m i is n n riri^ T S ^ ii Am im m ttA m s m m s

t^ t im wumm f m *•

A m m m m m it t m t& m t m^mAts. m t mm ivm & m m m ii m m m m , mMUMMmt

c# t m m m iM m m m fm, m s Bmm& or

m ^tm m iicisa^t

Iff m u m m

JIfliS I f i f

■%

Pmtm r* Ukio Utmtm Jalm % I U i

Vm emetity thmt th is chiwia

•ntf that in aor of»inioii i t i t MCiafaeterjr in

wcopm and civmHty aa a thaaia fa r tha datraa

of Haatar of Acianea in Agrawaay«

m u u o u m tm ti

eiMiitami

- , ■

' i. -

ABSTiUCt

th* fw tfow of th is thooi* vmm to ottsdy tho ehovintrr

•nd conotlco of flowor im £• nottdifleonoo S« fiojr*

£l«oon yaroetot llooot eo tw ^ r i « t i o »

froiR no«r«wliito to lin h t and dark pi»r|!»lo« war* aoloetod for

th is atttdy* (^roaaoa «M»r» attopptOMil aaKmg tlio llfioa* oaluK

«hit«N»flowarad ytaaits aa tho foauilo paranta* Ooa to d if f i*

etiltiaa atieotmtorod in ttvo eroaaiiHi* attrikt^a b lo to to«^«>

a i i ^ and/or aoviroiMMnta 1 eondifionat and/or ifdtaoront in

tko apoeioa^ no tm o oroaaoa ooro obtainod. Cenaoo^ taak^y#

tha inharitanea attidlaa propoaad nora abandonod*

fiimoRt idontification oaiiift mpmt- «iiro{Ratotmfb|r in*

ooivod too atapi# I ) analyaia of «fea atljreonaa pwwiant

and 2} anatyaia of tbo tlycoaldaa praaant* Aglyeona an<>

traota war* et^tainad ualng hydrolyalt of tbo patat tiaaoa»

and glyeoaidaa vara obtalnad ontraetlon v itb mathanol or

Mttianol«»HCl« BAV waa tha yrtnacr aolvant ayatao naadf

otbor mymtmmm woro HA««^1» tfSHAa, «4miAo, «&%Ut

C Ifo iaofToi^t ateolioDt For«atal» »wS IkW* tha oaiRp<«i«iita of

thoao aolvant ayatowt or* roadily availablo in tbo litoratoro*

XdontifliMitiofia nwro baaod ufion Rf«valu#a for id o varieoa

aolvant ayataoa and open eolor roaetlona In v la ib la and

u ltra v io lo t l ig h t» both with and w i^o o t anponla vapor*

SoffleioRt eone«ntratod ontraet voro d iff i*

eult to obtain.|, thaaroby hiisMgi^int eonoloalvo idontiflootioa

of tbo eonpounda proaont* It «Mia li^ieatod that tdbo phonolio

Iv

eonposlcioB o f tho p«Uilt sitch w>ro eomplmt thftn pr«vi«KUi»

ly r«port4»<l« An « « r l i « r •cxady d*nofi«er«««8 th« pr«»«Re« of

OB oaCtiocyoBldlBp eolvLdlai* oo woll os • ol»i!lo

onthoeyonlji» MoWidiii 3tS*dlslueooido* In odditlon^ l^o

proooBOo of oL«ht y m l l w yig«Nmco ofid ttiKoo oolorloos eo«M.

yoim4o« «d%leh flBOtooood io ulCKoviolot lights oos ^mmon

•trotod (A2, 44).

la tho prooonc otndyg o emwidorobly lorgor mmhmr of

eosipottoats Choa ttio obovo Indlootod. Toatotivo idonttfi*

eo€i<m of th# oglyconoo ond glycooldoo is prooontod Wlov*

AolycBBoot ao Wldltt forallc oeidkooaqpforol otlogio oeidqoorcotia

Loos eoactaotvo ovtdoaeo ouggostodigoooypotia j|*»eouMrie oeidaiaulotta oiaopie oeidpo to lot in ioorhoaaotin

£Jj£S£lJUba* »»lvidta 3,5*dt«i«ieooidoAt looot too glyeooldos of flovoaoo ood/er flovoaolot eao aoy booo hooa kooKpforol Swglttcooido

IntorroIotloBships oooag tho plgaMmts la tho prodaetloa

of phoaotypic eolor ooriotioa i » tho opoeloo aoy bo stnmsrisodoo fotloimi

DolohlBldlB*» blUOt 001(00

htltUto (MtelftUflBil* roddoalfii t^»A dA n.3^«disIofiaolda t3^5^1oIireooylotloB)t

latonolfylBfi

holvtdka lo o aothyiotod dorlootlvo of i^olpMfiidlm.

to««ihI» tgrUtloB*! dalllnt

C a -p ty iiM -U r. t» trm tmnc, „ f fU^rn— .UtllttfliJL*

bitMtmi

l> tiitmmmncit o f fUVeaa«. £lA1gBftaIi,* rm*m

or srollcw bodtgroinul, i f ttiooo plgssortco aro

l^roaant In hlsh cimc*iitr«tlottiai o trc i

2) gya&iaBftt,«-iMKPBfiiI,U» m iis« <»r ymtlem

baekcreundi mt*d/mp

3) bmek$roma mtfmet of

ptastl<S-bo«B« plsswite

antf tiflXCBiM* "boar* «o ohito

fMiColoi ss«r eMtriboto to oliltoiiooo

121* orotmo* or y l lw i m u

9 ig i> iia gU i,U } t, lotfegsrmiaa • in purpto

or iMismeii eoloro

t f l w w

D iow eft I roaaofiinsi or

2) hloh pH I bioolj^i eoiorloo* poooao liooo*

Tho ifitoroetlon of thoto offoeto roaulto In oorr oeoi*

plicMtoa Intorrototionohlpo in tho frodiiction of irihoootrpie

fleifor eolor in MfiftXiSfiOU* conetuoiro laontlfleo*

t i ^ of o il pigsMnto ofuS fdiMiolic eonpouiMlo prooont* ofMd tho

a«ton»iiMtlofi of thoir diotrilMitioci ««a eonowntrotion *00110

* fho poosihiility of ocriotkon ooo ongitootoa hr tho tontotivo iaontlfieotioci of etnnooie oniao*

v l

color ctovcocy wot»l<f tio ro to f^rocodo oluct^lociOB « f tt o

•boro Intoroctiotui*

.2-

Tk%ix cf Q tm rn n

A fts m c t, ..........

ht$t or ........... ........................................ ..

LIST OP ................................................ ....................................

ifmuMnjcTXoif .

...............................................................................•••,4

«m iw OP ........ •••••*•••5

U th « « f Ptem* CmXtft.............A* PLsvoetoitfs****************!)**************)

Hi« CcrotMioids***************.*************!!

It* thm Jht04mtim of

lit* Tito OMOfieo of rtm H »A« tlioB» 7h# CorotofioMo******.**********************??

IV* Thm ln€otrrotiitlo««4lr* Aactiocyonio «imS o ^ o rPiSflMiiss in tho Prodncvion of r t o w Color*S3

A* AnttMKyonlfi Colov**«*««***»«*c»*»**«««««*«««StS i* Ihm ContriSution of Fiovonoo «n4 floironolo

Co Flowoc Coloc««**««»***«»«i**«**«*«**«TS C* tho CottCrilMttioB of Quileofioo mwi Aooronoo

Co Fioooc Coloc*»»f 0* Coroconoido ond Ploooooido Oeotorrlnt

TosoChoc• • • • • • « • « • • « • « • • * • * * * * * * « * « * * * *T6S« IncocroUciofioliipo of Pl«»o«iCo In

V* THo ftoLoCion of Cho Soco«r«^ to Slniioc Voi4«*«*«3f A* lIUBMllttlfllB* ^ttHWjS5HMytiSg|8po^Oe*e#«e*e**e*#*#e#****#34c . ........................... ..

VI* Fopor ChcoMic^vooli3r*«*********«*****«*****«****39

Vll* Ulcrooiotoc Spoecroo«o|»y«******«**********«****Al

m TSSUU ABO »^£T^ODS,****.• • • • • « • • • • • * . * * * * * * * « * * * * * * « * *a3

1* l loCor ioU** .*** »** » «* . .* * «******** «** «********* « ,43

Zl* ............ ••••••* «* »***************«*****46A* Croooint****••••••••«••«»••••#•••••••*•*••**44

r i i i

§• •*•••••••••••••••47€• Colloecion of

ftlsyLYS AMO

1* F&old

B* Soodllfin

IX^ rigiMMirA» G>lTe— %j6— 9 9*9 9»999 9 9 999»9999»99m99t99mm99995% n. ^nlfem0m999»m9999999999999*99999m»99m99999\lt

OOHCUUSiOliS ANO

FiHtCMHiC l»c«rroUitiogiAhi|Ni*«•••••••••••••••••••• ••••142

atrSftSBCEB CVnm9999999. ......... ....................

T«bltt tm flmme mMrnrnltUmtlm o f ttoo fmwArnmlttm— mmmmmmmmrnmmmm,* * * * * * * , A3

Tobto tm ^moriptlois o f l^tmrlolo & »«i ta «lioFf>«i>or«€loa of Chromicotroi^a Anolysod for tho l^rooonoo o f HionoHe C lreoo idoo ,*.**.******* ,*** !!

Toblo 3« S f«o «l»oo mM Color Eoootleoo o f Sooto C;Oo(wryod on DiroROtotrtoH S» FittHro

toOlo 4« E f*y«lo«o ond Color Hooetlooo o f Sfotoi'OOOrrod on Chroootoftroptl Flritro tmmrnmmmmmmmmt

ToOlo S« Xf»ir«l«too ond Color Xooct&ono of Sporo ciMiorood on ChronociHSVo £>« fls i^ o

Toblo f* af*v«luoo «od Color HoooCloac of ^poCooiNionrod OR ChroiMeotrftfdi S, Fin««ro •••«,§§

toblo 7m OIMI Coll^ MoiOtllKtlO o f SpocoytMimnrod on Cferomitosropt) P, FItiaro

toblo ft« Uimwrnimim mm4 Color lioR^etiooo o f Stem Cbomood an ChrooetfiNiroidi C» rinaro

Toblo 9» Xf<»o«liMM ood Color lUMetiona of Spots Obeorved on ChroMMtci^rotbo H»l tfemoniiPliuroo 7 - I 2 , * . * * * , * * , ,83

Toblo I t , Ef«mlwwi ond Color loortiwBWi o f Spoto Cbootrvod on Chroeoti^ropho I » l l»6 ,Fttaroo I 3 « l t , , , „ , .......

Toblo I t , a f»m ti«io ond C^lor nmmttm* o f Anmoofonla Spot* Oboorrod on tmpmt Chromic<»8rotli» in Tbroo Solvmt Syotoaw, mw4 ^ f'rm onm tloo CfMBtorotim 8f*v«looft omf Color iSooctlotui,,,,,,,#?

loblo 12, Amroio ^ • o « l «o « and Color aoaottmia o f ttio re jo r Yollow Spoto cboorvod vn wtio«>4ifMNi«ioiuitl and 'Cbo«dlflMHi»lofMit Fapor C^uromti^iNi]^ l>mtopod Uaiuft SAi< CFirot ^ Im n il and Soeond Solmnt Aa l f u S io a t « d » , , » , , , , , . , , » , , , « , « ,| t S t

Tablo 13* DaacriptLon of ^ocorlals Uaod in thoIrotaration o f Cfcr«m*tc^ra{^« Anolrsod fo r tho ^roaonco o f A r l y o o n o a , , , | | A

Tabla Color ianttiona of Sp&t9cbaorvad on C^troMitosmph J » Flftwra

tmhlm |5« «fid Color loaoiri^o of Spotacbaarvad on ChroaMitotrafdi f lia ra SC*****.*,!!#

Tabla 1^, Ifwintliiaa and Color Boaeciona of Sfoca I'^aorrod on Chro«Mitoara|>h L and on Ro«»chroiMitograi!ili of AS| froo O^roMtoiroflk L , fitu ro

Xablo t f , lf#^altioa and O »lor Moaotiona of Spota Cbaorvod on Chroitato«rapli flsoro

'--I'tabto l i « lif«oolitoa and Color nyMotiona of Sooca

<tHi«rvod on C^roawtotrapti S , flftiro

TabU I9m af^oaliMMi and Color ioaotlima of ®peta ubaorrad on Chronototraipli F lfw o

tablo 20« ftfooolttoa and Color Haweriena of SpotoCbaowod cn Chrooatc^rafdi fito ro 29«ft«.«»«*t27

Toblo 21« Aoorat# af«o»ltioa and Color to o o ti^ a of tiio Yollov Spota obaarrod on Cno*d iitans Iona I rapar ChrooMCOira^ Dovoloood Ualng ro ro o «a l«* «««* * ««« « ,« «* * * «««« « « «* «« ,« » ,.* ,,« « « i2 3

Ibblo 22* Aootoiio M*o«luw and < ^ l«r loartiooa of tito Yollov Sf»o€ (?baorvod cm Cno^dioonalonol faf»or Cbrofsatoirapha Oairalopod UaicHI

Toblo 23, &f»v*loo and Color ^Motioita of Irobablo tialvidln Spmt Cbaorvad on Ono«dinanaiotiol fapor C^xtnHit««ropb Dovoloftod tIaloK foroataly and Coatporaclvo Rf«rtr«liHi «nd Color HoactlORa of Halvtdta*****,*****

lablo 2d# i .'wwogo sf»oiil«o and Color offrobablo e ila fii: Aeld Spot Cba«:ivod on Qoo* dliMmaienal lapor Chronatoirapba Dovolofod Using foroatal, mcd Gmtpasativo If ^ ^ lt io and Color Soactlona of Sllogio Arid**«tt«»«»»#«*«»«l32

labia 25* Color Maootiona« on l^por« of ClaaoMof Natural Flavcwtotd Ci^pwiftda, tbonolie Aoida» and S lU g le Aeid C5* It^ Soibol in 2C,Salbol in

X

Tobl# «Bd Color looetiono of l^laorSpots oboorood on lVo*di»ooolofi«l Fopor Chrooatogcooho Dovolopod BA (F lro tS«lv«nt> oow tf% M r ond dOlWAc (Soeottd

o i

, mernmt^ im c u #@sip, Soloont)^ oo lodiootod*«««*tfl

k U t Qf f tm M &

1*

Figttro t,

figttro 3*

Fitter* 4,

Fitrar* $•

Fittar* $m

Fiiur* 7.

Fitur* i »

Fitur* f«

a»ntt«l!*pr*pfe i t Shoriim R«|iitir«iFwiitlon* « f eit* Spot* m

Gtifit a s v , * * , • • • t i

Cl ismwmm ^oitifiii to ljittr* io tit io fi* o f th* Spot* Ob**rv«d c*i On** dt*«n*lon«l t*p*r O it0 ii* t* ir* ^ a*r*l*®*d Usint ••••••.••«••••«,••••••it

a »r«ii* to tr* ^ ©I rnmmlmm 1*1*t lr *Fooition* of tho Spot* f^o rvo d on too* di*on«i«Ni*l Fopor (%reeMtotr*ph iovoiopod 0*iai «A« (r ir o t So tro u t) mndl | S 1 ^ «(iotOiitS

11 Sboring i« t * t lr ofooieliMUi of tfto Spot* i:%*orr«Nil on ttw* d loon* ion* t Fopor Chronotosmph ©ovolopod U*lnt 8AV ( r i r * t Sotvont) « imS

io lr^ e )* * * •••••««••#••«•••••••«••« »#7

mrrnrntmmph f* Shorint 1*1*t ir ofooition* of tho Spot* OiNiorvod on Ono* dit*n*lon«l fopor ©«rotop«dL * if it ............................. ...........................................................................

Chromitott»ph C» Ctogtiim m m tim lo U t i r o Fovitlon* of th* Spot* Ob«otv*d on Too* dioonoioool Fapor Chro««tott**h DovolOfod Using SAW (F irs t Solront) and l« » C I (SooiSMil

C h ro in to tro^ 8*1# niMmtam aiooiog io to tlro Fesltion* of tho Spot* Cboorrod on Tim»* dia«n*i<m«l Fspor ChroootiHirai^ ©orolofMidi Uoing SAO (F ir s t Soirtm t) and i ^ l F io t r «n t )» 7 i

Oirootttogrspii H*ti ©lograsB S h ^ie g lo t* tiro Fositiioi* of tho Sr^t* Cboorrod on Two* ditt«a*iocuil F*por Ct»r®n*togr«^ l^rolopod y*lnt SAW (F ir s t aotrogit) and m m if (SotoeiS Solront)***,,**,••••,•,•••••••••,••••««77

Cferoostotrsfh ii*3f Sfeootni HolotiiwiFositiono * f ch* Spoco uiMiorrod on tOo* distmsicMMil Fspor C h »^* t«^r«p h INrrotopod Usinir S4W (r i r o t S o lrm t) «tii #a%lF CSoeond ^ i v o o t i , ••••••••••••••••*,•• •••••«•« ►78

I t i i i

Figttro 16#

rtgnro It#

r i i m t2<

Fignro ti#

F i t m 14,

Fiiwro IS#

Figwro 14#

F i g m IF#

CliroMfttogrittNb i « 4 i nim^emm iliovtog lotattvo Fogltioiig of tlio Spot* Oboonrod e» t^to* dlKNtfioiflool Fopoc Chromtogroidi Dovolopod Uotf^ B4W (F lro t Sotoofit) and «6%tF Cioooaid SoiVMit)#•••••#•»•••••••••••••#••#«««#7S

Cbteoata^topii tWSi 6 ta ita » Slioittei Rotativo Fo «iti«a a of tbo Spota Obaarvad oo two* dimaiial««Mil Fap«r CbriMMtoirapli 6««olof»od 6a li^ BAW C rira f Solvont) a i^ d t f lf CSaoood SolvoRt>#*#####«*#*#«»#«#####«#»«#####iR)

CNsoRiat^-tapb 8«4t 6la«ta«i Siiowls^ Bolativa Foaitlana of tbo SfMita ^ baaHraod on two* dlawiiaioa»i Fapor Chronatograpb IHraolopod Uatag B4W (F ira t Solwoot) and «6%1FCSaeof^ Ibatvont)*####**#*#####################!!

Onronatogi»fdi i* l i Ctagran ^lowtiig Rolatiwo Foaitiona of tbo Spota Obaoraad an tWe* dlnanaiaiwit Fapor Cbrowati^ffapb Sovotopod U a li« MM (F ira t Solvont) and dOBHAclioaond ioli^t)######«######«###*#«#»#«###«««Si

c^O M Oti^^pb l* t i Plagtaw ibewiy^ l^ la tlw o Foaitiona of th# Spota obaatwod on Tmm dlflManaiotial fmpmt Gbronotognpb Povolopad Uatng 84ir CFlrat Solwont) and dOSmoCioeond iotoont)#####*####*###••##•##•««##•###87

Cbraawtagrapb 1*3 • Plngmn showing Rolativo Foaitiona of tbo Stwta Oimorwod on tWo* dlnonaional Fapor Cbrenotogtopli Dowolopod 6alag B4M ( F i ^ t SoWant) and 4611^(ioofliid Solv«nt>###«###»####»######»######,»##fS

aiKOfsatogtapb l«4 i Slagtao' Sbewli^i Solatiwa Faaitiana of tbo Spota otNMoraod on tWo* dinonaional Fopor Cbaronatogtapb Bawolepod Ualfig U.M (F ira t Solvont) and dOltme (Saao«^ Solvant}##««###«####«*##«######«#«,,«*Bf

Clurwaatoipafii l«»9i &Sagr«« ^^Nooia ikalatiow Foaitiona af tha Spata Cbaorwad on tWo* dinonaioiial Fapor aitonatograpb Dowolopod Uaing UM (F ira t Solvant) and dO UbMr CSaeand SolvontI#«••••*«•#•••*•••»••••#••###««f0

Flgur* li*

Figuro

Figwro 20.

Figoro 2U

Figwr* 22*

Flgiaro 23.

Figur* 24,

figor* 25,

Kiv

^Chrc«ttt«4preph i*4i &iagriis StKMtiag Rolativ* Foalclona o f tho Spoto Oboortod on two- dlmwioiofMil fop«ar Chroootcgtor^ Doool4 p>«a Uoing BAW (f lr o t Selirmit) ond i^m m A Solwont) . . , ••••91

CliiinMNitogcopli J I 0logr«o ShGmia$ Holotim* Fooitiono of tho Spoto Qboorvod tm &no* aiAMmolonol Fapor Chroooitogropb &owolopod Doing F m o t o l . ,* .* * . .* , ..........,116

Cbronotogtoph «• Dlogmn Showing golotiwo Foflltlono of tho S|>ot« Obaorvod on c«io- dinonaiOiial Fa|>or CShroaNicograi DovolopiMS Uaing S A W ,, , , , , .•••••••••••••••••,,,•••••••••lid

(%ronatogr*{^ Li Diagraoi Shewing Rolatlva Foaitiona of tho Spot# OlwHirvod on Cno* dlnonaional Fapor ChroR^togtmph DovolojMid Doing Foraatali Ao-ehraoMitegraiN^r wf Spec Ag.^ Chnmatografh Li Diagioe Stowing

Aalatlvo foaitiona of tho Speta Obaorvad OR Ono-dlnonaienal fapor Qiirmuit<B«rapb Dovolopod Daing

Chroawitograph Mt &iagran Shewiag holatlco Foaitiona of tho Spota ohoorvod on mpm* dUumaional Fapor Chrnnat<^rtof4) IWnralopad Uaing Feroatat*******••••••••••••••••••••••••122

ChrooMitogcaph Si Diagran Showing ftotativn foaitiona of tho Spota cbaaroad on Mao- dlnonaional Fapar Chronatogra^ Dovolepod Doing Foroatal****•••*•••••••••••••••••••*•••122

<%ro«atoirai^ Ft D iagii^ Shcnring Sotatioo Foaitiona of tho Spota Obaorvod mi cwto* dinonaiMMii fapor ChrMBategi«0$ Dovoli^od Uaing tAV* «* «.•«••••••••••,••••••••••••••••••126

Chroaatograph Qi Olagran Moving Rolativo foaitiona of tho Spota Chaorood on 0»o« dlnanaicnMil fapor Chronatograph Dovolopad Uaing BAV«*.****»*,••••••••..••••••••••••••••12A

utaooucTit]*

Scndtot on flonHMP color Intiorlraniso hovo tnum in pftn»

tro*» for imnf poor** Sotooon, « t tho tom of tho continryi

invootisotod tho hohovlor of th# Ifthorltonco of flooor

color (27><, In hi®#s«rl#to to trofeo tut®

tho chontcol loolo fo r €!©««• r it h

tntor«9C hoint dorolopod In tho rototlooohlro hotooon tho

gonotiet and cho»t4Bol otmsctiiro of ftowor rioa»oiito (f 7 )»

Such otodioo hovo groot imloo in olueidoting P«o»o oetion Of¥S

latoroction* Struetnrol voriottono in nignsonto opncti d iff*

oront oUtnto oro tho roonlts of WKprooolcn of to rtie u lo r

g«ooS| op«t»ttng thr^^igli «ni^r»® roe tor* (3 l> »

Conoo ooro dooSgfMitod for c o l«r iid»«rit«neo hoforo tho

ehooteol of tlo n t flooor plissofito wmm

otodiod in ro U tio n to flmme color inhoritooco* Vpom choo»

Ico l ido ntlflcotlon of plioooto rooronsthlo fmt flowor color*

gonotte and chaoieal tntaroctiono ooro i i ^ o d oot im toroo

of apoolflc roiio action in tho prodiiction and oodificatioe

of chooicol atroetmfoa* Inoolodio of tho <mo».io«*y and nodo

of inhoritanco of flooor pigoonto to f«nda?«ontal to tho

olocidotion of tp o cific gono action in pigoant fsrodoction*

tha mnrpooo of th in thtnln oaa to gathor inforootton coo^

coming tho chaoU try and gonotlea of flewor eigoant® in

iimmmoaiMS: g, in hom* of toyiiii grooiidoorlE

for ftirthor atodloo of apoelfic gona action Inrolood In p i f

oont pro octian In thla apacioa* To f o l f i l t thia potroao, d

fmar oHjoetlvao of tho rooooreh war# pmposodi

I ) to Idoatify rH « i» i« » lly ttio yatlooF pttHNwitOf

3) to otoiy th « t^wmstte bohoviot of tho tollow

pinsaiitoi

3) to otody tho iotorrolotloitohipo botwooa tho

Sioiotleo and tho ehoolotry of tho yollew pig*

eootot Olid

to oltioldoto tho intorrolotiooiihlpo of tho

yolleo pigoooto ond tho onthocyooino In tho

prodftetlon of fhoootypie eoloo *mriotio«i*

of tho l^f<»flMitlen oootloblo todoy on tho gonotleo

ond broodlnt bohovlor of DoagsodltMB lo eontolnod In publlto*

tiono ond thoooo proporod ot tho Untooroity of flowoti l^ p o rt-

oofit of Agrooooy ond S oil Selimeo* Om otody roooomod

ehrooMQoooo eoonto of oooo PoorodtuF opoeloo <5?)i in

ssadxiceods# 2n • 33* thto t»«o boon eonfinRod threofh

otodloo by tho prooont oiathor* Chow ( n ) Invootiiotod tho

brooding ond intorbrooding bohorlcnr of tho throo opoetoo

la S O TM ( H i l l . ) iirb ,, im c,iagt» <Jo«o*> ®c*t ond

»• ooodvlconoo. fori. (43) ond forh Oftd Sotor (43, 44)

otudlod p rlB o rily tho inhorltoneo of pwrflo flowor rotor In

£« liadMlegaag- fork ototod thot fwrthor atodloo olong tho

lin o of bio vorh oro noeoooory to olweidoto tho notwro of

tho tnh o rtton ^ of difforont ohodoo and intonoltioo of

flowor eolor* 4o o fo llow o p to tho otudy In itU to d by forh*

thio thoolo Fooooreh propoood ro invootigoto tho inborltoneo

7

of tho rot tew flower plimonto i » sSSdSdtiBtiLft.

k* aandwlgffinoo wos ehooon for this orudr fo r oororal

roooonot « ) the spoelos is fo to s t ls llr ss s f m »

SfO crop In RswrI I i sim) , ronsooiMHiCly, stwdiss sro

noododt h ) mo siMKsiss had prowiocHitr boafi miod In s sini*

la r atwdy, prollaifiaffy roaolcs and data swsilsbts fer

purposso of conpsrison and supporti c ) k* •agtdtgigownn i#

Indotorrlnats In Its flo«MMrlnt, no ehst ftowom sro sssit*

Ohio s l l yoor lonp for both crosvlns snd snstyslsi

snd d> tho spmstos oshlbits s broad raitto of flower color

wsrlatient fro»e nosrly white to lig h t and darb parplo*

3

AClC.iNC^Le8QghmS

th* auchoi* wmld Itlco to o*|Hro*« hor trotttud* to

»<r« «olNwr ^oodo mm h r . Flofd tooiilo Oobtro for thotr o v«r»

roody ond bind osalatomro wttb eoro of tho ptonto* Hionlto

or# oloo dwo to h r. ^oont ««d Or* Gcmnm »ortbol«M8«v,

of tho OofsortBWfit of Atrofioagr ««d S o il Srioncog and to

©r. L# S# Sorgor* of tho D#|»rl&a«it of hlcroblolflia^^ for

v»m of mieh«fioodod laboratory apaeo and f a e llit io a , mm to

Or* «J* A . Himt» of ttio ©opsrtrsojit of Conotlra* for adrlco*

Tho author la «af>oeiallr aratoful to ©r, fwrnrnU >terttji» of

tho Oopartesont of Rortlrulttiro* for hi* pati«Kit« t«w»or®«*f

and Inaaluahlo aaalataneo.

RBVlSIf OF LlteitAtURC

Tha ffiiRttiitrr tit ilm m f 9ftlor

O|i«l0v, •• Mtrly «• 1414 otiadiod ^OMleol otmetoroo

« f plant piiipsonto in hlo Invaotlgaviovui o f anthocFonlno in

tho tordon onopdrogon <42). Stneo than* tho «^ «o ioo l «true»

turog of flooor plgoonto hooo boon invootlgotod ly fMrny

oorkoro (20, Povio* in 23, Oo«N!Nrift in 23, Soain in 23, Voodoo

in 23, 25, 22, 39, 41).

