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PROSIDINGsEltltrAE t!4g!or'tA!, Erolqq ?910
"P€Ep€kff aiobgi daLm PengElol.an Slhberdaya Hat€ti"
DlusunOleh:Im P€nwEln Pcjdi.s
Sodinrr Nrsion.J Biologi m10
P.nerblt:Faloltas Blologi Uni@ihs Gadjah Mada
Jl. Teknlta Selabn, Seldp ijra€Yogyaka.b 55281
Tdpfax, 0274580839emaft blologl{gm@lgm,acid
ceiakanl:Februad2011
ISSN i 07&979496$0S
sBto/BFnTMIKROPROPAGATI TANAI'.{N ANcGREK EITAM CEI,E '. WN&fri4 LiAdL
IIENCANPEi{USIPAN cEN PEN(IMBI'H TUNAS MELALUI.tgt rd.t ti"r
Edang s@6ni'. an lndridor, Eko A$! sulo@', rizqi. ungga N]llmlm", RadnR€stiqi'
1- Farals Bioloei, Utriv.Gitos C€djah M!da. I. T.l6ila Selataa Sctip Utd!,Yogltskrn! 55231, Indonsia,
2. Puat Studi Biotelsotos! Selolan Pdl?ftjd1 Univmilas Gadjah M L, Baek,Yqlakdts 55281, Iidd6ia.
A!STNAKAaigt*lntzm (Coelog,E panduru,4 Lindl,) tddtL sggr€k alao @ddik Kalittd.ntinu, t.ns d€ngm bos! bd*@ h1i libir nft.n. Salt id F.ldai useFl( i.i sMEBrnenffi ial]l4 [email protected] pua! kena pftdabutnn ee@ lid dri habitotD? miutnlw kome6ial. Urruk id pdlr lsna pei!8ystu balsal tamd egEr.k hjLdMildopoplgdi hnrnqn uss* !dt@ de.Ero pdisipm gd po@bu! nDd rnorredlItt Ambidopsh thaliae (XNln) &a$a Asobutetim tuieldc,?N LBA'W)4 &lahdilahi@. Pdelitis dilalota d@Cnr 3 *ali uleSaq m€ngeud&d .*sds p@tok@MgsEk lilam @lalui 3 talap: l) Pcma@ medie dAe terna& old Pha,laaoFis, Nl:Veir 4d Wen, \alv; Mus$is. ds Stm& MS; &! KNUDGON c, KC); 2) Podt!@stadim pertuDbrna piorokod unrk 6rget turfomsi gmetilq 3) Tmsfm6i sdetikplcBid 35S-KNATI dm plmid vecror pC@tr1LBA4404 lc dalm 8mn dggrek hitaD-IIsn peneltid dtuju$ai bahw peltuobr!@ 4 ,tta tibit deetet hilam &Inqil pd.trdi@ r,IP yaitu dird w.tiu 5 b!rl& sebula& l0% biii t h! hnbu! neljddi plald.t
Ghnih bu1h6 delg& se?Mg rlau). Prclokom .rge!€k nitan @u 2 bulu scatdloBif drgEqt"! lelsgi bobq tuod blset Brrt i&sf@tsi lEk!tui tlNfolmsied.rik d€!s!n vettor lGM 6eR46 32.9%, dd p35S-KNATT Mtp.i 432 %K6n!@i inLg@i 8a (llAT1 p!d! gBon r@@ t@f@d scltatrg diilahrt&dsse FCL Felodi pemb.ntute n@ p.da F.sLmd 35S::K{ATI l€bin tinggidit€ldingla t@foltd d@gd vckror sjo DNru tol]!l@ @n-tdEfol@
Ksra hrci: A4igr* nild, Coet sDdrad!/arz, nediM NP, tu@, tusf()!@i gd.tigAErobrluiw tun{rciN
pa,arda Lilfll) adila,b dseEk ald
elxlqrik Kslimtar tim* Aneerek inindiliki taEhenstik kelopat du
nahkok hllga 6wma hijau ceBh
ddgm 1a5.hm iernqnrt seperti violin
b6drm uEIl koniidm sampli hil8m
PENDA{IL'I,UANA"Sg"k hitlln
ddca beb@ps baejd bffia bij&
(Cullcc t99). Aqsr.k ini Erdislribui di
Malarsia, sln3na dm tfno (Cullfl,
1992), Di Indtnsia, l'lmm ini dapat
dijMpai di Cag& Al@ Padang Luvay
(K6'[ Luwa, di wilat n K'[lml'n
Tiou. ArgieEk ini berni( dicdi unrk
smiir N6imal Biologi 20io
dibtrdiday*d k rcM bureatrta Ymg
idah d{ bcrutw be6d.
