hill1981-sex pheromone of the saltmarsh

8/16/2019 Hill1981-Sex Pheromone of the Saltmarsh

1/14

Journ al of Che mical Ecology, VoL 7, No. 4, 1981

S E X P H E R O M O N E O F T H E S A L T M A R S H

C A T E R P I L L A R M O T H E s t i g m e n e a c r e a 1 2

A D A S . H I L L a n d W E N D E L L L . R O E L O F S

Depar tment o f En tomologyNew York State Agricul tural Experiment Stat ion

Geneva, New York 14456

Rec eived Augu st 4, 1980; revised Sep tem ber 22, 1980)

Ab s t rac t - -T hree comp ounds have been iden t if i ed as com ponents o f the sexphero mo ne emit ted by females of the sal tmarsh caterpi l lar moth,Est igmeneacrea Drury) . These are Z,Z)-9,12-octadec adienal I ),Z,Z,Z)-9,12,15-octadecatr ienal I I ) , andZ,Z)-3,6-cis-9,10-epoxyheneicosadiene III) . Infemale t ip extract they were foun d in a ra t io o f 1 : 6 : 25, respect ively, and intrapped fema le eff luvia the ra t io was 1 : 6 : 27, respectively. Co mb inat ions of1II wi th e i ther I o r I I e l ic ited susta ined upw ind f l ight in a wind tunnel , bu t

none of these com pou nds by themselves did so. There is evidence that theanten nal acc epto r site for II1 is chiral .

Key Words- -Sa l tmarsh ca te rp i l l a r moth ,Estigmene acrea, Z,Z)-9,12-octadecadienal ,Z,Z,Z)-9,12,15-octadecatrienal, Z,Z)-3,6-cis-9,10-epoxy-heneicosadiene, fa l l webworm moth,Hyphantria cunea,insect sex phero -mone,cis-9,10-epoxyheneicosane, trans-9,10-epoxyheneicosane, l inolealde-hyde, l inolenaldehyde, Lepidoptera, Arctiidae.

I N T R O D U C T I O N

The sa l tm ar sh ca t e rp i ll a r m o th , sti gmene acrea D r u r y ) , o c c u r s t h r o u g h o u tthe Nor th Amer ican con t inen t and appears to be unknown e l sewhere . Thel a r v a e f e e d o n t h e f o l ia g e o f a w i d e v a r i e t y o f g a r d e n a n d f i e l d c r o p s a n d a r e

u s u a l l y a p e s t o f m i n o r e c o n o m i c i m p o r t a n c e M e t c a l f e t a l . , 19 62 ).

T h e s e x p h e r o m o n e s o f o n l y a f e w s p ec ie s in th e A r c t i i d a e f a m i l y h a v e

b e e n r e p o r t e d t o d a te . T h e s e i n c l u d e m e m b e r s o f t h eH o l o m e l i n a a u r a n t i a c a

c o m p l e x , H o l o m e l i n a l a e t a G u e r i n ) , a n d I s i a i s a b e l l a J . E . S m i t h ) G u e r i n )

Lepidop tera: Arct i idae.2Suppor ted in par t by the R ockefe l le r Fou nda t ion and by Nat iona l Sc ience Fou nda t ion Gran tsGB-38020 and P CM 78-13241.

6 5 5

0098-0331 / 81 / 0700-0655503.00/ 0 9 1981 Plenum Pub l ishin g Cor po rat io n


2/14

6 6 H I L L A N D R O E LO F S

( f o r m e r l yPyrrhactia isabella , f o r a ll o f w h i c h 2 - m e t h y l h e p t a d e c a n e h a s b e e nr e p o r t e d a s a p h e r o m o n e c o m p o n e n t ( R o e l o f s a n d C a r d e , 1971). W e in i ti a te da n i n v e s t i g a ti o n o f t h e s ex p h e r o m o n e s y s te m o fE. acrea b e c a u s e i tr e p r e s e n t e d a n o t h e r a r c t ii d s p e c ie s . In i ti a l t es ts i n v o l v in g e l e c t r o a n t e n n o g r a m( E A G ) s tu d i e s o f m a t e r i a ls o b t a i n e d f r o m f e m a l e t ip e x t r a c t s i n d i c a te d t h a t2 - m e t h y l h e p t a d e c a n e p r o b a b l y w a s n o t i n v o l v e d , s o a f u ll -s c a le in v e s t i g a ti o no f t h e Eo acrea s ex p h e r o m o n e s y s te m w a s u n d e r t a k e n .

