Copyright is owned by the Author of the thesis. Permission is given for a copy to be downloaded by an individual for the purpose of research and private study only. The thesis may not be reproduced elsewhere without the permission of the Author.

A Tb Presented in Partial PUl.1'1l.ms'lt of the Requirements �or the �.egt"ee at Doctor oJ» Hdlosophy 1n Biocmmistry at

y University.

- 1968-

1

u

The bloeJD.,_is el 88l.ected cyanogenic glueos1des and glucoa1nclates dling shoots (14rg.e if'1 · tiB!jnue )

1'AII"'e aaed 4!Diduaively to a1Ddy 11nanarin biosynthesis while pl"Wlnsin biosynthe-aia studied in loth peach oots (m.u..ts pgmica, & tsch) and cherry laurel

ahoots <l• ) . Iaoprepylgluccainolate Old benzylglueosinolate aftac!l'ed ill 8CQJ."V)" grass eeedlings ( Cacbl.earia atN.cinal L. ) and garden

creaa aeedll"& aboota aatiwm L.) respectively.

� 8Dd 14 administered to plant tiswe. Thl cpantity of c�enic

gluooa14es dete.rminad by � �gen cyanide following eneymio h,yd-1"01ysia. The specific aet1 vi 14,y ar cU.lutian d the l.abel.led conpaand at'ter incorporation into the glucosidea

isothiocyana '

taiDIId by enzyme bydZ'olysis ot the 1nclates, identU'ied by cxmveraion

to ttd.Guree. derivatives. �-er. -and thin layer ohramltcgraphy enl elec� a

uaea to .-.arate �w:'l�tUe raclioaotive •

leolut7ftlc1DZtLme-U..14c. 2-cud.m1Doisava.leric aci4-U-14c. isobltJIZ'01'li tril.e-1-t4c trne-1-1'tc all teci in� l.iJiumrin

t 1M-u-14c. By at 1� labelled

t:4 .t.aobl�!ll&:alliae .-dA mtact t.lllr'1Ea' the calftlrm.cm

���������MW' �

l.ea&er extent.

Di trUe moioty d iAf."tA.uJ:WII.w...

tly CODVES"ted tD pi'UmSin.

�· Both � and --t,rar:d.tdle 1Jl abaota L-valiDIS Uv 14c.

w.

t with acid

aunan fiE tbm L-.a:U.Il0-U.1 ea" L-

pberw� '4c. �eri.c Nn�·14c waa not ai.gn1ti Uy

1n

Irxlus tria.l -�· I

tor the oppca:-tuni ty to s1nd,y in his cleprtment. I am indebted to

most h pf\11.. In pertiollar I \'IOUld llke to thank l. J. l'e

a1ao gratefUl �ar occasional techrdcal assistance f'ran !!l' • D. Bennett and f"or

his ccopen&tion in develop� a me od at detectin(; cyn.no[_'Cni.c C).,yoosides. Some d the eaperinents in this investi.::;atioo 'WO.lld not mve bt.-en possible

tbot.tt the w1ll.1ng aaaistance of iJr J. C. Hawke, l�y University, '\11th gas

chrGia�, er Ir J. R. fulston. Institute et �lear Sciences, far- 15N analyses. I am gratci\Ll tc DE- �· ::. Underhill and lroi". 14c labellecl cCX!pWJ1da and seede.

Not ta I woald liD to thank · .c:. Ccmn �or th intarest and attnul et1011 he

• Dr G. • Uer. nr

It has heal a pleawre to coopera tAt with

PAGE

i1

� iv

LIST OF TABTZ.S Zlii

LIS1' OF .FIGURES 1z

I lN'l'ROiroTIOO 1

1.1 G�id 1

1.2 General b\mction of GJ..yoos:ld 2 1.3 Gl�Uon 3

1.4 Cyanogenic gl;ycosi 4 Cbemistry 4 D1atr1blt1on and tunctioo 6

' tabal.i&n 8 . 1.5 Glucosinola '

Cbemiat;ry 9

Dlatrib.ltion 13

14

1.6 . cgeDet1c tionship be gl.ucoeinola 19

1.7 ide b1oayD i.a 20

1.8 2S 1., 27

II D JO 2.1 t 2.2 33

2.3 .38 M 42

s 1..1

11

2.6

2.7

2.8

9 Ill

J.1 2

.,

t.