sut ini poplLsi mggrck ini sdgat
n@llm bo&!! telocd PlnL Ad.
brbeqea seba6 ?rt@ lai! *ihat
keldl@ hu@, konvdi 96ssuae
lsb@, d@ F@cen!'Iid hl].J@ tecsa
rs ddi nlhiratl'F urillr nrjum tm6i.l,
K@<lala )mg dinadapi drlan laargl1m
ueg.k hitu ili ad,lat pdiodc
belbursFlye sancat leodck ([email protected] laF)
dd 6ugr rltlif $lit utd( disilagLd
al!! <tlbah.te. selai! ih!
pdlrmbs!!! u Btu hii dssck litam
jusa relaif l@bat du leb'h slit
dibrndinsko tugg.k l,imya, snireisE
dlpqlu&an rqlaioo-pnkLw khEus
(WiEro.M.t, 2009, lonuitai p.ibadi).
Unqrk itu pftlq saha F [email protected] mqql
raM &ge* nn@ unrt u5dIB
korsnlsi ba+ sq.@ , ,!r! nmplo d
rr,! dalnn mgkr mcnj.sa k!l6t rim
tliodivdit s KaliEaltatr tnrtryug
segat hcrhdsn tlebtt.
G.d IA',!fl (Knor€dl-l&. /&t
Aldbidopsir dalidn ) 1 @rtpt*d sala!
eb eeenr4 lnq h@eobot s* fMilr
ylrg diiolari ddi A.dbidops thatida
(Li!@ln er rl.. 1994). overrblrcsi 8en
tcEcblt prda bsb@p! tanal@ kmgdik
Eenunjuttd add}! oeristm ektopik
yans s@g{6n Padt Pobotukm
m'ntihu6, srebt! pad. t&nb*!|!
Giocoln 'r dl.. 1994). AEbidopsk (cnek
F kulb. Broroer UGM, Y@ak:l16 2}?5 s€r€ibar 2010
er 4t, 1996), iomr OttiIlr@ er ar,
2000). sel.<tr (Fngir er al! 2001) ds
mgset {Sdiani, 2007). Seniarti g' aa
(200?) nelapor*s P.Bliti@)€
neldo}!, llanstumali ger Ifl,{ry ymg
dikonE l oLL pFmtef tut jJs
caut'Iawr Mazait ,14 (355 C'MV)
ddgd p@t n ,1gmbtuium
tunehcid LBL4444 pada 6qst*
Phdl@Mpsis mdbilis L BlMc da
dipsoleh 31 nsa! ddi sstu Folokoo- Yq
cr rt (2001) jusr scnd4a&d
r&rnrlbuta! ntidn@ trc!$@ik
ranM D.ndobim "l't tlid. Tnds h
rEns di lrdsfomoi d6!io sq DOtl
(muTI nnoloe pan! D.ndbbib).
Pdbcntul€n NltitlB iro sagat
dds@hrgko d.hs nilmpop6edi
la.rt@ dssrk Eal ili noEojukLd
r.hh tcdadi pniletrtan btirotsi sl+cl
Fo@ uotuk d4al bsrcseqrfai 6.qjsd1
tam l@Cltp, *tingg. ssat
r@gu!tu!c}6 untuk dl.pltl*ik@ dal@
nitr@oplg6i L!@s &g€FK
Tdat dilalukm EikopoPagsi
talald dsgrct hi(,m .t tsu pdisipatr
gerr P€NFbuh Mlt Knotledl-like
Alabttop$ thdliN \KNAID. Drlb
lenelitian ini, modifikasi 8pledk rajmm
uegtt ni!fr dlltkukd itnsd
nqoJ6ilkm e@ l3r{fl ,E!e dikonEol
densd pMolor kotliiiulif 35S ddi vitu
C*lrlow{ Mddic ltr$ (CtMv) k€
d.lam protocod ulgr.k hitd ddsd
593
tn diatn ,lqtuh,denvn tundAciw
$ain LBA ,1404. P#liri{ bi berluje
arul trninek rto portri pdbentltu
tUM iLlm pqney.te mlal
(aildopmpcgcsi) qitsnar {lsslk bibll
d€qrn lenlsipd gen lMci Fnubuhnbs [email protected] A/abi.lopt$ kabai4(wATr).