M E T H O D S A N D M A T E R I A L S

S o l v e n t s w e r e d i st il le d t h r o u g h a 1 0 -p la te O l d e r s h a w c o l u m n . Te m p e r a -t u r e s a r e g i v e n in ~ C . G a s c h r o m a t o g r a p h y ( G C ) c o l u m n s w e r e g la ss , e i th e r 2m m • 1 .8 m , 2 m m X 3.6 m , o r 4 m m • 1 .8 m , a n d w e r e p a c k e d w i t h o n e o ft h e f o l lo w i n g : O V- 1 o r O V- 1 01 ( m e t h y l s il ic o n e , 3 o n 1 0 0 - 1 20 m e s h G a s -C h r o m Q ) ; H i - E f t ( H i - E f t 8 B P, c y c l o h e x a n e d i m e t h a n o l s u c c in a t e, 3 o n1 0 0 -1 2 0 m e s h G a s - C h r o m Q ) ; X F - 11 5 0 ( G E X F - 11 5 0 , 5 0 c y a n o e t h y l ,m e t h y l silicone~ 1 0 o n 10 0 -1 2 0 m e s h C h r o m o s o r b W - A W - D M C S ) , P o r a -p a k Q ( 1 0 0 -1 2 0 m e s h ) o r Te n a x ( Te n a x G C , 6 0 - 8 0 m e s h ). T h e c a r r i e r ga s f o rG C w a s n i tr o g e n a n d h y d r o g e n f la m e i o n i z a t io n w a s u s ed . M a s s s p e c tr a ( M S )w e r e d e t e r m i n e d w i th e i th e r a P e r k i n - E l m e r H i ta c h i m o d e l R M U 6 i n te r f a c e d

w i th a n O V- I G C c o l u m n , o r w i th a F i n n ig a n 3 30 0 d u a l q u a d r u p o l e m a s ss p e c t o m e t e r in t e r f a c e d w i th a n O V- 1 01 G C c o l u m n ( C o r n e l l U n i v e r s it y M a s sS p e c t r o m e t r y C e n t e r ) . T h e i n f r a r e d s p e c t r a w e r e r e c o r d e d u s i n g e i t h e r aP e r k i n - E l m e r m o d e l 2 5 7 g r a t in g i n f ra r e d s p e c t r o p h o t o m e t e r , a N i c ol e t 7 19 9F o u r i e r Tr a n s f o r m I n t e r fe r o m e t e r | l o c a te d a t t h e U S D A L a b o r a t o r i e s inG a i n e s v i ll e , F l o r i d a , o r a N i c o l e t F o u r i e r Tr a n s f o r m I n t e r f e r o m e t e r | l o c a t e da t th e U S D A L a b o r a t o r i e s in B e lt sv ill e, M a r y l a n d . P r o t o n m a g n e t i c re s o n a n c e( P M R ) s p e c tr a w e r e e i th e r f r o m a V a r i a n H A 100 o r a Va r i a n X L 100e q u i p p e d w i th F T l o c a t e d a t t he S c h o o l o f F o r e s t r y, S y r a c u s e , N e w Yo r k .

U l t r a v i o l e t ( U V ) s p e c t r a w e r e d e t e r m i n e d w i th a C a r e y m o d e l 15 r e c o r d i n gs p e c t r o p h o t o m e t e r .

E l e c t r o a n t e n n o g r a m s ( E A G s ) w e re d e t e r m i n e d a s d e s c r ib e d b y R o e l o fs(1977 , an d r e f e r en ces t he re in ) . The sus t a ined f l i gh t t unne l , o r w ind tunne l , ha sb e e n d e s c r i b e d p r e v i o u s l y ( M i l le r a n d R o e l o f s , 19 78 ) a n d w a s m o d e l e d a f t e rthose desc r ibe d by K en ned y (1977) and Fa r kas e t a l . ( 1974).