Tho n o jo rity of flowor pigiMMiro bolong to ono of two

grotipo, tho flovonoido (oop«>oolttblo plgfMRnto) or tho eorot-

oneido (plo«tid*bom o, M p-inooloblo plgwonto).

A . Tho ruironBitfi

Alooot o i l tho b r ilt io n t ohodoo of rod ond bltso

flowor eoloro oro duo to tho pro«oneo of ono or noro of tho

clooo of flovem»ido known oo tho ontheeyonlno. Othor clooooo

of flovonoid eooipotafido oro rooponolblo for yoltow eoteri

howowor, tho eorotonoido ttouolly oro tho eoitoo of yottow

piipooototion. S t i l l otbor flimm oido oeeount for tho oetuol

whitonooo of ooot idiito flowomt without thM o flowonoido,

tho floworo would porhopo oppoor oloeot tronolooont.

tion of flowonold and rolotod ?4ionolie coopoundo or tho

oceurroneo of thotr ootol rholotoo con load to ^ o produe*

tioci of oooo of tho brown ond btork pigwonto found In

Soturo (Swain In 23).

Golooewin ond Himrolnor f lr o t uood tho tanm flovonoid,

in 1952, to dotrribo o i l thoao OfMspoundo whooo atrurturo la

bated on chat ef flaweea leh ) (Swain in 23>i

11

5 10

PtnveiM CSntln in 73)

Pla ieKMi emiaitta ef ewe ^maana rlngat h and S, |eiiiad

eegathar by naaoa ef a tbraa«carben liirir feaiaad Inee a ^ •

pymna ring* Iba anldaeien atata ef tliia 3«>C HjOk dataminaa

tha elaaa to Which a |Nirtlc»tlar fiawoneid bolonga# tha min*

bar of eerwctwoa aeettally fotn^ in Raturo la tin iea d, and

thaaa a rt praaantad in tabular fom* ahowUig tho atm eturo

of tho 3«C jMMreion, hf ^m ln (ia iilii In 33)* tho dlhydtro*

mateonoa aiWi eha ehaleonoa, not poaaoaatim tho baaie f ta -

wono firolotofi, am inetadad aa flawonoida banama thay am

ehaniealty m d b io ayathatim lly etoaaty raiatad to tho echor

typoa of flawonolda (Swain in 33)*

Vlthifi m th etaaa, tho indiwidaat eonpeiinda am dlatln*

fuiahad p r im r i ly by m# mm^nm and o ri«nta tio ii of hydroiryl

ond m thoxyl gronya anbatitutod In tha two bonaono rinta*

In tha during of tho n a jo rity of tho ftawenoido, hydroryl

groupa a rt found aithar at tha Gy* only or at tha Gfm and

Cj^poalciofiat ganom lty, am unawthylatod, tho S*riti«

i® aulMititutod hy or®* two* or thmm hydroa^l or

fNitherrl groop® (Swiln In ?3>«

Piovofioid ctMvpoundt uoiMitly imi®t is tlo n t «• gijN*

eo®td«®i tti«t i®, ono or wmm of t^ n ir hydroayl groans® 1®

JoiniMS by • ®«Ksi«ocot®l iinfe to ® »o »«r« Aglfcm o i® tho

to r« o*®d for tho »tii«r*froo flovonoid®* tho priMiofico of

«o®ort in tho ooloooto eOnfor® ®«|>>ttoltthitity to th# flovo*

oold eoopoufido* ohi«^ or# fwoiwoliy inoolohlo* I » «?o e««o

of anthoeyoalfi®* tho ®tM?or® loport o to h lllty to tho «gly*

CORO in t$>«

tho s\^«r®* aliifty® Joiiiad at th o lr C|>po®tticHi* mnaally

hy m /3 olinltago* Intludo a if^ lo horo®#® and pontoaoi' if

(etonosldo®)* and di* a ^ tri^taoeharldo® Cbiooldo® ami trio *

•ido®)« I f two aogar® aro atta#^«d a t d if f « r « i» t f4ioiM»lie

hj^rojtyl®* ^ o ^ M u lt i® a di«oiio®id®* oloiio ©r

in a diaaeeharldo* i t ^ o » m t cmmm ot^ar* follcRNMl is

froouoney of ocevemMieo by 4<i>golaetoao atid Wrhamooo*

i«Mtrabtf>o»o* Ooglueuronlc acid* at«d 0«o^looo* ftio la tto r

throa baltif cothor rara« In tha anthooyanlna* Umrablnoao

haa ham found* in thro® plant §««#««* with tho mmmnX

0^*iitdEf iiylo»o ha® hmm found m ly in bioaldo®* ^ l y a

fan of tho p m tib lo dlooccharidos havo baon foinwt* and tha

trla a e «^ rid a a aro caro* Anthonyanidina altiay® ocevar In

flowora with at loaat ona atugiar attachad to tha 3*hydrmyl

groap (Horbomo in 73>* Atta^^aimt t® ttio $• and 7*poaltion®*

inataad of tha nor® u«tial 3* and S*foaitl«Mi* ia rat®

7

(llarhoriMi in ?3)« i « «h« im lf autar fnwnd llirt?ad

w il^ €h« Ciym and Cr*hydroayi grotipa of anthoer«ni»i> i^mtm

in 23)*

Xn tho flawonoa and flavonolat l-«ff«t»iiieoo haa Hmm

found la hotto atwl /^-lla lta to t© tho ^•^rdrofli^l $m m

Otiarcatln* Thoro aro nuworoua dlnonoaldoa af»3 di* and t r i -

aaecharldaa* In tlio flawonoa^ tho a ^ a r ia llfihod noat

oftan at tho C ^pea ttio ni othor paoitiena ato atni

Zn tho flavonola, tho iioat eemon i ^ i t i t i t io foltowod

by Cj and C^i« IWwt planta cm m tn mmm than ono ilreoaido

of any aglycomi (Sonin in 23>*

tho ehaieonoa atoS tho ainronoa> iluofino ia itho only

ougat found* IDho ehaloono giyeoaidoa uanalty c^oecitur with

tho eorrMtrondint gitteoaido of ftainifionoa and auronoa (iwaiii

in 23).

Aeeotding to hellcny CS9)f plant glyeoaiitoa in gimoral

■say oontaln, in addition to tho ahooo lWrlhc»o*

L-galaetoao* &« oumH»a«|| P-fwooaoy P-frtn^toaot ond j&M»mhin©aOf

tho la tto r onm ocewrelny to ro ly In Kafuro*

Iho c y c lic fmrn of tho ounara i t ionosfally rytnnoao ( 39)*

although i-arablnoao haa boon found to occur in both furano*

aldo and pyranoaido forau (Swain in 23>*

Acylatnd tlycoaldoa, in idilch a clnsianic acid r o a i ^

ia attachad to tho ougoct aro alt© linowst. In iftio Mithocyaw

nina* tho only acyl grmipo f ^ i d a© fa r aro j|-co«w »rie acid*

caffoic acid , mixmpie a cid , and fo ru lic aCtdi In oach caao.

6

tha « « y l sroup attaohad to tha at^Mir lifibad at tha

hydroxyl group CSaaln In 23* Hbrboctta tn tS )« Aatotm ^ a

ftavonola* m il of tha ahova aeida axeapt attiapio oeertxr aa

a ry l groupa (warhoma in 2S>* Aeytatad anthooyanina hava

baan found fa r *ora fr»o»««® 3y than acylatad ftaronolai

four aeylatad flaronaa or flav^aola hava hmm

(Saain In 23)* Harbema (tiarboima In 2S? atatad* hovavac*

that tha aeylatad flavanota vmy yat ba found to cmmm'mm

fraguamtly*

tha flavonolda can ba rathar broadly etaaaad into thraa

groups I 1) tha anthoeyanin piim nta* intttiding tha antha*

cyanidina (aflyeonaa) and anthoeyaitina (gl3reeatdaa}|

2> tha fUevomm and flavocM»l ptprasita* ineludini th a ir glfN*

eoaldaat and 3) tha alftor ftavcmotda* taeludUii tha ehal*

eonaa, tha auronaa* tha flavaeonoa* tha taoflavonaa* tha

b iftavonyla, and tha taueoafitho<^anidlnf • tha antheeyanlna*

haaad eharleally on « ia atructtara of cyanldln C3* 5* 7 , 3*,

A*«*pantahydr<nsyflavyltua cation) shewn below* are raaponai*

bla for naarty a l l tha ptnh* am rlat* rad* natnra* vtolat* and

blue colors In patala and laavaa* tha soeond greaip* tha

flavonaa and flavonola* ara ao^^tinaa hnown aa tha antho*

xanthlna* althoufd^ Harbotfsa (27) atatad that thla tarn la not

auch In use today* they ara m&itm h ighly oridiitad than tha

anthocyanlna * aa i » shown in tha follewtfnn sertietwoai

9

10

Fliivoiia r ia v m it C 2 7 )

Alfbowils fb# ftavonM aiMl flavonol* mtm e o llo e tivo ly

tbo neat abtKRdant traup# pravldlxm hmtmum SKMI and 500 kiiaan

a«d»ataneaai tbay ara laaa inportaitt than ^ antbaayaaina

fraat cba fMiInc of v io v of color* Thwy do* boMvatr* aoilto a

atip ificani; concribi^lon aa yollow p l p ^ t a o r aa co*pl|panta

CO tbo antbocfmilaa It? )* (^plipaM itatiiiii la doflaod aa tb#

focnatlon of a eoa^laa aalocola liy In tbo

proaoR«i of OBiOiocyafilii pljpoiita* cbangli^ ^lo color of tbo

afitbocymnin to a hlwmt ahado C4?)*

fba ntoor flavenotda oaia a Ito ita d coiitritMitifm to

floifor eolor and aro* ao fa r aa la kooioi* of raacrtrtad dia*

tribiition* Choaiioally, tbo oinor ftavoooida aro aolatod to

oacb oibmr and to tbo ooior flavooolda* Harbtnroo Ct?) aboood

rolacionshlp* 4i«iniiBiNicle«l.tp* Th* m troup

of poltoo piip»*ot« pt»hmhlf fmem^ l« to t » oooliitiefuirF tiiNit

•ppoar frocroontlF ooljp in tho flooor* of th* Coopottt**^ on*

•f th* 0M»*t h ighly <l*volepod plant f«* ili* * C27)»

Oth*r phm olie pi||*»*nt* inotod* ids* it*fttlionMi *<mI tho

golnono** 1h* xiuithoiuMi mrm jm llov pigiNMit** Hjpdroaiy*

gtilnono* and •ntht**tili«mo« naho « m ^ lig ih lo toaf^ilaition

to hlghor pUint color (27)*

1©,» JksMiaauiAB

Stttdy of tho ch««ei*tf7 of tho carotonoid *uh*t*ne**«

rad sfid yotloo plgiMnit*^ startod im tho l« t* 182©**# tho

•troctoro* of tho noco roadily im iila h l* carotanolda ooro

dattHnelaod In tha following yaora* tiia carotMtoida aro fat*

aoluhlo plgaaeta* Host of th* carotanoida hara a dO^eirhon

ah*latt»4 4mA naat poaaoa* an IS^Msarhon eantral «m it con*

peaod of aomn conjygatod deuhlo iMmMa and foiar aldoMdtaln

oothyl grenr* (Vaodon in 2 3 )i

I I

1 - 1

tha too and grotnNi attaehod to tho control n n it nay

althor bo tho cmro (lyeopono* /^•caretonot aed aaasanthtn)

or diffMrant C o(.carotan#t lotain» and achtaanona)* ioMi

earotanoida foaaaaa "ratre** pi»ty«na airataoMi, in which tha

l>oaitl(aia of tho alagla and dotibla bonda in tha ftolyaoa

chain ara ravoraad froa tha ’Hioraal* arat^wi afieaas abw a.

Carocaaoida ara known which poaaaaa fowar ! ^ n 40 carbon

•tona io th a lr carbon akalotonaf chaaa ara poaaibly natural

dagrad«ciofi products of war lout C^Q»earet«noid« (Voadon In

23)* % * chroaoi^oric, or eol®r»|w«dtect»i, group of a earot*

anoid* Which nay b® ra d , oranga, m- yollow , conaiata of th#

aariaa of conjugatod otafln ie doubt# bonda c o rp ria ii^ tha

canttwl u n it (Sawiaa in fS)«

Carotanoida can ba dlwidad into two naiji graura*

I ) i^ r® t«n «i, id i l^ ara hydrocarb^ai and f ) iia n th o ^ y ll# ,

Which ara ©*yg#n-co«tainlng dariwatlwaa Cl^wiaa in 23)*

Caret«toida in flowara a ra , c h a ra e ta ria tio iliy , l ) h i ^ t y

oridisad xanthophylla, aofN^ially S,i»«^50iiid«a <aur©«?

nanthin, flawonanthin)i 2 ) apaclaa-afiacific pigaanta

<M^-acholtranthlM» in SteBto&iA>l ard

3) carotanaa, occaatonally In coi^niriiiN B ly larga awotmta.

Fraetieally a l l yallew and la »(i«.y«ll® w flowara contain *

larga anounta of xanthofhytla and only tracaa of earotonWi

daap oranga flowara of tan roai^aaa larga at^oueita &t

carotana, at In tha d##p oranga frlngaa of {^rglaatia B ^ialia

Curt* and Mtailu® gugginia ^ a l . Or lycopana* aa in Caiondiila

o ffig ln a lia I* Colorlaat potyanaa ara alao oftao found* In

flowor rota la , ^rotoooida ara conc^cntratod in tho ^r^g o *

plaata, ^ o fo «hoy ara ptolwibly attachod to yrotoina

12

IS

(Ooodvifi In 23)#

Junta IvBla and Cara (Goodvin in tS i e^Mtaascad a tber*

ough atudy of tbo d latribotion of carotmoida In diffarant

flooor part* of Oniianin rnata Rafin*» Hiolff roaotta* pro*

»antod in tabular fora by Goodwin CGoodarln In 23)* doren*

ttra ta tha e<Mepl«iclty of tho plgiMMat eoopoaltioii of flooora*

PiNMity*«igbt dtfforont e«rot«wtoida aro llatad* tototh*r with

fivo onidinntlfiod onoa* utidor tbo tbroa yonotol boodtngaf

I ) earotano bydroearbonai t ) «B«wobydroigr xanthofAiylla and

aponidaa of carotono hydrocarbona afid of oonebydrosqr nan tho*

p t^ lla i and 3) dl* and poly*bydreay nantbopbylla and th a lr

apooldoa*

Ibieh oodr oith carctanoida oaa done by Coopor and

E l l io t t (12* 13) af«i Bu^or aitd Gavia Cl6)* la rlim r oocit waa

dona by tWnsloy (36)* but Coop«r and l l l l o t t ( I f ) dlaatrM d

v ith hia idantifleatloa of tbo carotonoid pigrtonta pr oducing

yo llo v eolor In a lfa lfa (Fadie*ao aativa L*) aa ea rotan^i

they idantifiod aaid pigttianta aa aanthophytl aatara*

Faach (41)* in a atudy of flouar color dovolo^itofit*

roportod that both earotanotda and flavonoida occur in osoat

yolloo*flo«rorad apacloa* In flovHura In Wbicdi both «ntho*

nanlf^ina and earotanold plgr««ita aro rroaont* tho doop«r

yolloo eorotonoida ofton dotomlno tho flooor color*

&ca»ploa of t h U aro Viola trle o io r t » and ro ra y th U i in

thaaa planta* colcnr ia duo nalnly to tho plaatld carotonolda

proaont* although largo ouantltloa of tbo antbiman^lfi*

<|ti«re«ti(i, « r « atao yroaont*

14

. ua- ■-

ftg fiftlag

Coepounda ara eolorad tMcatiaa thay abaarb Itid^t in tha

wiaibta ragiMi of thm apaetsrun (400 to SOO i y x } « ifhan lig h t

ia adaorbad in thia v ia ib la ragion and in tba ulttaw ielaf

r ^ io n (ISO to 400 ) , alactrMia tn lha wPlmemlm ara

axeitad. Otiaiaualy, tha aera t i i ^ t l y bound in ^ a nolaonl*

tha alaetrona ara, tha iMnra tha anatgy %rill ba ahleh ia

naadad to aoceita than* thua, f«m w»vm tigbely hound

a la c t]^ t , lig b t anargy ia adaorbad at abortar wavalangtba*

thia ralatlonahlp batwaan color and nnaaturatlon (paaaaaaien

af doubia or t r iv ia bonda) ««ia f ir a t ro c i^ ia a d In ISdS

Oraaba and tiabMmann (Swain In f3 )«

Tba tarn ebronoplunra %iaa apptiad, la 1888, to organie

groupa rreducing ealar whan praaant in a iM lacula, and tba

taro auoeoebrana tfaa apptlad to otbar groispa wbieb daapanad

atieh colors (Swain la 23>» L it t la waa undaratood about tbo

eolation of color prodtietion to tboao groupa « » t i l tbo o loc-

tronlc cboorloa of organie ebawiatry bad bo«n workod outi

today, i t ia agraod that tho ptxmmmi* of ccmjugation, chtoc*

oshoraa, and atmoehro^w a l l holp to atablliaa tbo roanttlng

oxcitod atruetwro, tharaby pronoting gro atir aaaa of alae-

tron tranaltion to highor anargy lavala* tha longar tba

eonjugatad chain, or tha groatar cha nunbor of chrotaephorao

and auxochro9«a, tbo oaalar aro tha aloctrMt tranaitioiMit

that ia , tha wawalanglb at which mtmerptlom and axcitation

occur ia longar (Swain In 23}« Swain (Swain im 23) axplalnad

15

in d a ta ll tha eonaacnaneaa of tha ahova axflafiationa to tha

flawonoldat Waadon la t 3) did tha aaea f m tha

earotaneida*

Iht-gflagjcici ,flg rigKtrnfialai;

4 . Aha fl8Y^8i,14t

Fcra i t t xwm aboot flavonoid Inharitatiea than

aixittt tha lhh«ritanea of any othar plant eimatlttjanta (27)«

GanatLe atudlaa hara ahetm i ^ t aany atapa in bieaynthaalt

of flaronotda ara eloaaly control lad by a lug la , Indapandant

ganaa, idileh produea highly «ab*teata*apaelflc antypaa*

taeant atodlaa of tha tidi«rltanca of flarcmolda hava poatt»*

latad diacrata atapa in flavonold bloaynfhaaia, aloim with

th* typa of a fo e lfic itio a whtdi tho ontytNNi night }>oatoaa

<77).

Flowar plgaant ganotlea ha* baan atudlad in about tOO

diffarant apaclaai of thaoo. 14 hava baan atudlad to y ia ld

tatiafactory infaraation on tho gonatlca and ehaaiiatry of

floaor color iaghoritaneo. flarbomo <27) praaontod a tabla

af thaaa apaciaa, and indicatad ahi«fi goiMM hava baan daaig*

natad aa controlling aynthaala or ateuet«irat aM>dlflcation of

flavonoidc. Ho pointad out that ovary plant haa a imlcu*

•yataai of gono Intaraetloni conaaouafttly. ganaa apfsarantly

controlling tho #*»a atop In th* bloaynthaaia of flavonoida

aor# glvon d lffarant aynbola In difforant apocioa*

Gonaa involvad In flavonold aynthoola ttmy ba divldad

Into thraa oat«g;oriaai I ) eimtrol of tanianRl frodauttioni

2) nodiflcatlofi of chanl^il atrtaeturoi and 3) d istrib u tion

of pignontation. Flavonold production and aodlfication ara

eontrollad by Indapai^ant gana action <27)i pattai^ing la

17

IS

controlled by « aerie* of etoaaly linhad aana* (3 4 ),

In neat caaa* ef single gene control ef flower eet<»r,

eha celorad flewara are yrodocad lay a denlnant ranei mtar*

rlaa are Kedlesae aativa (1 3 ), ia>S&iiaiiS iS m m ^

v ie ie oAtiva t* (1 6 ). Xaraly, tha white or nm»*colered

flever ia gevemad by tha deninant gana, aa in lanatiana

bataarilnA t , C|S) and red Cleumf (43)* In casaa In uhich

two or wore pair* of genea are involved, in t « r e l l « i ie in te r­

action takaa ylaca, reaultix^ to «M»dlfi«d ttmm ef *iag»le

Hendellan rule** SeiMi eatairylae are a* followat i ) reryle*

wentary gane action, aa in aoyhaana (3 0 ), flan (1 ) , and

sef*.rt€>cart^ua apaciaa (34) f t ) reeaaaive •yiam aia, aa to

sianthuR c«rY8ctiyIlm, U (4 f ) i 3 ) denliiant epiataaia, aa

to LgtUfgug adfiggttil (7 ) i and 4 ) dwpUm te getiaa, aa In

nutanc eyanldin-red flower of Trifo ltu B atSSSA (•)•

I t ho# baon auggeatad that a block to aynttioaia of

anthecyanto, aa to •oaso vh lto gwtanta which eentato tracm

ef pignant, aenotinaa tovelvoe on U d iib ite r rather than tho

loaa of an oaaantlal «i* y»# (27)* In aum maoa, tha con-

eantration of pignont vartoa to tho nunhor of pignont In­

h ib ito r gonoa proaont, to the ra tio of I i2 i4 (%rbocito to 20)*

figsMmt aynthoal* ia alee daeraasad aa a conaocnanea of

hotorogygoaity a* conparad to hommygoatty of tho deninant

fore (Horboma to 20). Althewgh tho accta^latton of d o « l-

nent pignont genoa w il l Increaaa thm overall arotant of

ftavonoid j«roaont, i t ha* bem fotmd that mm pigiNmt i*

fomad 1» bulk at tha axpanaa of anotrbar, at la dahlia (28)*

Utually a lln lta d aaount of praeuraor la avalla b la , and

diffarant pathwaya eocnpata far thla comaofi pcaetsraor* Aa

axaapla la ^tlC TfiiaU E. mJlML U , ia atileb tfia gaaaa I

(control of prodoctiim of antboeyanldina and flavoaota) and

J (control .of aurona ayntbaaia) eoaq>ata, aa that plants with

a high anthocyaala and antbonanthln eoncantratlon hava a lav

anrona eoncantratlon and vlca varaa (Harborea in 20, Alaton

in 23), 4 a in ila r rolattonahip, botwaan anthocyanin and

fiavona, has boon obaarvad in a of pl«sta (ftarboma

in 20, 27), In c o n tn a t, moem plants hava a na^anlan of

Incraaalng tha eoncantratlon of ona flavonoid class v lth no

affaet on that of anothar* for Mtanrla, in rcl«Mla

Sabina, ax lindl* in tha iSt (•©axalwr**) typaa, tha prodne*

tion of palargonidin is t r ip lad «d«lla tha eoncantratlon of

ralotad conpounda raanina constant eoaiparad to tha donbla

raeaaaiva (Harboma In 20, 27),

Lawranca conciudad, <m tha basis of axhaustiva studiaa

of pignant aynthaaia in M ilX A ©ntf • ««d

H,ttiPii8gir8Ulf tbat tha praaanca of doninant ganaa atisatUtaa

ganaral flaycmold production (Barbortia in 20, 27, 3A>, ^

poatulatod tha axiatanca of a **faadbacfc*’ i^achanian, vharain

thm daesand for a coaaMMi pracuraor stimilataa its Incraaaad

production, on tha basis of an incraaaad daaand fron an

anlargad nunbar af pignont panaa,

Hutationa at tha ganatlc lo c i eontrolllnit antbaeyanldin

19

hydroxyUitlcm «ra r«®ponslbl« for ooat ftowor color voria*

tlona in eoltivatod omatmntal plantt* tm gonwral* a wild

typo plant apaciaa with trouvo or blno flomnra givoa rlao in

cu itlva tie n to naganta, aearlat* pinB* and orango foma C27),

f alfiylation haa narar haan fman! to occt^ in flawenaa

and flavoRolt found with paanldin* natueidln* and fealvidlni

mmthylntion ia vauatly raatrietad to tha anthoeyaiiifis in tha

flowara of gardan planta <27). Svidanea for tim ganatic

eootrol of glyeoaylation la lin lta d to a faw y lm ta onlyt

glyeoaylation (attaaluEiant of avatara) and aeylation (attach*

mmt of a eitmaaie acid roeidua) haan found* ia tmxe

plant apaelaa* to ba aloaaly llidead <27). jterboma (27) want

into d a ta ll in a di«cua«l<m of d4»cljiant*rac«aaiwa ralation*

•hipa* diraction af autationa*^, and apacifie gana aation in

ragard to gm atie control of hydronylatim* nothylation* gly*

coaylatton* aiul aeylation.

Ganoa for tho c m tro l oa o x l^ t io n Chydraa^lation) In

tha b r in g of tha flawonold wolacula ara wall aatabliahad

(2 7 ). Saasiplaa ara carnation <M4aSteag. S2S£3£ti£i5$lZsi)• in

id i i^ tha 1 gana cantrola 3**hydro*ylatim of anthoeyanidin

and flawonol <27), and anapdragon* im tho U i«no ton*

trola 3*»hydroicylation of anthoeyanidin* flavonol* and

flawona <22* 27).

A factor haa boon ahown to ralao tha of th# colt ln»'

in tha flowara of tha awaat paa whan tha factcnr la in ita

raeaaaiwa farw <7, 41* 4 7 ). Ih ia ganatie factor* tanmd

20

•doll* (d>, charolgr prodtteas a bloar flawar color In Ito

roeMtiwo fone tboti it* dcwlniunt ollolo* l « is tmown trbot

nony of tho onthocyonlns bohowo •• Indieoeoro, tondtnfi to

tom bluo at altealiao and rod toidor acid ronditiono <t7)i

howovor, ehangoa in tbo pit of tho c o ll toy hovo boon dio*

coontod froo boing roorenalblo for color variation in floworo

having tho aaoo onthocy«nln« This o m tm d io tim vat batod

on tho foct « ia t tho pB of tho c o ll aap dooa not vary ovor

a largo onoogh rangoi tho variotton i t only about 0*S to

1*0 tmita ( i ^ i n in 23). I t should bo rao«nb«rod, houovar,

thot aa tho pH la ralaod to noor n o u tra llty , tvoro and i^ ro

of anthoeyanla plgisont la tranafecnod into tho colortoaa

p04Kido<»b««o form thoroforo, at p^ 3«5, tha nom al pH of

tha flowor, only a t iM ll part of tho anthoeyanln la pimtnmt

in Itc oxoniun, or cotorod, fom* Snail chongoo about this

pfi may havo a graatar affoct than «Rpoctad (Swain in 23),

In a atudy of darlnanco«rocoaalvonooo of aovMrol gano

octlcma, Scott-Honcrioff (53) » «d » tfio following t«R o m lii»i»

tlonai I ) anthotyonia and anthoxoniehln pr<^!uctton la

nanti 2) hydroiQrlotian aiWI oothylatioii ara diMBinontf 3)

3,9«dlglyeoRidic and aleylatod anthocyaiilna ara doninont to

3-n<moglycoaidtc and non^lcylatod anthoeyanina t and 4 )

unlfom plg«n«iitatlon lo doninant to flocking or narking*

Vorh^na was found, howovor, by Boalo £S iX* (A ) , to bo on

«Kcoptien to tho obovo gonoroliaotitma in rogard to hydixntyl*

otion and glycoaylation*

2\

6* Ib t

Thm lltfuratw o doaling with tho gonotloo of tho

oorotonoido io not « t oKtontlvo o« that daallnt flovo*

noida* Fario gjt &J,« (1 4 ) co9»piliw* flooot «?ol©r Ifhoirltanoo

charta of 75 diffaramt a^pacloai of thaao* <mlf t m » bad

gRmatlc factor# Idantlftod at affo etln i xanthOFhytl prodyc*

tlo n . Ona apaeiaa* .eagdjnalia 6ot^l* on i«mth«*

eontalnad a a in tla xanthophyli factor tdblOh t4^raj|«t4Kf In an

addltiva tnamiar* Hangatadorf and Frapa (37) fointd* |p a

atudy of yolloo «»doap«m In com* tliat coaaanthto m& coii*

tro llo d by a ainglo factor* X* ohtih aadiibitad m im l and

addltiva gono action*

Gottoratly* p ^ te ic tim of plaatid p lg iim t ia doaii*

nant ganotlcally ovor tha a ^ m c o of aald pl|^Mfi.t (41* S3)*

22

m n liittriM tiiiffn n iitfi itf IteittMCTiiii ma w tm i im m u

In mil r r g f l n f I f fX m m iJ M m

A. tetfe«yjaila.AJLfl£

Anthocyimtoft ara raspooaibla for thm rad and Mua

eolora of flw ara * lha prasanaa of 1N»tb anthocyiiiiina and '

earocanolda ray raault In «hadaa af cad, oranga, purpla, or

aaganta, or in wariagatad ftowafa < I2)* Carotanotda ara

uauatly taasooalbla fa r yaltow e o li^ i In yatloa fto«w r« in

which both carotanolda and flavonoida othar than tha antho-

eymninu ara praaant, tha oarotmiolda oftMi datarmina lb#

ftowar color* I t m f ba atatad, bowarar, ftiat tha fla w t^ ld a

offar tba graataat variation in colori thia ia rcMNwiH iabta

whact ana canaidara tha narrow rang# of atructtiral typaa

rapraaantad Iqr tha flavcneida (Swain in 23). M tfm tm t c a l-

orad flowara oftan contain tha aaaw anthocyanini ibaiNifora,

faetora othar than qtnm titativa and qttallcatlva affacta anaat

ba involvad (SiMifi In 23)* tha fo l lowing am ibaaa

nodifying affactai

1) tha praa<^c« of othar plgoanta, ma^ aa tha

carotanolda, tiaa hmm aantionM! abova*

2) So-pltaantatlM i w ii^ f la i^ a a , ftawom ta, a ^

flavana changaa cater to a bluar ahada* Co-pigaatitation ia

widaapraad, and ia ^ a only fcntnai nachanian to axplaln blua-

ing in flowara with aathylatad B-ring coepoonda, aa thaaa

caapotmda (a* §*, m lv id ln , pannidiii) casmot CM8p l« i with

m tala* L it t la ia hnown about tha choniatry of co*

23

pigffientaticm. Isut I t ia daflaai a« tfi# fa iw itii^ i ®f a r«s^

plmt t»atwa«n an anf^ocyanln and ftavonwi or ratatad aubatan*

eaa CHatbama In 23).

