Biorechnolosv anddts Future Potential for Ioprovemen~_of the Tpbacco Elant (A paper~ tobe_eteseated by hLI Heard at the TohaccPExhibition, Beijinp u,$eptember '87 fntroduc~a A-D It is appropilte to begin by identifying in uhat ~B~L11 to the tobacco plant are sa$ht. There are at least g~E~E~ced~9 [oF wboa the tobacco plant has iqortance · the fanet, the cigarette j C panufacturer and the eooslmer~ ~cr~t~e lain focus Of tokeco *vllapmt hu *l~dB) - cmrillrlti~u oL~5~.1 ~ibl BBi~ljhave dolinetcd, hla~a~e(y 14W hb~JIClh ~30 IrAs~g~ h R3 6~15 ~u*w4AL3jY~ A*3 OG~krmCI ~L~ET LH 4H )~~4ICCA' ~ctO( I; CL~UC r~ e~fc~ ia IQ i · _._J Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -2- it vill be of j~~:i~arii~ in the future development of tobacco cul~ivarP that the BIOYIPB Of ~,~L~!~ ~B~i~ii~E t$t iS fDf the production of cigarettes. Consequently if uo look to the future igrovementr in tobacco they ~,~VYI ~ (offerinr benbits largely m the grover) but ur can alro 1ERf ii Expect to sec dLvaiopPFnti tht de~i~e~by ~hp cilarlttl ~~i PyS A IF\X~3' Pdnufaeturer or O~E~"·ij~'i~:~t~:'~i~~. Ihesl 'd~vnrtreaol ~JI~R~P;G~~ attributes are recopnised as the consequence of growing, harvesting, curing practice, leaf ~aturation and blending of tobacco, ~L Ihw) I· ~Y ~D JBLrI U~ fk4'kaf Gku~O~, In this respect we must acknowledge that any attempts to achieve modifications in tobacco leaf genetics that result in downstream ''IB 'EpR 0C 4rov..~l uill C,r~nt ~B~ C~a, Q C~I P31~ by Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 I ·_ _ -3· ~s r~ere it 1 ellriler~lll lo$ oi ~P~E~E~4~F tO 'src'' tl~t I~iacd tobao ~(_t~ll~vijib~;~;ii~:~i~c~i~~ or t~e s~ ProltYI PIJ~Plh 10~( ICd ~IIDIIYe bLI1 t~t ~S~i~i~2~i~~~a ~ih~ ~~~ 3kl it ~n L I3C~PIPf dr ~i~F~~.~!i~l II~I~1J tlC IrPccsr ir flttllr EP~ljClfll L~ t~e infllence af ~c~f~ b~i~y~i~i~i~F·j~i~ 1Loql sa de dni~ oi ~ ~~lt to nl)o~ ~ it~ c~riroaclt ir ~cnctic~ll b~~~a. b, ~a~ct -w ~~1 9 'kbti~ 1~ ~~lrC~g~ ~ ~j~i~i~l~ rl a talum Il~lt Iaaticl real to ·Ic~il at 1I)IC~ ~ ~~DD IClf id J·~Ct ~i~· Idj~,~,~_~~ " ~L~a~~PYT. ~l~a~~i b~T!~ 10 I~ ClibPDF - v~~fastio.soni -k· ~r~ LdGY ~t ~ ~c~C~JDLbCICS. lor L1IC ai pnulb~PI ~ Ule;·~~ ~i~i~i~b~d~i Si~pd;lf3S~L~e~llldz' ~ ~R~~~I~ O y I1Pf Lretll~ ly ,,11,1,,,,6 tilsue culture tlebnoioricJ I ~c~k ecdl qhad~ O Idee~r ~ch~~kg Icle gpil I IpYdk rll darrl·ll e~ru~·. ~,~t;~tzc~r~75~p~t,,~ L~t~l~ lq l~d L arYr~ · · ~~d~l~ al ct alcal~~ril~ Ir di~crcte or i~olltel prrcl~el for pl~pt i~rapant, For i,~, ~i ~l~aa~td·;h~~~y~?.'~~ ~tiii~gs~8~g rcl~e in ~ clreialr 1II~ frAlti· ~~blt Ilcaul;Sc~Z~t~~~~ l~~s I~rl a~llte II ~E~S~f Ib~l ~ 'r'P1 tL~i··h'~h t~ ~t~ti~t~d ~q/ ClibPDF - v~~fastio.soni -5- biotcehnolosical techni~ur~ ~g'~ and that there exists jf723 that scientists villlturn tP these teehnipues in SifUltiOllS where further thaught might have shorm that "' ~c:~~ ·traditiwd are (M Tr;CNI rKL~tJ ~Y~Q1.SIS... .) appropriate.