I n s ec t s w e r e r e a r e d o n a p i n t o b e a n d i e t ( S h o r e y a n d H a l e , 1 96 5). T h e s ei n s e c t s o r i g i n a t e d f r o m t w o d i f f e r e n t s o u r c e s . O n e w a s f r o m a c u l t u r ep r e v i o u s l y m a i n t a i n e d in Te x a s ( Te x a s c u l tu r e ) , w h i c h w a s u s e d d u r i n g t h ef ir s t p h a s e o f t h is i n v e s t i g a ti o n , a n d t h e o t h e r w a s f r o m a s i n gl e f e m a l ec o l l e c t e d f r o m a b l a c k l i g h t t r a p i n G e n e v a , N e w Y o r k ( G e n e v a c u l t u r e ); th i sc u l t u r e w a s s u p p l e m e n t e d o c c a s i o n a l l y w i t h a d d i t i o n a l f i e ld - c o l le c t e d s p e ci -m e n s , a n d w a s t h e o n e u s e d d u r i n g t h e l a t e r p h a s e o f t h is i n v e s t ig a t i o n ,inc lud ing the w ind tunne l b ioas says .


3/14

PHEROMONE OF THE S LTM RSH C TERP ILL R 6 7

P u p a e w e r e s e g r e g a t e d a c c o r d i n g t o s e x a n d w e r e a l l o w e d t o e m e rg e a tc a . 2 0 - 2 5 ~ C u n d e r a 1 6 : 8 l i g h t - d a r k c y c l e. F e m a l e s w e r e c o l l e c te d d u r i n g t h ef i rs t p a r t o f t h e i r s c o t o p h a s e a t 1 - 3 d a y s p o s t e c l o s i o n ; e a c h a b o m i n a l t ip w a se x t r u d e d m a n u a l l y, sn i p p e d o f f , a n d e x t r a c t e d w i t h m e t h y l e n e c h l o r i d e .A l t e r n a t e l y, t h e f e m a l e s w e r e c o l le c t e d a t a n y t i m e d u r i n g t h e i r li g h t p h a s e ,a n d t h e g l a n d s w e r e p u ll e d o u t f r o m t h e i ns e c ts a b d o m e n s w i t h d i s se c t in gf o r c e p s . T h e g l a n d s o c c u r as a p a ir o f t u b e s a b o u t 3 m m l o n g b y a b o u t 0 .2 m mO D , w i th s e p a r a t e o p e n i n g s a t e i th e r s id e o f t h e m i d - d o r s a l l in e o n t h e la s ta b d o m i n a l s e g m e n t . T h e s e a ls o w e r e e x t r a c t e d w i th m e t h y l e n e c h l o r id e .E x t r a c t s w e r e s t o r e d i n a f r e e z e r. M a l e s w e r e h e l d 1 - 1 0 d a y s a t 2 0 - 2 5 ~ C u n d e ra 1 6 : 8 l i g h t - d a r k c y c le . T h e y w e r e r e m o v e d a n y t i m e d u r i n g t h e l a st h a l f o fthe i r s co to pha se , b ro ug h t i n to l i gh t (~> 1 l ux ) , and the n b io as say ed in t he f l i gh tt u n n e l s t a r ti n g a b o u t 15 m i n a f t e r r e m o v a l f r o m s c o t o p h a s e c o n d i ti o n s . T h eb ioas say pe r iod l a s t ed 30 -60 r a in .

T h e l in o l e y l a n d l in o l e n y l a l c o h o l s u s e d f o r s y n t h e si s o f t h e c o r r e s p o n d i n ga l d e h y d e s w e r e p u r c h a s e d f r o m N u C h e c k C o r p . ( E l y si a n, M i n n e so t a ).A c e t a l d e h y d e , p r o p a n a l , n o n a n a l , a n d d o d e c a n a l w e r e p u r c h a s e d f r o mA l d r i c h C h e m i c a l C o . P e n t a d e c a n a l a n d o c t a d e c a n a l w e r e p r e p a r e d f r o m t h ec o r r e s p o n d i n g a l c o h o ls , w h i ch w e r e p u rc h a s e d f r o m A l d r ic h C h e m i c a l C o .A l d e h y d e s w e r e p r e p a r e d f r o m t h e c o r r e s p o n d i n g a l c o h o ls b y t r e a t m e n t w i th

p y r i d i n i u m c h l o r o c h r o m a t e ( P C C ) b y th e m e t h o d o f C o r e y a n d S u g g s (I 9 75 ).T h e ( Z ) - 9 - t e t r a d e c e n y l a c e t a te w a s p u r c h a s e d f r o m F a r c h a n C h e m i c a l C o .

C o m p o u n d I I l [F i g u r e l , ( Z ,Z ) - 3 , 6 - c is - 9 ,1 0 - e p o x y h e n e i c o s a d ie n e ] w a sp re pa red by B . Ko va lev v i a a rou t e desc r ibed e l s ew here (H i l l e t a l. , 1981).

