~fsl Tobacco QwtmlslS Research Conlcrenco Below please type name, address and phone number of principal IIBSTRAeT EORii author, NOTE: The abstract must contain a meaning- g,~,,, p, Pjordjavie ful summary of the material to be presented. ~,,~ P,p~ttpep~ It must include the objectives of the Ee~ 11-2121 ~SCN search efforts, the methods used, and the results obtained, The editorial eotewittee Ilnivsrsity of Icntuelry 106·257·3309 reviews and accepts papers based on the ab- strict, Yery short or vague abstracts will not be accepted, TOCeO·S~ERFIE IIDRO~IIIIE lecmrrrmmff a DI~FEBPF~ OENDIIPES ~ B~BIEI TOBICCO 11 DIPFE~EF~ STAGES P ~MB AHD ~R·COB~NG lUrlana P. Pfo~lcvio, LowellP, Bush, Scott 1, Gsy, sad charles T, Yoc~oun, ~ronq Rpart~ept, OniPtrsity ol Ieotucl~, kdngfon, a 40516 ~1~ bnriel pnotgp# and a 1112 dithripa in ~lloid cPntat Mn grc~m uPdar reeeonnanded oultunl cwditiws R burlcp tobrco. Illtrdc·P~ltrog~n, 1~Peld and tr4lceo·aptclHo nitKlsanlpe (TSI) contelt vera ~nalJzsd in lseina end Ildrib tissw diriersPt stYIss or tobreo da~elep~ept and ~ir~u~iog. TSNI iO~Ptigp sad se~latk eere dct3cd in jPreniic tl*co pints and inerwd as plants nuhred and reaobed 1 nxlmm io onled ~~sl Bighsr leoels or TSHI IAn round in Line ~L~oid geaotlpes, and nicotfne to aofPiootiPI comerter IlnsJ than In nid~ib and lee o~loid Unes, rS11l Eoocmtnt~p wu ~tsr iree the Fsr~ph~rg oi.L leaI to lefiu adjacent to the ~idribl AUwloid ICCIPUktlDD pattern during growing apd ouFlng suggests tbat TSIA are prob~bl~ toned It apePss or tbs a]Jtaloida, Signiticsnt d high eorrelstion codriciwb between ·nltmscmolpieptinl [1) and tlsltrasomnrlbip~ (R(TJ end corraspsndins ~adid oorPicotine and anltahine, s~gsst that Piner t~aloids haoe I ppm rot In TShU rontioo them PiCothl, ~ the other Ld~ higher ~o~mts or in the tar ~on Iwar It~ pPsltlons iadkk that nltntl aight be oDnsiderei. w n iPportant tootor in TSI~ ronation under ~orable onpironeental and euFing oondltions, i, The abstract should not be lest than 180 words or more than 225 words, It should be included in the space designated above in form suitable for direct photocopying by the printer (if typewritten, use a carbon ribbon), 1, Please note: (a) Title all in capitals, (11) fioeation and postal/zip code follows author's name, if multiple authors are at different locations, (c) presenting author is Pnderrinad, 3, Mall original and I copies of the abstract to: GALE E, MATBlS, RES, LABORATDRTESI TENNESSEE EAST~AN COMP~JY, P,O, BOX 19'12, KfiJGSPORT, TN 376621 I1tSsAI 4, Abstracts should be received no later than Jtme 30, 1987, 5, Projection requirements: only 2" x 2" slides will be aeeon~mcdated, with maximum glide thiekncsl of 1/a". 6, Maximum talk time of 15 minutes, with 5 minutes for discussion, O ClibPDF - !:!r:!!:!.f3stlc.;:nlil tOBICCO·S~GeI~FIC IIIIBOSIWIHE ICCOIOIII~OR IX DIFFERENT CEHOTIPES Or BOR1EI rDB1CeD lr DIFFERENT STAGES oF CBOU;N IND I~B.CORIHC Slide 1 SYrRODOCfIO1 fba papar I as gcln~ to prlsent tDdal is tb, result or pur EOptipUing ItadlC) on tObae00.IpCCliiC PStrDSeIOna (TSHI) C I~~UHDPI LI(UFl]lllll.d dlll~llltlll 1D ~P~I((D J11PI (PP!II growth end CUP1Ppl Thare are different statePents In tbs l!ttnture