3) Matal €tialati©« (wttal-anifencfanin

wodiflaa •ntboeyatiifi color by catttlnf a blnaltif o ffoct* fhia

raculraa an stf^AihyArmty gronplfif C^rboma la 23. ©ipiiii In

23).

4 ) tho praaonra of organic aclda baa an af foot

upon tb# fom atlon of notal cbalatoa.

5) fit affiMsta bava baaw i>ravioii*|y diacnaaoMit

bliMtr color a ara rrodwcad at blghar fl? valuaa. Antbocyanln

axlata in ita colorlaaa pa«odo*b«aa fom a t alliatjheia i ^ .

ft) AlNiorftiim of antbocyanina lORto polyaacdMirldaa

alao la liwolvad in floaar color variation (narbmmo in 2 3 ).

7) 4 co llo id a l acato of ^ a aiitliocyanin^^^laiiia

color ebango froc bloa to rod aa «bo floaar dovolopa in

S,nc«&lffiCfe£S3ii (Warbomo in 23).

8 ) Antliocyanlii color ia alao aodifia^ by ssall nst

background, o r co*occtirr#nca and tatacactlofi o itb ©Char p li*

nanta. tbaaa o tb «r say ba otbar flavonold# or

carotanoida. Iba offacta of tbaaa bacbground pliaanta vary

with tha location in tho c a ll of tha pigaantai fo r anaapla.

aurono (y a llo v to oranga and locatad in tha c a ll micvota v ltb

tba anthocyanlna) ptm cyanldln (n«#<Mita) fi<iNNi oi^ftta^rad.

and aurcma ptua palargonidin Coranga*rad) givaa oratit#*

yaltevf hmorwir, ya llo v emrmtm&M < locatad In tba c a ll

24

pUaetda) pliia eyantdin glviMi brown (iburbemo tn 23),

Ihti CaBttltoitiM 9t ruyflBM i.m g E lanK w li .ta

n m w

laolfttod clycoaidiMi of tho ftavopoa and flavonota

aro eolorloaa to palo etmmm to doop yo llo v In colori hov*

avMt, tfcoy lo n a ^ l ly contrlbuto l i t t l o to f « ta l color d lro e t-

ly (Harbomo In 23, Svaln in 23), tb o lr contribution la

aoon In ro«pltcwKitation offoeta*

Tha v ld a ly occurring flavtmola, hsas^faret, ouareattn,

and n ^ lc a t in , ara rora or loaa eolerlaaa at pif valuaa nor*

aially found In tha c a l l , Fovaver, root v h lta , Itnsry, and

eraar flovara contain fiavona or flav<mol glycoaldaa, occa*

aiociaily flavenonaa and 3*hydr«a^ ftavanonaa (llarboana In 23,

dvain tn 23)• kaaapfmrol, ouarcatin, and o yricatin ara oftan

abundant in flovara of thaaa colora C29), Thtm, vhitannaa

ray ba dua tn part to tha praaanca of flavonaa and flavonola

(Svain In 23). Althotgih thaaa ptgsNMRta «ay ba ta iw d *cx>lor*

laaa,** In that thay ara not v la lb la to tha hiMNin thay

do absorb strongly In u ltra v io la t l ig h t , and ara *mmm* by

baaa (t^rbom a In 23). Atao, th a lr praaanco adds "body^ to

a patal to giva I t a craa» or ivory appaaranca Cl^rboma in

23). '«<!hita patala vhich lade fUvonoida eenaptataly ata vary

rara (flterboma in 23).

For a flavonol to contrlhuta to ya ilo v tlm m r co lo r, i t

miat poaaoaa motm atruetural faatora alx»ant frov thm thraa

coeeuionly occurring flavonola, IraaiBpforol, ctuarcotln, and

25

flifrrlcacin C2S), I t d«*w i»tr«tad tliat flavmtola

a ton* eaii produea v la lb la pigmantatlotii in sum eassw, wlilta

apaciaa lack tha flavonalCa) that color tho yollow fora C2f>#

Although ordinary flavonoa and flavonol* apfarantly do not

aetivoly contrllmta to petal co lo r, they n iih t ceoealvahly

do to i f praaant in vary high concantratiiaia ( 2 f ) , mmi

nethylatad, whan proaant in unuaual gljreoaldic fom a, or

when chalatad with iRetala <Har!«»mo In 23)# d e la tio n with

metal* can t»arkodly ehanga m* color of theae ceaipounda

(Swain in 23)#

Iha CflntrJ.hiitlen ef- Qialeonea and Auronee ce Fle>-m

Coloi^

Qialeenea are yellow to oranto In enter whan iao*

U ted (Swain In 23)# th e ir doop celora can ha raapenalhla

for tho color of flower* In m ich they ocettrt hewavor, they

ar* of lin lto d d iatrihution (llarhoma la 23, Swain In 23)#

Tha auronaa, aiiikltar In color to tho malc<mm, are

reapenaibla for the deeper eeler ef flower* in which they

occur* Thiqr, tee, are ef U nited d la trib u tio n , but betb

aurenea and chal<renoa hava be«n netad in Lofunineaaa (Swain

In 23). the aurtRiwt and chalcmaa fm euontly occur with

eareteneid* (Harbome In 23)i thia ia naarly alway* ae In

tha L4^unlno«aa (2 4 ),

Caretoneida and fUvaneida Oncuerin* Teeemwn

Manta having only flavonoida are re la tiv e ly few

In mcober (iiarhomo In 23, 24>* l^awover, while provioua

26

•tudloa hava Indira tad that earotanolda eontrlbuta fpora to

yallaw flowar color in plant* than do flavmolda* fotura

arudlaa nay tm ll ^ t n g flaronoida Into acoal prooinafica with

than (Harboma In 73}*

£« Intarrolationahipa of Pignanta In

Color w arU tion in g . « * n d v r a a a l t a mainly

from cuantitativa dlffaraneaa in tho conem tratim of tha

anthacyanin prasant* nalvidin 3*S*diglticoaida (47* 4 4 ).

MolwidiB ia a nathylagad dariwatlva of datphlnidin* which

pradoninatoa tn nanvo and bloo flowara (Karboma in 23*

Swain tn 23>. Oanarally* nathylation haa a raddaning affactt

howovar* thla affact la oftan not raflactad to any artant in

flowar color* baeanao of othar nodifylng faetora (Harboma

in 23)* loaaibla aiodifying factor® haw® baan diacoaaadf

thay w i l l ba conaidarad in ttm diacuaaion which follawa aa

they apply to £• lindWlCOOM*

Olyceaylaclon of anthocyanidina ia not an important

factor in flowar color* aa anthocyanidina do not occmr In

flowora aa aglycon<*ai at laaat ona augar ia alwom attachad

ta tha 3*hydroacyl group. Subatitotion of a attgar in tha 5*

poaltion haa a arnall affoet on colm* and tha 3*5*

diglttcoaida of nalwldio d iffara from tha Sflneoaida by baftng

flnoraacant In aolution. Ifhla prohahly affacta tha intanaity

of coloration in tho flowara. A cyU tion of thaaa glycoaldaa

with £*coumric or caffaic acid. I f praaant, way partly

quanch thla fluoraacanca* raautting in d u lla r flowar color®

77

28

(Rartpom* in 23)*

flavonoid and athar ptipiafita ahteh m y ba praaant in

tha patala probably bavo aeawi re la In narrow eolor m ria tio n

bot%roon tha varioua ealor eUaaaa obaarvod In thia apoeloa*

tho yollow ftavotioida appoarod to havo an Invaraa ©tsantlta*

tlv o ralationahip to tho ptnrplo anthoeyanln (42* 44>•

1* O A Y a nii iffli

Flavono and flavonol tlyeeaidoa ganorolly

eontrlbttta l l t t l o ta patal color d iro e tly (Harbomo In 23,

Swain In 23) i howovar, eo^pigpontatlon with tha anthoeyanlna

ran raaolt In bloor ahadaa* Ceopar and E l l io t t (12, IS )

roportad that In Hadteaao thoro waa a atrong indieation of

eo*plg»ontatliMi of kaoiipforol g lyco a l4 ^ with tho ati^hoeya*

nin proaont, anong than nalwidln. Co«plgiMficatiofi ia , in

fa ct, tha only knotm noehanlan to u ^ la ln blwolng In f lo w m

with nothyUtod B*rint eoaipownda, aueh aa nalvldln* H i l v l -

din I t ahowB aa tha atructora batew*OCHj

(8«nifi in 23)

Such eonnot eonplox with motaloi tiotal chotacion

tmnild eovmo « bluaini offoct*

Whitoiiooa, •• in tH« noor^whit* eolor elottoa, ooy bo

duo ill port to tho proaone# of flovonoa atid flavociola (Swain

in 23), thoao pl«i»onta can odd **body* to o po tal, i i v in t i t

a croair or ivory appoaranco (Harbcamo In 23>#

In a d d it j^ , flavonoa and flavtmoia n iih t concoivably

a ctive ly cocitrihuta to potal color undor unuauat conditiona,

auch aa proaenco in high concontratlona# High coociaitra-

clofia aoy ba proaant in tho fieor«ad)ito color clooaM , oa lo

htntod at by tha Inver*• ouantltativa rolatlonahlp found

botwoon anthoeyanln and yolloo piipoMita in £• aondvicenae

(42, 44 ),

Tho chalcoftoo and auronM, poaaibto cfxaotl*

tuenta of aandwtcaaea p e ta lt, dO COnttibittO to ftOWOT

eolor d lro e tly , Thoir yellow to ocvnio eolora can bo roopon«

alblo for the color of flowere in udileh they oceuri howarvor,

^ £• eandviee^ao thay would occur togothar with a o lvid ln ,

and tho ro a u ltin t affect la unknown, Tbo cholconoo ond our**

onoa, located in tho c o ll vocuolo with tho anthiKrywalsio,

could Influence flowor color aia^ly by produolnii a back*

grcamd effect. Far onomplo, ejnmidlfi (naionto) on an ourono

beckgroond (yellow to orange) prodtMoa oranga*r4Ni (Harbemo

in 23),

The chaleonea and aurenea nearly always occur with

29

carotaoold* In Lagumtfioaa*.

3« Carc^taano^g

Xe t« umsauftl for « plant tn contain only

flavonoid pigaanta. A ltlio ^h an anatyaia of tlio earotoneldo

probably praaant In £• aandvlgnaaa vaa bo5 d tha aeopa of

thla raaaareh, a eonaidaration ef tha poaalbla affaeta of

deair pTornmncm ia partin»nt to thla diaeuaaion* Bath antho*

eyanina and earotaneida occurring to g a th ^ tmn produea,

anong othar colora, purpla and neganta Cl7 ). Thua tha

praaanca of earotonolda could Influanco flo«*ar color In

£• ggRdiKlcgngg.

Carotanoida, aa v a il as tha ya lla v flavonoida, can pro*

duca background affaeta. Tha raaulting appar«eiC color of

tho flovar ia dopondont upon tho location of th# background

p lg M it in tho c o lt , and eyanidla (nagonta) on a ya llov badr*

ground of carotanoid givat a brovn color (Rarbomo in 23).

Tho earotonolda aro locatod in tho c o ll plaatlda and not In

tho vacoK>lo with tho anthocyanlna (1 7 ).

p¥i iMiy havo an off«Mct on color variation in

k* aandwiconao- «a tho color of anthocyanin variaa v lth pH,

This offset ia dubious, hovavmr, aa tha fdt of c a ll tap tanda

to vary only a llg h tly fron on# flovar to tha fiant (Svain in

23). Hovavar, at tha avaraga c a ll asp pH, 5 .5 , anthocyanlna

tond to axiat in tha colorlaaa paoudo*baao fora, and aaull

changoa about ttila pH could produca largar color offacta

30

than otharviaa aacpactad (Swain In 23)* Imnfanaa in eatiaoa

a blunlng affaet on tb# iaotatad pigaMmt, with a daermaa

eauaing raddar m lor*

51

V. the Relation of t h o - _to Stii^lUg Itertt

iangarteaM

As atatod in tha Introduction, thla thaal* uaa

daalmad to saicvo aa a fotto«N»up to tha atudy Init&atad by

Park (42) and Park and Aotar (43, 44) on tha id «n cifica tlon

and inherltanca ef anthoeyanina in flowar patala of

jihwaBfiilKt AAOiSMlCflaas* groupod parontal linaa into

color clattoa 1, 11, and 111 Ccolorad) aiwS IV , V*.!, «nd V*2

(naar-whita)* It* fotmd tha ra la t im concantration of tho

anthocyanin piipmnt batuoan tho colored and naar-vhlta flow-

ara to bo about flvo to onai h* Intarpratod thia diffarffioc*

In concantration a* baing undar ronogMiic control, with

tha dorlnant color gana, and sg the racoaaiwo gana for near-

«dilta» Th# function of gone Sjt woa to nodify tho amount of

pignant produead, with anthocyanin production bolnt tsaSar

tha control of a aoeond gano, Purthar, ha propoaad a

d ilu tin g factor, to aecoint for dlfforancoa within tha

colored and noar-tdilto groupa t the of fact of m i* gnco,

however, waa not aa claarty dlattngulatiabl* aa that of tho

Intonalflcatlon gana, ^ m y ba incAnp ataly dmtownt

ovm iiik* 4 fourth factor, umtaeed, wa* propoaad a* an intan-

a ificatio n factor or an in h lh itor to account for color d l f f -

•roncoa batwoon color claaitm IV and V| m it factor m y bo

aplatatle to and taaad on thaea propoaala, ho auggoat-

ad the following ganotypoa for tha ra rm ta l linaa which h*

uaod (42)• AACaCal^ for P3 and P« (co lo r eUaa X I) ,

32

AACaCadada for F2 and F ll (eiaaa 1 ), AA^caPa for FI

(etaaa I I I ) , AAcacafai.. .., for FA, F7, F8, F f , and FIO Celaaaaa

V -l and V -2 ), and AAeacadada* plua; tha fourth factor »a n -

tlonad abova, for FS (etaaa tV)*

Farb found M ily on# mnrpliati anthocyanin, idiicb ba Idan-

t i f lad, by pa pay cbrcHMtOfirafliy and almorptlon apactra, aa

auiWidtn 3,5-dtgttraaida* ^ a atatad tiuit tbia pigm nt waa

prim arily raapenalbla for fl<»»ar coloration, with fanaral

color variation raaulting nainly fron ouantitativa d if f a r -

ancaa in pigasant concMitrationt elaaa 1 pat* I* wara found

to contain nina tlnaa aa cnjcb af Iba plgnant aa eiaaa V-2

patala*

Flavonoid pigmnta othMr than anthoeyanina wara ob-

aarvad on cwo-diwanaianal papoy cbronatMiraf^a t Fax5t atatad

that tbaaa appaarad to hava a aaeondary ro la , both g u a lita -

t iv a ly and quantitativa ly, in tha prodiictlon of narrow color

variation in flowara* Ha foimd aight yallaw arota In v ia i ­

bla and u ltra v io la t t ig h t, atnd thraa flueraacaiit apeta in

u ltra vio la c l i f h t i thaaa ha praoMitad in tabular aiWf d ia -

graacatic fom* tbaaa yallew pigoenta wara not id a n tifia d ,

but a diacniaaion of Farh*a Invaatigationa and obaafwati<ma

waa includad in hia atudy (A2)* Ha concludad that tha pra»

duction of flowMt color in JEU iff dtia to tha con-

cantration of anthooyanin, hut la ralatad to ttia praaanea or

absanca and Intanalty of tha yallow pigwanta* Fa fe«nid an

inwaraa ralatlonahip batvaan tha contant of anthocyanin and

53

ochar. yallew. ftavoaolda* and tm auggaatad that thara waa

eai^titliM ti far tha lla ita d a^ tya t of tha mmmm praeiaraora

fa r tha antharya«if»i and athar flair«aaUa« 4 cmmem ayn*

ehatle pathaay In aarly atagaa haa baan aufgastad by tha

a ln lla r lty In atruetura bataaan anthocyanlna and othar flav*

enoid ronpounda (5 0 ). and la aupnortad by avldanoa af con*

patition fo r coaaion pro«nxraora bataaon d lffarant rtaaaoa of

of flavonoida (Harborno in 30. Alaton in 25. 2 i ) . Portbor

avldanea la found In caaoa in vhl<h a aianlo gono eontrola

atR»atleution in two rolatod atructuroa (2 4 ). ioaa pXanta.

bovavar. do not ahov avldanea of eompatitlcm botwaon fU vo*

noid coopouada (Rarboma in 20. 24. 27).

In hla F j peputatiena. Fart fotmd eoaflieatad aagraia*

tiona far f t o v ^ oolori ho atsgcoatod that ihoao aight ba

dua to tho Intoraetiona of gonoa for antboeyanin and othor

ftffvonoida. i f aavaral i« io a ara involvad In oontrolltng

tha production and Intonaity of tha yalloa ptgaanta. thla

Intaraction say raault in a eooplieatad ganatic aoda toe tba

production of flowor color variation (4 2 ),

Park (42) and Path and io ta r (44) raportad m tho

fUidinga of Gaacoigno afi (44) and of F^a yth and

SUnmnda (4 5 ). id»o atudiad anthoeyanijta In patala of

SsisadlMB* ilo o flowara of 1* latifoiitan mm vara f o ^ to

contain nonoglycoaldoa of a a lv id ln . and tha rad*blua flovara

vf &* t?Qlvgacrjift OC. vara fowad ta contain both rantoaa

34

glyeoatdoa end dlwexmoittreoaidoa of m lv id lA (46)* tho

hluUh*rod ftoworo of £>* frutaaonno S«hlfidl* eotitoifiad dal*

phlntdin (45)*

Baeo*foltti (5 ) , In a awsnary aoeowrtt of tho phonolic

eonttituonto of taavoa of dleotylodonooii plants, f^intad out

that d ofln ita rolationahira m y oxiat hotwoon tho rang# of

anthooyabln typoo found in potalt mnd tho fhonolir typoa

proaont In tha plant body* lio atatod, hmiovor, that tha

patal pignonta* bolng vary atutooptlblo to gonotle m r la t lo n ,

aro not ao ^ ^ ra e ta r ia t ie of tho apoeloa or aa in d im tlvo ofiIta taxononic poaition aa aro tho f^anolie conatltuonta of

tho vogotatlvo tiaauoa* dovertholoaa, a oonaidoration of

tho anthoryanlm proaont In tho vi^ota tivo porta of

l aaiaodttffi! snacioa nay ahad l l i ^ t upm id o ranco of anthocya*

nln typoa ishleb night bo found In tho potala* * a m * ^ ith

(3 ) rofortad a emaldarobto m^sKoit of lmten»*mtb0trym i«» of

cyonldin and dalphtaldin in tha groan loovoa of canadanao

PC* In hla atnrvay of tho phonolic conarituonts of loavoa,

Bat#*Soith (5 ) tndieatod that tho coB»«ci«r oonatitum ta,

eyonldUi and dolphtnidln, at loaat, woro proaofit in 22 and

16, n a iro ctlvo ly , of tho 54 apoeloa of U^nirlnoaao lia t id t

jjMBfitiJUas arooiaa voro not lla ta d , howovor*X

c ,

Coopor and E l l io t t (12, 13) found that atudioa of

diploid Hedlcaao aativa and 2I« faloata L« had a la rlfia d tha

Inboritanco of tho anthocyanin plgranta (36, 55, 52 ), but

35

that much Immn known about th# ysltow plftmanc# (1 0 ), I t

wt# known that thara war# two claaaaa of yollow pl|^®fita In

alfalfa* tha plaati4*hom« carotafiolda* and tha yatlow* acid*

aolubla anthonanthlfia* L lt t la waa known about th# rolatlwo

ii^®rtaneo of thoao plftnonta to flowar color or about th a lr

ifiharltanco, Coopor and f l l i o t t aonaratad and Idantlflad

tha cooponant pl|t»a»ta In a lfa lfa flowara and dataralnad

th a lr ralatlwa Inportanca to flowar co lo r, Ihoy found that

tha eamtanoid plgnanta vara p rin w rlly raaponatbla far

yalltw flowar color In alfalfa* and thaaa oatatanolda wtra

id a ntlf lad aa xanthafdiyll aatara, tho nanthoiN^lla fltta d

Into Intanaity elaaoaa* and ^aaa claaaaa %*ara corralam d

with tha intanaity of yallow co lo r, Coapar and S ll ia t t alao

found aeld*aolubla mthoaanthlna* ain cptarcatin and thraa

kaaapfarol dm'ivntiwaa* aagragatad c{uafttitativ»ly and

<pialicaciaaly, Iteweo wao mmm awidanca of a yotlowix^ affact

froai tha ouaveatin darlaatlvaa idtan xanthophyll waa abaant

or in traea amounts* but kaampfarol* a pal# yallow pifnant*

had no yallowing offacti I t d id , howavar* thow awidanca of

co*pi«rwmtatlon with tho anthocyanlna.

In furthar atudloa* Coepar and C tlio tt ( I S ) raportad

that fr«ai ona to four factor# had baan poatulatod for control

of yallow pignant production in both d ip lo id (10, 34) and

and tatraploid (34* 34* 37) alfalfa* Hodring with diploid

alfalfa* thay hypothaaicad two ganaa f&t tha control of jtan*

thopliyll aatora raaranalblo for yollow flowar color* and

34

vieh addit&va affaeta, raaultlxii la a

l i 4 i i » 4 i l twtia* thaaa flv a la ta a a i.^ etaaaaa yrnwim

trim d iic a to tha Ititanaa yatlaa of Fadleaao XMlSSMStL* No

Inharttanea pattaim aaa oatahtiahad fo r tha t lx e^Mreatia

p ltrofita . two of tha tstiraa kaarpfarol mhmm&

iaofio»faetorlal a i^ratatlon , v lth aaeh balng eontrollad hy a

dlffaront dooiiMiat facia, or ! ^ ,

37

V I. lacttc, filMMffliSBgiashi,

f «p «r ehrooMieosmphy, actNnrdlfidi to Skoa^ and (5 4 ),

m fara to "proeoaMKi that i w ^ i t tha raaolntien o f m oSm*

ttxra aa a eonaaotianca o f diffaraneaa In rataa at idiich tha

Indivldttal cc^pimanta of chat nlntisra nigrata t h r ^ h a

atatlMiary nadiun undar tha inf liHKiea of • asohila ph«aa«**

Tho proci^a waa f ira t da«<hrit»ad by Conadan gg g| . in

1944 <S4), f rotsHP yoara latmp, SataNi>^lth <4) daworihad tha

applleatioo of thio tUKhfilqno to tho idontiflcotion and

aopuration of anthocyonldina and rolatod flavonold plgnonta.t

Sineo that t ln o , tho itoo o f popor chreootoixophy in tho

f io ld of ftovonoid ehaniotty haa Imroaaod at an alooot on*

plooivo ro to , ond today tha tochni<mo la ooad roittiaoly in

noat invoatigationa of ftavocioid eai^»oiiiida In ria n t iM torlalo

(Soikal in 20).

tho oolvont oyoton 84V <n*botonol, oeotlo a cid , and

o n to r), ahotm by lataN*Si8lth <2, 4 ) to roadlly ooyaroto antho*

eyanina, «naa ^ o aolvant ayatoo f ir a t uaod In popor ehroaio*

tography of flavonold caatpounda I t mm Intredueod tn

1948 by P a rtrid f^ , otio uaod tho ayator in tho chronatography

of atigora <4, 8oto*Snith in 4 9 ). to Ita oxcoiloiit ro*

aolvtag poooro (Solkol in 2 $ ), 84H rapid ly ^memmo p^ntla r,

and today I t ia a t l t l tho aolvont ays tan noet coaaaonly

ployad in flavonoid veck.

tho popularity of papor chronatogra^y oa a nothod of

aoparatlng and idootlfying tho flovonoid# and othi^ phanolio

S8

eoeipoucM!* ia dtia to at laaat four faetora, ia tha U it r in -

t ie eolor of atoat flavocioid ceaipottnda in v it lb la i i t h t , or

th a ir appaacanca In u ltra v io la t lig h t (Saikal in 20)i in

u ltra v lo lo t l ig h t , aoat rhanola appmr colored, atid

reeee v iv id ly , mr elae ebamb lig h t and appaar dark ptnrpla

(SaUcel in 25)* let addition, eolor noy bo changad, anhanead,

or aaally dovolopad, i f lacking, by varlooa roagonta, aoeh

aa notai aatta and ae«M«itogo hydrcatldo (16, 19, Satoal tn 20,

29}« Sacofidty, papar chromatography proaanta a rapid caithod

of iaoiaclng eonpounda in tho mlcromcKninta and free tha

riaturoa in whim flavonoid corpounda aotlat in plant aator*

tala (Soikal in 20, Swain ia 23, 26), Accordtag to Swain

(Swain in 23 ), aovaral glycoaidaa nay ba aoparatod and ttioir

atructuroa dotMmiaad, ualng <mty a graw or ao of p lm t

m t o r la l . I f a coaaaonly occurring aglycono only ia to bo

onaljrcod, 100 a tilllg ra m ef froah tiaaua m y ba awfflclant

(Sata-Swlth In 4 9 ), th ird ly , flavonoid coapotMida aahibit a

wid# ranga of a o lu b lllty ehacaetoriatlca, m k liit ttioa ideal

for paper chromtograidtic analyala (1 9 ), F in a lly , tha

tochnlcwa la m a ily loamad and givoa roawlta nolcltly aaing

ra la tiv a ly inaxron*ivo apparatua (SalVal in 70), Conaacuont-

l y , the application of pmpmw ehromtograpliy to roaoarch an

flavonoid and ether phenolic cowpotawSa haa baan wldoapraad

and laro llflc (&ata-iN»ith in 4 9 ), and today Rf-valwaa of o i l

tho m ln elaaaoa of flavonoid coepownda are a m ila b la ( l i ,

19, Sotoal in 20, 26, 29, Sato-Snith In 49 ),

3f

Ta nta tlm idantifteatieii of an unknown flavonoid eoie-

pound can aMAatinaa ba wwda by obtaining ita Rf-valua in a

variaty of aolvant ayatana, and gatharing data cm Ita

color raaetiona in v ia ib la and u ltra v io la t liid^t with and

without aavaral raaganta. Hoca concluwiva Information mm

ba gatbMrad hf rtnnnine tha unknown conpound alengaida Imovn

atandarda, cm tha aaaa chronatograjpbie papay* ^ ^ ruRning

tha tmkno%oi alana and than in a nixtura with tha Imown atan-

dard to datarwina i f thara ara mwy diffarancaa batwaan tha

two runa« Both ahoutd ba parfomad in a variaty af aolvmit

ayatana, caking uaa of aavaral array raaganta* Oftan u ltra ­

v io la t apactrat data obtalnad for pitta unknown contpounda

w il l pravido avidonea for a concluatvo idantification*

40

v n , E l t t i v i s l i t iB n cttg tsg tiAltheufh tha f ir a t ralLabla valtiaa of u ltra v io lo t ab*

aorptlon maxim for ooo claaa of flavonolda* tho flavotioa*

woro ohtalnod aa aarly aa tfSV, by % a rx fn »k i, i t waa not

u n til tha mid i930*a t ^ t u ltra vto ia t apoetroaoory baeamo

widaly uaad in atudlaa of flavonoid eompounda (Harhomo in

49)« I t waa at thla tin * that throa groupa of wofk«ra*

among than .Goiaaaam* Hirh«ma* Horowita* Hordatrflfe*i'

Swain* and HSrhaowMr* bagan rocordin|i anaw tho apoetrat «^ar*

aetariatlea of a l l th# naior claaaaa of ohaRola CHarhoma in

49)* In a d d it lm , daiaonatratad tha Incraaaad watua of

naaauring apaetm In tha praaonca of alleaii and othcur diag*

noatle roaganta* aneh aa alunim jr F lo rid a * aodlmi othylata*

fuaad aaditiBR acatata* and boric aeid*aediuR acatata (Jtsrd in

20* Saibal in 25* Barbema im 4 9 ). I t ia now peaaibta* with

tha uaa of auch tachniouaa* to dotoraina tha atruetnraa of

aoaio flavonoid eomsioyfida uaing apactra alena (JNacd in 20).In racant yaara* aaif*cacordlng apactroptietMHitara hava

bacoina availabla* thoraby a t e llf y i* ^ tha taak of ra*

aaarchar* and twSny a larga aaMMmt of r a f « r « ic « Infmwation

ia avallablo conciMBniwi tha apoctral chatactarlatica of

plant flavonoida (Jord in 20* Harboma ia i f ) , Vbm uaad In

eanblnatlon with aoparation of conplam nlxturaa by papar

chromatography* araettal 4mm ara of inaatlnabla valuo in

tha chacaetarixaticHi and idantlf tea tion of flavonoid conati*

tuanta* avan vhan praamt In anall anoiHita (1 9 ), flowavar*

41

■ Ineo alxturoa of dlffarant flavotield pigaantr tmy intaraet

in ra lu tlo n . ttia uaa of a pur# »a«pla of tho unttnown la

oaaontlal I f obaorvad fsaxlaa and othar apaetrai data ara to

ba ra llad upon «dtan eonparad v lth raportad valuaa (1$.

harboma in 49)*

Spaetral eharaccariatlra. ataeh aa naxiiM and ahifta In

narira brought about by tha addition of diagnoaele roagonta.

can ba uaad ta idantlfy unknoana by coo^riaon v ith rafaranea'

data avallabla for tha flavonoida. In addition, tho apactrun

of an unknown any b# coaparod with that obtainad for authon*

t ie aatoria la . both v ith and vithout tho mroaoneo of diag*

noatle raagMita (Katboma in 49 ).