0

I . Z , Z ) - 9 , 1 2 - o c t a d e c a d i e n a l

0

~ \

I I . Z , Z , Z ) - 9 , 1 2 , 1 5 - o c t a d e c a t r i e n a l

~ 0

I I I . Z , Z ) - 3 , 6 - c i s - 9 , l O - e p o x y h e n e i c o s a d i e n e

FIG. 1. S tructu res of I, I1, and Ill.


4/14

6 8 HILL AND ROELOFS

T h e t w o e n a n t i o m e r s o f c / s -7 , 8 - e p o x y - 2 - m e t h y l o c ta d e c a n e ( d is p a r lu r e )were prepared by Dr. K. Mori (Mori e t a l . , 1979) .

M ic roche m ica l r eac t ions were usua l ly ca r r ied ou t in 1 -d ram v ia l s hav ingTef lon- l ined sc rew. -caps and rou t in e ly invo lved the use o f a sub s tan t i a l excesso f the r eag en t s invo lved . Som e o f these r eac t ions have been desc r ibed in de ta i le l sewhere , such as ozo no lys i s (Beroza and Bie rl , 1967) an d a ce ty la t ion wi thace ty l ch lo r ide (Hi l l e t a l. , 1977). Ca ta ly t i c hy dro gen a t ion was w i th Pd /C aC O 3i n m e t h a n o l o r e th a n o l a t I a tm o s p h e r e o f H 2 a n d r o o m t e m p e r a t u re .E p o x i d e s w e r e c o n v e r t e d t o d i o ls b y t r e a t m e n t w i t h 0 .5 H 2 8 0 4 i n 5 0a q u e o u s t e t r a h y d r o f u r a n [ T H F : f r e s h ly d i st il le d f o r m l i th i u m a l u m i n u mh y d r i d e ( L A H ) ] f o r 4 h r a t r o o m t e m p e r a t u r e ; t h e p r o d u c t w a s re c o v e r e d b yd i lu t ion wi th H 20 and ex t rac t ion w i th Ske llyso lve -B (pe t ro leum e the r 60 -68 ~ )(Ske l ly B). Red uc t ion s wi th L A H w ere ca r r i ed ou t e i the r in d ry E t20 o r in d ry,pur i f i ed TH F fo r 10 m in to 1 h r a t room tem pera tu re ; p rodu c t s were r ecoveredb y a d d i t i o n o f d i l u te , a q u e o u s N a O H ( 1 -5 ) a t i c e -b a t h t e m p e r a t u r e u n d e rN2 a n d e x t r a c t i o n w i t h S k e U y B . H y d r o x y c o m p o u n d s w e r e c o n v e r t e d to t h ec o r r e s p o n d i n g b r o m i d e s b y r e a c t io n f o r 1 h r o r l o n g e r a t r o o m t e m p e r a t u r ew i t h t r ip h e n y l p h o s p h i n e d i b r o m i d e ( T P P D B ) , f r e sh l y p r e p a r e d f r o m t r ip h e n y lpho sph in e and b rom ine in CH2C12 (So nne t and Ol ive r, 1976); p rodu c t s werer e c o v er e d b y e v a p o r a t i o n o f t h e s o l v e n t in a s t r e a m o f N2 a n d e x t r a c t i o n o f t h e

p r o d u c t w i t h S k e l ly B . H y d r o x y l g r o u p s w e r e t ri m e t h y l s i l a te d w i t h h e x a -m e t h y l d i s i l a z a n e ( H M D S ) a n d t r ic h l o r o s il a n e ( T C S ) i n p y r id i n e (S w e e le ye t al ., 1963); t he p rod uc t s were e i the r ana lyz ed d i r ec t ly by GC or wererecovered by evapo ra t ion in a s t ream of Nz, add i t ion o f water, and e xt rac t ion w i thSkel ly B.