~u·. :u~;t·' of bi6teehnologg vithin the ccntut of plants there is an f~:4~~D~'S~s importance, thalof S~96~i~h~N~f~-~and tbeir interaction"nq6nnNnr6n? between plant and thilasphere for efSt~t:upt~ke of trace m~triea~sc~ill ~lliP~ dil~UPICd~ ~i~R~is CuRPe~i P~I(I~D~I! PL~~~ ~RE~PING~ ~~ ~t~B~ i' ts ertablilhed, ~ntiqna;geiePce wll ~ObLU4 e~t~rollllhout· ,his~tp~ for the igtove~ent of agriculturally Lportant P USE FLI P craps, The process relies on ~p~o~~d ~2l~and thl ~e~p~ 'deoiiSli:itti;ibutes o ~~ dains ~e~ual:rgrphuetioa ·of pflPtq~~ O ~J~·.'. 9 Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -6- IJ~ii~j~E~t~ lihiea I·~e~h~6~t~o;e~ Often neCeSliilteE the use of Ikd vithin the ~dt~~:~nial:~~~oeic;II: vari66~ii~iithin n species, The ~lllb~n~sauL. . process can be r~y~C~F~:~?p~'~e~l~Ltf~lFI~!~Fiitc~IF~I~~·pe ;i j applied or ideotified at ~i~p~n~p~9 as is tie care viib IO~fl~P~j LI~'C~, ~ P~aivrBt~~hysl:~'~ Thelituation is~;~;e';c~~ ~bi?cSg where some traits can only be assessed fplloving th~tC~t~ ~j~E~;i~B:.pl~p~c~r~5o~!~~~~bfBc~·e, Under these circllmstlrceJ, the ~ft~`apd~ti?e~s~ for inprovements in aenetically based product attributes Llean~s P~Sq~P;~S~n~3~F~T~;d~~pe~ii~a k? LAN*LtiOWI ~u~y, 44 ~ ~~~ilillan 'O Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 ·7· ~tl~i~~Tt ~e~t~P~l~i~i~i~J ColPlbb~f$i~evcca:faictia ·~~rcn~a~:lbai~MI:n~C ~YI I~~'-"'''~*'~~C..; · ..~b~~phl~C:*~ ~%'bt'ii~lll ~ introduce in r~cl~lric~ rhe ·jor restriction for the use and application of plant breedi~ in all crops is ~~rci~ti~r; fo~b~t)l xaald take L the adQ! !_ I 4 CO brsp iTui Y ~~14 lhl 4~L ~(~ ~hy NoT ~~e~S, ~G~,. Ib gj~i~i~i~i~i~i~ IPr tobacca iSracrlt L11 iccn f ~ ~ir~ ~P~E24 ~ ii ~t ~j lecl~5p!:lr4v~~lla~ta *( tO the last Itletfen I~a ai tilt tccblaa, ~E ~1~$ P~qCF\1(~~Ci ~ ~d~ ~i ~ r~R rC ~i~t~F~Zi;cb~i;~$~al~ot tba voild t~~t ~~iiP~BI5l~"i~TU7~i~;~5~~r~r ClibPDF - v~~fastio.soni -8- ~ost notable being ~e~stal~,t~~1~91 USA, Bhfj Cafuda ad~f~q~ag ~u*· limllbuL. the coPtribution pi the ~i3~S~:~~~-~;i~j~:ii~i~ in Ipplyin~ ~i~ror ~h~ ilp~oal,nt·of ~f and gtljpiJ iOil(lD, LU been ~tstand~, T~:-e -"' ''" .·· ;arc.rcspoasible foi'bost eo~Pc~c~i~"'c~~~i~'d~i~~"~rh : i~~:Z1~~ kPr~dQ CbonIt, Thr j~ of ~htsc$i~ih~l funded prPgraPIPeS for the reasons outlined have been to ~th the gjpl (PPhSil bCiPI in term~ of 1C~~f~p~ pulialarly ~;tole~j~ aa~rsLt~g. In addition there are ~i~s :4~i~t~9a~·F:*e~?t~i~ Llh~('iA'i;anlS,, (i) j~jj~j t~llthcr vitb intervention support by Cwetnent tends to F~d~~eoa~t!~g on the intmlletion of P~-tapo~So ?Il~iv~f~~:(particu~arly those serring the interests of a specific end-user), Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -9- (ii) ThetP~E~sens$bf tobae~ ~relcir~bSor plant Lprovcpcnt, ~uvu ~ h~ ~ V 55~1 .. There would appea to be a need t~_fo~eff~.b~:~*~'i~.bf~t~q~l~:~~ in the developpeat of iuhre tobacco breeding progran~es, ii t~ ful~lj rfle~l~ product improveaent is to be thieved thrmyh ~wep~il uc CconS ~n EJZY 'i~tZcr~LoLDtl3. ~ ~~ ~t~"a;.i;~*~~~o~~l*~p·~~ prosran~s aal rrratllier in thl developPcnt of new cultivlrs ~4t~ 1) provide the basis l~p~BI F \·~ot CR6~lt~ O O Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 __ -10- B ~tfl~ It~lcr~Yy ~t h nl~ f~ a ~i~'~i;';;Y~~~~t~:i;t t~~un6~ ~%p~CC~ , 1~5~ ~I La ·d I atad·b· ble~~ ~*rd* II (s8q~ d ~·t Y~rbL~a L ~db '~6~5~~i ~ -1-1--n3-11 ~d proccJlls (1). ClibPDF - v~~fastio.soni f_ _ -lr· r~ Wl~kl*C ~i~'~tN I d~ I ~ ~~c8 cMi R . ~ .., It I&J~BC~Yh' $~eO hi Isd~J r I ·~1 ~*t L t)·rl ~i~b;b~iifilaq r 1~1 c~, · QOliS·( hSt~r ~8( T~dz~i;a of rqil.,i ~~i~i~to ~Bn~c~~* mi ~i~i~i~o~ ad -~~3· ~I II"""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""'8t~~~~~i~.iF~il~isirl~f;~'~.~,-,-~;,'~_~~1 1 d ~iiidolWli~ (eQl~i=*~i~i~E~ai~clIr j~ (1)~ ~lsc~w k~ ~LMkd ~ w krdl ~ Ct FL~ ~htiL*r,~ ~ gQ~ IN CLY Hb ~llcbMG ~) P All ~ ., ~ii3i~j~t~ii~-~iJ, tlL~a·YnY~~·~· ~t ~LI I.1~ ~· ~tB · f~tl O O O N ClibPDF - v~~fastio.soni -12· ~ dec terhnnloaier by pe~wi~~ Th( iinitLd ~q~Oir the reported uJe oi a~Plt~ clrisl pllPtf iP ~b~~Eii~l=Tti II·;· ~~T-F~P~ 3), Of a Chinese cdtivii~~cTah' tha~:;lpls~~~~irb7Isfa~~P~~.~i~Tf:~S1-~P~'df:~lbCtiC~ly:~MiP~U~e~ pi~pt~, Plltlbb L~iEi(i~E ILli;t~Ersi] 1Pd BTfeoeil~sfcrl(r) F~t~ uere achieved initially usins ~i~ep. In~Ij~F~ did the companies eonceroed d~to have a defined c~-~l~:~P ~O yt I, These innovations ~e~S~e commercially uh~i~~gol is a major probb fPr tobacco or indeed vhere~S~Y~c~i~id~ 8~I~;9t ~dl) it 9*UI L"lt ~~;;E:~;~-~n~;4 processes, a~]i~]i~jtiibl~:·'fiij'n~-~ji~n'ii~atiiioltP':'g~iiet~ii, have not been i~;-~aa~c~dl~:~_t~coulCi~a~rtab;b;~i~fl~i~~~,~b~!ij~·i~i~e~p crt~F~9~g~e, of ewroe, that this work is being progressed in th~!~e~l.~~Le~dl~;~hb-ai;;6~0ltbese cc~p~k's). r Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -13· k~T ~ ~~ 91~ ~c'Zti~l~V~ thrw cP~ r~y b~ ff491~ ~ Ta~P~c~o 8a2~fii~Ti~r~F~~ or art r~'C~o~"~o"i~~~;;~~~r~ 1, Plant Breeding i-~oc~c~r Ft~ Re~s~O. ,, The k~;i~o~t~;i~~~l~~:il'·t~ikF~.~S~'~T~~ in the developlcnt of nex cultivars using new technologies, The key and ~t;~j~tb~~:~Yel6lii~q::~l~;e~;r~iotioasbip knd~d~jjt~nding tobacco blends (filler and flavour grades) and product quality, ~g~i~f~c~ ~TiS9UP dture irwinit~i~ Ih) Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 ·II- ~q~i~f~i~l the teehnique uherebg apieal or lateral bud or even other plant structures can be I&2li31~Eidand aseptically groxn on 1 ~~ has ~k~Eiderabl~.~P~ff~EI id slu~e~a~f( Bfff~i~it-:~fl~iiiCIll~ut7~1 Its primary benefits are: (i) Ig~.~P~.~;~~getion':~~bj~j~j particniarly applicable for the ~toPasation, lit is ~·L ~Paintena`nce of 7-~Lle an indivrdwl plant ific characteristics are required (psually identifj~d from a 'plant population), ~Ta `\i ·--·- ~4?6~1· rolu~$naPental C~i~oatrt~Jtc. Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -15· (ii) ~he Pte55Prer ~9C~c~b~i~0[ 'L'S fm ~ 2) glOJY1II L iot ~e crldicltion of plmt liselle~, ~d ~ui ·I· heat) and ~ihr. '·'·.i ~ a, LI bet~ qplird to4 p·t ~a h de L~~ ~ ~cl LI~F~O~i~F~'~t~l~?