonte resistnt 6). Reltive trnscript undnce of (A) LOX3, (B) AOS, (C) JAR4 nd (D) JAR6 in coroll, pistil nd stmen of 3 to 5 replicted wild-type (lck rs) nd line 1 (white rs) nd line (gry rs) plnts of (left pnel) nd (right pnel) plnts. Asterisks represent significnt differences from vlues of ech ccession (s determined y ANOVA followed y Bonferroni-corrected post-hoc test).
10 α-kb [nmol/min/mg/protein] line1 line line1 line Supplementl Figure S1: α-ketoutyrte (α-kb) concentrtion in uds of 4-5 replictes from nd (lck rs) nd their respective inverted repet () lines 1 (white rs) nd lines (gry rs).
11 (A) (B) N HypSys_1 Nt HypSys_A_1 N HypSys_ Nt HypSys_A_ Sl HypSys_ Sl ProSys St ProSys_1 St ProSys_ Sn ProSys C ProSys Sl HypSys_3 Ph HypSys_ Ph HypSys_1 C Sys Sl Sys Sn Sys St Sys_1 St Sys_ Ph ProHypSys_1 Ph ProHypSys_ N ProHypSys Nt ProHypSys_B Nt ProHypSys_A Sl ProHypSys Ph HypSys_3 Sl HypSys_1. expected chnges per site 1. expected chnges per site Supplementl Figure S11: Phylogenetic reltionship of (A) N HypSys nd HypSys s well s Sys from other plnt species nd of (B) N ProHypSys, ProHypSys nd ProSys of other plnt species (sed on deduced mino-cids sequences). GenBnk ccession numers: Cpsicum nnuum (C ProSys: AF375), Nicotin ttenut (N ProHypSys: AY4567), Nicotin tcum (Nt ProHypSys-A: AY33148, Nt ProHypSys-B: AY33149), Petuni hyrid (Ph ProHypSys: EF5548), Solnum lycopersicon (Sl ProHypSys: AY91, Sl ProSys: M8481), Solnum nigrum (Sn ProSys: AF375) Solnum tuerosum ( St ProSys_1: AF373, St ProSys_: AF374).
12 line (A) (B) (C) (D) Supplementl Figure S1: nd flowers t different developmentl stges. Smll uds (A), uds efore nther dehiscence (B), uds fter nther dehiscence (C) nd fully opened flowers (D).
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X p -lu c ct ivi ty doi:.8/nture8 S CsA - THA + DAPI Merge FSK THA TUN Supplementry Figure : A. Ad-Xp luc ctivity in primry heptocytes exposed to FSK, THA, or TUN s indicted. Luciferse ctivity normlized
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. Norml Physiologicl Conditions. SIRT1 Loss-of-Function S1. Model for the role of SIRT1 in the regultion of memory nd plsticity. () Our findings suggest tht SIRT1 normlly functions in coopertion with YY1,
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