A l l of tha flavonold eonpounda axhiblt Intonta abaorp*

tion in tho u ltra v io lo t rogion of tha apaetrun. and thoaa

Chat ara celerad aba orb alao in tho v ia ib lo rogton (Harboma

in 4 9 ). In ganaral. aaeh elaaa of flavonoida haa d ia tin ctiva

apactral eharaet«riaticai tha dlffarant elaaaaa nay ba

diatlngutahad freat ona anothar by obaMTVlng tha ganaral

ahapa of tho apactral eurvaa and tha poaltlona af aartiea aa

v a il aa of ainiaia. thouldora. and infloetlona (Rarboma In

4 9 ), Harboma (fiarbotna In 49) praamtad a tabla givliMi tha

ganaral apactral eharactariatica of tho various elaaaaa of

flavonoida*

42

H4TCRULS ARC HStRODS

1* m S iM U liTha parm tal m to rta la uaod ware mong cboaa uawd ^

Farit (42) and Pattlc and iletar (43,44)* thaaa fNtrantt war#

daalgnatad hy fmtfk aa F| through F l l , and wmtm elaaaad by htai

into color elaiaoa aecoming to tha Kortlealttaral Colour

Chart of tho Ikoyat H orticu ltura l Seciaty (31)* Aa eontawina*

tion of tha o rig in a l parantal tinea with feraign m ta ria la

nay havo takan plaea prio r to tho oonssianeawwnt of tbo proaont

atudy, parantal eolor elaaalflcatiofi waa rarado* Aa a ro* < • '

• u lt, tha parantal planta, havixMl ftowar eolora Mmgyimg, frota

naar*«<hlta to lig h t and dark purplo, wara plaeod Into on# of

six flower color claaaoa, X through V »l and V-2*

Flowar color claaalfteatlen waa d<ma oarly in tha ffdm*

tng of a sunny but cloudy day* tha H o rtle u lt^ a l Colour

Chart (31) prowldad tho baaia for tha ctaaalfteatloR* Only

flowara m i ^ wara ontrippad, with tS a atandard potal upright,

wwra uaad for elaaalfylng tha planta* trippod flowwra wara

not oaod aa tha eolor of auch flowara fadoa rapidly follow*

lag tripping*

Raaulta of th is ra «»cU fa lf leaf Ion, givan in Tablo 1,

d lffo r frow Fatk*a o rig in a l elaaaification* tha diffaraneaa

Miy ba duo to contanlaation or to individual dlfforaneoa In

Intarprotatlon of colors* o ftm uhan a parm tal lina waa

ropraaantad by two or leoro planta, thaao plants wara ptaead

in d iffarin g claaaoa* This accounts for tho dual color and

43

claaa lla tlnga for f4 and FtO.

A4

♦5

FI Cohalt v io la t l i t

F2 klnaral v io la t I

F3 Roaa portla 11

FA Canallia roaa or V .l

Faatal aauva IV

F5 *.♦* —

F6S^ ftoaa purpla IX

P2 Faatal ivnxva IV

— —

F9Sj Faatal l i la c V-2

FIO Faatal aauva or IV

Faatal l i la c V.2

F ll hinaral v io la t I

* Aeeordtsig to tha Rorcicultwral CaUnar Chart af tha Nayal R ortieultural Soclaty (31)*

** Fatal* not avaitahla for d a ta i f teat Ian at t i^ aat»a tina that tha ahova elaaaifleatlon waa aada. F5 via plaead in Fadr^a elaaa XF| Ft waa plsead in Fark^a etasa V*2« Fact'a ctaaa IV was fudiata plde, and hi* elaaa V*2 vaa vanatian pink <42 . 43 . 4 4 ).

A* ^ggiMlat

Croaaaa wara attaaptad anonf parantat liiiaa ualiii

o«a of two cochBlpuaa, eoRVaiitiofial aoaacolatim and rrlf^ittg*

Thaaa nathoda follawed tha tachnlana oaad by Park, with nod-

iflcatlo na (42, 4 3 ),

x« (aaBiMttondl je aiC Tilitlga

Unopanod flowara wara aoaaotilatad in tha lata

aftamoon, batwaan thraa and aix a^eloek* imaeulatlaci waa

aeeanpliahad by palling off cha t ip of tha bud, tharaby rmSt-

iag Cha anthara aceaaaibla* lha anthara wara than earafully

ranavad with a pa ir of forcopa* A fib a r pollination hag waa

plaeod ovar tha raeaosa baaring aautaeutacad buda, and a n o ia t-

anad cecten h all waa inaartad into Cha bag at ch# baaa of

cha raeaiMi* Pollination waa parfomad tha naxt naming ba-

^waan aawan-thircy and alawan o*eleck* Pollan fron tha naia

parant waa oallaetad on foreapa and earafully appliad to tha

atigna of tha aanaeulatad huda, and tha racanaa wara eowarad

mm bafora*

2* TriPBteB

Plowaca wara !uutd-trippad uaing a pa ir of for­

eapa to rub tha kaal of tha flowar, prafarably ona with an

araet atandard* I f rtuhbing tha kaal did not produea tha

tripping raaponaa, tha atanana wara u m a lly rataaaad by

apraadlng tha kaal patoU with tha foreapa* Tha anthara wara

ranovad fron cha ralaaead ataeans with foreapa* tha fmpeapa

46

IX* t^rhoda

vara vlpad v ith athyl alcohol to avoid ei»ita««io«tieii of tha

atiipo v ith undoairad pollan* th# oxpoaod atigaa vaa

fltMhad v ith vator to rafeovo anchora and adhartf^ pollan

gralna. ainS/or to c»tiao to hnrat any adhorlng pollon* Tho

flowor vaa drtad. and vaa thm pollliiatod and haiuiad aa In

tha f ir a t oaChod*

In both nothoda. emro vaa takan to aaatiro that tho

flowor color of tho walo pamnt vaa alvaya dackaHr than that

of tho fooala paront. udiieh wta ehoaan fra » tho noar*vhito

group (IV . V *l. and V*f)* ^ tho baaia of Fark*a findtnta

that purpla flowor color waa doninant to noar«whito. eroaooa

nould bo diatingwiahod from aalfa by obaorvlng tho flowor

color of tho offapring*

ho racognlaabta trwa eroataa vora obtainod in thia

atudy* Aa a conaoowanco. tha ganotie otwSy propoaad in tho

outlino of objoetivoa waa abandonad*

SwHlliai groMtti

Sooda w«ro acarlftad by puncturing tha aood coat

with a dtaaactlng naodla. and wore gorcinated in p o tri

diahoa or dtroetly in a o ll in f la ta . Sooda vora inoculatad

v ith tha appropriata AhlaQblun. Ccwlnation in aoit vaa

vory poor* ybma aoadlinia vora about ton c^witlnatora high,

thoy vora tranapiantad into peat potat whan ibay vora about

tvonty eantlnatara high, thoy vara tranapiantad to eno*

gallon cana*

47

<2* Sullttctiaa aX,

i« fIatMc„Bi4Kcnta

Flovara for pignant axtraetian wara

eotlaotatf in p U a t ir bags in tha morning fttm aavan*thirty

to alavan o*cloeit« Flovara v lth an i ^ i g h t atandard, aithar

triprad ar untrirpad, and flovara jm t bagienitii to raiaa

th a ir standard, vara aalaetcd* Patala only vara uaad for

pitecant axtcaetlon, Oftan tha avpply of flo var a vaa Itm itad,

aspaeially during tha autueai monttMi, Mid conaapuantly only

thaaa paranta baaring a aufficlant mmbar ef flovara vara

aalaetad for aaalyala*

Oom of tbtmo mathoda of axtraetian vat uaad, dapacidlag

upon tha elaaa af eovpounda undar anatyaia* tha aathoda

ara rafarrad to a* tha mathanoUKCl mathod (42, 4 4 ), tha

aMathanol nachod 0 3 ) , and tha hCl cathod (8ata*Saiitli In 4 9 ),

thla mathod of axtraetian, uaoful

for tha atudy ef anthocyanlna, aa v a il aa othar elaaaaa of

flavonolda, fotlovid tha tachnipua tmad Farir (42) and

Fark and Rotar (4 4 ), v lth coma aodifioitiona* Famla vaara

cruahad, using a mortar and paatla, in a smalt amount of

cold mathanol ro nta ln ii^ on# pareant hydrochloric aeid (v / v ).

tha raaulting aaah vaa vaahad into cantrlfuga tubas using

tha M tra cting so W ant, and tho mlxtura vaa cantriftgiad to

aaparat# tha patal raaldtia fron tha aupamotant detract.

4$

Tha aupamatant waa pif>#ttad o ff , v ith potrolauv athar

to ranova ehlorophylta and camtenoida, and eoncantratad or

drlad In an avaiporatlng diah undar tha ho<»d«

2 ,)

Thla natliod mm usad for tha axtrae*

tien of flavonoids othwr than anthoeyantfia* fatala mtm

aoalrad in abaoluta trathanol u n t il th i^ haeana tranaloeweit,

or for about ona and <ma*h«lf hoora* tha asctract mm dacant*

ad, and tha ratals vara vashad twica wildi alMioiuta n a i^ n o l,

Tha vashaa vara addad to tha o rig in a l (oitraet, and th# ra*

aulting nixtmra vaa vashad with ratrolainn athar to ranova

ehlorophylla and earot^Mioida* Tha mittmet vaa than anraporat*

ad to drynaaa at in t^m t»athanol*HCl nathod*

3 ,)

Tha purpoaa of this csathod woe to

obtain by hydrolytia tha aglyoonaa of tho glycoaidoa praaant

in tha patala, fOtloving tha tochnioiio for hydrolyaia out*

linad by Sata*Smlth <Bata*Si»lth In 4 9 ), Aftiwp hydrolimla,

tho hydrolyaoto vaa doeantad o ff , and tha roaidne in tha

toat tuba %aia voshod with abaoluto nothanol, Tho waah voa

oddod to ti^o a x tro rt, ^ o eonbination vaa f ilta ro d , and tho

f i l t o r popor mm vaahod with absoluto nothanol, Tho axtroet

aolutlon «Mi8 than vaahod by aciding an approxliMtaly apual

voluna of laoanyl alrO hol, tho uppor, ncm*aonoona layo r,

containing tb# aglyeonaa, vaa vashad tviea v ith va tar, and to i t was addad an iaoaryl a t c ^ o l waah of tho IcoRsr,

49

•quaoua tayar* tha roaultlsig laeanyt alccMttol fraction wa«

thon ptacad la aa avapocatlim dlati and driad aa la tha

pravioua tvo ^thada*

An a lta m a tlva nathod of obtaining tha aglycanaa waa

also uaad* Ixtracta from f»«thasiol»HCl mthod war* hydra*

lyaad by ad'lng 2 R HCl to tho oxtracta in oathaaol and

placing taat tubaa containing thia nlxtura in a belling wctor*

bath for tw«nty oinutoa* 7h# aglyconoa wore than woahad out

of tho hydrolycod extract uaiim anyl or laooKyl alcohol*

The (ia o * )m y l fraction waa p la cad in an owaporsting dlah

and driad*

la a l l of tho above oxtraetlon nothoda, tha driad

roaidua waa radiaaolvad in Ita raapaetiv# extracting aolvant,

oxeapt for tho aglycono roslduo, which waa rodlaaalvad in

abaoluta oothanol* Tho purpoaa of avaporating tha extract to

drynosa and than radlaaolviag In tha aolvant waa ta ronova

watm frow tha axtract ahleh could have cauaod intorforanco

on tha chrmaacograpti* following the rodtaaelvUig, tho « k*

craeta wora applied to tho ehrooiatograrhic papora*

6* riKSdflt gyg.u;.gg|*l8n

Purification of tho axtraeta, whoa done, in*

volvod proporativo chroaotography, uatng Rhatssan nimbor

Chraa (#3) paper and tho BAR aolvant ayatoai* A fte r tho

chrowatograpbic papora had driad, tha antira run waa cut

into ai!«all pioeoa and placed into f i f tom o i l l l l i t o r a of

abaoloto aothanol for olutlon* After about twelve houra.

50

tha aloata waa tfacantad Into an avayoratlng dlah and atlnwad

to ovaponkto to drynoea, tha raaldua ma radtaaelvad in

abaoliita mathanol for analjrtie ehronatogmFhy,

gao«e ..mrQ'.-.■atoagarhw

% Analytic paper e^rom tm tam y waa

uaad for pigewwit idoncifieationi Xf-valtiaa obtalnad tn tha

aolvwnt ayatooa oaad «NMra CMiparod with Rf-w»luaa awallabta

in tho tlta ra tu ro , and tha color r«aeti<ma of tha apota ob*

aarwad wara cowpatad with thoaa racordad for known compoonda.

Doaeandlnii chctwaitogvaphy vaa parforwad m iRg Whatmn now

bar ona (#1) papor, 23 by 43 contlwatora, In glaaa chroawte-

graphy ehawbma 29 by 33 by 49 conticotora. In aema caaoa,

tha chanbora wara kapt aatoratad by placing aowe of th# a o l-

want, or tha lowar, aouaoua layar of a two-phaaa aolwant aya-

tan, in ttia bottom of tha charbar, Ocm aionally, tho papora

wora aquillhratad p rio r to coeonaneiiMR th# run by allowing

tho wspora to aatucat# tho paporo* Tho t i» o of tho chroma­

tographic run dapondad upon tha aalvant uaedt a<aea ran fa a t-

ar than othara* thm tamparatcar# waa f a ir ly conatant, varying

frow 22* C to 25® C and avoraglng 24® C, Kany of fho runa

wora mda in atdwluad l ig h t , fapora wara altowod to dry

following tho run, and thon thay war* obamvad tmdar v ia ib l#

and u ltra v lo la t lighk*

Spota wara mrkwd In poncil, th a ir celora racordad, and

a iw ila r obaarvationa ware eada after onpoauro of tho papor

51

to tha funaa frcm aaamwiiua hydraaida ( 1^3 vapor)• Tha ftf*

valuaa of tha apott wara ohtalnad by avaragixig Bfrnvaluat

arrlvad at by {saaaurins fram tha mtddla of tha o rltU i apot

to tha alddla of tha apot* atuf from tha tasadtjig adga of tha

o rig in apot to tha laadifig ad«a of tha apot. Op cvo*

diflumaional chromatographa, th* Rf*valiia in tha f ir a t aol*

vant waa ramaaurad aftar movaaant in tha aacond*dl]»analonal

aolvant.

Tha aolvant ayataaa daaerlbad balow war# twad*

£4^1 n*butanoltaeatic aeidiwatar •• 4 t li5 (v/v)*

ttppar layar. thia aolvant was uataally praparad fraah* hot

on oceaalon waa on* or four days old.

ro ra a ta lt aeatic acidihydtoehlorie acidiwatar i i

SOfSilO (v / v )

liAc^KClt acatic acid 1 hydrochloric acidtwatarit

15i3t82 (y / v )

i41ii toloana*acatic aeidiwatar it 4i t t 5 <v/v)*

oppar Uyar* Thia aolvant waa prap«rad fraah,

IStifili watMrihydrochloric acid it 97i3 (v / v )

iStHAei watariaeatic acid i i 85it$ <v/v)

watariaeatic acid t i 40i40 <v/v)

I watariiaopropyl alcohol 11 40i40 <v/v}

2 .) y i t m ig ld t , BttttcttroisoBT

Fignanta wara alutad for apactro*

acopie analytia by cutting out tha apot fron tha ehroaato*

graph* and than cutting tha apot into anall piacaa.

52

IhM * m r* pUe«d t"to •>>••»»«• for or loort

twanty-four h«mr»t « felai^ peapiy^ W a blank

p i«ra of papar In tho aaesa faahion* Fa|?or roaidua uaa ra -

«»ovad uaing « alntarad gtaaa cruelM a. Tha u lcta vlo la t

apaetrun waa obaarvad uaing a B a ck i^ lHr«2A aeamitni apaotro-

pbotometar* Ifomwar, a ltraw iolat a ^tro a e o fiy mm i»M ior«aa-

f « l dua to Inporlty of tbo apota*

53

4 . (;c c u to i

QfenemittonmV ansaaeulatlcm vat tha aathod of ereaa*

Ing f ir a t atta«&f>tad« Ihlm tnathoa yioldod l l t t l a aueooaa in

ohtaltiing poda. parhapa haoauao tho flowora ara ao fto g lla

that they aro daaogod diartog tha aeaiaociXatiOfi or aioply fey

hoing handlod* fho buda oftmt droprad off ahiio tiolfm eoaa-

eulatod. f a l l o ff dorlag tha a l# it . or mdsrwoat abaeiaalen

folUinrlng pollleMitloti*

Tho trlppUig oathod yloldod poda with taora auoeaaa

tha aoaaeulatiim aothodi tharofora. tr lp p ln i worn tha aathod

tsaod for tha g r ^ t o r part of tho eroaaing* Unfinrtooatoly.

howavar. non# of tho offaprl»t. producod ia thia atudr wara

raeognlaabla aa trua croaaaa* %y tho tlma pzogaaiaa war#

id «n tif ia d . I t waa too lata to raoMko tha oroaaaa la otdar

to follow t h r o i^ om tho gaaatit atudlaai propoaod*

C ifflcu ttio a aneountorad la tha cbtaiaing of truo croaa*

aa wara dua to faulty toehnltiua. to poor anvlrotsstantal oofidl*

tiona. or to both* fa rt of tho d iffle u ltla a wmy p«rh«pa bo

iahorm t in tho apocioa* Haay of tho aathara ahotiad avi*

danco af airaady having dahlacad vhan tho f l ovara vara

trlppad* Thia vaa alao indieatod In obaoevatlofia. by fark

(4 2 ). of tiva of pollan aae didsUMsarMKO* In thia oaaa. aolf*

ing could hava atraady talcan pUea bofora tha flovara vara

opanad* Altho«yh tha atparianco of othor vorkora ( I I . 42.

UMSUVtS ARC CISCUSSIOfI

43) SndicaCad that aandwlearAe tiaa d if f ic u lt te eroaa,

aona auecaaa at craaatng haa b a ^ attalaad Clt* 42, 41, 5 t)«

B* Send lina Growth

CarelnatloB d lra c tly ici a o ll waa vary poart me-

caaalva nolatura, darplni o ff , or wtahlnt away of tha aaada

eauaad graat loaaaa af potamtlal aaadllnga* Dhaii aaada wara

plaead f lr a t In p a trl dlahaa to gar^inata, garwljuiticaa waa

about ona hundrad parcimt*

55

M m U U m U mA* dlvcpaidoa

I . EoBulfca

ISxtracta war* abtalnad ualng aithar tha

mal^«nol<*HCl mathod m: tha ftathane 1 rathad* tfb la f daarrlbaa

tha praparatlen of chrecvttogtapha afialysad* thaaa

graphs ara praaantad d ta g ra ;^ ties l ip and daacrlbad In

flguroa I threy^ii IS and tahlaa 3 th re a p 10« ^re «w it© * r«^

A is not incfltadad aa Rf^valwaa fron thla chr<nMteprpah wwa

not rat labia dua to tmt^nUpm*

•• ^nii>ftCTaaAni

Chroma togrspha i^aparad ualng tha mathaaol*

MCI natbod wara analyaad for anthocyanlna* Anthocyanin spota

ara rafarrad to aa Ao| and An^*

*>• ,S^A«r,

A l l chromatofrapha wara analysad for tha

prasanea of othar phanolle eonpounda* Speta of othar phan*

o lica are Isbalad and rh i^ through tha mxebarlng

ayata© vaa darivad fron a grouping of apota, frop aapaeata

ch^MEatographa, thought to com Nipo^ to mm anothar* fvst^

that axplanatlon followa in la tar dtaouaalon* Othar apota,

which ara not dlaeuatad in tha taatt hut which appaarad on

tha chrMMitographa* ara lahaltMi with Arable fnaaaraia, start*

Ing with tha nuebar mm for aach individual ^r^*at«^|«a^*

3* Elg,tail,Slim

•• AiBluBsarAiiiiis

56

ChrosMtotrarha woro f lr a t run to vorlfjr

roaiilto obtolnod by Fork (4 2) one} Fo*k oimJ Rotor (44)* Fork

•totod thot m lv ld lii 5,5*dltlueeaido imo tho only ontho*

eyaain proaont* According to Kartin (38) and Swain (Swnte

in 23 ), i t i « unuouat for only <mo dariwatlva of a o i«« lo

otlycona to bo proaont In a fw rtlcu lar apociooi ofton,

anthoeyanlna ara acconpanlod by cloaoly ralatad ccnpimnda,

auch aa, ii| thia caaa, natvidla 3*tlm oatdo, oolwidin f*

glucoaida, and/or othor nalvidin tlyeeaidoa*

In a l l tha chromtotropha, a la m « rooo<»colorod aroa

waa obaarvadi howovwr. I t waa d if f ic u lt to dacortBine I f

thia araa raproaontad eno anthocyanin or aavoral anthocyanlno*

Color v a rU tlo n waa obaorvad, under a l l lig h t eonditloito

(w iaibla and u ltra v io lo t l l f ^ t , with and witlieut anwonia '

vapor) olens tho entire aroa, awklns I t poaaihla to divide

tho aroo Into color aonoag one of thoao aonoa, hewavor,

appoarad to bo wore pronlnont than tho othora* I t waa

pointed out by Hunt (32) that thoao aonoa could bo alatply

orooa. In fdiieh tha eoneantratten of the anthocgronln waa loaa

Chan that in tho woat prooinont aono, or aroaa roproaonting

breakdown producta of tho eowpound fom lng tho neat pront*

nant aono* I t waa aaauRtod, thm aforo, that tho ant ire roao**.

colored aroa roproaontod ona conpound, an amdimyanUi, Ao|,

(8ao Fiflgiroa 1, 2, 3, and 4 v «) thia aaauaiptiofi waa aop*

ported by tho fact chat the roao«^olorod araa rfOMiinod intact

* Sm>t Ani waa alao o^acvod on dbroootmraph A*

57

98

toblo 2, CaacriptioB of Hotariala U««d in Cho Froj^oration of Chro«Mit<^rai>ha Anolysod for tha fraaanca of fhotiolle

Qlyeoaitfoo

ChroaMtograph faronta Uaod extraction SolventSvataoa

A n . 25 hothonoloHCl 8AV

8 h«i»t*nol*f!Cl BAV

Cr y V r S s j / n i *

Hothanol*HCt H4c*liCl

D P5 Hathanel-HCl 1) BAV2 ) 158H4C

8 n o 1) 8AV2) BA«*HC1

r r t , 2 4 , m j .n o , n r

Mothanol 8AV

0 m hcthonol 1 ) 8W#2 )llR C i

H .l 22 hofhanol 1 ) i4W2 ) « 0 « 2

H -2 211 hotluinol a

H*3 211 MarNanol a

IU 4 24 Methanol a

B -5 m j Hothanol a

K .6 n o Methanol a

1*1 22 Methanol 1 ) BAV2) dOSHAc

1*2 211 H«Mdianol a

X -3 Fit MaChanet i8

1..4 24 Mathanol m

59

Tabl* t , PmnmipMim of Ha«d: i » mm Fr*poratl.«« of Chromtographa Analyaod for tha Fteaanea of fliatiolle

Qlyooaidaa (Contimiad)

CSuroaatograph Faranta Uaad Rxtraetlofi____Mathod ..

SolvaotSvmnmmm

1 -5 m , Kathaaot 1 ) SAW2 ) §mikm

1*6 n o l%thanel m

Figora 1* Qmemtograph i i WtMBmm la la ttva faal«€ lm a > f thm Spota Oliaarvad an <bia-dtmn«iof»al fmpm CShromtograpti Oavotopad Ualtig 9AV

flginra 2, Cltreffatmraph Ci Plagram toc»vifig ftalatlva Poal« tiona of tha Spota Obaarvad oo C|Mi»dlsNmatonal Papar ChrooKtogram Pavalopad l^aing

-V .'-

-sS BAWFRONT 80 60 40 20 ORIGIN

FRONT 80 60 40- HAc-HCI

20 ORIGIN

Stot

Aft|

Fh.

62

JSsI ia .

R f-v «lu « In V lsibln \)i i>Uht

U ltra - v io la t Vlalbla,lxXtbl, J JL it il.

f r a -v lo la t.h U h l ,

0.23

0.82

roaa f l . rad, b l« , f l . roan* b l .« g r . tU pink

d u ll p*-rod-

y» fl* y* brt* y. f l . y.

* b rt . - b rig h t, #i> » f i uoraacant, bi* - gr* *a ra t i, p . - Burala. r , « m lio w

Tablo 3. Rf-Vfftoao and Color Roaetiona of Spota c^oroad onChronatofraph 8, Figura 1

An,

Ph,

Oi37

0.05

Rfwvalua in Viaibla££81 .. laftcsiiSI- l iU h t

Color Raanrian» iM

roaa,f t .

roaa

roaa- b r. Co y . -b c .

U ltra ­v io la tLlthS

f l . b rt . ra d -ra»a ,

f l . roaa

f l . y .

