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&OHJOFFSJOH)JHI4UBSDI$PSO-FBWFT

4FBO&8FJTF ;BDIBSZ++BSPV 5IPNBT%4IBSLFZ


.JDIJHBO4UBUF6OJWFSTJUZ
%FQBSUNFOUPG#JPDIFNJTUSZBOE.PMFDVMBS#JPMPHZ
3BUJPOBMF 0WFSDPNJOHUIF:JFME1FOBMUZ
5IJTTVCQSPKFDUGPDVTFTPOJODSFBTJOHUIFZJFMEPGFBTJMZEFHSBEFEQPMZNFST  5PBUUFNQUUPPWFSDPNFUIFZJFMEQFOBMUZXFXJMMCMPDLTUBSDIEFHSBEBUJPOBU
TVDI BT TUBSDI  JO DPSO MFBWFT 5IF HPBM JT UP NBLF DPSO MFBWFT B CFUUFS BTQFDJmDEFWFMPQNFOUBMUJNFQPJOUMBUFJOUIFMJGFDZDMFPGUIFQMBOU&BSMZJO
GFFETUPDL GPS B DFMMVMPTJD FUIBOPM GFSNFOUPS5IF NBKPS BEWBOUBHFT PG UIF QMBOU HSPXUI DBSCPO  GSPN TUBSDI  SFJOWFTUFE JO MFBWFT DBVTFT FYQPOFOUJBM
BQQSPBDIJOUIJTTVCQSPKFDUJT 
QPUFOUJBMGPSIJHIFSZJFMECFDBVTFFOFSHZJT HSPXUI 0ODF TVöDJFOU QMBOU NBTT JT QSFTFOU UP DPOUBJO UIF DBSCPO 
OPU VTFE FYQPSUJOH TVHBST GSPN MFBWFT UP NBLF DFMMVMPTF BOE 
 FBTF BOE NFUBCPMJTNDBOCFTXJUDIFEUPDBVTFBDDVNVMBUJPOPGTUBSDI
DPNQMFUFOFTTPGGFSNFOUBUJPO5IFNBKPSCPUUMFOFDLJTSFEVDFEZJFMEJONPTU
TUBSDIBDDVNVMBUJOHNVUBOUT 5P EP UIJT XF XJMM VTF 3/" JOUFSGFSFODF 3/"J
 BHBJOTU UIF HMVDBO XBUFS
EJLJOBTFFO[ZNF (8%
BOEUIF4FYQSPUFJO5IFTF3/"JDPOTUSVDUTXJMMCF
$PNQMFUFDPOWFSTJPOUPGVFMT DPVQMFEUPBTFOFTDFODFJOEVDFEQSPNPUFS 4"(
PSBOBMDPIPMJOEVDJCMF
QSPNPUFS 'JH
$POTUSVDUTGPSUIFWBSJPVTQSPNPUFSTBOEJOWFSUFESFQFBUT
DFMMVMPTFDPOWFSTJPO XFSFNBEFCZ#JP#BTJDJO.BSLIBN0OUBSJP$BOBEB XXXCJPCBTJDDPN

UPOTFRVJWBMFOU
DFMMVMPTF TUBSDI  Sag12 Sex 400bp Intron
SFEVDUJPOJOUPUBMZJFME
    

UPOTFRVJWBMFOU
35S alcR alcA Sex 400bp Intron

UPOTESZNBUUFSIB ALCR
Sex 400bp
'JHVSF)ZQPUIFUJDBM*ODSFBTFJO#JPGVFM:JFMEXJUIB1BSUJBMDPOWFSTJPO ALCR E
Sex 400bp
PG$FMMVMPTFUP4UBSDI Ethanol E Dicer
added
Sex 400 bp
*O NPTU QMBOUT USBOTJUPSZ TUBSDI JT QSPEVDFE JO UIF DIMPSPQMBTUT PG MFBWFT E Sex 400 bp
EVSJOHUIFEBZBOECSPLFOEPXOEVSJOHUIFOJHIU*OUIFQBTUZFBSTPVS
MBC BOE MBCT BSPVOE UIF XPSME IBWF NBEF TJHOJmDBOU QSPHSFTT JO
VOEFSTUBOEJOHUIFQBUIXBZPGUSBOTJUPSZTUBSDIEFHSBEBUJPO 'JH 8FJTFFU Target Sex mRNA RISC
for degradation
BM   -V FU BM   4NJUI FU BM 
 8F DBO OPX NBLF FEVDBUFE
EFDJTJPOTBCPVUXIFSFUPNPEJGZUIJTQBUIXBZJOPSEFSUPEJTSVQUUIFTUBSDI 'JHVSF4FOFTDFODF*OEVDJCMFBOE"MDPIPM*OEVDJCMF3/"J$POTUSVDUT
EFHSBEBUJPOQSPDFTTUIFSFCZJODSFBTJOHUIFBNPVOUPGTUBSDIJOUIFMFBWFT 5IFQSPNPUFSPGUIF4"(HFOF BDZTUFJOFQSPUFBTF EJTDPWFSFECZ:PP4VO
/PI BOE 3JDL "NBTJOP 
 JT BDUJWF POMZ BU UIF UJNF PG TFOFTDFODF JO
Chloroplast Cytosol "SBCJEPQTJTBOEXJMMCFVTFEUPESJWF3/"JUBSHFUFEUPUIF(8%FO[ZNFPS
UIF 4FY QSPUFJO "O "MDPIPM JOEVDJCMF QSPNPUFS XJMM BMTP CF VTFE UP ESJWF
3/"JPG(8%BOE4FY5IFBEWBOUBHFPGUIFBMDPIPMJOEVDJCMFQSPNPUFSJT
Starch Sucrose UIBU UIF EFWFMPQNFOUBM UJNJOH PG TUBSDI BDDVNVMBUJPO DBO CF QSFDJTFMZ
DPOUSPMMFEBOEUIJTQSPNPUFSXJMMXPSLJODPSO