Synthesis of cis- and trans-9 10-Epoxyheneicosanes. 1-Decyne (1.7 g,1 2.3 ra M ; F a r c h a n R e s e a r c h L a b o r a t o r i e s ) i n d r y d i o x a n e (5 0 m l ) w a s t re a t e dwi th l i t h ium a m ide (1.1 g , 48 m M ; A lpha Inorgan ics ) fo r 3.5 h r a t r e f lux , a f t e rw h i c h i t w a s c o o l e d t o r o o m t e m p e r a t u r e a n d l - b r o m o u n d e c a n e ( 6 . 4 g , 2 7

m M ; C h e m i c a l S a m p l e s C o . ) w a s a d d e d d r o p w i s e o v e r a p e r io d o f c a . 3 0 m i n .T h e m i x t u r e w a s k e p t a t r e f l u x o v e r n i g h t . A f t e r c o o l i n g , t h e m i x t u r e w a sd i l u t e d w i t h w a t e r a n d e x t r a c t e d t w ic e w i t h S k e l l y B ; t h e S k e l l y B l a y er s w e reco m bin ed an d w ashe d success ive ly wi th d i lu te n i tr ic ac id (ca . 1 ) , water,s a t u r a t e d s o d i u m b i c a r b o n a t e , a n d w a t e r. F i l t r a t i o n t h r o u g h a m a g n e s i u ms u l fa t e b e d w a s f o l lo w e d b y e v a p o r a t i o n o f t h e s o l ve n t . T h e r e c o v e r e d c r u d eprod uc t , 9 -hene icosyne (6.1 g ), was f i lt e red th roug h F lo r i si l (pe t ro leum e the r ),a n d t h e r e c o v e r e d m a t e r i a l s w e r e c o n v e r t e d t o t h e a l k e n e s w i t h o u t f u r t h e rp u r if ic a t io n . R e d u c t i o n w i th s o d i u m - l i q u i d a m m o n i a ( W a r t h e n a n d J a c o b -son , 1973) y ie lded (E) -9 -hene ico sene , wh ich s ti ll con ta ine d l a rge am ou nt s o ft h e a l k y n e . T h e E i s o m e r s r e q u i re d w e r e p u ri f ie d b y G C c o l l e c t io n f r o mHi-Ef t . Ca ta ly t i c r edu c t ion wi th Pd /C aC O 3 and qu ino l ine (Green e t a l ., 1967)y i e l d e d ( Z ) - 9 - h e n e i c o s e n e . T h e e p o x i d e s w e r e f o r m e d f r o m a l k e n e s u s i n gm - c h l o r o p e r b e n z o i c a c id .


5/14

PH ROMON O F T H E S L T M R S H C T E R P I L L R

R S U LT S

659

I n i ti a l ly, a n a l i q u o t c a . 1 0 F E ) o f a c r u d e f e m a l e t i p e x t r a c t f r o mE. acreaw a s c o l l e c t e d i n t i m e d f r a c t i o n s f r o m a G C c o l u m n a n d t h e f r a c t i o n s w e r ea s s a y e d f o r E A G a c t iv i ty. U s i n g a n O V- 101 c o l u m n , o n e d i s ti n c t a r e a o f E A Ga c t i v i t y w a s a l w a y s e v i d e n t , w i t h a s e c o n d , e a r l i e r , a c t i v e a r e a s o m e t i m e sp r e s e n t. T h i s e a rl y a r e a w a s m o r e d i s ti n c t a n d r e p r o d u c i b l y d e t e c t e d w h e ng l a n d e x t r a c t s w e r e u se d i n s te a d o f w h o l e t ip e x t r a c t s. F e m a l e e f f l u v i u mr e c o v e r e d f r o m P o r a p a k Q a l s o r e p r o d u c i b l y s h o w e d t w o d i s t i n c t a r e a s o fa c t i v i ty w h e n c o l l e c te d f r o m O V- 1 0 1 . W h e n e a c h o f t h e t w o E A G - a c t i v em a t e r i a l s w a s r e c o v e r e d a n d r e - c ol le c t e d f r o m a H i - E f t c o l u m n , t h e e a r l yO V- 1 0 1 a r e a o f a c t i v it y r e s o lv e d i n to t w o E A G - a c t i v e m a t e r i a l s , w h e r e a s t h el at e O V-1 01 f r a c t io n s h o w e d o n l y o n e E A G - a c t i v e c o m p o n e n t t h a t h a d al o n g e r r e te n t i o n t i m e o n H i - E f t t h a n e i th e r o f t h e o t h e r t w o c o m p o n e n t s .T h e s e c o m p o n e n t s w e r e d e s i g n a t e d I , I I , a n d I I I , i n t h e ir o r d e r o f e l u t i o n f r o mH i - E f t . P r o m i n e n t G C p e a k s c o in c i d e n t w i t h e a c h o f t h es e a r e a s o f E A Ga c t i v i ty w e r e e v i d e n t in G C t r a c in g s o f t h es e e x t r a c t s. T h e i r e q u i v a l e n t c a r b o nn u m b e r s w e r e c a l c u l a t e d t o b e t h e fo l l o w i n g u s i ng a s e ri es o f n - h y d r o c a r b o n sa s r e f e r e n c e s t a n d a r d s ) : o n O V - 1 0 1 , 2 0 0 ~ , 19 .7 3 f o r I a n d I I , a n d 2 2 .3 5 f o r I I I ;o n H i - E f t , 2 0 0 ~ , 2 3 .6 f o r I , 24.1 f o r I I , a n d 2 5 .3 f o r I I1 . T h e a p p r o x i m a t e r a t i o sb e t w e e n t h e t h re e c o m p o n e n t s w e r e c a lc u l a t e d t o ~ e a s f o ll o w s : f o r a w h o l e t ipe x t r a c t 1 : 6 : 2 5 , f o r a g l a n d e x t r a c t 1 : 6 : 10, a n d f o r r e c o v e r e d f e m a l e e f f l u v i u m1 : 6 : 2 7 .