reaerd!rg the stege oi tobacco ds~c~opmsnt end faring It which TS1A etaFt to lora cad ICCIcUIIta is tbs IsaPas, Sole raports slain (8acbt 1915; Boitlannst a1,, 1916; Bcehr at a1., 1911; Bccht st 19f?l; Bleht at ~1~, 1918; Idans at a1,, 1983; ebanhcrlain ct 111) 1984; Boihann It all, 1915] that then are se ncasurab2t aPounts al TSYI in IrecP tcbuca , Iceordin( to thess raports ~I Il~aloids lad P!tnta llin rise to FSU durirg toblece EPringand procaasipg, Bouc~crf tblra are also rcport3 (8ush ct a1,, 1919; PjDrdJa~ic ct al,, 1985) vbicb rcoognize tbs prsscnca c~ B'. n~trosePornicPtin, (XN1) in green tobeeco, fbi slip objeftites of thin study wits; (") to tnollre loditidual Jtllofd, rSsl end pltratc· nitrolcn EOPtsDt in Iallpa and nibrlb tllsllc of 5 BuFleg gcnat~pss end fI·1112 differing in ~ltaloid cPncentrations In order to get the profile of their Ice~ulatlop and d la t r Ib at ion wifhln the toblleoo slant at d~lhrcnt,tyea oi deteloplent and ···--~ Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 lir·curing; (b) to detcmln, relationships b,tween pncunora and tobacco·spseirle illr~loid·dsrind nitrosaninss in kPIPa and aldrlb tissue and the whole Ici~ respectivell, WlrEBIIL IFD IETBODS laterial Slid a 2 Burleg 21 and three lines isogenio ior lowered ~l~iloid content developed by t,lg and Collins (19'11), as well as I Burlc~ nicotine to norniootine converter breeding line i fe3~9 Ind 1I· 111? were chosen tcr this study, fobaeco was grown ardsr reconPendsd cultural conditions for BurleJ tobacco in'1983 end 1984, Slids ~ tear aacplca were t~ep IrPm iour sr~l~ pealtionl dllring n~ctlti~lgrovtb, It tOPPIDpl at blrveatl at tDd·Pf- Jrlloving during thlcpr~mg proeela, and alterair·curlPg as oo~pllta. Ildrlb was rempved Tron IlllinP;lnd all ~qles vere i~Pediatelf ~rozsn and sPbss~ucPtll Irseee-dried, Slide P Tobacco need in this study had a wide Il~lloid nngc~ The highest eiounts or total ~lsloids In eured tobeceo oi BY 21 genotgpe series were nessPred in high parent BI 21 both in leniae and Pfdrib end the lowest in ii BY 21 (shown ~liluts Ire the oeePs oi 3 reps and )1 PosltlonS), Total ~ll~aloid content in Iamina and Pldrib or converter breeding line was lower than in fiP and BI BT \P 2 O Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 21 rtapccfitllt, iUlclcid 1Ite1 ip Illina ol rI-1112 as aeedvtlt higher then in genot~pss o~ BT 21 series while total ~l~alo!d content in Pidrib tissue was lover then in lr 81 21, rf·1112 oornieotlne ~ceau~itor) was chosen ior this Itud~ bselusl if contl~ns blgh enouots ct Eyoamiol, PerPicorine, anatabioe and anabasine, Slide 5 letbods The artbods used it thh stPdg tor nitntc·nitrogep, Ilr;lPid end iSWI dcterainotlep were the slpe 13 we bed described earlier (Djordjs~le st a1,, 1986)1 BESUUS IWD DISEVSSIOH /Itllaldds Iltbough eight tobsceo ~l~~loids were oonitored in this studl en11 data on nlcetine (Ae), nornieotine (BBIC), anatsbine (1)1~), and IPebaslne (IflB) accuauletloa I~ tobacco It ~~rious stages al growth and curfof will he presented nov ~or these tour Il~aleids Ire oi 11Pportlace 13 tSII1 prwPrsors~ Tbs results ot ~Ualold deterPinaticn shoved that BIC cootent in Isnina oi raur genotlpes