Viaibla-L U ftt,

vlolaCJJLshL

b it , y , diall w m w

n . y .

* b rt . o b rig h t, l i . w ^ iuoraaeant , # t. m b i . -I* hr* o brown, v* ■ valluv

tbbta 4* Rf-valoaa atid Colmr Raaetiona of Spot* <^haarvad on________________ q>r— tMwm c. riKur* »_______________

Flgura 3, Olagtiwa ^ ^ i n i « a U t lv « Foal*tloiMi of tha Spot# OhaMwad on tbo«Mllm«HMiotMl Pmpmt ChroMitoirarh Davalopad Ualnt SAW (F ira t Selvant) and I3%li!Ae (Saeond Solvent)

FRONT 80 1 r

FIRST DIMENSION

60 401 \ I----- T

20 ORIGIN_T------------ 1--------

Ang Aril T ^Ph, o

BAW

SECOND DIMENSION

65

Ta bU 5, Xf-valtiaa aiid C o l^ Raaetiona of Spota CMbaarvad on Qiromtofraph D, Flgora 9

Rf-value in MW l3«?Ae

Viaiblao u « .v io ta t V ia ib la v ie la t

! h . 0*81 — y . f l . y . - g r « , f l .

y*

b rt . y . f l . y .

An| 0*20 — p tn k -p ., iMuva

d u ll to f l . reaa, b rt . n . rad, f l .

roaa

f t

t s i 0*10 — d o ll pm or Ro

apta

aicifflfl. gtoiniton

IS L 0.23 0*64 BwganM f l . roao* P*

f d o ll■aganta

0,St 0,04 ft* F* b rt . y . b r t . y . b rt . y .

! h 0.22 0.72 Ro Ifo t b t.

!5 a 0.58 0.44 Rot Ret Ref y.*p.

"*15 0.76 0.50 Raf y* Ito? Ret

0.62 0.50 Rot Ret Ret y*• bet, m b rig h t, fl* • fluotm eoRt, ft* » fa in t ,

eolorlaaa. t • eolor taieartalii, b l« m bloa, gr*•

« greati.

Figtiro 4« ChrcMaocogvofh St Dlogrov Shoviiii RoUi€l,v» 2o«l« Clocui of tho Spot* OiNiociPod on tVo«»dlnon«iotMil fspor Chromtogimph Dovolopod Uolng B4W (F i r t t Solvent) «fi<S BAc*BCl (Socond Solvont)

FRONT 80

FIRST DIMENSION

60 40 20 ORIGIN\ -J I I I

AngcpC^SszksCl)2 I

• BAVV

SECOND DIMENSION

68

XabU 6, Rf.valtiofi and Color Roaetl^a of %»ota Obaarvod on Chrcnatograph I* Fifur# 4,

Inn* tnM ^V a n o r

Rf*valua in ViaibloLioht

U ltra .v lo lo t Vlaiblo

Lixbt

U ltra .v io la tLU K t

i2 Z 0,06 OaOQ roao*br,.roao

f l , roao mm tmm

0,84 «»4» y« f l . y . mm mm

1 0,12 mm b r , .y . oUvot r .

mm mtm

1 0,18 mm I t , t i r , .y#

«tf. mm mm

An, 0,00 0,40 roao? d u ll roao — d u llp**y*

Hi , .1*2 0,82 0,04 fl* y . y* fl* y*

!2a 0.60 0,61 4>«k mum « . f l . p.

H i 0,00 0,06 or It* b r , .y ,

O .lf I t ’r .br*.y#

d u ll ro d .l«p,

l t* r , d u ll rod.

b r.

mm

mm

«»d»

mm

H i

H i

m

m

tt

I*

«

W

m

m

9

•

m

m

• f l , m ftuomnnnBt* I t , • lith t* I t ’r , • eolorlaac* br« • brown* gr* m ttoan* p, i

lifhto r* Bo • i purplo* y , t*

»;■

iKi'-.'s'V'--a-.‘' ■-. \ - 4-' \: -"' :■

FM^ro 5. Otr«NMitogr«fh f t UimMmm mrnw^_ io la tlv o root* tioea of tho Spota cbaarvod on Cii#«dlie!anaioiial fmpm Cturooatotrapli. D^ovolopod Ualng 8AR

BAVJFRONT 80 60 40 20 ORIGIN

71

Toblo 7, Rf*wniiMNi iiatf O m lm f Rooetlono of Spofo oboorwod «mChroMCotroph F , Flguro S

Spot RAWin V U ib U

Cltra*v io le tLlobt

Vitro* V U ib U v io io t

0.71 jr. with fm Oldoo,

7*

f , b r. mm mm

1 O .lt ®40o*F«tpinkf

i t . V . , V.

mm mm

O.tA piidtf F«f y**F*t i t . p .

mm mm

0, 2« p . , b l.* v», nogontn

1 V , , v .* b l . ,9km U v .

mm mm

i 0,30 t y . t ibV .f mm mm

0.A6 Hot u . p. — * .

& 0,56 Not dll* p. mm mm

z 0,93 F «» f - •*

bt« m b liM , br* « brown, U iv , « Uvondor, pm Mnmla^ w V vio late w* i* wwllaw

Figur* 6* e i«g ra » mwmim Solotiv#tien* of tho Spots otmorvad cm tbm^dtieofMiioiiol tmpmt CiiroiMitotvopb Cowolopod Us in f 8AV (r in o t Solvont) snd tSBCl (Sseond Solvsnt)

FRONT

FIRST DIMENSION

ORIGIN

BAW

SECOND DIMENSIONQ ORIGIN

1% HCI

FRONT

7*

Tmhlm 9• Hf-valu4Ni Color Roactima of Spota t^ o rv o d on Cbrenatograpb G, Fititro 6

Color Reaction* inNH« Vapor

UmmtiRf -va lue in Vlaiblo

Llcht

U ltra -v ie la t Viaibla

v it r a -wiolatLlcbt

i S u 0.72 «Mb y* f l . y . b rt . y . f t . y .

(emCMr) d u ll dk. p. « d u ll b r.

X C .ld mm- troao d o ll y .-w . I t . b l . Ma

z 0.26 roa# f l . p M turn. b t . Ma

z 0,34 mm do I t . f l . y . mm y*

i 0.54 mm y* d u ll b r.1 FS4__ — 0

b r t . y« d u ll lar.

0.59 0.17 r* g r . -b r . b rt . y . f l . y .

0.71 0.21 m M « •

! h . 0.50 0.020.22

or bo{to

y*y*

Hado

fl* y . f t . y .

!2H 0.43 0.86 SoT t t d u ll p.

IS4 0.30 0.220.02

or Ho Ko

y*y*

MoMo

f t . y . f l . y .

X o.io e .ii plnlt do I t . b l . d u ll w.

2 0.2S 0.20 rooo n . rad in t . b l . w*

2 0.23 0.54 pinltt f l . b l . m f l . b l .

2 0.28 0.58 pittk f l . pljdi b l. fia

• brt* « b ri«h t| i f a in t , int* • is eelor uncartatn,

— s-a!atai„B«at,j

dtf. m d «rk , f l . m fittoroocont, f t . » tonoo. I t . m l ig h t , Ho • colortoga, f m

b l» « bluo, br« « brawn, g r . • groon, p.« v t o w ^ J K . . 1 ,1 1 ««

rigitr* 7* Ghromatoitaph K*H ihowing llolativ# Foal*tioiia of tho spota w^baarrad on tUo^ljKooaiocial Fopot Chmotofvoph I^avoiorod Uatnt BAR (r i r o t Solvont) and 60%IF iMmmd iolvoolM

•« GHroo«totm|>fe lt*2i ^tagraw ihoMtCNi Rolativoroa&tlem of tho spott Obootvod o» Tvo*d|jftOfialofial Fapor Chtooatogpopli Oovolopod Oa&iia BAR (F ira t Aolvoot) o»d 60%IF iSooofsd i# lva «itl

Fiittro 7* Chrowaeogroph H*3t Oiaiprao §tw»viiis BolativoFm itiona of tho Syata « ^o rvo d cm tOo«»dtaoiiaioiial Fapor OKrooatofmpBo Oavotopod Oain« BAtf (F ira t Soivoot) and dimiF (Socood Bolvotit)

Figuro tOt OuroiNitoirofh l^ii^rac^ BolativoFoaitiona of tho Spota CRuiawvad on ‘XW-dtoanaiooat Fapor ^ranotoiirafiha Savolopod Oalfti BAR (F ira t Solvont) and BOttF (Socond Solvont)

FMrtnro l i t QhrmotOAtoph Oiatr«iK Sh^ltig io la tlv oFoaitiona of tho Bpota t boorvod on lUoedtswiialonal Fapor Oiroootoaropfea Oavotapod Ualni BAR (F ira t Solvont) and 6 ^1 7 (Socond Solvont)

flco ro I2« atronota^Niph. &iof|tfaci '^^ovljig BolattvoFealtiaaia of ttu» Bpota ciworvod on 'AaawdiSMfiaional Fopor ChrowotoarapM Oafvolopod Oaing BAR (F ira t Solvont) ond S O ilr ( Sooond Solvont)

Th# six flguroa which follow, cm pagaa 76 trhrooih 51,

are rigttraa 7 through 17, raapactivelp*

FRONT 80FIRST DIMENSION

60 40 20 ORIGIN

SECOND DIMENSION

FRONT 80

FIRST DIMENSION

60 40 20 ORIGIN

SECOND DIMENSIONORIGIN

6 0 %IP

\l

FRONT

FRONT 80

FIRST DIMENSION

60 40 20 ORIGIN

SECOND DIMENSIONO ORIGIN

60 7oIP

FRONT

FRONT 80

FIRST DIMENSION

60 40 20 ORIGIN

SECOND DIMENSION

■ 0 :0 ORIGIN

6 0 %IP

FRONT

FIRST DIMENSION

FRONT 80 60 40 20 ORIGIN

SECOND DIMENSIONO ORIGIN

60%IP

FRONT

FRONT 80

FIRST DIMENSION

60 40 ?.0 ORIGIN

SECOND DIMENSION

Q ORIGIN

6 0 %IP

FRONT

82

Tsbls 9, Slid Osior AssetIm is of Spsts Obssrvod miChrosstoftmdM H*| tbrsimb IM l, Ftgwrss 2*12

Ultrs*Af*vslus In V U ib ls v U t s t V is lb ls v is ls t

£fis& m — m u _____ U itis -------------U8H£_______ l u b s . is m h i^

rifftt gtetpiiCT

2b|^ 0,70 •• y « , y » t ft* P»§

F*i br**y., die* p . ,

dk* br»

X 0,04 •* y*. Us d u U p*,F*-9*t y*»y «* 8 r., V ,

X 0*14 ** rsss*p«, f t . y « ,do, y . y .* v , , V , ,

y . t r .

X 0,22 •• y . I t . V . , V , ,y .* v ,

X 0,23 •* y » , y .*p . f l . b l .

X 0,29 •• dk« V#, f l , rossdk, v ,* p ,

t, 0,30 ** I t , V , , y , f l , V , ,y ,* v ,.

I t , b l,* v* ,U , y.*p,

2 0.38 •* y , y . « v , , y«*gr*,y « i y*»F** y»*

V ,

X 0,50 — y . I t , p . , y ,* v ,, **y*-9*

X 0,90 -M. y , f , »o I t , f l , V , , •*f l . V , , I t , V ,

•3

toblo ♦, &f*v«ly«0 and Color Roactiooo of Spot# i^boorvod on QiroMtographo K«l through H«6, flg u rM 7*13 (CentimMd)

im

__SifiSJL-.Rf*valuo In 9AV bOllF

Ultra* Violbla v io la t Vloibto v lo lo t

^LlJrht Ll.ho

Xfi 0*9S - T* n . b i . , f l . b l . . v .

«»0i mm

0.78 0.86f t . y .

g r .* p ., t y«*gr.*p.

y* y.* g r.* p .

« - u * ‘6,61 0.82 y» y .* g r.* p .,

y .r* y.*gr.*p .

y .* g r« ,y*

0.78 0.89 y*. f t . fm y.*bUt.or y .* b r ., t r .* b r .* y .

y . y*. y «* g r., y#*br.

0.72 0.87 ha dk ■ f>. , bl.owh.

!ia dk. p . , b l«*«h.

H i 0.34 0.004 Ko f l . V.g I t . f l . v . f ,f t . f l . V.

if# f l . V .f I t . f l . V . , f t . f l . V.

H i o .io 0.76 ha dk. p. lia dk. p.0.86 0.72 00 n» dk. p. «»«» dk. p.

H i x 0.33 0.82 Ro7 bl.*vh.9 Wot b i.«w h .f

S i l l 0*29 0.68 No bl.*ob. 0040 bl .*oh.

I h i 0.42 4 0*64

0.68 4 Ro 0.69

• «• m

O .l i 0.70 No y .* b l.* v .,pink

bo I t . V ., pink

! h i 0.68 0.88 f t . y , f l , V. v .« y .

H i i 0.26 0.84 bo dk. Pm mm dk. p#

84

Table 9. &f-valuea mud Galea Xeaetiocui ef Spota i^baaevad on Cbroeatograplia iU| throogb K-6, Figuraa 7-12 (Contlnuad}

U ltra - ’Xf-valu# In Viaibla v io le t Viaibla v io le t

■Scat,. „ M SL, &S111 i .U M , U t iiS .

0,65 ©,75 f t , y , f l . v , y , t fl# v ,t

t\k2\ 0*65 ©,82 ft* y , f l , v , y , t f l , w,

X ©,24 0,68 Re f l , b l« -v h , , pliSt,f l , b l , fl* c u r t ,.

turcs,-wfi

X 0*3© 0*77 roae y * -« h ,, b l , y , -a b , ,y,*p,* y ,» P ,t

8 r , -m « t r , - y , - m ,4-.

1 0,23 0,86 t r e d -b r . -t r , f t

^ die, • <ia«k, l(l,'• "^^le e re m e t, f t , • ^alliit, i t , • l ig h t , Re • eolerteaa, t - color oacerttin , b l , « blue, biV, • b u m , b r, e beemi, g r, » ftawy, P« e porple, te rg , • tereeeiae. v , e v i e U t . ab, - white, y , e ve ila v

Flgura 13. OiroaatoiWirli 1*1 • momiwm S a U tlvaFoalelono of tho spot* t‘hwmry<mS. m fopme OtroiratoarKfk tlaing lAV CFIratS «iv«ftt) and iClRAa CSacaful io lvn n t)

figura 141 OiraiMitagffapk 1*31 Ctagran Ihawtiii la la tiv a Foaiciofia of Cbo Spot# c«baaraod on two* dloooalonal fapor ^rowaconrapli Cwv»|op«o Calitf B4V ( r i r a t Solvant) and U « (Sacoad Solvaot)

Figwra I9 i c^hroaateogvafdi 1*3• Clagra# Showtiii gwiacivwloaitieaa of tho Spots ( hcorvwd on two* dlnoaaiooal fapor Chtonategrapli Gavalopod Calai BAR (F ira t Solwont) and 4<TONA© CSaroed Solw«Rtl

Figuro I (Si Obrenatotmfb 1*41 tiagrar Showii^ i^ la tiw «foaiti<ma of tho Spota Cbaorwod on TUo* dlsMMiaiooal Tapor Chroxvtograph I^awwlopod Utiog BAW (F ira t Solwastt) and AOtHAe (Soeond Bolwm t)

Figwra |7t Ci^raaatograph t*S« Clagraw Showing iw iativaloaitiona of tho spota rbaorwad on Two* diaianaioiuit fapor Osromitoiiraph CawolofHid Caiaa BAW (f i r a t Solvont) and dOYHAo (Soeond Solwwnt)

flgnra 181 Chroaiati^raph 1*81 Plagran Showing l^tatiwapFoaitiona of tho SpcMCa ha«rwod on IWo* dltMMBaiofwit fapor Chro*'*" totswph Ihrwolis^^ C « i i » 8AM (F ira t Sotvant) and A(5T(me (Soeond Solwont)

Iho aix figoroa whieh follow , on pagoa 84 th ro n g 91.

ara Flguaraa 13 thwtmofh lA. raapactivwty.

FIRST DIMENSION

SECOND DIMENSION

FIRST DIMENSION

60 40

C D C Ph,

7 6

■BAW

SECOND DIMENSION

FRONT 80

FIRST DIMENSION

60 40 20 ORIGIN

SECOND DIMENSION

FRONT 80

FIRST DIMENSION

60 40 20 ORIGIN

SECOND DIMENSIONQ ORIGIN

6 0 %HAc

FRONT

FRONT 80

FIRST DIMENSION

60 40 20 ORIGIN

SECOND DIMENSION

FIRST DIMENSION

FRONT 80 60 40 20 ORIGIN

SECOND DIMENSION

92

Tsbls IQ» Itfovstuuw ond Color S#««rflont of tpoto ibsonrod on ChroMtofr«)!hs 1*1 chrotigb I*#, figoroo 13*18

'*oot .Hf*v«loo tnaaw ftcmiAo

V lo lb ULiahr

---------M laa a »»

Ultra*violetL l s h t

aihL 0,75 mm y* dk, b r.

X 0,02 mm do d u ll p.

z 0.12 mm I t , r , , do f t , f l . V,2 t, y , , y# -v .

I t , y ,*p .

1 0.19 mm i t , V , ,do

f l , b l , , f l , b l,* v ,

y .-p .#

y .w *

X 0.24 mm It* V , , do fl* V . ,

f t . b t.

1 0,30 mm dk, p*, do

f l , roao, f l . V . , I t . V ,

& 0.32 mm p|j*,rlnit*p«, I t , p.

f l , y,*.dt , f l , V .

I 0,35 mm y , , »o ,i t , p .

fl* 7 . . 7 * .

f l . I t , V ,

A 0.39 mm y , , do 7***9.

1 0.52 mm y*, 3o dk, far,,dk, p«

Itm* v u ib to v io u eJ O t U . U l U -

n

tablo to* llf*v«liiot and Color Moaotiona of Spots <£baonrod OR Cbrooatograpba U l through 1*6, Figocoa I3 * li (Cootinttod}

Im

. I'OOt .Rf«vaIuo in SAW gOtHAo

tla lb lo1U&8.,

y «» No, f t . y , t

Ultra*v ia lo t Liaht

' ^ = ■ 1

n a ib lo

M i p g iUltra*v io io tLlabt

m 0.93 mm I t . p . .I t . V , , f l . V . ,f l . b l .

** «id»

11 0.98 mm y ,* o r«ty*

f l . b t . , n . V.

** 4|H»

0.79 0.79 ft* y . dit. b r. in t . y*ty*

y * tr *

0.63 0.80 y« I t . p . ,y*»p*t

dk rt P»Vdk, b r.

y*8r*»

y . t r .

0*82 0.83 y* , tar. . be.

y . y . < ^ . , dk. y .*gr

3 2 0.31 6.42 Ho T* •I t . f l . y . »

It* o.

t y , Cfadoo) y*.

y .* p .»goRo or I t .

V.

H i<■. ■

0«68 0.79 y .» W dk. p . . b l . t , 7 t. y . .

dk. p . ,P**pr.»

H i 0.2t 0.89 I I t . f l . b l « , I . It* V#, f l . m m . , y.w ch..

I t . v . t . I t . V .*t».| I t . v ,* g r ., f g r.«o h .

H i 0.00 to 0.A6 *•O.Ol

I t . f l . bl,V*

»• **

V4

7«bl« 10. and Color Roarci aa of spot* Cb««rvodon ChroMitoiropba 1-1 through l -$ , Figuroa 13-18 (Contliittodl

5oot Rf-valtto illUM some VlclbU

Dltra- 1 vlolat

Xll£JL-wIt. fl. V.

VlaibULiahtolSpa-violotLirht

O.Oi 0*62 Ho t U. fl. V.0.00 0.02 do fl. V. T T0.09 0.81 mm fl. turq. «• r.-oh.

«»10 0.88 0.69 y* V., dk. hr. — P»^11 0.32 0.29 mm dk. c. or

bllc». It. V. It. bl. •r v. dk. p.. It. V,

ih z 0,88 0.94 mm ft. V. —*

!Sii 0.24 0.42 — mm m>m Pm0.38 0.46 mm mm — dk. p.

I 0.14 0.68 pinkf, ft* rooo

tl plnk- V* It. bl*. bl.

T

z 0.12 0.74 ft* roao dk. p.t, ft. sr.-bl. t It . fl. bl.

1 0.18 0.8t ft. rooo It. pm ft, p.i 0.23 0.73 n«k, ft. roao

fl. roao-v., fl. bU. p.-v.

It. bl.«turq*bl.

fl. bl.It.

n . bt.1 0.29 0.78 int. roa*. fl.

roao roa*.fl. rod

bl«.torq.bl.

dk, p.

95

Tabto |0, afid Color Booeciofia of spoco Cboorvodon Cheonotogroplio 1*1 through 1.6, figuroo 13.18 (Contimtotf)

■Cato, ftM g tiu a lla

.-SaftL-

A

M «^ luo USAW- SSOEt£

VioibloLlaht

Ultra.wUlotLiaht

0,33 0*78 pink, ft , rooo

fl* V,

2 0,27 0,88 It, w,,V ,

dk* p*

V U ih Ut r o .

v io lo tLiaht

I t , h i,* I t , V,* tu rq , b|, o h ,.y ,

b i,* I t , p,g tu rg , b l , t dit, p,*bl,4Hir* I t , V ,

* ih , m iorit* i i , m ^iuorooeotit* i t , m #ola»* tot* mintomiOf I t , « l i ^ t . Ho • oolortooo* Y • color uocortota* b l , m bluo* b ib , m block* b r, « brown, g r , m grcon, o r , « oxongo* P* * purplo, furg , . turquoloo, r , • w tetot, wh, m wbtto, y , m yollow

when run in any ef tha thrmm aelvaat ayseam* Tha ftfevatuaa

and ealar raaarlaiia af tha yrttRinant tana vara thm a mad

for analyala ef the eeaq»o««d.

In additien ta tha aheve-daaerlbed reaa««elered^ar«a,

on# ethar, aefum ta, antheeyanln aana, An^, waa ehaarwed,

(Sae Fiinaraa 3 and 4 * «»)

tha poaitlen ef the roae-eolerad area on Chrenatograph

8 CFigure 4 ) waa ineanaiatant with that ehamwad an the

ether ehraMategrapha, Thia tneenaiat«eicy way ha hypettui-

aimed aa fellewa* Far aom raaaen, tha najer antheeyanto

eenetituant, An|» did net neve in SAV| the apet mewad stem*

m e t in RAe*!tCl (Rf « 0«40>, with an Rf«walua eanpamhl# te

that ebaervad in Chrenatograph C (HAc»PCl) (Rf • 0*37)

(Flgtire 2) for toe naior antheejwnin apet* The apet whiali

had an Rf-walua in MW of 0«06 did not nova in RAe-hCt*

thia «pet had an Rf-value eonparabla to that of apet Aa^

(Fignra 3) (R l • 0*1O)*

Table 11 preaeota the Rf-m lwea, in th m « aolvent aya-

cena, and eeler reactleaa ef the nnjor antheoyanin apet,

AOj, alang with eerraapeiuSiag data obtained by Park, tmwk

and Retar, and ottiar werlcara (A , 17, Saiket tn 20, 26, S3,

42, 44} for nalvidin 3,5-difluooaido, Alae praaantad are

date obtained far e peaaiblo niner antbeeyanin, apet Ang,

alang wich eerreapendtng data for nalvidin S-glwceaide and

aelvidin 5-glweeaida (4 , 17, Seihel in 20, 26, 4«> , Oata

96

T F Spot Any m a ataa obaorved on Chroautograph A,

tmhlm tt* and Colee Smetloiia of Antho^anln Spot* o i««rv «d m fmpmt Oire*M tetnfdHi Is Throo Solvonc SyattOMr, and Raproaastativa Cenparativa Rf«valiNNi

md Color RooetioM

■Coi ow 'Roao04oni.ini

Spoc

An.*

Vltxo* Ultm*Rfovalno In tiolbto viotoe Via lb to vinlot

M S L . i s m t L itfc t,, .L U h l J k L t iU U JL W iX .

6«27 0.64 6.38 f li roaeonv

rad*roan

btno doll nagonta

Fark (42) and Fmk and Rocar

(44)

katvidtn 3,5* diglneoaido

0.22 0.51

Dtma and 4 r d it t i <17}

a 0.31 0.42 flno*roacontcoriao

Jati»*4oith (4 ) ' a mauoa blna orbtnoHiroon

HM^Mrao <26> • 0.31 0.42 miaroa fino* roaeont eoriao

todorar and Sodocor (35)

m 0.22

Soikol (M k o l U 20)

a 0.31

tmhlm 11, aod Golimr RMietrioiHi of Anctiecfontn Spota Cboorvod m Pmpme Cliro* MCographo la tbra» Solvont Syatana, and R*r««a«Rtativo CiMnfnrattlva Rf*v«l«MMi

aad Color Rooectono (Cootlnood) .

ad^JboorSpot

ar Rfvooioo ia V io ib loKAr*HCl Llaht

8 ttf»*v io lo tLleKt

’ Oltro* 9 U lb lo v io lo t

Liftht Liehr

An^aa 0,08 plait d o llpwrplo

poltoo* fltta* broott roacoat

blao&UMI and A rd it t i

<17)holvidia 3*

glocoaite0.38 0,29 dotl

porpU

Saro*Sotni (4 ) • tNHOOa bluo

»iarboma (26) » 0.38 0,29 oauoo d o llnaovo

Focfcoc (48) « 0,34, 0,38

SoUcol (SoUcol to 20)

m 0,38

UwHi mm4 A r d it t i (17)

K atvU ia 5m ftlnroaido

0.43 0,22

Sato*teitii (4 ) m OOOfVO btoa

Tablo 11* RfoVftluoa and Color Roac^rlocia of Anthocyanin Spots Obaorvod on Fapor Chro*Mtogra]i4ka in Ihroo Solvoot Syatooo, and Roproaaotativo Ceoforativo Rf««alttoa

and Color Roaeticoa (ContUiood)

Cclar &

. . Woirfc.«c_........

Spotor Rf*val«o

. M IL 122il4fi 0.51

inPAg. ^ 1

VlaibloLlabt

Ultro*v lo lo tLiiiht

iimm and A rd it t i (17 )

fbitvidin porplo nonvo

Bata*Snltti (4 ) m naovo

iiarb om (24) m 0.S8 pnrplo 1MWO

litre - ▼Islbto v io io t■ JLU © t„, J k U h t

firay-blno

Ladoror and lodarar (35)

0.40

^ Ao« ia ahovn in i^igorao t , f , and^ I* ahoon In fignroo 3 and 4.

to

fo r «ho oflyoono, m lv id ln * oro oloo orooontod <4, t7 , 24,

3S)« It coy l»o cooii from Toblo tl that ctio major anthoeya.

Clin apot ia m a t lik o iy aatvidin 3,S«dtglocoai4o, Farit *a

roaulta aro thoroforo fupportod*

Sfiot An^ waa «PoaJc and did not appoar ropoatodly imdor

difforont m poriaontal aitoationa, and tma not obaoroad in

tho aococid dinanaiaii* Cn tho baala of ita Rf^^yalua and eolor

roactiona, apot Any waa lifcaly noithor nalvidin 3««tne<niido

nor ataWidin 54kglttcmido, botiaivod to bo poaaiblo eea^oaian

conpounida to naW idin 3,5<»difIturoaido (Swain in 23, 3$)»

(Soo fablo tl*> ninor apot alao roproaoetod a poaaiblo

indiaation that aa additional anthoeyanidin nay bo froaoot

in tho potala of isnftslCSBSS* 7f i t ia proaont, i t awat

bo tn traeo cencontration or in a highly tmatablo fo m , and

nay bo roaponaiblo for fdtmotypie color variation obaorvod

In thia apocioa* According to iiarbomo (Harbomo in 23 ),

flowora with nixttaroa of th# aatho^raftidin i^^poa Cpotaryimi*

d in , cyan Id in , and dolpliinidin) uonally ara of tho wapoetod

intocnodiato color ahadoa, with polai^onidin roaponaiblo f « r

pink, a ca rlat, and orango«rad cotora, cyanidin roaponaiblo

for crinaon and naganta* and dolfhteidiii roaponaiblo far

nauvo and bluo* I f ona typa prodcaiinataa, aa apparantly

oalvidia 3,3«<ditlocoald« (a dolpliinidtii d orlvativo ) would,

thora ia good ccnrrolation botwoan tho prodoninato typo and

color*

I t nuot alao bo conaldomd that tho additional anthoeyanln-

lik o Afip, m y oljRply bm m dogrodatien proApnt otthm

•f teho m jo r anthmyonin ahawn to bo proooot. m lv id lA 9 .5 -

diglticeoid*. or of aono othi^ cooptnmd. I t m y bo aom fvcm

tablo I t . hooovm. that m W id lR . tbo ogtyeono of tbo m |o r

onthoejranln praaant, is not Indiootod as on# of tboao broak-

down produeta.