Iden t i f i ca t ion o f I and H.C o m p o u n d I w a s i d e n ti f ie d a s Z , Z ) - 9 , 1 2 -o c t a d e c a d i e n a l l i n o l e a l d e h y d e , C A S r e g i s t r y N o . 2 5 41 - 6 1- 9 ), a n d I I w a si den t i f i e d a s Z,Z ,Z) -9 ,12 ,15-oc tadeca t r i ena ll i n o l e n a l d e h y d e , C A S r e g i s t r yN o . 2 4 2 3 -1 3 - 4) , a s d e s c r i b e d b e l o w . T h e s e s t r u c t u r e s a r e p r e s e n t e d i n F i g u r e 1.S a m p l e s o f I a n d I I w e r e p u ri f ie d b y c o l le c t io n o f t h e c r u d e p h e r o m o n e e x t r a c t

f r o m a n O V- 1 o r a n O V - 10 1 c o l u m n f o ll o w e d b y r e - co l le c t io n o f t he E A G -a c t i v e m a t e r i a l s f r o m a H i - E f t c o l u m n , w h i c h c o m p l e t e l y re s o l v ed t h e tw oc o m p o n e n t s .

C a t a l y t i c h y d r o g e n a t i o n o f I a n d o f I I y i el d ed i n d i s ti n g u i sh a b l e p r o d u c t s .I n b o t h e a s e s t h e p r o d u c t h a d t h e s a m e G C r e t e n t i o n t i m e as n - o c t a d e c a n a l o nO V- 1 01 a n d o n H i - E f t . T h e C I - M S m e t h a n e ) o f h y d r o g e n a t e d I , h y d r o g e -n a t e d I I , a n d n - o c t a d e c a n a l w e r e i d e n ti c a lm / e 269 , M H +, a s t h e onep r o m i n e n t i o n ) .

C I - M S i s o b u t a n e ) o f I a n d I I s h o w e d M H § i o n s a t 2 65 a n d 2 63 ,

r e s p e c t i v e ly, i n d i c a t i n g t h a t I i s d i u n s a t u r a t e d a n d I I i s t r i u n s a t u r a t e d . T h e s es p e c t r a w e r e v e r y s i m i la r t o t h o se o b t a i n e d f o r l in o l e a l d e h y d e a n d l i n o le n a l d e -h y d e , r e s p e c ti v e ly. T h e G C r e t e n t i o n t i m e s o b s e r v e d f o r I a n d I I o n O V - 1 0 1 ,H i - E f t , a n d X F - 11 5 0 w e r e w i t h in 0 .1 r a i n o r l es s o f t h o s e f o r l i n o l e a l d e h y d ea n d l i n o l e n a l d e h y d e , r e s p e c t i v e l y.