CARA KEFJA
Behan tca@on daa baHe4
Bshn E mu jes dicealdr
6np! pbtotm (le*eabdgd @brio
tucet l) c pantututd Liadl (JuE
dipruleh ddi &l*,rd &d I@ Ostids
Ns.ry, Jaw. lidu). !N 4 lutar.
Pbrotm dit@bulhr pada nediM
VIlt dEsd p@@ba!!! k€.t tg 150
nei4" pimg 150 bg/L,lcbp6 150 dg/I-,
d.! NAA lppe Krliu diid.rbsi d.nsE
crh$E prtft 1000 du 6ffi tontinlr
B.k ri ,mg digun la h€rop.
bnary vNtt AsrcbactqtM tunef@im
LBA,|404 )mg mmbaw plsmid pG|H
<redor dlt\, pIOlAn, .br ESOU}(h.bd pta.h"t). llarnid dlr bolteri
dipmleh dEi Prot Yastuon M.cbi&,
Univmii's NrsolE, Jelee, Ko6tul5i
g6 I3L{?1 dapa. dilihat !sd. Cuba l,
cmbd 1 skesa 6agEa 35s:Jc{.44r,Alak pdan nmujiL*.! lanjmsnzgaen }!rg ilsspliEttsi cnzim Taqd€nsd priM 3!.6ifit lw,4n F1Rl
Fa.litia {,ildota dcnsd , tali
ul&cu. tMeigld.tatr clcplar poiotod
dBsEk tild nclalui 3 l,b,p:
l) P€nenre Eedi@ d.sr.r!.i}
Unt* md!@leh pem@bunm
opd@l pbtotom deed< 14
dilslml@ lllsfer Folokot4 lesllct l!9 4
@ h.di& d.&i tuudsd c (Kc),
v&h ad w@t (vlv), M@hige. de
sk@s (Ms), dd Ns Phll3moptb (NP).
Mediu diltdilisGi d.rBd dtoklaf pad.
l2l"c, I etfi, !rl@ 15 Es'lt Digunak@
p.hidbh direrd 6 d. sctidg p.tddis!
itibuai tiga degd. Diilti pe*€obdlgu
prototom lmpai telbdt* tu@,
P@CUh@ ti4gi tiN, pejdlg rbu ke
tiga dibhrLs lcliap mirggr
2) Ie@tu@ !t diu pettdbu!4
ltolokom @b! brg.t tsslom6i
Pddtud fi.c poiok m lat'lik
rl,lahl@ dslcan g}poubll6t?r
poiokom d.lm mediu NP- l<lltur
disifpd pada p.ncanq@ 1000 flw d.!
suhu 25'C, ?6gdatrn dil.hrtan
tahadlp fdc lertdbmgm poioto@
3) Ttusf@si goetik plsdid l5S-
KNAT1 dtu plasmid wctqr
pOd&4LBA,1404 ke dald emof,
T@fodsi gacri! plmid
35$;KNATI de rccror pc&o t! dalarn
8dm eesEk hilam dilatutd d.nBe
4€rods s@isni et a|. QwT) d@s$
E6difitai ncdiu nrs$uL ScrdrnpElTltE pBde ncdiun qM (cdrl6
Id/*'dio, Medntii vliv+2.4D lng/L)
elanr tier h.i, petotou di@dan
.Lle hihr b!&tqi -irr,6@t,im
tunda.td L844,{}4 Fng nebd!
!'l@id 35S:lO{ATr rtau v4ld pGFo
s.lan 30 tmil t Fudia! dft.f,nddn
slama 30 nRit Poldom dita,m
tflbQli pada ordiu4 CIM s4lF!3 ? hBi.