~. C~L~1~~~ ~re te ae~LIlT ~t~ii~f~ ·~r tYd ~b tc~ala la ~i~f~l~Y)!~~ rl tl' i~lelle l~ditiY ~·:J~ ;nk~' "~1~' u~r*? f~j~I ~glC Clt~e ~nl~~doL I~c~tfl ekalha uhere tisac E~tUTC p101YOtiOn ~ I~tfb~ldil ubcre Ui~ Il~tp~~ll I~YIBIILd ~ylb~n~~ ClibPDF - v~~fastio.soni -16· need or cost basis, tlowever, it has~l~ei~t~Jthe ~A nt-b~Rnec(·r~~~.. ~Erb~8~arereueh ·too~ ehat cwld rrovide tha baris of an ~i;ii~v~:~ib~~ fol ~61~36i~i~~saa~·i~'~ regional comparisons and on the development of cultural practices for product improvement, I) 9o6sl~il~ls~i~a~:~ ~i~3~f Cloned enctic stability across a plait population would provide an ucellent basis for studying and optinising harvlstifig and curing processing practices. 1~ 'O ... .j Nw Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -17· ci ~i tb~t~diute~t~ md tblir e[i,ct DP. talateo (ulliliea. d) 3~fs:ofldif ferent~aiid';norinalIj·es~abli'shea cPrtns praeticcs~ ~C~'1":~ ··· -····· ...r··~"~' hvp FmZ PAP~I·TPh Llrvd ii~-Y~ It~O..~ ~id~Jrl~, paralleling the practice uitk other crops, tissue culture tCC~mi~uc~ co~d LL Utililld rO~i~l~n &~L~·nlr~f~hi~;ia.~~~. Ihis ogy cad and has been used under two scenarios: (i) to short-circuit the ~election and baintenance of plants fr0lD a sellreaatins pop$lation that have rrlgance for hybrid plant seed production prior to utensive i CO baih~ col;PaL~,... (ii) use of ~HS p~pts via tissue culture aaintenance in the production of t~s~iiol~bi~ally based hybrid seed production. Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 _ __ -1B- L BtCC~hP~~~L~Ua~ci~dfj~T~u~`~t~ O kt~cr edtre tot Lploid pl~lt prodPction) Icthololoa tclic~ or aci~iel rl Irc~~~ti· ii ~f~E~t~p~d~t;f~ ~lt ice ·Lq·e ~?C~il dr ~Si~e~f?ll~i~ii~El~ IC tclcl~r~t~d liplo~d (In) yl~ntl cia rdcr ccrtlin circ~tPnccs be I·~a ~·a~j~i~g~Xg~ ~cte ab e· L ~La~ B~8~4~dO~P~EP~ 8~· '~~'~ ~·pp~i~g) ~tt~·~C~C~Y~ Cr~i~i~ (i) idcltifie~tioP ~ Irllctfol oi hlloil plrtr ~i~ 'Iclirlle ch~rletcriltiel' th~t er bl cbrololow arll~d O~d~~ I b ~~~r lolola~ (h4n0 ~lb hr Lnclll Cplla or lircet aa, I~ 10 i9 ClibPDF - v~~fastio.soni -19· (ji) for use in the Bevelopmcnt of breeding lines fot hybrid plant production, Plants produced in this panaet should in theory erhibit greater beterosis [vipar) because haploid derived in-breds are truly holozysous Whereas those produced by conventional breeding are not. This is probably not highly relevant to tobacco, a plant that shows limited hybrid vigour, suggested to be because coscreial tobacco cultivars are derived from a naturally occurring hybrid (that gives rise to an arppbidiploid plant). (iii) tool for chromosnmal additions and substitutions~ (I~iil~o w~~ ui~j· w~n;Rlaeli. 3~ lee2aavi~o~ ?pbe~otypie:sclret~iE~L~?L~.~;;oe. i the ~~IL~UI pld~t·:?~fl bl~ll~O~Ykl~P. pressure can be applied such as ~~rr.~tre~t~i~'salt ~4utrie~ES~s~cl, C: I i I; PDF -!::!!::!!::!.f3 StlC.i: 0111 ·20· r~e Irrhf~ bl Yadf~~t·b d~~t~)~hl~a il·r ~~~I IOIIPC~F~-~B?hl~l .