6/14

66 HI LL AND ROELOFS

S a m p l e s o f I a n d I I w e r e p u r i f ie d a n d e a c h w a s s u b j e c t e d t o t h e f o l lo w i n gs e q u e n c e o f r e a c ti o n s: ( 1) tr e a t m e n t w i th L A H i n d r y E t 2 0 t o p r o d u c e t h ea l c o h o l s , ( 2) a c e t y l a t i o n w i t h a c e t y l c h l o r i d e t o p r o d u c e t h e a c e t a t e s , a n d ( 3)m i c r o o z o n o l y s i s t o p r o d u c e a c o m p o u n d i d en t if ie d a s 9 - a c e to x y n o n a n a l .A f t e r e a c h r e a c t io n , th e p r o d u c t s w e r e e x a m i n e d b y G C o n a n o n p o l a r c o l u m na n d a p o l a r c o l u m n ( 6. 7 m i n a t 1 70~ a n d 4 . 0 m i n a t t 7 0 ~ r e s p e c ti v e ly ) a n dm a t c h e d t h o s e o f t h e e x p e c t e d p r o d u c t s w i t h in 0 .1 m i n o r l es s. P r i o r t o th eo z o n o l y s e s , t h e a c e t a t e s f r o m I a n d I I w e r e c o ll e c t e d f r o m O V- 1 0 1 . A u t h e n t i c9 - a c e t o x y n o n a n a l w a s p r e p a r e d b y o z o n o l y s i s o f ( Z ) - 9 - te t r a d e c e n - l - y l a c e t a t e .T h e C I - M S ( i s o b u t a n e ) o f a l l t h r e e o z o n o l y s i s p r o d u c t s w e r e , f o r al l p r a c t i c a lpu r pos es , t he s ame (m /e 201 M H § wi th o the r ions a l l le ss t han 20 o f t h is ) .T h e s e d a t a s u p p o r t a 9 - p o s i t io n d o u b l e b o n d i n I a n d I I.

O z o n o l y s is o f p u r i fi e d I p r o d u c e d n - h e x a n a l , a s d e t e r m i n e d b y c o m p a r -i so n o f t h e G C r e t e n t i o n t im e s o f t h e p r o d u c t w i th s y n t h e t ic h e x a n a l o nXF -1150 , 100 ~ (5.2 and 5 .3 m in , r e spec t ive ly ) . These da t a i nd i ca t e a 9 ,12d o u b l e - b o n d s y s te m in c o m p o u n d I .

P u r i f i e d I I w a s al s o o z o n i z e d to p r o d u c e n ' p r o p a n a l , w h i c h w a si d e n t if i e d b y i ts G C r e t e n t i o n t i m e o n X F - 11 5 0 ( 4 .4 m i n a t 5 0 ~ ) c o m p a r e d t ot h a t f o r s y n t h e t i c n - p r o p a n a l ( 4 .3 5 r a in ) . T h e s e d a t a i n d i c a t e t h a t t h e e n dd o u b l e b o n d s o f t h e t h r e e i n c o m p o u n d I I a r e in p o s i t io n s 9 a n d 15.

A U V s c a n o f II ( c y c t o h e x a n e ) sh o w e d n o e v i d e n c e o f c o n j u g a t i o n . T h eG C r e t e n t i o n t im e s o f II o n t h e v a r i o u s c o l u m n s a l s o c o n f i r m e d t h e a b se n c e o fc o n j u g a t i o n i n II . T h i s le a v e s o n l y p o s i t io n s 9 , 1 2, a n d 15 f o r t h e d o u b l e - b o n ds y s te m in c o m p o u n d I I.

A n F T - I R s p e c t r u m o f II re v e a l e d th a t t h e d o u b l e b o n d s in I I a r e al l Zd o u b l e b o n d s ( F i g u r e 2 ). T h e r e w a s s tr o n g c a r b o n y l a b s o r p t i o n a t 1 73 1c m 1

Iden t i f i ca t ion o f l l I a s Z ,Z) -3 ,6 -c i s -9 ,10-Ep oxyhe ne icosad iene .S a m -p le s o f I I I w e r e p u r i f i e d f o r t h e v a r i o u s a n a l y s e s b y c o l l e c t io n o f th e c r u d ef e m a l e t ip e x t r a c t f r o m O V- 1 o r O V- 10 1 f o l lo w e d b y r e - c o l le c t io n f r o m

H i - E f t. I n s o m e i n s t an c e s , t he c o l le c t io n s w e r e p r e c e d e d b y c h r o m a t o g r a p h yon F lo r i s i l, f ro m wh ich II I e lu t ed w i th ca . 5 -1 0 E t2 0 in Ske l ly B ( thee f f e c ti v e m i x t u r e w a s d e p e n d e n t o n t h e a c t i v i ty o f th e F l o ri si l) . T h i s w a sc o n s i s t e n t w i th a n e p o x i d e s t r u c tu r e .