GWD
Sex4
3/"J JT B NFDIBOJTN UIBU JOIJCJUT HFOF FYQSFTTJPO BU USBOTMBUJPO PS CZ
B-Amylase Glucanotransferase
IJOEFSJOHUSBOTDSJQUJPO%PVCMFTUSBOEFE3/"JTHFOFSBUFECZDPOTUSVDUJOH
Maltodextrin Maltose Maltose Heteroglycan BOJOWFSUFESFQFBUXJUICQPG%/"JEFOUJDBMUPUBSHFUHFOFJOUIFGPSXBSE
PSJFOUBUJPO BOE  CQ PG %/" JO UIF SFWFSTF PSJFOUBUJPO TFQBSBUFE CZ BO
JOUSPO5IFEPVCMFTUSBOEFE3/"JTDMFBWFEJOUPTIPSUEPVCMFTUSBOEFE3/"
D-
En
zy
m
e
NPMFDVMFTCZUIFFO[ZNFEJDFS4JOHFTUBOETPG3/"BSFUIFOJODPSQPSBUFEJO
Glucose Glucose
UIF 3/"JOEVDFE TJMFODJOH DPNQMFY 3*4$
 5IF 3*4$ UBSHFUT UIF
DPNQMJNFOUBSZN3/"PS%/"UPQSFWFOUUSBOTDSJQUJPOBOEPSUSBOTMBUJPO

'JSTU"SBCJEPQTJTUIFO$PSO
'JHVSF1BUIXBZPG5SBOTJUPSZ4UBSDI%FHSBEBUJPO
5IFSFBSFNBOZQPUFOUJBMUBSHFUTUPDBVTFTUBSDIBDDVNVMBUJPO8FUIJOLUIF
HMVDBOXBUFSEJLJOBTF (8%
FO[ZNFBOEUIFTUBSDIFYDFTT 4FY
QSPUFJO
BSFUIFCFTUUBSHFUTUPQSPWJEFUIFHSFBUFTUJODSFBTFJOUIFZJFMEPGTUBSDI
XIJMFNJOJNJ[JOHEPXOTUSFBNDIFNJDBMBOETJHOBMJOHFõFDUTPGNBMUPTFBOE
HMVDPTFBDDVNVMBUJPO
6TJOH"SBCJEPQTJTmSTUQSFTFOUTTFWFSBMBEWBOUBHFT
854UBSDI&YDFTT
5IFTUBSDINFUBCPMJDQBUIXBZJTXFMMDIBSBDUFSJ[FE
,OPDLPVUTPG(8%BOE4FYIBWFCFFODIBSBDUFSJ[FE :VFUBM ;FFNBOFUBM

5IFTFOFTDFODFJOEVDJCMFQSPNPUFS 4"(Q JTDIBSBDUFSJ[FE /PIBOE"NBTJOP

"GBTUMJGFDZDMFXJMMHJWFVTBOEFBSMZiQSPPGPGDPODFQUw
"SBCJEPQTJTJTWFSZBNFOBCMFUPHFOFUJDNBOJQVMBUJPOBMMPXJOHVTUPmOFUVOF
XIBUXFMFBSOGSPNJOJUJBMFYQFSJNFOUT

$PODVSSFOU XJUI UIF "SBCJEPQTJT XPSL  3/"J DPOTUSVDUT UBSHFUFE UP UIF DPSO
'JHVSF8JMEUZQFBOEB4UBSDI"DDVNVMBUJOH.VUBOUPG"SBCJEPQTJT IPNPMPHVFPG(8%BOE4FYIBWFCFFONBEF5IFTFXJMMCFQMBDFECFIJOEUIF
" NBKPS PCTUBDMF UP PWFSDPNF JT SFEVDJOH PS FMJNJOBUJOH UIF ZJFME QFOBMUZ BMDPIPMJOEVDJCMFQSPNPUFS BOECFIJOEUIFDPOTUJUVUJWFQSPNPUFSPG6CJRVJUJO
PCTFSWFEJONPTUTUBSDIBDDVNVMBUJOHNVUBOUT mHVSFGSPN/JUUZMÊFUBM
5IFUSBOTGPSNBUJPOTBSFCFJOHDBSSJFEPVUCZ%S)FJEJ,BFQQMFSBUUIF6OJWFSTJUZ
PG8JTDPOTJO.BEJTPO

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