·Y.i~ Ild~d d·t d~a. c) Ccllllor reehl~aic~ P' "L~~Y · 3 ne rjor rcJc~rcb cifort it t~il ~rc~ ylin Iliq toblefo li~pl~ ~s ~ ·u ILt ~ ~a Il.rl t~E~cn~i~-~E~id Lr~l ~~~~ )di~rb jh~i~i~·~g~Cr~p - ~ b lo~al!i~9~ff'~to~b~iI:or~~s~e~n~i~ -rrcs a 1~;9p~~e fr~atl:~~~,9,10). In ~ddition, effort h~ ken fccusad on rtilili~ tL IIOPtlPIO~I OT il~ld Illilblli~ thlt ell be rhie~cd i ar~L dal·.· ~ ~~t r~l dt.5 I~i~z~s )riaL O O O N W ClibPDF - v~~fastio.soni -21- There are essentially two types of somatic variance: (i) S~oln~aill~~_pene~atian of somatic Under certain conditions Jt plant culture, the excised, oreanised tissue becomes ~j~j~ ~ir~~i~i~a~jd Pia~le cell culture. PE~t~ic ion, cells are ene~r?i~t~ djafq~?iwp~f~!:~s~6iIc cr116 an event that leadl to a dPa~5~;f,~?~i~q~f~!E~,~v~tiat,ig the frequency of which can vary with culture conditions (11,12,13), The experimentor uill select eulRre conditions fo"qphbp~·r~.the*l%pee~.ifieYje~~~Dca ~F~clli~li~llrvarittioP_:1 P:t~e ..~d~sPitt~.~f~w~'~~tll~:~I~l~,~i~$4 ~ji~g T~t~pl~~e spontaneous variation or somaclasal IJAC(rO)~JUL ;F ~ L*r( variation t·ht~-~·t~ ;· ~ ~LL-:a;hm;dtioab~t~ "bi~:taljePF~f~aimprovePent , In tbe Ease of~S~ its value is Ihitrd ~Z~f~:~.YS~~li~ti~i~:Ith~t can take place at thc~ltiarei~j ,b Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -22- (iiibL~RIPr i~tra arilailirv In essence this is analogous to the above except that the variability is induced or enhanced by the use of chemical mutagcns. This approach 4~pl~:~·:~i~!~~~:th~:~'i~:e"s~·n~.'F-1~5 ~iF~p and ~aiiru:* P~i~,-·st-~L~PP~·L···pt·~l~.~iRm?m~ IC~mY~ln. ~m~nrntr .YL,LUL~r~ ~'i`) P~I~P~T bh~hl The following eonclusipm can be oade repardins the~iil~~ acid of the u$'.offii~.'iHrir8~t~chaolo~ia , i~~· -·· SiRFIC;ilO IUgi~ynk? . ~ . i) ~overeomeF.aay,EMat berriersr ii) its dor benefit is that it can pctentidly pravidl~eE~deneti~ ~'~:ilbili~SeSc~t~:~j~fsince regenentipa ~otcntisJ~has been ~I~d5KP~ 0~ Q· Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -23- iii) t~Je~E~I11~-~2 is b IC~acFT~!~~ again-st Yhich c~inwre~ly ~iFl~eant~faprove~c~$ can be identified (cllircntiy this approach ha relevance ~PI~J~E~B~P~PZP~ where s~tibn:lerft~ia can be applied directly to the culture epolitions ruch as ~Bnd Putri~iooal Itrelser and E~e~ ~mtit~and resistance.) iv) ~dd~~I~CLe~d~ 6~:·Iiidaeed vsriabilif~pl~fs nor~always_st~ab~ generation to generation liMi -, F~i~ his ~inu~*h ~naol,e ~ ~bll~?!th the cellular ~pptoMq,:brHdiq~strt~~have to Le identified for ~i~!Tpo~~il~~~~*p:~'attriliiitis';i~the *hew i~so produced ~~I Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 __ -26- L(~1; e~l~ r ~nrz~\~ In Iractice, gD1~4~clC~i~I~e al~holah capable of ereatinr new variations in the plant is ~P~tcd~t~rentl~~b~~he"~e"~~2~ttion -pfterL; plant L~r ·- "" '' I ISf!