In eoRoloaion, m tv id la 3,5-diglucosldo uaa found to bo

prooont in tbo pokalo of ill* sanduiconaa f lo m ro i Idila find­

ing eonfiroad tbo rooulto found by fork. A aoecind ontboeya-

nin uaa indicotod, but u itb tho lin ito d m owit of infoem tion

obtoinod eoncoming thia oin<Hr apok» i t voa iopoooiblo to

mko * tontotivo idontifieotion of I t .

Clooa te tbo idontifieotion of tho minor ontboryanin

apot m y bo goinod front o eonaidoration of anthoeyanina pro-

oont in oidior Coawedluo opocioo and in other oanbwro of tho

Logusiinoaoo. holvidin (4 4 ), doiphtnidin (9 , 4 5 ), and

cyanidin (9 ) uoro roportod aa eonatltuonta of fiouora md/or

laovoa of Doogodliio* Cyanidin and doiphtnidin, at loaot,

uaoro indleotod aa eonatltuonta of loovoa In nany LMtuninooao

apoeioa. Doiphinidtn 9,5*idlgttteoaido had on opprepriato

Af-valuo ia BAW (Af • 0.15) to b# eonaldorod oa tho eddltion-

o l ontbocyonin (1 7 ). Ib io anthoeyoaln, bowsrmr, ia fluoroa-

cont furplo In u ltra v io la t lig h t (1 7 ). tho d u ll purpto ettlwc

of opot An^ eould hov* boon duo to lou eoneontrot ion, i f tttia

opot did iadood roprooont thia onthoeyaoin. Ovor-fwnrifieocion,

on tho port of Fork (42) and fark and Rotor (4 4 ) m y hava

101

eauaad tost of this olnor eonstitosnt fre » th n ir oKtrsets*

ysfa«,.ribiaallc„cttBBawatii

I t tiss found that tha aoat d iatlnot

spots wars obaorvad on ehroaoto«r«f»ha in which mony (OOa)

fiowara had btto twiwd for pifptofit ortraetion* Oftan tho

aopply of floMtrs waa lin ita d . thoraby UtpoaSjiig aooy lin ita *

tlana eaiuiad by waalt ayota. U ltra viotat apoctroacopy could

04^ bo uaod offoctivo ly und«r such conditiona. Htny ai>ota

wwra obaorvad on oach chroauitiu(rayh. oftan ovorlapttng. and

dalinttaclena of tho spots woro d if f ic u lt to dotarolna.

With aueh l i n i tat lone, tho p«Hrity of tho litdlvidtMii apc^a

waa doubtful.

Iho author obaorvod <mo or two najor yallow spots, and

about twmty*ono oiner apota of vtryiag colors, tboao aro

diaeuaaod In tho foilow int aactiona.

7«> taJag

Caa najor non*anthoeyanin spot was

•««B on oao*dlo«iaional runs. A ta ilin g offset of thia spot

waa obaarvad. tho spot wta m farrod to aa spot Fh| « r

dopaoding upon whathar ita color In w ltm vio lo t lig h t woa

flu<Mroaeont yollow or dark brown, roapoctivolyi ovidettco

indieatod that thia aojor spot raproaantad two conponanta

althor suporinpoaod or oworlapping. Xn tho aoeond dinonaian.

spat fhj^ raoainod intact aa spot rh|*1.2t howawor. spot fb|^

althor roaainod intact aa spot fh|^*|,2. or aopamtod Into

two horiaotttatly adjacant apota. fh|^*| and fh|^*2»

102

oeeupiad afproxlm tely c$ie — sm arm aa apoca

Fh|, -| Mid ibi| <i»2 eaatbiiiad* tha two apata tmparimpmad or

ovarUppIsm In apota Ph| and Ph|^ ara rapraaaotad hy apota

and yh|^-2 in tha aaeond dlaaMialan* A tmmmry of tha

maraetMriatlea of thaaa apota ia praaantad in Tahla 12*i

tha aaparatian ef the n a je r apet in to two apota if id l»

eatad th at apota rh| and lb|^ rapraaantod a n tx tn ra o f two

conponaata, Gn tha baaia of ro to r In w ltra w io la t l i g h t , ono

o f thoaa cowpmanta, f h | ^ - l , corraapondad to usaior apot 2 t -

w hita rh|^»»2 aorraapondad to tha tm io r apot fh | ^ , Ferhapa

va ry in g anparinantal aonditiona eauaad <aia o r tha othar of

tha m j a r apota to bo naakod la tho f i r a t dtnanaion, u n t i l

aaparotod aut la tha aacond-dinanaional nta« th ia hypethaaia

waa aupportad by tha f lra t-d ls a n a io n a l nm of Ghrmatogtwph

<3, In which tho m j o r yo llew apot appaarod In o lt r a v ia la t

t ig h t aa a flvMcaacant yaltow apot w ith a brown apot appear-

tng a t Ita lowar rantar* Stieh an a ffe ct could hawe be«e

produced by tho oupariagoaitlen o r overlapping af to# two

apota fh|^^-l and fh | ^ -2 , F u rth «f atgpfsert e f th la l^pothaala

arlaaa fran tha ccm aidaration th at tha brawn c o lo r ef apota

Fh,^ and lit, ^ -1 ,2 in u lt r a v lo la t l ig h t could have rapmaanCad

m

the vlaual appaaranea of a nlxtura of tha eolma yallew and

purple (3 8 ), which for aono raaaoti appaar«id aa yallew alone

In apota rh| and f h | -l ,2 « Tha two mnpaaianta, rh |^ -l and

£hj^£-2, nay thaa hava bean yallew and jnirpla, raapactively,

or aech nay hava boan a eonbination of thaao two colors, aa

104

Tablo 12* Avorago llf*valuoa aiul Color Aoaotiooa of tho Ma* jo r Tallow Spota Obaorvad oo Qna*dioaiiatonal and IVao* dijM M ional Fapor <%roaiacoira|)ba Oovolepad Ualng BAR ( f i r a t

Solvont) and Soeond Solvant Aa Indleatad

Z laJU B lasa iiaa

0.82 0.05 7* n . y . 7* f l . 7*

0.72 7* d t. b r. 7* fl* 7.

Th.m 0,82 0,04 0.04 7« n . y . 7« f l . 7 .

' i j r0.78 0.82 0.79 7. dk. b r. 7# 7»*8r.

10.80 0.82 C.80 7* 7.«P »

7 » t r .7* 7*-«r*

0.78 0.87 0.83 0.21 7. y .* b r. 7. 7 » » i r .7*»br.

>b|t fliraroa t, 1, 3, ond 4i fiattroa 5«18|n » f l ,2 i Figwoa 3 and 4| fh,^*|.2i fi«nroa 10, 12,l i , l i , and iSt and Fb^^*l and Fh|J*2t Pignraa 6, 7,8j 7, 18, 17, and 18.

** V U « • V la ib ln , wt « U ltra v io lo t, I t . • l l t b t , b r. • brovn, t.r* •» groan, p , m p u rrlo , y . « yollow, m . m d a rt, f l . * ftnorooeont

*** Rf*valttoa yroaoncod for BaV or# thoao ebtalnod a fto r ra* naaanrooiant in ti»o aoeond dlnonaion.

indleatod by tho eolor rooctlone shown tn Tohto If*

A lto m o tlvo ly , In view of tho ro lo ttm ily o in llo r tf*

vslooo obtslnod for spots end tn tlAW (0,8? end 0*7?,

rospoetlvoly), i t woo poosiblo thot tho two spots roereoontod

tho s«no compound t tho color in u ltrs v lo lo t lig h t stsy h«vo

veriod froo ono chroovtogrsitih to onothmr duo to difforoneoo

in oxperlmontsi eonditiims* Intotootim ily, spot fh| oi»|>eerod

on thooo ^ro o o t^ro p h s firoporod froo ooth«nel*lfCl OKtrsets,

and spot i l u spposrod on chronstogrsrhs pronorod ooinp

oothonoi ojrtroets* tho tsothod of ootrsetion, ond thus tho

a cid ity of tho aatraet, night hovo aecoontod for th# d iffor*

ing color raoctiona of whot nay havo boon o alnglo coopoond*

Although cho yotlew plgsHmta aro not as sanaitivo to mm

ara tha enthecysnlna. I t ia known that color chongoa with pit

do oecwr anenf thoaa eoopotmda (Swsin in 73), in thio eaoo,

tho two d lotinet opeta obsorvod ia tho soeond dinoneion

oould oetuslly bo ono largo spot o xh ib itin i dioeontinoittoa

duo to oroos of low concontrotton,

tho hypothoois thot opots Pb| and roprosontod ono

coopouad doos not, howovor, aeeeunt for tho d lffo rin g color

roaetlons in w ltro vio lot lig h t of tho two spot# soon In the

soeond difconsloa* tho ovldoneo is thoroforo otrofigor for

tho hypothoalo thot spots fh| and rh|n both roprosont tho

suportopositlon or ovorlapping of tvo e«mponont«, thoao

spots probohly raproaontod difforont oppooroneoa of tho sow#

eiaturo of conponcBts* th is dlfforoneo in arpooroneo could

105

liavo duo f dlfforlfig Mrporlaoiital cmditiosio.

Zn tko «•••• «h«ro only on* largo apot. fh|*l,2 orMMMIgMWOnaO

Pfc| e| ,t f waa oloorood in ttio ••cend*di»osiaifloal moa, chia

apot rcvorad tho aaoo roglMi aa tho two aoparoto apota,

and Foatiapa thl* largo apot woa la roollty

two apoto, wlii^ for aooa roaaoo ladiod dlfforootlotloo* In

only an# mn. Chromatograph Z*S, waa tho largo amt India*

clnot, eovMtlng only tho aroa of afM»t Fh,^*t. In thla eaao,

tho proooeeo of apot Fh|«»t ««o oooationahloi it night hawo

boon oithor proaont tn mmII eoneacitratlon or obaont. SInco

tho appoacanea of apot Ph|^*l mao oooattoiMbla In anty thla

run. It woa aaanmod that thla waa doa to tow coooontratlao,

and thla mn waa Inolodod with thooo in which both opoto

woro proaont and aoparato*

In Ibblo 2S (Appandiat) aro ontliskod tho color roactiooc,

oo papMT, of twoiwo claaaoo of flavonoid eoopowndo (Soikol

in 20, Soiltol in 25), oloiii vith thoco of ph4Moiic oeida

(5, I f , Soiltol in 23) and oilaglc acid <3, Soiltol In 25)*

Ihia chart ma nacd to r tho paatpomo of t«ntatlvoly Uhmtify*

Ing tho elaaa of tho j^Moiotic coepoondai howovar , it could

only bo wood ao a row^ gwidolino in phanol idamtificatlaii*

Ita oao waa also llnltod by tho fact that color roactiona

and dollnitatione of tho apota war# eftoa difflcwlt to dofino

accuratoly, duo to aovaral rosaonai a ) low concontvationa

af tho eompownda prooonti b) dtffaroneoa In IndivldtMtl

Intorprocationo of colorst and c) tho vioiot bookgrovmd aoon

106

t o r

undwr u ltra v ie la t light*

On thn bntin of tth lo 25 (A|i»pondi]c}. tho eooyotnidt fqmi«

ing tho M ijor yollow tpott noot lik o ly awy ho olniiood on

f Uvonoo « nd/or fUvonoU* Boeowoo « eonptoto l in t Ing of

Rf*v«luon In vnrlowo solvontn of o i l coopotmds fren thooo

two oUtnon wnn not owniinhlo. i t won inponnihto to idontlfy

tho vnUenown eonpownds from inforwotion to bn found in tho

lltomttaro*

7 «) r,ism i r a t i

Smcopt for tho oppooronoo of ontho*

eyoain and tha najor yntlaw apot(a)» tho aawoml ehronato*

graphs anhlbitad faw a in iU r it ia a with on# anottiar with

raspact to eolor rooctiona and nunbiMr af apota* I t was

d if f ia u lt to corralato apota fren ona chronotograph with

thoaa froR anothar. a noeoaaary atop i f on« ia to eoltoet a

aariaa of Rfwwnluoa In difforont aolwant aystana far an on*

known conpound. Sueh a eolloetion would pnwrido an inwnln*

abln old tn tho idootifiootioo af tbo unknown*

i MMro pooaiblo. tho apota fron tho vnrloua rbronato*

grapha woro groupod aeeordlng to th o lr a ln ila rltio a In Rf*

wnluoa ond color roactions* tSbla 24 (Appondia) proaanta

thoaa graoplnga. for pmrpoaos of eoiu>aria<m« and prowidaa

th# baaia for tho nwnbar lng of spots diagranad in Figuroa

3. A . and 4*14* Siaco tho colors wora oftan wook or d if f i*

cu lt to doaerlbo. color rooctiona twura not glvan aa strong

a conaidoratlon aa Rf*waluoa in grouping tho spots* Thoao

•p«t« mrkod «d.tK an aatariab could ba oithar plaeod aiooo,

or lato ofio of sovoral groupai fdio otiMMr fioacibto groupa

aro dooitoatod uodor tbo eoluaai ontittod "Rooarka** Som of

thoao apota ocourrod eo noro eban eno ebronatografhi otbora

appoarod to bo diatrtbotad randooly fron ehronacograph to

cbrmMtograplni otbora waro obaarroS oo only eno ebronato.

grai^« lha quaation aa to whotiior ctio ninor apoto ropraaant*

ad original ccntp«ma«ita of tbo axtraot or produeta of

diHiradation waa inpoaaiblo to anawor without furthor

onporinontation•

Roouita ohtalnod for ono paront war# Ineonaiatont with

thoao obtoinod for tho aana parant on a difforant ohrooato.

gxaph. It waa onroaliatie to try to difforontlato anong tho

paronta aa far aa tho proaoneo, abaoneo, and eonoontration

of apota woro coocamod, Ih«ro waa, in faot, no nood for

auch a diffarootiacion ia tocna of tho gonotle analytia, aa

no offapring woro producod fron ottonptod eroaaoa* Such

lefomatiee n i^ t havo bo«>» uaafui, hx«wwver, in dotomintng

tho rotativo eentribtttima of tho conpounda proaont to

phoootypio flowor eolar*

3* Aaaoaanmt of roaulta in m la tio n to orovtmitt

yorl^

Facit (A3) and Farb and Rotar <A4), uaing two.

dinonaional chroiMitographa dovolopod wich phmot aa tho firat

aolvant and BAW aa tha aoeond aoiVMit, obaorvod altogothMr

twolvo apota* In addition to an anid ecyanin apot, 4, thoy

toi

obaorvod aight spat*. X I through Xft* war* yaitaw i »

v ia ib la l i | ^ t , and thraa apata. JX through |X» which vara

not v io lb lo , but which flucroaeod, with oMoonio vapor, ondor

ttltro vio lo t light* Foi^ did not atato tha color of thia

f liaoroaeaneo* Tho roaulta of tho abovo atudy woro ctoa r-e ut,

and eorroaponding apota wora d ia tln c tly aaao an hia aavaral

^ t o m togra pha •

Aoaulta of tho currant atugy woro oot oo c lo o r-c u t, and

apota woro prooont on tho chremcogropha which woro not fo«md

by Farit* Thooo diffMroncoo asay bavo bo«n dhio to difforoncoo

in toehniouo and to tbo oatount of purification ritirit Fotf;

uood in proporing hia oxtraeta* Ho nay wall havo oinut-

purifiod hia oatrocta in tho oooao that ho loot oano of tho

traeo coaipouBda found in thia otudy* A a in ilo r implonotioni

m y account for tho fact that ho oboorvod only ono onthecyo*

nin opot. vborooa chio otudy rovm lod o poaaiblo minor antho*

ejranin in oftdition to m lv id in 3.S*diglueooido*

thaa. a Ruch nero epmtplmt aituation than chat doacribod

by Farit ia auggoatod by tho prooont rooulto* Hlo cloar-cut

findinga in uaing a oinglo cwo-dioonoional aolvant oyoton

rovoaiod only part of tho phonelic coopc»mda proaont* havar-

chaloaa. hia toctmiquoa noro e lm r ly rovoaiod tho yotlow

fUvonoid pignonta not wall ooparotad by tho ayatooa uaod In

tho e u rrm t otudy* Tho aolvant ayacooo uood in thoao anpori-

nonta rovm lod thooo proviouoly undooeribod pbonolico. but

intoroetiono omng coopounda and poor oatraeto iatpadod tho

109

n o

rmolttCioa astd i4«ntlfle«t& on of Cho ovorobiaidoiit conpononto*

Sovorot footoro coaiptlefttod tho pcodvrtioo of oKtrovto

oopoblo of pioldiag mromtogrorhii obleh ooro roodlty Intor*

protod. for OKonpto, obtoinlfig « a o ffltlo n t ctMfttltr of pot*

•lo for phonol oKtroetloii woo d iff lo u tt duo to ttio oooll

■l»o of tho floworo «od th o ir Uek of otnaidoaeo on tbo plonto*

Aloo, eocMoatrot&on of tbo p&gjRonto followisg m trte tio n woo

eoo^lleotod by tho oforoodias offoet of tho nsthonol tn tho

ouoporotittf dioboo.

I t lo oucgootod thot ia furthor work in tblo fto td ,

nero plonto ohould bo grownt oloo, floworo obmld bo

gothorod ovor o pnriod of oovorol d«yo» thoy cMty thon bo

d rlo d , and tho oxtract nay ba obtalaod fron an arennolation

of driod potala. A lto m a tlvo ly , tho potala nay bo pUeod In

tho ro frigo rator in nothanol*HCl or m tbanel, dopondlng upon

tho ontroetion nothod to bo oaod, and tba oxtracta nay bo

accnnulotod ovor o pmiod of aovorat dayo.

Socondty, tho o xtn cta ahould ba eo(maiitvatod to dry*

ttoaa in o eontatnor u itb a anall bottan aurfaca aroa, auch aa

a v i a l , cm trifu g o tuba, or a anall ourclbla. Ibon whoa

tho oxtroctlng aolwont la addad to tbo driod oxtract. In

proporation fo r application to tbo papora, tbo dtaaolvod

ontraot w il l roania eonfinod to a anall araa.

Froparativo ehronatograpby, tmod for inn’poaoa of pnri*

ficatioB of tho o M o e ta , aorwad anly to loaooa tbo concon*

tro tie n of tho pignonta. Tbo R dhroaotographa (R*t through

tt«4) were prepaid®© followiiig prafMicativa diraiNitofraphy,

whlla tha X elir«mat<^pc«mt C «*l threttgh l> 4 ), udiieh aahihitad

Rf-m luaa ra la tiv a ly a in ila r ta thoaa ef tha h ehronatographa

for tha Btajor anthM^NHain and tha najcar yo llov apota, vara

praparad vithont i t . lha only d if f ar mcaa vara • ! ) m a

diaappaacnnca, with t in a , of tha najor anthoeyaniti apot froai

th# R ehroantogra^Ui 2 } tha appanranco of d latlnet ya llav

^ la * ^ *4d Ph,^-2 In runa of porpla-f tovnrad plant# in

H (H at, H*2, and R »3 ), hut to vhlta-ftomMrad ptm ta to t

( t « 4 , t -S , and l«4>i and 3) a a m lta r to ta l nunhar of apota

to tha M chronatogxapha than to tho I mrooatographai aoat

of tho oKtra apota to H wora aluatarad naar tha najer antho*

eyanin apot* thia altoination of apota nay ha tntarprotad

aa a purlfieatien i f i t ia aaaunod ttiat thm apota aitotoatad

wara artifaata or topuritloa, Qa tho othor hand, tha a lt o i -

natad apota nay atoply have rapraaantad eonpenanta of tho

m tra at proaant to anall eoncontratiomi aa auch, thay oould

aaaily hav# Imhhi toac durtog tha pwrifieatlon ptoeadnraa*

OiffM ont tr la la appaarod to yla ld dlffarant raaulta,

and d iffie u ltio a wora ofieomitarad to viauallaing a l l of tho

eonpounda praaont, duo te the problona dlaeuaaed provioualy.

I t waa unreatiatic to a ttm p t to pair tha oon»«»theeyanln

apota obaorved with tho yollew or fluoroaeont arota aeon by

Parir (4 2 ), for aovoral roaaona. the lorgoat 3f*vatuo he

obaarved to BAH waa 0.9€i to contraat, aovon apota olwiarvad

in tho preamt study had Rf-values groator than 6«56, thla

I l l

M j h «v« ba«n duo to tho ordor lit « d i l^ tho oolvontt woro

opptlod to cho tvo*dioo(nologi«l ehrcwotiHiropho* Soooodly,

tho color rooetlofio of cho oi?«co ohoorood in thio otudy voro

d if f ic u lt CO dofino* f ln o lly , i^o oolvoot oyotooo uood in

tho prooont otudy ond in F«rlt*o work woro d iffo ronti Potlr

proporod two-dloonoloMit ehrofwicoiiropho uolng phofM»l ond BAW,

in thot ordor* fhoaol woo not uood in tho conront ocnrtr duo

to poroofMtl hoolth liosordo*

Idoncifieocien of tho tndlviduot yollow ond minor opoto

rocuirod ooro doco ond roforonco infomotioa* Spootrol doto

ond flcnro oactonolv# lla to of Bf*v»luoa for thoao spots free

o nunhor of solvonto umto noodod* Iho loportsnco of pwrity

in Cho spoctrol onoiysio of ioiknown eoKpounds hos boon dis*

cusood by oony work ora <19, Horbomo in 4 9 ), On th is basis,

spoctrol OBolyolo «mo ioproctlcol os on old to tho id o ntifi*

eotien of tho nen*ontbocyonln spoto* hero concontrotod,

pure CQopounda oro rocuirod*

B. Biuggflm

I* ABittJLSB

Aglyeonoo woro obtstnod by oithor tbo oothonol*

HCl oothod, followed by hydrolysis, or by cho HCl nochod*••

In both essos, the oglyconos woro wsshod fren tho kydrolysoto

uolnt a ry l or lao*anyl alcohol* Aglycono apota aro roforrod

to oa 4>| tbraugh Aq4,

tabla 13 outlinoa tho proporotiim of chrosotimrapha

onolyaod for oglyeonoa* AAV, forootal, and TAV woro l^o

112

•elvwit «yat«M ehoaon for ebr<MNitogra|iby off tho ogtyeonoo*

Tho chrtHMitogropha ara diagramad in flguroa 19 fhroogh 251

tha apota ara daacrihod in Tablaa 14 thrcnagh 30»

8AW waa aalaetad boeaoao fif•valuaa fo r many of tho

aglyeonoo aro availahlo i«i thla aolvant ayatow. Bato^Salth

(5 , Bata*Smith i » 49} doaerihad tho eharaetoriatiet oM oao*

fttlnaaa of Foroatal and tMt in aglyeooo cHrowatMiivi^y* Ob

ehroaiatotrai^a davatopod with foraatal, glyeoaidoa rwi faatar

than aglyeoBoa, and tha levar half af tha papm, naaror to

tho o rig in , w ilt ho fro « froa any ucdiydmlyaod roaldtsaa,

naking datactian of agiyeonoa in thia rogian ornrnfm Ovroan*

tegraphy oaing a aacmd aolvant, nm aaparataly, la naadod

to roaolva tha eowponanta of tho uppor holf of Fforoatal

ehroantc^rapha, whieh tanda to ho erowdod with lo«a highly

hydronyiatod agiyeonoa and roaldnoa of uidiydrolyeod eonpianoa.

In ^ i a aoeond aolvwnt ayatan, TAW, tho aglyeonoo w i l l ran

in tho aana ordar oa thay did in tha Foroatal aotvant am*

tarn, tmt glyeoaidoa, oatora, and cenatltvonta with gf*valoao

of looa than 0,90 in Foroatal w il l f a i l to novo fron tho

M rigia,

!• CllgMlliaB

« • XAllflM, IfilTBimtt

tn tho foroatal ehronatographa, two

yollow apota, a nsajor apot, Ag|, and a aaeond apot, Agg, wara ohaarvod. <Soo P|g«troa 19, 21, 22, and 23,) Iho

eharaetMrlatiea of thaaa apota aro aunnarlaed in Tahlo 21,

113

114

Tabla IS* Oaaeriptian a f Fo ta ria lt Caad la tha frmpmtmtlmt of Chroant^raidia Analysad far tba fraaanea af Aglycanaa

Chraawtatrapik Faconta baad SntraationMothod

SolvontAea CAM

J F t , r s , m , , F7, r « l j . H i

Hithaiiol*IH31. follooad by

bydroljraia and woahiiic with anyl alcohol

Fomatal

It F l . FS, FASj. P?; P ^ , , f \ i

• BAW

1. m Forootal

MTp :

m Foroacal

P I . P4, PS, P i » l » P io . p ft

net Focoatal

o » P I . P4, PS, P tp j. Plft. p fl

mx BAW

r F I . P4, PS,p 9 «i* n o , p f l

»C I 1AW

Q n . m , . RGl 8AV

* Chranat^rapb 0 la not ahaan in tha ftgnma aa ita apota oautd ba tdontlfiod*

nano af

FLguro |9« a»jreiiiito|ic«{>h <21 Ulngnwi Show lug t « U t l v « fo «it io ik « o f tho Spoto ci»o«rvod on Coo* d looo ti oaol Fopor Chrooototcopb Dovolopod UoUig Forootol

Flcwro tOi (%f*oiMitosK»pli l i D i««r«o £%e«iii|t ftolotivo Fooitlooo of «bo Spoto cteowod on d^o* diMNioiomI Fopor Cbrooocoiirofdi Dovolopod Uolnt SAW

FRONT 80 60 40-FORESTAL 20 ORIGIN

FRONT 80 60-s£ BAW

40 20 ORIGIN

117

Tablo 14* Itf-valuoo nod Color Roootioiia of Spota Cbaorvod cm Chreowtcgtoph J , f lg w o 14

Salat.Jk

U ltra -Rf-voluo ia V U ib lo vioto f V iaiblo v io U t

Xsax Jgagta.uL.,., .XUfat., Llabl, , JSUaiL, kUfet ,,

Ag. 0,50 So y , y ,f »©ro ia t ,— A y ,

Ag* 0,58 I f , p, roao t yo llo o -lab (tofida te fado)

« ia t , • tataauao. I t , • l ig h t , ho « eotorloaa. p, m purplo. y , • y l l o o

Tablo 15, df-voliioo oad Color hooetiooa of Spota i^ o rv o d on Chr omatograpb t . figuro 20

SoeaAfwvaloo ia Viaibla

Liahr

U ltra ­v io la tLlahe

V U ib loLiabr

™ R i « r -violocLlatif

Ag|t 0,91 y , -b r . int, y* ia t, y.

MBttHMbmmaoaa*

lo t , y .

1 0,2? So y , -o h , -v « .v ,-id i.

y,«v,*«dt,v , —ob*

2 0,38 Ko y .-v . ho y ,-v .

1 0,98 y , -b r . ?•»y , -b r .

y , -b r .y ,-b r* .

* i » t , « iatoaao . ho « oelorloaa. be. - wrowa. V, • vloiot

‘igur* 21 • i t Ciagcom Sbowinc R oU tivoFoaieiona of tba Spat* Obsarved on Qfio* diaMiiaionol Fapor csirooiitoitiiph Covolopod Uaint Foroatali nombromtography of Spot A g ., Chnwatosmrb L« UlatraB ^howiag *— * fU lotlvo foaitiona of tbo Spota Obaorvad on OmMdinonalotMit Fapor Cbronatogcaf^ Govolopod Uoisui 15%«ac

FRONT 80 60 40- FORESTAL 20 ORIGIN

I I I I I 1 I T5 3 2 _______

A g i ^ Q A g g ^ ( )

FRONT 80 60 40- 15 7o HAc 20 ORIGIN

12®

tmhlo }6* &f«valu«a «imS <^lar Raactians of S|»ot« Obiiwrvad on OiranntogTOpl) L and on fUH^hroancografk of Ag| fron

dimnaCoAmph L . Figom 21

af*v«lua inSoot

V ia ib ULlniit

o im *vta la t V U ib ln v ia la t

Llaht Llaht

0*54 y* fl* y# In t . y* in t . f l .y*

H i 0.38 y* y» y* In t . y .