in Bl 21 series increased rrca ~cg~tlti~e Irowth to and·oi·~tllowing but decreased Iligbtl~ in eored tobseoo, eOPC6rPIDg nICOtlPe to DOrDiODtiDB COPVerter breeding line and T~·111~ the highest snouots o~ ETC were secuaulated in lulne It hsrrest, I tsPdeoq or al~aleic concentrstieos to decresse during curing is well ~nawa iircft Clit; PDF -!::!!::!!::!, f3StlC, i: nill (ho, 1972; Istsusbiai st 11,1 1983) which is sttributtd to dr) wright loss, Slide I Is can be seen on this slide (the results are expressed in Igs of %TC per ~ra~ or dry tobacco weight) tbs BIC content in l#ini frop the tips or four Burlsy 21 BIDOt~PCS grown in 1981 increases Iro1D L1 to 88 88 21 111 stages or tobacco develcpnent and ~ir·curing, I1IC content in C0 and ~I·1112 was ~~ higher than in the geootgpss or B1 21 series until harvest, Iller blrtast 8le eontentipk~ip~ ol thess two gsnotgpcs decreased as ~ir·curiog progressed, Especially prooaunced decrease of RIC content aftsr harvest occurred in lamlns or Il. 1112 due to high conversion of 1TC to tllC, Cured Tf-1112 contained even lover uouPts or DC in luiol than tf 8'8 21 Depending on gepotype PaXIIal IPHZC accunulation In luinll occurred at topping (Il), It harvest (88), or the and of tallowing (82, BP, Co, Tl) what was alp,ctad since fbe hlghe,t nicotine to norPicotins conversion rats is either at tapping and hsrteat (pkt sstPrati00) or during curing (Yarnsesp end LtzlngrF, 1968), eon~rrsion or BIe to INle is gePetielllg controlled and tal~a plier in the leaf, Slid, B llle levels in luioi of all sis genotypes were relatively tbs sane at vegetative style, It topping tbs highest alount or 11IC was Peasured in Ll BT 211 It harvest an tnbaoced ~ceu~ulaf!oP of tHIC occurred in lanioa or BP 81 21 and Il. 1112 increasing through lellowlngt as well as in converter, in onred tobacco TNle content in Iaiina or all genotgpes decreased, Ibs Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 highest sIDUPts Or HWf in cured ~l~iPI were measured in TI·1112 snd then in coolcrt~r. )FIC content in cured faoins or 88 hi 21 was epcn lover than in fT Sr 21, INIIT and 1118 ucu~u!ition in laPina of tbs genotlpss in hi 21 series basicslll followed the pattern oi 81IC accumulation, The highest amounts or lllr and 1RIB in lanina or conertrF breadiPg line vcrs tsasursd at baroest an was the osse with nicotine, The bighcnt arounti oi IRIT and ItlB in rI·1112 were neasursd It aPd·o~·lellcwiPg, Slide 9 IIIT and IRIB content at earlf stage ol tobacco derclopPeot was uniform in all sit genotlpes, It topping tl and 88 BI 21 ieeumulstcd the highest amounts of these two ~laloids, It baroest, lod·o~·glllowing and alter lir·CPripg was EODP1Cte the ~plounts ol lnr and I8lB in 1aPina oi g~Pot~pcs in BI 21 series were increasing rroP ii to BP, 1BIT and 1888 Ir~els in luioa oi converter breeding lint were lower st 111 stages cl tobacco growth and early stage ol ~ir·corlng comparing to BP BI 21, The accumulatioo ol Illf and 11118 In lalpiol oi Tl 1112 was PUCh hilhCP ct )Id.O[.