~. P~i~F~s~ps would ilprovements post growing ie laf and final n'~%jj~·l~;~:~;~l~:~t~-~ly:tii~b :plant :populltianslfrol which improvements Cn4 ehg Pi~ac~c~ ~ ~D~P~Y~., would have to be selected,' ~avinsbeen cured, p8ed.·~md processed to 1 ~UBN ~ 1*0~ P~t~ki RIPRB~CN 3. ~tlt~,Fnmrlic~6~eej~apg, ; F;, I~reb 'io PNA - ~Lr*cJ c~ ceS Fnd;a4 C~r ~ba~-; In broad terms, p~F~e~t~is the process whereby I~·~m~5~"~PP~:~~'nE~iB "d t,cgh~e~ ~iir~iidi~~twallr'~iirbdnt:plantsnfot;wa~El~·s~:aeTF"~'d~ii~-~ii~s~i~'d ~!f~i~ This process usually follows thL outlined ~la~ ~S~g~B) Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -25- Sms Is ~'ilii tirS~v~lut' .Sym~'.N.SCi~lm·T~a(· r\ia-d) (i) ;iep~ii~i~ aod eharact~rinri~n of ( ~b~Fb~fJ1~-aiit~t or genes, -"ol· rb ~rb;L.la(*·*J Y ~cr· Pc·-c(' u~U i IJOLP~ , , ~i~ii~~bl~·r~p~~~j~jlikpyPd ~U~i Cbh'5reCICCit. O de~;~C~tl~~b·,c;,,,tI~Jd b~~t~a~aP4~c~onai Thil vill include 'preparltion' of qpropriate rt~atg~, in~ and ~f$ P~e~L~ in ardtr that t~e d~S Ope'dcJ ant ii e~EgBl~dfifie~tJs, Vc~L~Z "Fu r~ukt~ , (iii) the p~t~.t0i~-c~!'"SF~e~ ··~u~·c.,: . ..~ . i~fh~i~iCPt'.q~p~ uslag ~lpprPprl;lte~v~l~s~q, This can necessitate the developpent and use of appropriate ~ spd t~j~ to *hieve ~,~ii~';i~:~n:?''n~b;i~:~h"t~nsfdi~d plant ·c~Y Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 ·26- l~~b(i~;E II ,~ (iv) iY- Y~e(6~E redl to ~ ·~lldSiiatb~·iio~2aod~t~Ylr~ E:_ ··IF~T~i"r~l k~. 81a ~ IS Ci ir ~ NC le e. ~Ylig~ ~ir cy!4~9~iI~·C L R~Uidr tL +~~lall~i~i~~e~]~n. Lsl Idir .~1Biile r~ · iul Iii a·· ad Icld ~ direct uprefrion of the ~ene in the Iclt ThiJ ~ an Eli~P of dbLhrv SPtcr~~ II i~rlllfnl c~plbni~ of ~~~iC Ilecitie·~jiiiq. lo~rt~rl h~LaXla~m~l~r~,nr~c~~iF:~~ Ft ~ai~d~ ~c·~t~l~ilJt~Lrea~e~~lr,~ UL~L d~ ~ ~-~~d·h· ~ t~ htrl bl n~nt ~~ a~ ~:~le ~e14 tlLlt *lirllltiOll B 111~4 lid ti~;~l~SC P~i~' ilailt( Illfll IIIIIEltlOL ClibPDF - v~~fastio.soni _I _ -27- 4fi"E~I~c~-~P~d~ itlcrtfltac~fr o~ inSyc~ri~ t~e, p·--~~ ~'r~t~t~F~Y ~ LH5t~ ~t~ii~if~ ~f~C~ftr~ It E~E~l;r\L;~~~· Yllp~j51~ b b ~ I~e~ I~i~i~i~il~l ~L?I Ir~~~ll~~nne~ Ihdl~e~ttf~~ de dtP~II lr till dtR I· )II 1 ts~j~j~e~~ ~r~e~9p cl~Ja~a d tie 14c~~ YQq~t tp o~ty~or I~k~tldid~ P~~~~!olllfc~pi~oche~i~$ lad ·Icedlr )loeclsll tla5E~pl~t~.~ull.gl~cNI~BI o~.kP~L~ 111~ it ,~rtiE$1Y~tl· td aYs· ~I v~ ~'"' Y 9E~P~ ·' 1l~jl ~U~ECli~FO~b~C~ti~ LI~I~I~I~I~I~I~I~~ I·0·i 1~~.~L~ii~ICinr~Lj~i~I F~-~ ·d '~ ~·t ~ r·C~ld~, I~~ to ~qiu~t:(s~-Irclil c~c~·~Ll'tablClb yldiit~L li~illr Ippro~eh bl Lsll rcftrrsa to ~t r. r--·· LbnLLnl~~~la~aa~~nb ~~i~~~~l~!~ ClibPDF - v~~fastio.soni -28- lethadoiogies have been used to aehie~ B~lg~q~f~ resistance and to P~a~ce plant~~of pT~Fp4·gT-~i,n. Naithtr of ~ ac~iee~ents needed a funda~pmtal uaderstaPding of p~ant ntabo~BP;rjrptl rather are relatively s~te single sene~aits idePtifiedn:bactcrib~j: ~f S~StLPS that verL vill'.stadied and~a~e~i;l~~a i~t~ inveltieation ·stxl .~.~i.i~, a f~~Ytl p~L tutLb.:P~P~C7iill~· sienifi~ant ~it~ Howe~jl:lh?-b~'S~'~fa?i~i'c'en~in5;~-~;;~daiC~F~il-~B~I potFnr~to I~ay$a~ti~!r in~!ur~e unfilaelur~l eplt~, prodj~litics and pianl attributes that uill p~-~ne~..