D !.� CctnD •••

Chl'oma 44. 4.7

�t of rodioaet1v1ty 51 Al1alyais of 15ri 53

In RE.:rul..T:S

tioo t1on cl into

.A and B) Mministration c4 1� la ......... _ u-.UIIil"' to t'laz

( imant C) I tyramide and isobu t,rmitril.e as pro-

54. St.

60

oursars c£ limmarin (�ts D and - ) 63

2-�oln�trile-1-14c aa a .. eurser d l1namarin (uperiment F) 65

� of pr:umain (aperimnts �· to I)

� oi.nistra tion (:£ pbenyla ton1 trll 1-14-c and phalylac� 1-14c to pe&Am Ober.ry .&.Bi� ( J ) 77

nrtli'Dr!ler or 79

' 8)

labelled wlaUie intettlediatcs, I (tD:pet"i­ment 1')

lo.belled vola tile intermediates • II ( ezper1ment ')

&opartles am stzucture or carvwmd IV

( ezperi.nx;ln R to tJ) 3.4 GJncosf na1 te bJ.oayntbeaia

lrec:ursc::ars of isoprop71glucosinola nBD1&S V and . )

1 Glllcosinola te b1.osyntbesis

4.2 Cyanogcdc �e biosynth

100

108

108

112

116 117 124 127 140

I n

Ill IV

y

VI

VII

VII�

IX

X

xn

Qldmea aa proeat.II'8QI'8 Oft l.im.!!Brln

�pc:lfttiCID f'.l 1� into l.im.marin

lt:JDarparatlon at iaobl�trlle into l.i.mJ:::m:':ln

Iaota-pc:n.tisll or 2-h�tyrcmitril.e into 1.1m.aar1n

Od... .. precursors c:£ p:nma.sin 1n obelrty laurel

Ozl.mea precu.rsars af � in r>eac::h �tlon <£ phenylaceteoitrile-1-14c and pbeeylaoo­

th;gdxoc'D"dc acid-1-14u into prwaasiD Ma.rdelord. trUe as a precursor of pz:unasin

Reaul.

�feet of D.L-2-a&�ioDeldO&ime propioni trUe

XIII Sft'ect � D�tbreaDinr!t an4 iaobut)Taldo'dme an

J.V

l.inaJIBr1ft bioa;rnt.bea18 ant:� ti ea- !Dbi'W.tar Cl'1 l.t.r.amar1n bioeyn� ...

tzaa x...u..-u-14c

PAGE

5

"

62

61..

68

75

76

10t

1

2

'

4 s

6

LIST OF FIGURES

Em�� �yaia of gluoos:l.nola tsof.llioc;yua,tes aD1 Di tril

to gi thiocyana

I'DaarpOI!'at.ion o£ radioa.ot.ivi ty haD I,.. ......ul:lU tlaz

IDoQrparatt.on c:4 rad1 cti.v1 ty ac14-U-14c 1n linen n.a.x

•

7 lnoorpar-aU.on fl radimct.i.'fitQ 1aolu 14c

1n l1nen nax 8 �oration c£ rad.i..c6ctiv1 ty &am iaob.&�eh.Jde-U-14c

in linen flax

Incor--rzro.tion cl radiaa.ct.i.Tity t'.roiB L- :me-u-14-c. l.eo-btt t;:troni tri.l.e-1-14c &rld 1aoblt.namie-1-14c 1n lineD t1u

10 �at.ion ar tlvitijr �tt.Jranltri.lo-

p

10

12

18

57

61

"

1-14c �ea c,anl.d 14c 1n l.1Den tlnx 69 11 lDDorparation t1 radioactivity frail � cynn14e-iltc

12 �����m� �� llltril.-1-14c into am'1110 --� in l1Den t"l..az

ia

15

71

7l

17 glycol su.ccinAte oolumn wl th the 1nre ni'DarA:IIID8

indicated

18 Rad1oactivi.1;y in 'VOlatile ca:rpormds .t'ftHn l1nen t1u treated th L- ine-tJ-14c and other tiiJl.e.belled

19 Raatoaoti.Yi ty 1n ted volatil COIIpOW'lds from linen f'laz tee! with L--val.ine- 14c and other unlabelled conpomds

wlat.U ra41mcti"fity horn� rl at"ter trea12.alt

X

with 0. � ao1cl at !!)0 10.. Radioaotivity in latue coq:omds obtained by tretJ. ...... , ... �

cl ODqiO'mcl IV 105 22 Iaobu\Yl'aldoxi.m&-()..gluooside; a posm..ble structure fer

OCiqO'Dd IV 23 lnaalpanr.tlon ot radioactivity trom L-vali.ne-U-14-c in scun:,y

25

26 27

gi'U8

lnl»rpca-ati.on ot radicect1Y1ty fro iaobl� 14-c in acarvy �-as>

Incorporation cl radioaot.t ty ia,..u-14.� in ClU'Yy ---

I-ropoeed path-.y of cyancgm:lc g&.lliDQaG'1!1 bl.osyntb..s.s rd.trilea

107

110

111

11J

121

122