PFlot@ di@ci doge ncdie ll VW
eir + Cdobr,4 3008s1 uottit n@ke
Frimbohd agobatlltih, ldrodiu
dipiodah dal@ oediM SIM ($r@t
t dlct , rsdthj 0.15!M NAA+ 5uM
ziP) rsne dihn&n dcnern Ksl4isi! 100
Es/L utol *Lkei hlcfol@
Protokord $ns lqlas *Glsi, dinihDs
ft.k'rdsi FrlsfoffiirlE d.o8d
oQlghiths jqol!! pototoF ,nng ic@
Faldr.3 sroroor LIGM, Y@*nna 24?5 ftDbmh€r2o1o
hidup dArgi detrge jmkn iotll
probkom teg liit{sfotrMi,
EASIL DAN PEMBA.EASAN
!G6*&6a! d@ efisiorri
tri4foldi bdgdhDg Frla metodc !t
{,ro }@s dieru.kt& K tu t ,,/obcrpm pdting drld !.nyta{h sl
sipieD. pdqlilrtm Fo$lpu! sllski
n!6fodai l.'@ud m.dim lqrlhr
,@g srsu4i in€rqald h.t t aotilg d$i
londisi kftr. Mcdim tulftr ]sng banq
lid.k hanF B@d'iMg kshidupa!jdi.eu hli ,hjl Me&trs
lem@butd dm pelilmi $@ i,
,rtd. ll.l tcEcblt b@;r& urhr! 3dnp
Aesi€d (Slenllt 1989), Dalao b!did!t!
t dno mEgE&, b.hdp. f,.diu baal
(dsd) y@s niN distlral(g! a!i@ hir
.d.!d ncdiu Kru&on C (KC), vrcir
6!d Vsrt (VW), da! Mu?shiep md
5t6s (Ms) ( rdini dd Es4 1993).
bLe ,r ot (I98) t.In6il neobuartdpodsi n diu bm 'dg lebil
dibiur.! u
Idar !a@d lo€l3p @t@'ter) ditcntuk n
5 tanep pskenba.3u ddgd kitsia
seiaga bdilo! lae l. ?rclokotu
hcdScltul shr dlu utufu pejeg
qeidfiak! t iieria td*!l hmilpd.litim Bdujutto tunw
pcrhdhrta! id r'td bibn .ngisrek nis&
lclbai& padi ndi@ NP Filr rlalrdgrh! 5 bds !&nat'!k lq4 biji 16jtlt
nnbun d@jadi pl.ldct (stldib bllbG
sarina N.Eid'ar Bidogi 2010
dtu; fa$ 2, a@ dsEE 2 dau; nse 3,
hlus delsd ! .tau: &se 4, hss d€3r! 4
dNn k 3; fae 5, plmd.t dflcat 4 .!tu
cmbar 2. Fa.e pe.EEbubm rF* dltt pl?FilEl Ctelag\e paadwqta tr tto pad. madiwl
d6eu reprsg .lau). Hst ili dlebtlks
t,l@ NP ndslrdu.g as nitbgs
de pndphor yeg lcbib iilggi
dibdrdiqbr &cdlm hJtu deet !
Tab.t l. P.mnbuiit! tD* dln llantlet mgs.t hitsn dala$ tuhu ft s-lro pada We4inedi@ tuobuh. set Lh6buratr@
JnbLh i@ dr! !l'!dd Fd. *r'lp fr* td@ndr
'Pdbdblcn lbrdd lerc.pat
59€
t
Pminbm .kdm }tsaC tepat dd
opti@i lddi.i tobltivGi sdnsal
n@batu dalm sistem F,rDf()I@i yd8
.fisid utuk ll.@e eCigr.lc Pomlus
LokulriGi tm
kzl@ tulup ldkdbe8amya
46uju*kr! baltu Ptda s@t itu
perkmbesm ndi,tcm uj@g batdg
ssin ddd rdE! aE€I, s.hileg! Prdcs
epreg@ D@st@ lrscbq| detrgrn
orekpcsi Fdcsm diha.dpld dtt*
Ildil F.sfo':tri Fbtokotu
arssfk bitao d.!8Fn P35SKNAT1
nal46 d@gEn w.tor pcr@ $jt
odujuklo! trtwdi ilasfousi '9ng
c1lltrp siglinre, y.itd 32,96 % utuk
vettor 08 dai 54 pmtokdm da!6t
tuDbtih Ddb.!fttk tu6 9ad. h.dim
del€tli edhotik Kr!sFrsi!), de 4125%
(15 ddi 35) (t bd 2).
