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 •
leolu
t7ftlc1DZtLme-U..14c.
2-cud.m1Doisava.leric aci4-U-14c. isobltJIZ'01'li tril.e-1-
t4c
trne-1-1'tc all teci in� l.iJiumrint 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 201.8 2S
1., 27
II D JO
2
.
1 t2.2 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"i
ment 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.marinlt: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
ac
14-U-14c
1n linen n.a.x•
7
lnoorpar-aU.on fl radimct.i.'fitQ 1aolu 14c1n 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 t1u10 �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� ��
lll
tril.-1-14c
into am'1110 --� in l1Den t"l..azia
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