~CI1OYIDB Lid Llt~r curing was conplete than in gP Si 21. Y~tr~te Besplts of nitrate dettninstiep are qrprsssed in ngs al nitntc·nfrrolcn (803·1) per gnm ol dry tobleao wtlgbt. Dcta presented on the nest two slides are the lpesnn or three reps, Slide 10 Shown data illustrate thst there are small dilrereocss Clit; PDF -!::!!::!!::!, f3StlC, i: Olil ---. .--- in xa3.n eontent in IaalPa or d!rrerent gepo'.ypas It ~c~eiltiYL Irovth, It topping the Ibaolute ~iluis Ire a bit higher than ~or tobacco iuplad during vegetative growth, It herveat R03-W lerils uere lover than st tOPPIPgl a PbCDOICPOI vhlCh vaI Observed by Hamilton ct a1,, 1982, and which was associated with the ao!; aoiaturc, P03·f ecPtl~t ialaln~o~dirrcreat Ienatlpesst harvest ras pretty unilorn again elespt in tI-1112, During air-curing Slids 11 oonceatratioP o~ N03-ll inerusrd and reeehed I oaximap is cured tobaccol however bili1lr 1 the highist asouPts o~ H03·H were eccuPuIatad It 1!1 Itages or tobafeo dctalop~eot and ~ir·euripg in la~iPa or lov ~l~leid lines in B'I 2lgenatgpe serta, Fbe higher Il~sla oi nitrate in reduced ~llloid Icootgpes colpared to co~Ecrei~l ep!t~nr Burleg 21 way occurred due to their lover yield potcnt~om! associated uith the Iow allraIeid genetic factors, The lovest IDOUDtS oi nitrate were ieuuFid io ff-1112, Tobsceo·spicitic oi~rosadnes fhrce tobaceo·specirle DitrDIIDiDCI Yere IDDDitDrCd in this Slide 12 study: I'-PitrolonorPleotinI (BWtl), 1PI·nitrosPapltlbhl (BIT), nnd 5-i )I·nethyl·8lnltrDllpipo ).1.(3-pJridll)-l -buranone (191), The GC oethod wblcb wee used rer TSNI d~tcrPfnstion dig not allow the separation epd puantllicatlpn or B'·Pi:raaoaa~baslne (HdB) in tobacco, The results or our study shaved that TSNa lon~tlop and Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 .------ soeuoulatioP in luiPI of 111 six genotypss started in tbs ~Srld during ngct~tlpd Irovth end Incressed thoroughoot 111 atyra of dllllOPICDt and ouring rsaching enexinup in curet leaves, Decreese or IUa~oid and iPcrcue or TSIII coneeotratlons in cured leaipe suggest tblt latter eight he ~orPld It the eIpense oi corranpondlag ~l~lloid precursor, Slide 13 The results of TSRI detaninltiop sre e~presscd In Pgs per Llran ol dry tobacco weight, Is do be seen iron this slide INN sccuaulstiPn in lseins of sil sin genotlpas during ~cgetl?lte growth end at topping ula snell (nInogrsa levels) Opd IpprcximatelJ the sane. Ihe differences between $enotlpes stlrtsd to show UP st hanest though, in BT 21 series the highest uounts of INN were o,uur,d~ in 8P B1 21 end tbs lowest soooots in II hi 21, The oenbentrstions ci IPWI In aowcrtsr breeding line iron the harvest on were higber cooparsd to BP BT 21, The eceunulation oi I1P in TI·1112 coincides with INIC scctllpulatlon ··· trend, Slide 15 while the highest levels pi IIr in lenIPa ci 011 tour Burlet 11 genetJPes were mcuurad in BP BI 21 It 111 stages of tobecco developpePt end Euring the highest RI1ICPUIDUlltlOD 00CUrred in Ienina of 80 BI 21 oPil during curing, The highest Ilr ecnwntntlcas in the ssapies t~ca during tobscec develoPnent (rclatdt!oo through hsrvest) were eeesursd in 81 BT 21 though, CoPpar!ng the conrerfer breeding line end TI-ItIP the higher seeueuletien of Ali~ wee in the gePotyPe which Eantllnbd the lover Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 ~ --~- amounts of corresponding ~lalold precuraor, Slide 15 rbe higbsst concentrations of ISllt in lanina or hurler lines were measured in hi BI 21 atall stages of tobacco dc~clopmcpt through ~od·or·Jt~lovingl ggg accumulation in coPPerter breeding line was higher than in TI·1112 during tobacco development while the greater amounts of hwg in cured Ismina were found in rI· 1112, a trend vh:ch was entirely opposite from ILC accuPulatioa. fl ~ Proiile lecording to Indersen at all (1982) chemical composition of ~ir·corld tobacco changes during storage in controlled environmental conditions. eomgaring to other nitrogenous compounds the greatest cbsnges occurred in the levels of tobacco· specific nitrosamlnss and their precursors, In order to atoid misleading conclpsions about formation and acoumulation of TSBI In green tobacco since tbs samples analysed /·· in this experlmeot had beeP atored for two years lanina and midrib tissues of standard burley eultinr tI 14, grown in 1966, were analysed for al)taloid, ~03·Y and TSBI content iaDediatefl after aanples were ta~cP from the field or the barn when ~~r· curing wss complete, Il~alold and nitrlte·nitrogln accumulation in kamina and midrib tissue of f'I 14 during growth and air- curing and tbsir distribution within the plant were as expected rob~cco·speciric nitroaaminr profile or IY 1S gites tbe proof that HHH, 111 and ppg formation and accumulation starts in the field increas ing throughout all stages of tobacco 8 1 Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 dllllDpllDt IDd ePrlPtl Stido 16 doni rSll ICC1II1LtlOD OCCPrrId 1I1111 in nidrlbI dPriil tobIcoo dIlllopllnt to inorlIII ~ilnliie~cfll It ~~d·o~·llllovlnl lad rcnch thl pit in aPrld rillP1 (dlfl 1P( not IhOlP)I fbl ooaclntrttionl o~ fJ11 in Ildrib tinIrl rorl IlntrIlll lerlr tbIp in lIlinI, rbe hilbest IccpiplIrion al I11 Ind III 1! tobneco pl~Pt oecarr,d In IIninI Iron th, ItltlI ~rol lov,r po~itioa~ SIidI if rhiPb ooPtliPld lotlr IloPntl or Il~~loldl Ipd LiIhlt Inouotl or Ditflt)· PlfrOl(D I ~I 00D01PtrltiOPI in tbl 1II1II frO) dlrlirint po~~tiens wOrs r~fhir IeIttorrId wit hin tlo plant I tbi hilhlIt IloPnt oi III in l~~ip~ or Onrol (r 14 win in t)I iron thI top poslrloP, Illdt II fSII l~i Irotilr It IhoVP (lr11lr (OIJ It 11,1 1916) tbIt Il~~loia oonooItrntions loP,It in tbl ;loIt blIcl in li(rib Iod IdjIeInt lIlinI IDd bilhoIt It llIr tip ltd odll, thl II1I Slidt 19 pIttlrP Ipplles ior othor tObncoo ~l~~loid~ ILioh nrl tolfi~tll Ionitorod In OPr Ilb, ODll IfOltll( 1I1I)I l(r) lD0o01ialDt protlolI liPdinl~, fill d,rsmin~tio~ in 1l~1p~ Trot Ildbll lpl Ilripbort IIQtion oI bPrId toL1C00 1OLf (ff II I II drown in 1986) C I I bP D F~w w .f 3 s t I o, c o nl revelled the sane pattern of n it rosamine eecuoulstioP and distribution within the leaf like Il~~lold precursors, Slide 20 Statiatlca_l etaluations Ye had presented earlier (~SordjaPic at al,, 1986) tbst inditidual toblceo·sptciric nitrosaPines (HW1, IIT and II~) ip~ cured laaina and midrib tissue were significantly and posltlpaly C Eorrlkled uith rllpcctla altlleld prterrlorlllfiIel mt lpa )IC nspict~al~) but Pot with nitrate, Correlation coefficients calculated over all data obtained in this erperiment (6 genotypes, 2 tissues, 2 positions, replie~tcs, 5 dates, 2 fears) rs~~83 the following: (1) leitber of monitored tobaooo-speclfic ~llcllo~d·dlrind nitrasapines in lamina is significantll correlated with either Slide 21 oitrate·oitrogcP OF nicotine, On the other hand there is positioe and significant correlation between Indipidual and total ~SII and minor Ililoldil (2) there is no correlation between TSWI and their precursors in midrib tissuel Slid, 22 Generall3 low, oonsignltL~nt and often nsgatla correlation coefficients for oitr~tt·oltrogcn and Illy, lla~ and IllfI: in lalina and midrib tissue suggest that nitrate concentration Itself is not the liiiting factor In tob~cfo.specille nltrosamlne formation, Tbs higher levels of TSWI were round in Ianina whicb 10 Clit; PDF -!::!!::!!::!.f3 StlC.i: 0111 00Ptlnld, depending on g1no1fP) Ind the stage Or tobaeco allelOpltDt and OPriDb, PP to 10 tiler 1CII nitrate than lidrib tiasPeI In addition the higher OonoentratioPs of rSll were neIslred In genotJPes rhiab aoptlh,d looer anoPnta of Dltrltl- nitrogen, Pithin single fobIcoo 1I11 higher nitroaarines were found on p,ripbtrf where the 1e1e1s or nitrate are lover eolparlng to ~id.~~ctiaP ldjaosat to nidrlh, Bovlttrl rSll Ir~~l~ in tobaoco inereaIed fron IeletatiOn to curing 10 11 Pitrate. nitrclen, Bl~idlt, vithip toblccP plllt the bilbsr ~SNI iertla doripl plIpt dltllOplePt and lir·OPrill Xlre fOPDd in the ICaIIs Iron loter Positions Vhioh oontain higher InOPntl Of nitrate and loYer anoonts of alllloi(i than PPPlr Ieateal 111 these obsIrtatiopi aPggeIt that not nitrate 1OlO)s alone hot in sonjiagtion Yitb other tariah)ea night he 00Dlidered II ailniflcanr faotot in ellralold nitroIation, hoist ooot weather diring tobaeoo I~~~lopitot and suringI aonscpoentll the higher population of nitrate PedPoinl organists on tobaaeo 1111111 1111 PPIIIIOP 1P rllltiOl tO the lrDUDdl pB Dr roblopc, naturitlf stage of 111110 at then teehnololl or b~rtlst and orrlPI are i)PDPtlPf 1aotorI rhiob deternine the extent of nitrate oonterlioI into nitrite, I nitroIatint agent, Oae or the rats to linirlrl nitrosafioa of tobacco ~l~~)old~ is to isbibit nitrate rldpotlon to nltrlte oaPsed )t nierollora eorrelation Ietveen IndlrldPeI tollceo·~p,oi~te nltroaalinel aid respectiie allraIoid preoprsora II /Insr~fll high ~ilniiic~et, These results indicate that ~l~~loid oonointratioa eod EOL~OlltlOP bt th) tOblCCD tielUI 1F1 IOrl IlpOFtlDt 11 C ClibPDF - v~~fastlo.som ---- --- ..,. 1 · ___ indicators ior TSNI lornatlon. genetic oanlpulat:ons, agricultural, prectiees, and curing wbieb allow control over the levels, coaposition and distributioP ol naturallt occurred Ilhloids witbin tobacco pilot could be a powertull tool to gain control over the levels oi tDbaCCO·SPlelrlC nitrOlaPiDeS in tobacco products and slo~e~ ggggLUglggg i, fSHI accunulated In juvenile tobacco plant, increased as plant aatures lad leaves begin to senescence and reached I 1axIDUP in cured leapesl 21 TSII formation Is Pot correlated vith nitrate concentrations neither in laiina nor in Pidrib tissue oi tobacco. 