cdbdr 3. P.rtrnhunan pFrd(m !6da bdb.gii \didi medim pddauu 6 bulo Sldld:
Il@pdtimbu*n loses tue. 8dy$s d.tri6al l.Fi d@sau tlelat
nclcb dapis tlng mtuimal, Htl
l!6etut da!6r dilakik@ ddss
ncruiFnai ko'ml8i rsdr&,.rrt
dd laEa tohltnEi. wrth kokrldBi
FrC pdjes nainskatldn cfi.i.ni
tnfff@ai (i.u ef al, 2001). D6i hsil
r.4dar!A pfot6kom m8g.k njl.d
lou 2 bdo ssrt Bpruif disu.kd
s€basai lrhe h.'i'n tqgct u!tu!
tanrfmNi. ltorokom lffibd dipilih
t&sfo@si pGM dlr pKNATI pad.ke.mb& C. par.lututa Li\dl
597
3 45 91% (24/62)
l 54 32.96% ltq/54)
P.r5S::F{AT I l 4t.25o/a (t5B5l
Fardi.5 Eobsi uGM, YqFkari. ?4-25 Sspbmba 2Ot0
semirN.sloE|Eio|ogi2010
Hasil kdfi@si idtFgEsi sEd KNATI pad. gdm tiM tadid.i tdafdd@
Gdbs 3. tl6il elcLtrolmrisabplifklsi ge! KNATI {hlqnploiolom kddidrt t nsf(md,M: mr*a Lrabda DNA/styI. P:pl.nid 35S::KNATI, NT: No!
Ttmfme no I dd no2.
M PNTTITZ
FEbfls Frnbrmikd nlm pdz
tulfoll@ 3's:fNATl l€6in ti.gsi
rlibddingkr tusfoma dened re(tor
ej8 mupu ian't@ ea-ned@
Etspl@ d@ dtuohlte pade Brdiu
12N? NAA 0,15 hiledolar, ptdt
atsph! ddi hi.n'n m FesfoM {t!i
G@bd4. Pdt nttrt& 1!@ in vitre dail!!al@ rl6fod& KNATr ladancdi@ j/, Nl dit Ebah NAA 0,15. {A)Tede m-!@fm.., (E) Tua ddi.tlplu tt!!,l@ ltntfolMj5S::B.IAT1. @e 5m}
KESTMPULANMcdim t 16.& uhrk indrki nMt
cnccrlt bita6 C pc.dsar4 Urdl. adaldbcdie NP. !e pFiotnm r.rb.i! teg
diglnald unt& e*8plm taefolrui
59C
t ebdm nmhn stu tuas edmgke
lda €L.plan d.@
lrtufomdfis:: I(NATI nmrbuA 7 trG-
Pmghinusa &elomi p@b@lu!&
had bGih Llu lEias d@ @.ih lsus
&Lh*n Fdurssu @p'i pcrfinblha
@o€!di dortu nos Yatg n'!'rni
ad.lat pmlotm urM 2 bultn. Sbt€m
trNfoll@i s@tiL de!'ge m€diator
AsFbrkr.riM tDcfsoiffi dq.t
ditqapt& pada dsgEk nitu. Fektdst
t@dl'l@i plda poiokod ut*
pC'@ &b$d 3296% dd ubr*
pKNATI selss 4315%. FFh@i
pdbdt !t! n44 Pad4 tiastobfl
35S::(NATI lcbih linggi dibedingta!