~wil~iod~:t'~k~l and iiiil~h'it.4 ~S~~e~fi;· set'sw?;b~;a~;i~;i~;·'·Tad~i~;~:~h~s: ·P Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -29- (i) ~aniPulation of the nlttatetPduetasl, coder (ii) Modification of tar PicotineSatios Ibv Re~ic ~npineering (iii) Ms"ifiEa_tion of taste and flavour smthetic·pathwavr (iv) Iffid~x~m in ude,sf a~r~ult~al inPuts ~~ "' 7L~~''L' ?CL-~tl- i~·~Y PIC~~ ~t~ hsdl Llttti:~n~L~d~ad ptib~r~f~i~ ~i~i·di~tP~ ~i~mannn~o~ect ·r~sistlh~e:·in-pli~$;ind theue~ak~eriid' iili~t~ CI~oT~t`rfluTL~s ~ada fer til i~i~:~s~iiS~'i~Id'?evcl op 'Stiatelies foi' tb~r DLIIE j c~CT ~· tn i., ?;~ · IPiiCfeii~ifi~LbfioP,~ In qr_dcryto~l~i;;?i~~k~'d~;l~St and develop the appropriate techno~ogies it~ir~l~~b~ ~i~BLa~.f:the~S~dustr! ta~;it~i~tld;*jinC~`its' co~ercial sdaI~f~i~b~iitiliZ~~ Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -30 - 4. Mciecular Biolopv:?Klne ~ppifillteehnim~es ~ILe r~gru ~VIPn~i ~L1EF RtFc~Ox~t~ Tp f,, ~MPu~~5u7 N~J 7Sc~~ By i, yi'i b a~ C~i ~cQPlceg ~d c~hBZ~PPdt~ IM4 dlffl4~ C~ , Thir aren has been r~ed froP:iptcific ~eneti~'~jp~yj~g~-ta·islicPts some of tbo56pia;off bcn~if~i~' fuadaPeatal~leCplar researcbl that are ~lifli~~co~e~ia applieltionT The ~b~e~pri~ariiy to the usL dnd ~~licaim,~(a~p~i· ~l~i^s~P~i~~a na~a~ly.oce~rini Cban~ea in the DIUSti~sthl oc~n7'ai ;;ii~l Itl~ ~t~sib~ v~b iq;coP~in~tiaP witP:hilbli~Cifi~--ciia~jT~i~esj ~6b:t0 an arrajr of fras~jlts i~n`l'i~ue~'~o·an dndiq!dp4lr:pl~dp! A sefies of can b~developep) to cha~i~d~ In this a~ru~er it DFI be polrible to DbtdP 1L~tglf m ~ti~al pllqt $d~~o~i~portandy thro~~h~h~·~Rl~~i~~te'~P~fibt!~~ oE5ihbri~dinp~and ~cleelilii7'i~i~tab~i~f~''r~.~i~onshfpj 7et~c~l a p~ant-penetic infonatidP~ad observable't~g~ Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 __ -31- Such an approach will provide the basislfor: 1) dl~iii~:p7bf't~t~ii~g in Pol;edar~rerP~providins a b,si~ for trait enhanfe~ent b) facilitate the~i~bi~jition ~C··~·'~'~ (1 ~n~la~ intl~til a toils inro·to~c~ai blti~ 11 to s~ ,,,.,, ,~~~ ,iII~~ in ,,,,, ~gc~iPl~fn~,piPail'ii~i~f In. W~n~l~i~ i.linatioa e) Lesin to provide focus for specific Ilenetic epsineerin~ where more pragaatic approaches fail, f) of eP~fY·iPPO't~4 ~~ a 5hOTtenills.bf~p!ant.i;t~]iii~i.scales in ilproving aeronolic or product haled traits, O Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -32- pr~~ide-a·ppt~`ln~cdidh'~lc~gtionXfor· noledlr · based ICQ~PPCE ,thlLt .------ ·· I..,.... hbs· relevance fcr,improving*the tobacco plant~ij~i~th fpfor PpronoPic dnd da~mtltreu beoefi~, 5. ~8~J6~o~t3~h~~lif~b~'si~i~'~;~~ for Plant Benefit S~unpi~, l~,m- ~I1C~P~. There has been increasing ~Tae~e"that B'ranse of soil picro-prganismg atb~:tha~~the rhi~obi t~ean ~aeilitate the optske of nu~~nfJ aad i~abe cares rte~t~lr protcct~iirhinlt di~~~?