3. positive and signilicant correlation or TSPI vitb alltaloid pncursors indicates that allaloids are deternining factor in TSII ~onatioP, a, Indct~nln~te variables snob as conditions during various stages oi tobacco growtb and curing wbicb increase the availabilltp or nitrosating agents and subsepueotlt the extent or rSYI ~ce~ulat ion should be studied in greater detail In tba httlrel ggFEREllegS ldapa,j,D, BrunP~~ann,I.D~, eecbt,S.S, and BcTrmann,D, 1983 1 ICS SLIP~Ser. lobsrseP,8.1,, ~aspcrbiuer,~,J,I BurtoP,B,R,, tlanilton,, and ~odar,E,B. 1982, Cbaogis in cbsmical coiapositloo ct bo~ogco~zid·l~ar cured and iit·cured burleg tobacco stored In controlled )EPlronEeDts, J,lgr_ic, Food eben.l 163·668, Bush,~.~~, Builton,J.Z., and DPt!s,f.D, 19791 eheiical pualitl ol Burlc~ tobacco oodii!cd by curing reglpe, TeRC, Ibst I Cba~b~rliio,Y,J,, Severson,R,F, and Stepbeuson,nlG, 1984, Lirals ol P·oitraaaooraieotiPe in tobaccos grown under tarJing agroncmic conditions, Tob~Sci,, 156-1581 Djerdjevie,l,VI, eay,S,L,, nPc(ovo,C,T,, lad Bush,llP, 19851 Illloids, nitrates and nitrosaPIPos in lamina and stec oI Burlcf tobacco dil~erLg In ~l~loid conceatrstion, ~9~4~~!bst, ksl~ D jo rd i evic ,1 ,O,, gay ,S ,1 ,, Bush ,I ,) ,, and naclouP , 0 ,r, 1 9 86 , 'Correlatlon betueen tob~eco·spteitic nltrosaPlaes aad their precursors in iidrib tissue oi burlep tobacco, COth TeRC, Ibst~ ~ Id, GaJ,S,, and Bush,t,~, 19861 r:nter].eai variation in ~lt~lo~d colpoaitlon of burl~~ tobacco Hi~9~14P~ Tab sc on I 40tb rCRC, Hall~ton, J ,t ,, Snab , t,l,, and towe , R,B, 1982, bit rat e concentration chnnges during seaescence aad air curing oT bprlcy tobacco, ~ob.Sri,, XXPI, 133-13'1, BecbtlS,S,, ornar,i,n,, ehan,el8,, eorn~ao,P,, and Tso,r,e, 19'15, On tbl) origin Or ii' rDit rOs onD rnieotipe in tob~ceo~ 29th '2020, Bcc bt,S,S,, ChsP,e.B,, Oraa~,R ~~ ~ i Boihuo,D,, and rso, T,e, 1911, Cbnfcal studies on tobacco ~io~~ t01t Tob~eeo a p e c i I ic ·n it to sarin as: origins, c I re In og e nle i t y and altlbclino. ~IRe SF~~rubllcati9ga, ?e, 395·1131 HechtfS,S,1 Chcn,e,B,, Doog,l,, Ornar,A,n, Bo~[PsnntD,I and Tso,T,C, 1911l1 ehcmical studies on tobacco spoke, t~: Studies on ooP·tolatil~ nitrosamlnes In tobacco. Britrog~ zur Tlbl~roraehueg, P~, 1-6, Clit; PDF -~::!!::! . f3StlC, iOlil Btcbt,S,S., Chcn,C,B,, Birota,N,, Tso,T,C,, and BeTIIPIOP,P. 191e, loblcco Ipsclr!e n~tro~~alnr~; rorsatiop iron nicotine In titro end during tobacco curing and cateipegrnicit~ instrain 1 Piccl J,Hatll ~ fpst, (PS)I 819 · 829, BoThnannD,, Bicbt,J,S,I Ornal,B,n, and Y~nd~r,E,I, 1316. ehePical studies on tobacco s~o~,. IbII, litrcsonernleot~pc: presence in tobacco, ~oroitieP and clrclnogeakltl. IIRE Scisnt_ll~e lubllc~tions, ~, 30'1· 320, Be~haon,DI, taPoie,E,J,, and Becht,S,S, 1985, Bicotine: a precursor Tor carcinogens, Cancer letters, ~, 6t·151 ICgg,plOl, and eOtlinSIgIBI 19111 Inheritance Qf percent total allraloids in lieoti~pa tabacup I, TT, genetic sfl,fts or two loci In Burlep 21 a Ii Burleg 21 populations, Can, ~ etPet, Crtoll, 2Bt*~31, nstsushlo~, S,, Ohsumi, T,, and Sugauara, S, 1983, eonposition o~ trace ~lilolds in tobacco leaf k~ina, IBfiel Bool, 501-910, Tsa, T,e, 1912, Phg~olcgJ cad b~DOhCbi~trt 01 tobacco P1IPtl. 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