X 0.77 y**br. b r. y.*be. b r.

1 0.87 y . -b r . nbr.«nr.

y .* b r. y .* b r .«v « .b r.

1 0.91 y .* b r .t It* y*»I t . y.vw., y .*b r.* v.,

y*-ad».

y .* b r ., » *1

I t . y . .

y .* b r.* v .

i 0.93 y »* b r.. y .* b r. f y.*bt**v.,

, F . i I t . y .

y .* b r .. 1 ^

y«*tef«,y «* b r.* v ..

I t . y*

1 0.995 y .»b r« br**v*. b r.»b lli. .

b r.

. y.*bir. Iti ''* * 5 b r . . bllt«.

b r.

^•.ohmaat^raph af Ag|

XSmxs.Yallaw 0.10 e«

o .isyallaw f f t

Fnrpla 0.90 ca 0.95

parpla f I f

Mha g,,liCUt> .Ti w YtliUnf

Flgttr* 22* C2immiCotrfipli Hi Uiotram IM^owinji Rotativ* Feaitiana of tho Sf»eta C-hooraod oo 0 »*» dlJMmaiottal fmpm ChroMitoitraph Oovolarod Vatng Foraatal

flgura t3« QiroiKaeatraph di Diagro» ShMrtng la la tiv * Foolttafia of tho Spota Chsorvod on uno* dloanaional Fapor Chrenotegmfh Uavotopod U«lnp Formital

FRONT 80 1 F"

60“T~

40< FORESTAL

20 ORIGINT T \ I T

-FER U I SINIAPiC ACID ?

^ F E R U L IC A N D /O R

( T ) ( )

FRONT 80 60 40-C FORESTAL

20 ORIGIN

123

tablo 17. 8f»v«luoa and C^lor Raoctiona of Sftota €%aorvotf on ChroMtc^rajdi K, Flgttro 22

Rf*valoo in V U lb laUltro* ^ ^ ^ t m * v io le t Viaiblo v io lo t

t i lMe p. f t . p . r*

At4 0.2ft He v*«pink Mo v«*plait

X 0.0ft Mo V . Me V .

1 (fMT* 0.74 n lic oeid and/orainapie aeid)

Mo v«*ob.f Mo v.*wb.f

^ f t . * fa in t, y . « pollaw

Mo » oolcarloaa, v* • v io lo t , • cdiito.

tablo Rfovalooa and Color Roaotiooa of Spota Obaorvadon Ckreewtosraph 8, Flgoro 23

C tJ .« k

Sootifwaluo in Viaiblo

Ultra*violotLlebt

Vioiblo vielot

t i l0.54 Ho,

ft . p.fl* y* int. y* f l . y.

0*45 Mo y.*v* Ho y.*v.

0.3t Moto

V**«ll*y.*vb.

Mo v.m b* to y.*vii«

1 0.89 It. b r.* y.to

bll(**p. dk • p»

It . mr • *y . blk. p. to dk. p.

Z o .a s It* br**y* «r* It * br**y* f

- J „ 0.97 It. br.*y* t y**br. b r.

124

tablo 18. ftf.viiliMMi ««4 Color Rooetiono of Spoto Obtorvod on Chromtogroph 8 , Figuro 23 (Cootifwod)

fi8lM r,ai4«LBat..lJ

Rfmvrnlw In VUlblo vlolot Vlolblo vlolotHaOi ..gfigiittL... ito h t L lih t llite t Liobt

1 0,99 dk. y ,.b r* b lk ..br* dk* p . . b tk ,.b r .br*

^ dk* w dork, f l , 1 f l oorooooot, it7 • fo lo t, ii«t* *intoooo. I t , • l ig h t , ko • eolcNtlooo, f m eolor oncortoiii,b lk , • block, b r, m brown, gr* m grown, p* • porpto, v , «v lo lo t , oil* m id iito , y , m yollow___________________________

Flgur* 24« ChroMtograph Ft Dlagraa Showing Ralatlv* fealtiona ef tha Spot* Obaorvod on Ona* dlnanaloffial Paper Chronatograph Devatopad Ualng XAW

Flgwra 25* G h re m o ti^ffk Qi OjUtgcan Miowing itaiarlvn Paaltlena af tha Spota obaarvod on Qna* diaianaiafial Papar Chrm m ttvm ph Oavatepad Ualng BAU

TAV(/FRONT 80 60 40

- [ T"FERULIC ACID?

> r —C

20“ I 1----SINAPIC ACID ?

A

ORIGIN

Agi? p-COUMARIC ACID?

FRONT 80 60 40- BAW20 ORIGIN

127

Tabla 19. R f»««lo *a and Color fcooetlono o f Spot* Obaar»od «Mi Chromatograim 2, Flsora 24

„S«3LtC ftfiCtlUltra* ■......

Rf*vatoa in Vioibla vLolot Vlaibta v ia lo tXfifiX U li iitfe i fctobt , IrViBl Llsht

4 «|t 0*03 8a y . , y* y .T F lr a t )

Soeond O.IO 8o y«*y.* ft* y* y.*P«fy**wh* y*«adi*

Third 0*24 Hof ft* v**oh* ? ft* v**wlii' i t* ~o ia in t , Bo o CO iw »l«ia ,' f w eolor naaor t aXn, y* <■

g » l a * 9. w J t ia U l . vh* w o u rn iU m ^.

Tablo 20* Rf*vsttioa and Color Roaetlona of Speta c^orvod on Ghreaiatoaraidt Q, fljtoro 23

Jia l

Scot

AJXI

Rf«nrsluo In V ia ib UU ltra* ’ wlolot v ia ib loLlsht L lsht

U ltra*wiolat

0*90 br**y» y* br**y* y .

I 0*22 br**y* p**br* br**y* P**br*

1 0*42 br**y* p«*br* br**y* p«*br*

1 0*94 br**y* br**y . br**y* br**y*

i 0*98 br**y* y* br**y* y .* bsr* • brown, p* • pnrpio. y* • yoilaw

128

Xablo 21* Avemgo Af-valuo* afid Color lUMCtioiia of ttio Yollow Spota Obaarvad m Ona-dioaiiaiMial fmpmt CbttMato-

grapha ©avalopad Ualofi feraatat

■ ,a n w « 8i g U 8^ l | ^ ^ _____

It ta -Averaga Ultra*Rf-valoa in Viaibla v ie la t V iaible v io le t. I f lg i l t iL . , UiifaY itiifein i i i k t U i l l I

Ag| 0.S2 f t , y* f l , y , in t , y , f t , y ,

Ag^ 0,40 y , y , y , to t , y ,

* Ag| ia abem in #lg«rea 1 ^ ,1 1 , 2:^, and I I , 6igemte* I^ R ^ a J , L , H, and 8, Ago ia abewfi to Flsnaea 21, 22, Chraaetegmm® I* and K,

** y , • y a llo v , f l , • flnereaaant, f t , « fa to t, to t , • totanaa

Speta Agj and Ag^ wmt# neat lik e ly kae«ipf«tel •

0,58 CS, &ete-S«itti to 4 9 ), 0,S5 ( 4 8 » and anercatte

^*^^Poraatal * © vM ), raapaetlvaly, an th# baaia ef 8f-vatnea

praaanted by Bate-Snith <5, Bate-Saiith to 49} and Feeket (48)

fer tha thraa cmmm— t flavonel agtycanaa to F e rm ta l,

Exentoetton ef a lin lta d mwber af Rf-valnea fren ether

aoureea (S , 29, 8ate«»Saiich to 49} todtoatad that at laaat

two othor flavanola, iaorhafeietto • 0 ,5 S ), alao

a camon flavonel (Harbome to 25), and patnlatto (Affereatal

• 0,48} eewld ba rapraaantad by apet and that at laaat

thrae ether flavonela eenld be repreaanted by apet Agy,

Saaaypatto (kfforeacal • ©*41)# ninntotto (Rfrereatal • O .A t),

and, again, patnlatto (kfforoatal • 0*48), Rf-valnea of

elaooM of eoopoiHMit othor th«a flavoaol woro not oxooinod

duo te loefc of owoilobility, Zt aheuld hm noted, howowor.

ti®Mit rolotlvoly few plont opooioo cento la aoro than two or

chroo elaooos of flaweaoldo in glpeooidic raobiaation

(Karberaa in 29)« Aatbocyaaidiaa and fUivonolo arc cleaotjp

roiatad atruecurally and biogmoticallp (Harborao ia 2S)i

tborofora, tho fiavoaoia night ba mwxt aatpoctod to accoinpaay

tho aathocyaaiaa* For tho aaao roaooa, aooo aloilaritloa ia

thoir glycoaidic pattoraa oight bo oxpoctod (Harborao ia 25),

i t waa d ifficu lt to aacortaia which of tho apotv oooa

on chroaatogrofdio proparod uaing tha othor two aolwonta.

Xaw ond lAtf. cocroapondod to apota Ag| and Ag^ oa tha Foroa-

ta l chrooatographa* A major follow opot woa anon near tho

o rig ia on tho TAW chrooatogro|d>j pot. according to

Soto-Soith (Soto-Soith ia A 9 ), apota Ag| ond Agg, twariag ftf-

waluoa of loot thoa 0,80 ia F«roatot. ahould hawo ohown ao

oovoaMOt ia TAW, (Soo Figaro 24 ,} Forhapo tho low Rf-w«luo

of thio pallow apot in TAW • 0, 01) ia iadicotiwo of

noa-oovoawot. in idiich caao thia apot eould bo apot Ag|,

A nojor yellow apot waa ao*n on 8AW Chronatographo F

and Qi o corrooponding apot eould not bo locatod on BAW

Chroowtogroph O, Thio atajor yollow apot, proooatod in Tobto

22, wao thought to porfeopo eerroopend to apot Agt § i t i «

ohown diagroawocieolly ia Figuroa 20 and 2S«

If thia apot woro iadood apot Ag^, ita Rf-waluo io BAW

indieocod that Agt waa awro lih o ly booaqpforol (Bfggif • 0,89)

m

Tabla 22* Avamga Rf*v»ltia and Color Raactlona ef tho Tallow Spat Obaarwad on Qa«*diMan«io»al Fapar Chraaatograpba

Omalapod OnlRi BAN

■— ■I,.................— .„Q2l8g„ftNlgUa[

Avaraga Ultra* *®tJltra*Rf*valwa in Vlalbla vialat Vlalbla violat

kstaxL — m ------------------- k itu L . k u a i - „kUfet u t it iL ..Ag| 0«90 y«*br« tet. y, iitt* y« tat* y*

A Ag| la ahowB tn Pigwroa 20 sad 25, Cbraaat«upP*l>fet *» » br* m brown, y, m yallow* int. • intonao

130

than aithor iaacbaanotin (Rfggw * 0*7|) or patwlotin iRfggv **

0*73) C18, Soikal in 20, 2f>* tboao eonpotmda «»y a t i l l bava

boon proaVnti I f ao, Mioy wore not raadlly rocognisablo on

the MAU cbronotographs*

b. i^aJaJLAiA

Duo to the known proaoneo of aalvidin

3,5*digloeoaido in P.aandwlaanaa petals (42, 44), i t was

oKpoetod thot tho aglycono awlvidia soutd In> feond an tbo

chreawtegrapbs• hslvidin, with an Rf*valuo of 0,40 in Foroa*

tol (17, 48), abould have b««n tomA at tbo aano positisn or aoatowhat boyond tbo tea or yellow spot A ^ • 0*32)*

Oa Cbroawtogroph J (Figuro 19), a apot <Ag ) waa soon with

aa Rf«valwo ef 0.58* In TOblo 23, ita Mwirootoriatiea aro

prtttntod, along with tho eharaetoristleo of aalvidin, for pmrposoa ef comparlaon*

Omor onthoeyanldias, dolphtoidin (Nfgeroetal * 0*32)

Toblo 23« R f«v«l«o and Color fUMetlono of Froboblo Holoidlfi Spot OtNiorvod on Ooo«>dljMHi»lofi«l C9ir<»Mitetroi^ Doool.opod Utiog forootol, ofid Cootporotlvo t f *0x 100 oim! Color

Koxetionx of KxlvldiJi

m

Ultra* **Pftr«-Sf*v«luo in V U ib U v io U t V U ib U v lo lo t,.l«rg g tt l iclih l iilihs JiUfeK,, iJUOUs.

illo o i cooda fxdo)

Ag« 0«5S lig h t p* roao f jro llooii^(tooda to

Katoidln 0,40 p« a(17 , 48)

Bo Wid in noovo tray*bluo

* ^ 3 io ahoon in Flgnro |i, (ihronototroiMh Jp» m pnrplo

and cyanidin (Sfy«roatat ** (1 7 ), woro aulgoatod aapoaaiblo eonatitwonta of J2* aandwleonoo potala* 12toro waa

no afoidonco fm thoao two aglyeonoa on tho Fcwoatol chroma*

togmpha*

ao-chromtegraphy of apot of Chrenatograph L (flgwro

21), in addition to nncow^ing tho proaonco of naWidin,

previdod additianal ovidaoeo of tho proaoaca of kaanpforol*

Tho Sf*walno of tho yollow apot aoan ia tho ro*ehronatography

uaing ISSMAc waa 0*10 to 0«t5| tho Rf*waliio of kaoa^forol in

iSSitAo ia 0,10 (14)* Tho othor two poaaiblo ftavonola con*

aidorod in tho Idontlflcatioo of apot Ag , iaortiannotin andnMalL

patulotin, woro not, howovor, olininotod fron cenaidorotioci.

tha Rf*valua of patulotla ia alao O.tO in ISIUBMii tha 8f« valun af iaarhannatia In I5 l^ e «aa oat awaiUbln. but aach

of tha ftavanala far which tf«w«lwaa in t5%RAc wnra awailabla

had a walwa of about 0.10 (ti)«e* g lk lifiu tfilfl

ISrldanca far tha praaanca af tba caag»tax

phanalic caapaund. altaglc acid, waa obaarwad om Ghreaata*

grapha M and R <apat Ci** Flguraa 22 and 23«) Yha

eharactariatica of thla apot ara praaantad In Tabla 24. along

with tho eharactariatica af allagic acid*

132

Tabla 24. Aworogo M*waiuo and Calor Aoaetiona of Frobablo S llagic Acid Spot Obaarwad an Ono*di»anaionai fapar Otrana* tographa Dawalopad Uaing Paraatat, and Caoparatlwa Rfwalua

and Color Ronctieea af t iU g lc Acid

Oltra*Af*valua in Viaiblo v io lo t Viaiblo

ItfiSSt .gaPMtdl lirUM fctoht L irh t

Agg 0.28 bo w«»pinlr. Ro w.»pinlt.v»«wh. to to

y.«wh« y«*wli.

S lU g io 0.33 Ro aoft w «. Ro lig h t w «.acU bluo y.

(S . Sotkoi in 25)

ia ahawo in Piguroo i i aiU H . Chroaote^pha if and

Ro ■ coiarlaaa. w* m w ia ia t. wh» m whtta. y . m ymtUm

d . xoota

Escopt f*r tho M jo r yoltew ojpot toon on

Chronotofvoplio E ond Q, tho BAR chroootogropho voro d if f ic u lt

to cwrrolftto v ith tho Forootol chrenotottnidtt. ttm cfeiof

noofnlnooo of tho BAR chronotogtophc v « « in dononottotlng tho

cnecooo of tho hjNtrolycift of tho glyeooidoot tho BAR mrornt*

togmpho of tho oglyconoo did not rooonhlo tho BAR ehcooMito*

gropho of tho glycooidoo, thorohy indicotiiig thot

glyeooidoo, brokon doim during hydm lyoiof voro ohoont fron

tho oglycono chr onotogropho.

With tihm Forootol chronotogropho. It voo d ifflc iilt to

d if forontinto onong opoto %rith Rf^volnoo of 0.77 and grontari

a«Md) apota, according to Boto*teich (Bato^teich ia A f ) ,

ahottld ham boon raaolvad by ehrcMBatogmphy in tAR. Chrean*

togmph F ClAR) (rigtiro 74) ahowod only l^roo apeta, along

vltb a eoneontration of lanobilo conatltuonta «dilch raanlnod

at tha origin. Thooo throo apota, tha fira t of which waa

thwught to pooaihly roprooont opot Ag|, hod kf-voluoo of 6.03,

0.10, and 0.36, roapoctlwoly. 8ato*Snlth (5, Bato*8aiith in

4 f) praofMitad &f*valum in EAR for throo coaneti phonolic

aeida, thidi could wall hnvo boon rc Hroaontod by th# throa

apota obaorvad an Chranatagraph F| thoao varo s*eoamric

acid • O.OS), alnapic acid m O .lt ), and faralic

acid « 0.2S), roapactivoly. Xn tho ahoonco of

furthor infomatlon and data, hoifovor, aum poaaibllitioa

roproaantad apoeulatian at boat, tha colwr raactiona of two

m

ot thro* apota. In fact, did not auppore ttooo idanttfl*

cations, as comparison of ths data In tha Tablo 19 with tho

Tablo 2S (Appsndlx) indicatod* Factsllc aetd, aa tbo idontlty

of tbo tiblvd spot (Bffgy • 0*26), was porbapa awpportad bp

eonaidmratian of tho eolor roaetlons* Tho color roaetlons,

howawor, wora not takan aa pesltiwo rofutatien af tba abovo

spoculatioos, in wlow of th* proviooaly omtionad d if f i*

eoltioo In col«r daflnitlon*

Such spooulativo Idoas, ooroowor, bring to li|^t tho

too*ofton nofloetod guootiMi of tho proooeeo of phonolio eon*

p<n«Dda othor than tho coonon fUwonoldo in flowcnro* In tho

past, workara howa ehoaon to Ignora thoao eonatltuants, par*

hapa boeauaa thoir proaonco tonda to eonplleato tho alroadf

eonplon task of idMitifying tha flowonoid pignanta*

Svidacio* for tho proaonco of tho idionotlc oeida sian*

tionod obavo on tha Foraatal chronatosrapha woa d if f ic u lt to

oatabItab, 6um to tb* aforo*»#«tloood crowding of conati*

tuoota with Rf«violao* obovo 0*80, Chronetograph K, how oor,

did snhlblt o spot with oppropriato eolor resetions to be

elaaaifiod oa oithor ainopic and/or fo ru lic o d d , which oro

not aoporotod in Forootal • 0*84 to 0*88 CS}}«

Tho spot ebaorvod an Chronatograt^ H had aa Bf*valuo of 0*94*

<Soo Figaro 22*) Thio opet nay hava also boan prooant an

tho othov Forootal chroaatographat i f ao, i t woa aoakod by

tha aany colmrad conatituonta obsorvod in tho uppoijhmli of

tho pspor.

m

la conclualoa. It 1« llkoly that kampfarol and guorea-

tin, twa af tha thraa cocMMaaar fiavoaala <B«fan$Kith i » 49),

mmy ha aglyceaaa that ara praaaat la hydrolyaataa of £•

aandylcanaa patala* Additional aglyeooaa my ha amnt iao-

rtiaMsatUi, toaaypotin, niatolatin, and patulatla, wich iao-

rhamocia boing amng tha four mat eonmn flawanola

(Karbama ia 2 5 )* Iho conflrmtion of tho praaanaa of ona or

aor# ef Choa# flawonola would not olLainato tho pmaibillty

of too praaanco af tha othara, aa nora than ona conatttuant

eould hava boan praoont at ona location* In too provieuaty

•antianad aurv^ of tho phanolie eonatltumta of Imvoa,

dato-Uiith ( 5 ) atatad that, amng tha Laganincaaa, hydroxy la-

cion in too 8-po«ltian ia fraouant* kmmnft tha flavonala,

toia aopparta toa praaanca af gaaaypatin, ana ef the flava-

oela nentioned above aa a paaaibla idaatifieatlon af apet

Agf* In toa aaaa atudy, ha found otiercetin and l(«aapferet

to be proaant In 33 and 27, raapaetlvaly, ef toe 94 tpeciea

of Laguwinaaaa llatad,

Tha praaama of m lv ld ln in hydrolyaetee m a tantatlvaly

daaonatratad* SLlaglc acid , a cm rla* phanolie, m a indicated

aa a pm aibla eenaticuant*

r in a lly , th rm phanolie acida, g -coum rie acid , aisapic

acid , and fe ru tic acid , wora indicated aa peaaiblo cenati-

tu m ta , with evidanca being the atraiMiaac fer toa praaanca

af facullc acid* I f tha apot with an R f-va lm of 0»93 on

Chramtograph f (tAW) i « j^^c«n»Mirie acid CRfy^if « ©*©9),

135

thou tho proboblllcy that i t ropcaaaiica spot Ag| la alieiiiatodi

thla dooa not affact tha likalihood af tha rraaanea of

kaaopfaral in tha patala.

!• ) tho flavanaid alycoxldao

It oaa cofiaidarad prababla that tho

najor atlyaatio apot. Ag|. waa tha atlyeana af tho tiajar

yollow glyeoaidie apota. and/or (Soo paga 104*}

thMo apota nay tharafera ba kaaag>forol glyeoaldaa. Kf*

waluoa iM BaW wara awallabla far thrao kaanpfarol ilyreaidaa

(18. Baikal in 20)* Of thoao. kaonpfaral 3*glwcoatdo

(Rfg^y o 0«70) woa th# noat likoly idantifieatlao af tha

glyeaaid# apat Ph|^*2| kaonpfarol 3*glacaaida la tha neat

caonaaly occurring flawonol glyeoaldc (harboma ia 23).

Hovowor. mra eaaclualwa avldanea waa not avollabta Ui that

tha Hat of Bf*v«iuoa ia tha othar aalwonta (paga 104) waa

Inauffielant*

B ii i b 4 fiamlwi

kbgagiigB

Fhanalie carbanylle aclda (banaaie.

ciiwianle. and phcnylaliphatic aclda) occur widely in plaata.

being aacotid only to tha flavanalda in iiRpertanea (Boikat in

25}« Baeauaa hydralyaia with diluta acid (hCl or HAc) wuat

pracada iaalatlon af tha fraa phanalic acid (Baikal Ui 33).

i t wna aapactad that thaaa aaidt night ba found in tha

hydralyaataa af tha petal tiaaua*

heat of tha alnpl* phanoliea (Cg*C|i ^ a bonsoic aclda)

134

aro widoty dlatrlbotod to aitsioaporata (Harborao ia 2S}« Oa«

or laoro of tbo eoatoon eianaoie aelda (C g -C j), j{:»eo«Mrie acid ,

caffoic acid , fo ra tic acid , aad alaapic acid , oeei«r,ia cca-

hiaod fora, ia p ra ctica lly ovory highor plant (Rarborao ia

29)* Rarhomo (Rarborao ia 29) diacuaaod tho variooa i ^ a i -

bio coabiaod ferao, aacag idiaat acgar dorlvativoa and

acylatad flovoaoida, rhanyialiphatic acida, fiach aa phanyi-

propionic ocida (foraod by tiko rodaetica of tho o liphotic

doablo bonda of cinaMiic o cld o ), hovo boon dotoctod la

!4otaaroi a o lilo c ic a c U (ft-dtydroaQrphaaylSMPOploaio a c id ), tho

dihydro dorivotivo of o-cotmocic o d d , ocrcaro ia tho

Loguaiiaoooo (Borborao in 25}*

Aa odditioaai phoaolie coastitaoat of dicotylodoaooa

plants ia ollagie acid, a cooplax phaaolie; £ltagic acid

ia a coagMnant of allagitaaaiaai taiailaa aro proaont ta

aafficiaot aiKHiBta in tb» tagaaiaoaaa for coairorciol aaoa

(9, Borbmao ia 29)« J|* camtwleonxo ia high la taaala, with

a eootoat ia toavoa of fowr to aix pwroont or mro (9t>*

Acoordiag to Bato-Soith (5 ), tho coaoionly oeoarrtag

phomiic eoBacituonta of pUata iadada allagic, caffoic, and

^•couaaric adds* Sabataacaa diffodn^ froa thoao phoaolica

by oao wait troaofemotiea oro tho aoxt neat cooacaly foiaid*

Ha roportod caffoic add and fi-cooaaric add ia |i aad 29,

roapoetivoly, of tho 54 iogantaoooo apodoo iistod (9 ),Siaapic and foralic adds oecm-rod ia 20 oad IS of tho 94

opocioo, roopoctivoiy*

tS7

Tho oxooiMtloo of tho hiooynthotle pothooy lootflxig to

flovoiKiid product ion jirrovidoo on oxplotuitlofi for tho wldo

occurronco in flouMTO of tho ocido dlocuoaod to tho proeod*

log porogropho* Flovetioldo oro hiogonotlcolly dorlvod fro*

tho odditlo* of throo oeototo uoito to ciimooic oeid or o

rolotod eempouod (Orioohoch in 23)*

y ittg y lg lg t ggtgirmgutnr

Spot Ag, of Chromtogroph L (Figuro 21, Tahlo

18) iioo oitttod for opoctrol onolyoio* Tho roouito woro poor,

prohobly duo to iavmvity of tho opot, a« iiroviottoly dio*

euoood, co*chm«otogr«i^y of thin opot in tSlBAc yiolded two

•poto, ono purplo ond ono yottow, thoroby indicoflng tho

prooonoo of ot looat two oglspconoo in tho oluoto* (Soo tobio

18, Figuro 21«) Spoctrol onolyoio of tho yollow opot woo

poor, ond olthcugh i t did not ohow pooko indicotivo of

koonpforel, i t woo not tokon •• ovidonco ogoinot ito

prooonco* Thoro my o t i l l hov# boon noro thon ono oglycooo

In tho oonplo, oo ooot of tho flovoaol oglyecwioo wmor con*

•idorocion howo oppronimtoly oguol Sf«volcMMi in 15tHA« ( I S ) ,

138

OQ»QLm i\m AM? Sl»flARY

Tho purpeoo of thla thaaia %Nia to atudy tho choailomy

and gimotica of flouor plgaaoita ta P .aanidvleopao aa a foitav*

up to a prwlous atudy of anthoeyaaiiia couduetod hf Park (43)

and Fatle and ftotar (43, 4 4 ). tho ohJoctlvMi of tho roooar«^

umpo aa followai

t ) ida iitlfleatiM i of tho plgawntat

2) dotormtnation of tho lahoritaneo of tho pig*

amntai

3) atudy of tho l»t«rro iati«a ia iiipa hottioofi tho

gonotiof and ehoniatry of tho pMcMmtai and

4 ) atucidation of tha IntorrolationaliiiNi hotwoon

anthoeyanina and othor plgnonta moaant*

It ««a found that tho acofio of tho paohlon vaa too

hrood for tho tin# availabloi conaootmitly, ttui objoctlvoa

woro not coaqplotoly fu lfillod*

In tho proaont atudy, flowmr colMra woro groupod Into

als eolor eiaaaoo, X through V-1 and V*3, %rith noar*«htto

fleworo bolng plaeod Into otaaaoa IV, V*l, ai^ V*2» Croaam

woro attouptod anong tho patontal linoa, but faulty tocd%*

niguo, poor oovironnontal eonditlona, dlffieu ltioa Intioront

in tho apoeioo, or a eenblnatlon of thooo faetora pr«w«at«d

tho obtaining of any rooogniaabl# truo eroaaoa, Thua, tbo

gonotie atudioa propoaod in tho ebjootiwaa had to bo abandenod,

Papor chronatography waa usod in tho onalyoio of tho

pigiMmta, vich idontlfieatlotia bo tng b«««d apen «ftd

eolmr rMCtloog. Tbo iavaatlgatlm of tbo eonpoondo proooot

Ijivolvod two gtopg, •• fottomi

1) onotysio of tbo glyeooitfoo prooont ond

2) ooolygio of tbo ogljreoaoo proooot.

A fxroliaiaory stop oadortokon woo tbo eotifirmtloa of

Idooclfleotioa obtolaod by foi^ of tbo coapooodo proooot.