-~'i~,iir~jtresC Tho~L of primary interest are of thei~j;"re~c~i~t~i~i~Fi~I~ of which there are essentially Bn~i~ppeJ, ccto- add';n~c;Piihoiri~~ Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -33- it appears that ep~l~8irtJf'r Of p~i~penefit to fa~st~bc~s xhilst P%l~?ycorrh~~rtm~g~ilp~t~l4cia~ed~;ith 1 vide:variety ~ Il~d~l~t*nqllrllW~*Culuuj Alt~iFiThaJ proved po~siblel~ cultore~K~t~o~cor~liJia in-vitro using continuous culture conditions it b~2~ot~ea pqb8*ible to bulk P~F~;.V4~ F~g~lc~Fht~In~th~b~'va~, production Systems for9 WPlrhilL pave been devlloped Ptilizing ~o~a~st~incb`n~~~~~iroPwqtr . " $'i"l for use of this systa for c~tdial"bmefit depends:op: i) bde Mteria castl ii) .~h·1 specifid'plaitl: types UlsrJ ~'~fiah nh iii) cb~·~aviniis 'bn'~iduecd 'fe;~iiiler ose" Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 -34- 6iSlsl eoli~iti~ll ~~to'~,~;fo~4~-~aeeo~ To date, the full benefits of using such rpbiotic soil licro-orllanisms ha pi Ip ~( lolly rr~.f~l int udl Ibwj. df ~prtntlll under defined conditions and might provide a basis for engineering additional beneficial changes in the organisms for mutual benefit, Concluding Remarlts The future is technically challenging, but the serging technologies can and should be utilized to develop novel tobacco breeding strategies, desibned to ret the future needs of the Industry. There nill be 1 need for the Industry to carefully define its goals and to maintain a commitment to fundamental research effort to achieve substantive improvsents in tobacco~ ISAr ~wa~xi~) hL ~br~LHofi~ e~A 01 HnlW ~W~J~VIPl~h~~ Lhr ~b~LSY~ t~ Ih C~NC h*I~M-SUgittCO~ i~ re rM, O Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 References: 1~ Bicotiana, Procedures for Erpaiwntal Use. USDI Technical Bulletin Bo. 1586, 2~ J~ Schell ct al, Plant Cell Transformations and Genetic Engineering in Plant Improvement and Somatic Ceil Genetics. Ed, Yasil Icadaic Press. (1982) 3~ Vatanabe, Y. (1975) Jap. J, of Breeding 25 (1) 79, 6. Blakeslee, A.F., Belling, J~ (1924) J. Hered. 15, 195, 7. Chih-chinp Chu, Haploids in Plant improvement in Plant Improvement and Somatic Cell Genetics. Ed. Yasil ~cademic Press (1982), 8. Yelchess, G., and Labib, C. (1974) lolec. Gen, Genet, 135, 2ii. 9. white, D.Y,R,, and Vasil, i, (1979), Theor, dppl. Genet. 58, 239, 10. Cocking, E,C., George, D., Price-Jones, n.J., and Power, J,B, (1977) Plant Soil Lett. 10, 7. 11~ Sacristan, M.D., and Herehers, 6, (1969). Molec, sen. Genet. 105, 317. 12, tarkin, P.J., and Scowcroft, W,R, (1981), Theor. Appl. Cenet. 60, 197, 13. Burt, L.G., and Hatzinger, D,P, (1976), J. Hered, 67, 381. 11, lidholp, J.H. (1977), In '9lant Tissue Culture and its Biological Applications' Ed. Barz. Sprineer, Berlin. 15, Scowcraft, W.R. (1977), My. Agronomy 29,39. 16. Thomas, E., King, P,J,, and Potrykus, I,(19i9). 2, Pflanzen, tuchtg, 82,1. 17. Proceedings of 'CORESTA' conference, Greece (Sept. '85), 18. Second international Tobacco Exhibition, Richmond Virginia (1986). 0· Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111