lesfond &ngd Ello. sja muPa
emd m&Easf@ rtitu t.66e 7
UCAPAN TERIMA r,{slgldiM l$ih di!.tpla! r4sds
Batak lviFkns@a! (?eEiltk Fms dd
Edwrd Orcnid N6ery, Pligd tea
TIIN) yds tebn tt@bdl Prctokm
P.r!rr.!EiohliuG,Yoqyrid2+2sse0Lnb42!10
IdggEk bitan sbagni sal,L sanr b6hu
lq€litim, Pfl€litim ini dilshma*a .16
biata Angetu DIPA UPM, Fiba! Mulj
Tah@ gi6ah Bft.ilg x\'[ ddgm s@t
P€.jejitu Pelaklm.la Poelitia Noam
LPIM-UClvt604/2009 la!8gat 3 Apiil
2009 dd LPPM-UCM/10452010 t!4sal
4 Mci 2010.
DAFTAR PUSTAKA
tudin! J- ud R E nc 1993.M itupmFaEatio n of @titk t$awiLy & Sor!, Ift. Nw Yo*
CtwL A, c. Lia@h, &o S. Eak€. 19e6.rq%I/ Inlfued Lbcd Laveswin E ropic Matuld whflOve'*pNed in,'{utir4prtr'Plantce A: lt7-1249.
Cltrei, t, 1992. nE AftJtid Eoot.Cskidg€ udveBity Pr*6, GrBt
F!usj6, A, P. Gi@i@, G. Mtt , cNi@Iodl, A cbiapp.fa MB.iiodli A.M. IrD@61i, w.Daitt , II, w Oacteld. ed D.Meion! 2001. OveHplsim ofKNATI i! letile $hitu kafDet.rmln.lc Covt! io 3 sh@cLikc Ldet.di.,t Go'rh&ffii.i.d mth d Ae@uldioaof LeporteryLTne c),totinis.Plaat Physiolop, valt26; 1370-I3t0
Iilrii! M. O., S. Icbbashi dd s. Matsui.198. cdr6l of cFwtn edDdclopnat of Poiocom Lik.
Body non C{tt\s 6y Ca$dSffi in Pral@,pstr, PlaBiotednolop, 15 (1): I8rl8?.
Lilsln, C., t. hle; t Ymo8shi, Ksdit *4 ud S. fialc. 199L 4Kaou.tdl-lite oreobd Ge,e bi..4artlo?Jir is ErP!F ed itr {,.!veg.latirc Mdittcd ddDEmlicaUy Alte6 tr.veMorFhologJ t!fl ovdpt6scdin T@sgdic Pldk. Pi.,r CetlR"?.t|s 6: 18591876.
Nishioutr,4., M. T@oti, T. sahsolo,ed M. Mais@ta 2m0. Ovd'ExpNion of Tohqcm f,etedl-We al6tl Haitah6 GaqAI1l€ VeiN Iaf MoDholoBY.Plaat Ce! Phitiologt 1l (15):
S6iani, B., a- In&imlo, a Pumiok!,S. lsnilidglin, N, S!!@, TBh*.wa Y. Yoshioka, Y.Mrchidr, C, M..bid& 200?.AsnDacterim-n distedt d4f@arimf rn. vild od!1d.Fci6 rbdmop6b aoabilisPlmt Bintahtolos 2, 26t-272
Sl4d4 t 1989. OEtld lroldead@ bYTisu. C trre Tc.nniqus- la!P6dl tnd Fuiurc, In Prit tt!4lt.\N. 19A9. Modn Methods tnOrch t Cnena{d, Tte Rolc olPhrsiotog, E@log), atd
Univdity !c!. Nw Yo&Yu tl, S, H. Yeg, dd C, t Con 200l.
,lgmbr|riutuMaditllATtusfmation of A D€t mDimOEhid Wilh tlE Cls I KtorAebe ImEl. Plont CeI Rqoie20:301-305.
Fakdr6 Bdooi UGM- Yoo€kab 2+?5 $ebmber ?or0
; . _
;;;s- : i . !€€ElsO\E EEEis 3 i:Ns=,qq= i. i -. ,91:, :€- H ui lo
9 i.;c!aO '< q! o 'I
, -5 : T =: ! . ! !E :=\S f i ; X9\ =: b?^$E 5 tz:=t9d"s E i -q{} I ri iz-i"E; t'=--€ i<S=-) j - 'gu, \E : o n; +F.?
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