Fork found ono oethoeyonldia, mlvldln, <aio of ita

glyeoaidaa, natvidia 3,S*dt.gtaeooida, la bia atadloa* Fark

atao found flavonoid pignonta, othor than atithooyanin, idiieh

ho did not idontify, and atatod that thaao pigaxmta oppoarod

to havo o aoeondary roio in producing narrow eolor variation*

Proaont atudioa eonflmod tha luroaoooo of nalvidin and

nalvidin 3,5*dIgtucooidn. tho preaoneo of «n odditIonai

ottthocyonin apot waa indlcatad, although an idontlflcation of

thia apot could not bo inido*

tho erlginal intant of thia thoaia waa to atudy tho

yellow pignenta* It waa found, howovor, chat tho phonolic

cowpoaition of tho potala waa far noro coatplaa chan oxpoetod,

and tha ehronatographa warn erowdod with colarlaaa eoekpoanda

ffhioli aahibitod varying color roactiona utidar altravlolat

light and with aoaMnia vapor* for thia roaaon, tho atudy

waa axpandod, and an actanpt wma nado to identtfy othor

phonolic conatltuonta praaant, auch aa tho phonolic acida and ooaiplox phanoliea llko ollagle acid*

Bocauao of Idto proaoneo of ao nany conpotanda, and dua

140

to d ifficu ltio t Mieoiaitorod in obtolnlxig ouffieioiit onouoto

of cooeootrotod oxtmeto, tbo Ixtorprototlox of ebroMito*

gmpho wfto d ifficu lt* Oiffmofit trtxlo oppoorod to yiold

difforont roouito, ond difficu ltioo uoro oneomtorod In

viouolUIng o i l of tbo eonpoundo prooont* fmt cbooo roooono,

opoctrol onolyoio «oo of linitod opplicotion ond uoofulnooo

in tho idontificotion of potol coootituonto* Ono Mijor non*

ontliecpnnin opot woo oboonrodt ovidonco indicotod tbot tbio

opot roprooontod two eonponoato ouporlnpoood or ovorloppint*

Tbooo conpononto woro tontotivaly idontifiod oo flovonolo

oad/or flovonoo* About 21 ninor opoto woro oboorvod

oltototbor, oltbottfli tbo oovorol ebronotogroplio oidiibitod for

oiniloritioo to ono onothor with roopoet to eolor rooetiom

ond nonbor of opoto* Idoatifieotion of tbooo opoto woo in*

poooibto without IMMTO ccttcluoivo doto* Bo otton^t woo mdo

to oorroloto tho prooonco, oboonco, or eoncontrotion of opoto

with tho flowor color of pormtol linoo oo roouito wwro

ineoaoiotoat*

A conporiooa of tho ohovo roouito with thooo obtolaod

by fork <42) ond fork ond ftotor C44) ouftoootod ttiot ^ o

phonolic oitUOti«W in oaadwteaaaa potolo io nucb noro

conplox thoa thoir roouito indiootod. It wxo unrooliotic to

ottonpt to poir tho ninor opoto obowvod with tho yollow or

fluoroocont opoto ooon by Fork (42)*

la tho prooont otudy, kooopforol, guoreotin, fmrwlie

ocid, ond oliogic oeid, ia oddItion to nolvidin, woro noot

141

atrofigly Indleatad «■ tho «ilyeon<m procxmt. Additional

oilyeoooa ooj hovo bo«o aoong iaeehoonotin. teotypotin.

n lM tlo tin . and pa tu la tln . with iaovhaonotin bo Ing ona of tho

four M»gt ccNwsoa flawonol aglyeotiog* Ceggyyotio ha* boon

Indieatod aa a poaaibta eonatltuant af lofiiwniaoaaa baaad on

taaoneoie atudlaa ty Bato*Sieilth (S ) .

It waa prababla that tho aojor aglyeoeia praaant. id«aiti*

fiad tantatiwoly aa tcaoayforol. waa tha aglyeono of eno or

both of tho najor yollow (awparlnpoaad or oworlapplni}

ilyeoaidlc apota* Linitad awallabla avldanea indlaatad tha

poaaibla praaanea of Icaanpforol S«>il«eoaida (Harboma In 25).

ono of tho eenpoRonta of tho aajor yollow apot.

Additional phanolle aelda indieatad aa poaaibla eon*

atitnanta %N»ro UTowwrte acid and tinapie arid, thoaa

ptianolia aeida nay hava baon praaant ladopondontly in tha

potal eoHa. or in eonbinad fom i anong othae poaaihllitioa

(Harboma in 2 5). thay nay hava axiatod aa acyl grewpa af

tha flawanaid pignvnta.

Idoatificatian af tha caratmaid plgaMwita wna bfyond

tha aeopa of thia thoaia.

r t o a w t l » Y t r r t U Y l i n i r t ! l n t

Htividin ia a nothylatad darlvativa of dalphinldln

(nawvo. hlwa)i nothylatian cawam a roddoniiig affoct. al*

though thia affact My not bo apparant dua ta nadlfying

factara aetiwa in tha ca ll, tha prinary eanaa af eaiar

wnriation In aaftdwieanno ia cwantltatiwo dlffaraneaa In

m

M lvid is i 3,S*digt<teosido eofieentriitlcm* 3,5*D|glueo»yUtlc«t

eiiusos Koro Intsno* colors and could account for tho totan*

a ity of color t « th# yiarpto*f l<m«rcd eolor ctsaaoa*

InfonBatloo woa not avsilshio «m tha influaoea o f tha

praaanca af phanolle aclda and allaftie acid m flouor color,

uoloaa tha phaootie aclda oxlatad In scylatad for# with tho

flavcmoida* Acylatien of tha 3,S«illj|lucoaida with x*touosrte

or eaffaic aclda «*ould roault In dollar flowara*

harrow color variation could bo influoncod by tho

praaonea of othor pignonta* It «ma tantativoly auggaated

that flavcotolo, oapeelalty Itaospforol and cuorcotln, and

flavotioo olght ba prooant In £• ggg^tKUmit potala.

Carotanoida, probably irosant aa volt, wmt* not Ijivaatlgatod*

Tho proaonco of flovooos and ftavenola probsbly roaulta,

through eo*pig»ontation, in a bluoing offact on oolvidin

3,S*diglueoaida* In tha naar*«diita flawsara, thwio pignanto

nay contrlbuta to tho •body" of tho potala* I f tho flovonoo

«nd flavoiiol* sotiafy any of tho unuauat eeciditi<mo ison*

tionod proviouoly, ouch aa rrasonco in high eoncantratisna,

thoy mmy actlvoly contrlbuta to potal color, probably in a

bockground offact of antboryanin on craan or yallow eolora*

Choleonaa, i f proaont, atao could produco a bockground

offoct of nolvidin 3,9*dlgluco4d o on yollow to orango eototo*

fVrplo and nagonta colors ara producad by earotsnoida and

aathocyanin occurring togothar in potala* Csrotonoida olao produeo baeVgro«md offacta*

w

fividcaae af pH affaeta aaa net iarvaatliiacatf* I f praaant,

pH affaeta eeitld eauaa btuar (with high pH) ar raddar (with

low pH) eolara, wnlaaa affaeta ara eewplieatad by tha

tandaaacy of tho anthoeyanin ta awiat in ita colorloaa paawdo-

baaa tmrm at high pR«

Ih# affmta diacwaaad nay ba awawariaod aa fallow*•

ImlBhtoltila rfiliYiito. .,(Mth,Y lit tea) QlyeoarUtloii (3.S) bloo, aawwa roddonlng iatonalfying

ft9*Pll8tgnttitifla Uckarcwnd offocta

144

dulling bluaing

HdYtaftU iiril CUitaatt

glwa *bady” to whita potalai

•ay eahtributa to idiitonoaa

anti n a M M iJ L li

orango- ar yallowlng

g fc m w tU t U i t lattBCTtiilai

roault In pwrpla or awgonta eoiora

oH offoora ( r i

1 ) low ta

roddaning

2) h ilb tH

bluaingt

calerlaaa paoudo-bato

fliYM iflUo lU xm nM

craau ar yaHaw bamgraund, i f thoaa plguanta ara ptoaant in high eaneantrationa, ate,

gtial€onaa> aurotiaa ( f )

erango or yoltow background

filggttapMl, (? )

baekaround affaet af plaatid-bam a pigwonta

Tha intaraeclan of tha abovo affaeta raaulta In wary

eawplieatad intarralatloaahlpa in tha produetion af toanotypie

u s

flowor eolor lo oondwleongo. tho eeneluilvo idooeifieo-

cion of o i l piioonta ood i^ooolie eeopoundr laroooot, ofid tho

dotoroioocioo of th o ir d iatribotion and eoneontratioo aooog

eolor elaaooa, would howo to procod* olocIdatloti of tho oboro

intoroctiofia*

r > , v . . " ,• v/• . ■ 4 - y-,.: V- v»- ' -y -

Afpm otx

to iio iS . <*oior R— etiona, on of IW iv o Ctoumn i f w i l ' fla iwim ail_fta5pf t m i u , A f i n tijM f r i u s i s A tit in (2 <

Claoo of Coa^oifl^ or ItioflMilta Aeidi M t a m n U ..........

AathocyaniA^

Ptaveao*

rUoomoV

kmromm(Reaaalmmmoll

eooMoro*

Qioteoeo’

n o

Y i i i M t k U k t y i t t iY iu u t h m t J t i t t i i

PlaitoroegoRod^pmrlo

Ottli rod or purplo RtiMmmy Piidt n «M ,

M l b W

Polo yoltow

Polo rottoo

neorseeoet yollow , rooo, briid^t rod, ptnn>lo|(3,9* diglyeooldm )

M l brewR^*^ Rod*browo Yollow*fMrowi)

Yollow

Bright yollow***Y # liow*gr»ow*>*Browao

Yollow

Bright yollow to ormorc^oi*

Yollow*!* to Ctongo^

*.* *•7Bright yollow** Oroofi*yollow**'

OraagoOrango*|itirit

brow I dark

Colorlooo

Black*! dark black*

Yol lew brew

C olorlw o

YellowCroftgoRod- oroag*Pink

Colorlooa

g s B n g g :

Btaiah •Octl blwa^

Bright Yollow

Y o llo w g ro w Otttl pmrylo

BrM ht yollowto lla wNgroonCroon

Yollow^oraogcCrangoRod*orangoBrown

S 3 ! ! ! ; * !Pnrplo* **SUek

Colorlooa , « Palo yollowl** v £ l!a c i£ jts ,_ _

Tablo cifolor im e tim a i'o n i ’aptnr, of ^ioolvo dioaooc of' liotum l if^iovMioid CowpMmdo,ITlffigllfi AfiidBa illtf, f t i l i f l to 29> ^ t t t i l te 22*Jl

9tigy fliigtiwii, to.clooa eor fbaoollc: AcidXtotocwBRSJLMM

laoflavono'

Ytotoit U i l i t tfUnnrtoltf ..fctoltf J t i i lt r l i liiiti;

C olor loaaColorlooo « Foio yotlour^

Loneoantboeyofiia , Colorlooo (F la v a a -3 ,4 * d ie l )*

FlavaaoeoU

loaf lavonaiM^

Cihydrochateoaa^

Catacblii , (P la vm -5 -e l)*

Boasaie a e ld a '

Clmiaoiie aeida^

rhanjrla t iflMic io a c ld d

Colorloaa

ColMTtoaa

Colorloaa

Celorlaaa

Colorloaa

Colorloaa

Colarlaaa

fmint porpla^*^ Faia ya lla w Yallaw (a lw rt X )

Calortaaa

Cask pwrpla

Colarlaaa to fa lo ya llaw

Colorlooo B U ck (a b o rt X )

Colmrlooa to Falo yo llow

Colorloaa

attm ato,totoB ^to .w tam lif

G onorollycolorloaa

Flworooeant bloo

Calorlaoo

Cooocollycolorloaa

Colorloaa

a b S S S J S S £

Fa lo t pwrplo Falo yo llow

Colorloaa^

Calortaaa to Folo ya llo w

Flooroacofit fwilo blwo

Black

C oooratlyco lo rlo a a

Flwor aa raot blwo

Calortaaa

Fopor, o# m ritin iil—

i«n of or Fhooolie

Y liU L iJ i lg Ir t ttitM Y to ltt hXakl JU mM mJ.

SBtclgte fftittalU Afiitft

Caffoir «c Colorlooo Sloo CotorloooDovo-eolorod <oeid

popor)i Bright , hloo*oialot (d ry )

BtooLight. groMiiirti*

hi«o*

g^eooMrie ocid^ Colorioas Colorlooo^*^ , Block (abort >^)

Cotorlooo BlwoBlodk (ohort A ) . Doop M «o*wiolot^

Foxwllr ooid^ Colorloaa B lmCroon (o d d popor)

81oo*oiolo€ (d ry )

Colorlooo1

Blwo , Vory hright him *

Sim ple ocid^ Croon (m id popor) Bloo«iwiotot (d ry )I Vory hrlgbt Moo*

Hydroeoffoic•cid3

Colorlooo Yollow Colorlooo y«ltow*«rooB

4 coomric aeid^ Colorlooo Yollow Colorlooo Bright yottow

g ^ o o m ric aeid^ Colorlooo Colorlooo Block (abort X )

Colorlooo Cork yollow ^•locfc (ohort X )

2 ,3*£»imtliOKy*. eUuMMlc aeld'*

Cotorlooo Yollow Colcnrlooo Yollow Bright h im

(oher*>s ) s

ilir'llRoaetlomi. '<m f«p «r^"o f ■nSBm

S J S i S T B f -cumor Fhonotie JLS iltaessD

3 ,4*!>l»otfici«y*« eiaifumlc «<UI

jj^umthoQcy- , ctaiuuBie actd^

E lU ftU Acid^

Y U ili lt iiiliht mi3EMiiaXi£.„.ktofat ~ iSE BSSEiCotorlmo

Color loM

BlttoBlock (short A )

Colorlm o Block (ohm t X >

Colorlooo

Colorlooo

ComtlBH ItlBXMlUB

SoftyVlolot

BImBlock Cohort X >

Colorlooo B io ^ (short X )

ilg h t solloo YollowT

^ tofawroBwroo oro oo Indicotod o fto r todlvidool clooooo or coopooodot oarcopttooSr «ro notod oftor IndivldtMl color rooetiotio* Footnotos l» d l« i t o roforoneoo ••

, follooot * Soikol la 20 I Soikol in 29

I r

ife. R f-m lfim aiMl Color Spota <^arvod m TWe-dfamPapar ChroMCegrapha Pawolopod Uaint 84V CFtrat Solvaot) and t5H<4e, BAe*SCt, ............„ . , . -§ f i3 U L i ..i a i L l ^ ^ l ^ n i g i C g B f i , S f l l Y m U •§.

Rf -va lue in 15% H4e- 1% 60% 60% Ultra-

C h r w B4V HAc net HCl IP BAe V U ib t# wlatat&ati iM L , X£Jl ifiJL l l U , X U . ilth t, ■ Usfet...

v i t i b u_L U b s_

tra* wtelot U tfcl

Roaarka

0 0.22 0.72 Ro Ro f0*1 or 0.3© 0.02 or Ro Yollow He0* 0.30 0.22 Ro follow Ro

1-1 0.30 0.42 — Yotlaw mm-U 2 0.36 0.44 — U r. f l . mm

y*1-3 0.32 0.42 — Yollov mm1- 0* 0.27 0.40 Ro Lt« w. T

4W . !•« 0.33 0.43

Rtua r i . rmllmm F t . jro li Yaltaw< fa Tallow

ew rit« ow M o d o a i^

y*-p* Oona ar I t , w.

AW . 7** 0.30 0.72 0.02 0.43

Pb. 0* 0.3S 0.44 Rot ROT Rot Yotlow-p.- J X* 0.40 0.41 mm mm n . p .

G* 0.43 0.84 RoT f t fiwll p .1- 1* 0.48 0.72 Yotlowl t 1 f1-S 0.48 0.83 — Ok. p . 81. ft 0k . p.1-6 0.48 0.81 Y .f,R ot Ok. p. F t. y . P * -^ .

4W. 1 0.48 0.82..^ 1 g>M M l f iiM ..... ___ fU U _________

and Color > tnar Syotm Ohaorwod on iw * 4lltxNwi » t<wwil'Chromtoitracha Dovolocod Calng (Ftoot Sotvofit) and |5%me» HA<HR€t. tlWiCt.

if d r (ilMMtf t e lY tn f f )i . ,i i . , .te la r 8rf.(^ftiiriiiiiii)

M -U l i H i t i v i o i » u « i t

ChroM* SAff RAO n c i »Cl IF HAe violotm t Xia, W l xgl. iiO, la , ...LkM., UtiiL.

• ^ r r » -r >m "r>L££ar

T C m I o “ l l f c rv io la t

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0.2S0.200.200.200.240.2C0.17

O .t l0.83

0«800.880 .f l4.940.92

MoMo

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L t . f l . b l . — F l . tu rn . —

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^ 2 7 pa 0,82 0,50 Bof Hot Oof Yottow fht or

4 claaaili^ileatian WMcnrtaini apot baleagad author iln thla atona, or in annof tho groupa doaignatod imdor JlinydUl*

** AV. t w avoxogo of valoaa w ith m t oatwriaki Ao, 2 « avomgo of a tt vtt«NW o«o P I. « hlno, h lk . • hlaek, k r , m hrovn, g r , • groon. Bo • eoioritHHi, p, « pnrpio, ^

tnrq, • tmrqnoioo, v , « v lo tm , wh* « o h ito , y« » yollow, • dark, f l , m v»

U h U 26* Rf*mli Oirom totca

I — t t — of bCnor S fm f Oboarm d «wa> ‘iTwMHStomotmoi ltaiti« BAR IBHCt,

(eofiriamd) fluornocont, ft. m faint, int. m intetuia. It. m ligbt, lt*r. m ligbtar, f • colmr 4|iwNitiooaliU» «r oneaHrtalA, ** • coI«mt iK>t racordod

- _ 4.Mar.b0 ,

Ul

M Tta W C U c m o

t . Bomoo, e* it*, Jm 0 , CttlbortaoA, ond J* «* LaiRbwrt*Inhmrltoiieo mi arnrnm ond tlmmmw oolora In ftax* Anron* 4* 921494*459. 1944,

2, Baco*iNsltli, S. C, Anthoeyanina, flavonoa, aiu) oi^or phanolle cenyoonde. Bioehae. Boc. Byap. Bo. S, pp. 42*71. CaaiNrldsoi Caolnridiio U n lv ira ity firoaa. 1990*

3* ----------- -... Loueo*anthoeyani»a« 1* Ootaoticm andidoatifioacian of oothoeyooidlfio forood froo looco* onthoeyontno in plant tiaanoa. Biaetiao. J . S8i|2t*tt9. 1934.

- ......... -J-... . - .......-» fapor dHrmmtmmwmpby mi anthoeyoninaand ralatad auhatancoa in patal ontraota. Battnro 1411 835*838. 1948.

4.

9. .... ....................... .. tho iNbotiolie ean«titiMotto of plontoand th a ir toatonooie slgnLfieaneo. I* Oieotylodcmo.J . LliiiiMn Smt* Land., Botoay 9B»99*|73.

4. Boolo, 0 . M«, J . E. iN^ieo, and B. Bcott*Monerioff. tho gonotlca of Yagbona. 11. Ghoniatry of tbo flavor eolotir variotiono. J . 41t49*74« 1940.

7. -j-i-x Bobtnaen, B. toblnaen, and R. Beoct*honorioff* Oonotioa and chaoiatry of flcwNMr eotomr voriatiaii In Lathvnia odarattta. J . Oonot* 37i379*388* 1939*

B. Bloak, R* J . , R* loBtrango, and 0 . c ^ t g . Fonog ChrcaM- IR S I^ K c A||j|HfyT.r^RiM8l* 9ov York! Aoodaoic

9. Srovbakmr, J . 1* Cyanidin*rod vbito c lovor, a ditplieato raoaaaivn dsotant tn Y rlfo litin rot»ona t . J . Morodity 93# 143*147. 1942.

10* R. 2. , and R. L« £ovia. riovor eolor inhoritaneoin diploid a lfa lfa . C tw Sol. lt437*A40. 194!.

I t . Oiow, R. Intarapaeifie tiybrldisatiofi In tho ooctnai^oa»MiiJo. M. S. thosia, Univaraity of hawnll. I^ U I.

12. Ceopor, R. t . , and f . C* E l l io t t . Flavor plgnonto in d ip U id a lfa lfa . Crop Sei. 4i347*37S. 1944.

13* Coopar, E* i* , ond F« C« S llio tc . loborltatieo of ftooor pignonta tn diploid a lfa lfa and chair ralationahip to flowor eol<nr Inhoritaneo* Crop S «i« 9id3*d9. tfoS.

14* ihtd.f eitiait Faria , C* R, J , Hanoy, and 6 , •• Rilaan*A aurvoy of tho Intoraetien of gonoa for flavor color* Fiidi* Stato Univ*, Agr* %tp* Sta*, Tocdi. B u ll, 7B1*m o .

159

IS. i^ v ia , £»• V*, l»* A, Ta ylo r, R* f* Aah* JUmmJUjEUI balaanina L,*«tho Isdtoricaneo of f lo ra l eolora* Cm o^ca 43i|#»3A, 195B*

16* i^aanaolly, C* 0. M o rtta n co of ohito flovnr color tocoonen vatch, XLsJA aativa. Agrou. J . 50i7d3*7#4, 1998.

17* Ihaaa, A*, and J . A r d it t l . Iha anthocyanin pignonta of p l o M . pp* 138*194 in Eooorinyitol thoalnlnavi

18. Oago, T* 8 . , C* 8* Cooglaaa, and S. 8 , Rondar* Idontt* fioatien of flaoonold eonpounda by f i l t o r papor mrouu* tography* Anal. Qion* 2311582*1585. |95t*

19* Coiaanan, T* A* Anthoeyanlna, ehalconoa, aaoranoa,ftavanoa and rolatod wator*aolublo plant pignonta* pp*

(K* Paom and H, ¥* Tracoy, oda*) B m lin , 08ttingan, Eh»ld«lborgi Spriogar*V«rlai* 1995*

21. « and 0* A . L . iKnhloui*^* InharitaneoIn tho oam ation, earooeitoilya. Vf. Th#idioniatry of flouur color variatio n , I . aonotioa 32• 410*433* 1947*

22* CoiaaaMin, T* A*. £* C* Jorgmaon, and B, L« Jmnaen*Tho chaniatry of flovor pignautation in Antlrnhtnan MJhlg. Color ganotypaa* I* Tho flavoncld conponmta of th# hoaooygoua F, H . t eolor typoa* Arch* Blochan. Blophya* 49i36«*388. 1954,

24, Hagon, C. W*, Jr* Influonca of gonoa controlling flonor color on ro ln tlvo gtiantitim of antbocyonlna ond fUuonola in potala of jg ia ilija i balaaatna. Oonotlea

w

25. Karbom*. J . ^pamida. Maw Toakf Aea^Nmtc Jhraaa. 1944.

24. ■ lha chraMtegrapliie Idaotlflaatiaii ef

4 4 i7 8 7 -m * 1959.

22.**** CHI UWaXOgXaynEC XCaWaiif laaXMR OX

iwtoeeyeitoplgeaete. Jm ef Chronatot. I •473-481. |958.

a a s s i m . a S ” i £ 3 J iS l g t ! ! " i l l yy

74* I b l ^ i eitlji|^JLaa^ V. J . C . , and R. Seett-gafiaxiea and ehamiacxy ef flauar eelewr

ta dahtiai a mnt toaofly ef apeeifie pIsamitatieM. J . 6 m m . 30«t59-224. 1935.

29. Rarberea, J . B. Ptm t pelsrmaiiaia— XV, Ftavamla aayallaw flawar piceanta. fbytechm latry 4«447«497. 1945.

SO. Hartwig, S. I . , and X. Rtaaaa. lahar&fcaaaa ef flawar ealar af ■aybaaaa. Cxep t e l . 2itS2-153. 1942.

M ppvtaht}: M a tw y . Ea«tmrfi 1 ^ 1 K « r t U a ln m tSoeiaty. 1938 m*4 1941.

32. Hwae, 4 . Paraaael eeemoitieatian. 1948.

33. H ytla , J . fwraanal eaaamnieatiee. 1948.

34. Lawranea. V. J . C . . and V. C. Staargaea. Stwdloa ang.tS»,89MUrBIMl* 711. Oamelea and chanlatry af flawar colar In tha gardan fa rm , apaelaa, and hrbrlda.Raewdity lti303-334, lOSK

54. iappwr. R «. and I . I . Cdland. I^ m r it 4uaea af flawwrealar In a U a lfa . J . An. See. Agrwn. 3|i209-214. 1939.

37. hangatadorf, P. C «, and ©.* t . frapa* A dlrwet anantlta- tiwa raU tianahip batwaan vitanin A In earn and tha nmbwr of gonoa far yallaw pignontation. Selanea 78 • 241-242. 1931.

38. Harti% f . Pmaanat eenomleatlMi. 1948-1949.

♦0.41.

42.

43.

44.

39.

44.

47.

44.

50.

51.

4* J* tondofi#Arnold and Co. 1951.

141

Edward

Odland* h« t . lidiorltanea of flowor eolor In Caoaigun — L . fro c . An. Soe. Hort. S « i . 74i4 7 5 -4 ll. 1940.

Foorh* 7 . Colour dovolopnant in flowors. Ami. gov.F U n t Ptiy«i»l* 4i273-29«. 1955.

Fork, 8. «l. Itdiaricofieo of flounr eolinr in £ioa«adtmtS. F «y . k . 8. Ih o n U , Onlvoratty ~ R a u i 11.

- I .> and F* F. Ratar. Qanotie atudlaa in Spanlah elovor, OoanodltMfe aandvteonao I . hoy. I . lnliori« tanao of flom ir eolor, aton eolor, and laa flo t narkinaa. Crop Set. 8t447-47C. 1948.

SfMwiaK e ia n ^ , lation and Idont potala. Crop Set, 8*470-474. 1948;

• Conotie atudlM iniSmSM. 4* Fay. I I . lao-

cation of antkoeyanina in flouor

43. o itin g foraytli, W. 0 . C « , and 8. V. SiaxMuida. Aatwvoy of tho anthocyanlna of aono tropical planta.Frac. Roy. Soe. (London), Bar. 8, 142iS49-5«4, |954.

ii8l8L*f t i t Ing Catcoigno, A ltch la , and 0 . E« Whitn.A aurvoy of anthocyonina in tha Auatralian f lo ra , J , Frao. Aay, 8«c. 8 , 8 . Walaa* 82*44-70. 1948.

Foekot, 8 . C. tho fmmra of tha variation in flowor colour U tho gonoa U lta n M * FhytologUt 59*138- 144. 1940.

J.- - - I * fhanolic conatltuanta of loavoa and flooora in tho ganua Lachyrua. pp. 119-124 in lit FU nta in lloalth and RiaaoMt J^ o a d ln a a a?_w'iirrry i r r E ^ r r r r s i f T T r T i r T m f F T * -n i m m r m i( J . 8, Fridhan, a d .) ffao Toek* FMgimnn Fra«a<

49. Fridhnn. J , 8* (o d .}«

^ r T m r m x m x i%

Sohiaaon, 8. Fomatlon of anthocyaniiia planta. Batura 1§7*172-173, 1934*

Rotar, F . F . Faraonal eoaaaunicat ion, 1949.

u t

52, aotor, F* U« Dmt«. Cycolofleal •ciMfioa in th«g«mw DeejBodluxi soma chromovoM Am* «l*Botany I Z T l I t r * W *

S3* Seott-Hanerlaff, 2* A biachanlcal turvay af cms*Fandalian faetora fo r flemur eolotur* J* Qanat* 32«117- l70* tS36*

54* 0. A*, a ^ ©• H. aBitiiiBmtill,,gLdlMllytiTc|^^ChaBlatnr, Maw Yorki Molt, ainahart and Wteatan*

5S* Stanford, £* H* Totraaooie inharlcaneo in alfalfa* Agron. J* 43t222-225* I f S l .

54* Xwamlay, B* £* Flowar eoloor isdioritafiea In diploidand tatraploid a l fa l fa* can* J* Agr* Sei* 35t46t-476*1953.

57* Woihing, B* h* flowor color ii^Mritanco in a lfa lfa*J* Aa* Soc* Agron* 40i76B-75('* 1948,