A m a s s s p e c t r u m ( E l, 7 0 e V ) o f I II s h o w e d a m o l e c u l a r i o n a tro le 306,w i t h a n i o n a t ro l e 2 8 8 ( M - 1 8 ) , c o n s i s t e n t w i t h a n e p o x i d e s t r u c t u r e .H y d r o g e n a t i o n o f I II o v e r P d / C a C O 3 p r o d u c e d a c o m p o u n d w i th a s h o r te rr e t e n t i o n ti m e t h a n I I I o n O V-1 01 a n d H i - E f t ( e q u i v a le n t c a r b o n n u m b e r o f2 1 .2 ). T h e m a s s s p e c t r u m ( E l, 7 0 e V ) o f h y d r o g e n a t e d I I I h a d a m o l e c u l a r i o na t 3 1 0 , a n d a n i o n a t m / e 2 9 2 ( M - I 8). T h i s w a s c o n s i s t e n t w i t h t h e p r e s e n c e o ftwo c loub le bond s i n I I I .

To e s t a b l i s h t h a t I I I i s u n b r a n c h e d , h y d r o g e n a t e d I I I w a s t r e a t e ds u cc e ss iv e ly w i t h L A H i n E t 2 0 , t r ip h e n y l p h o s p h i n e d i b r o m i d e ( T P P D B ) i nC H 2C 12 , a n d t h e n a g a i n w i t h L A H . T h i s s e q u e n c e r e d u c t i v e ly r e m o v e d t h e


7/14

P H E R O M O N E OF T H E S LT M R S H C T E R P I L L R 66

3 2 1 8 PPM

WAVENUMBERS

I I

3 2 1 8

WAVENUMBERS

FIC. 2. F T- IR of II and III (as films).

e p o x i d e g ro u p a n d p r o d u c e d a c o m p o u n d w i th th e s a m e G L C r e te n t io n tim e s

o n O V - 1 0 1 , 1 8 1 ~ ( 2 0. 7 m i n ) , a n d H i - E f f , 181 ~ (9 .1 m i n ) a s n - h e n e i c o s a n e . I na d d i t i o n , t h e m a s s s p e c t r a ( E 1, 7 0 e V ) o f n - h e n e i c o s a n e a n d o f t r e a t e d I I1 w e r et h e s a m e ( r n / e 2 9 6, M * , a n d s e q u e n c e i o ns c h a r a c t e r i s ti c o f a n u n b r a n c h e ds a t u ra t e d h y d r o c a r b o n c h a i n) .

T h e p r e s e n c e o f t h e e p o x i d e g r o u p i n I I I w a s c o n f i r m e d , a n d i ts lo c a t i o no n t h e 2 1 - c a r b o n c h a i n w a s e s ta b l i s h e d , b y t h e f o l l o w i n g r o u t e : t r e a t m e n t o fc a t a l y t i c a ll y h y d r o g e n a t e d I I I w i t h 0 . 5 s u l fu r i c a c i d i n 1 : 1 a q u e o u s T H Ff o ll o w e d b y p r e p a r a t i o n o f t h e tr im e t h y ls il y l d e r iv a t iv e u s in g H M D S - T M C S -p y r i d i n e . T h e m a s s s p e c t r u m ( E l , 7 0 e V ) o f t h is t ri m e t h y l s i la t e d d i o l s h o w e d

t w o p r o m i n e n t p e a k s a tm/e 2 1 5 a n d 2 5 7 , c o r r e s p o n d i n g t o t h e f r a g m e n t sp r o d u c e d b y c l e a v a g e b e t w e e n c a r b o n s 9 a n d 10. T h i s t y p e o f f r a g m e n t a t i o n i sw e l l -do cum en ted (C ape l l a and Zo rzu t , 1968; Eg l in ton e t a l ., 1968).

A s a m p l e o f c a t a ly t ic a l ly h y d r o g e n a t e d I l l h a d t h e sa m e r e t e n t io n t im eon Hi -E f t , 170 ~ (25.2 min ) a s a syn the t i c s amp leofcis-9 10-epoxyheneicosane.


8/14

6 . 5 2 H I L L A N D R O E L O F S

S y n t h e t i c trans-9 10-epoxyheneicosane h a s a d i f f e r e n t r e t e n t i o n t i m e 2 4 .0m i n ) o n t h i s c o l u m n t h a n t h e c o r r e s p o n d i n gcis i s o m e r 2 5 .2 m i n ) .

T h e l o c a t i o n o f t h e d o u b l e b o n d c l o se s t t o t h e e p o x i d e in I I I w a se s t a b li s h e d b y r e d u c ti v e r e m o v a l o f t h e e p o x i d e g r o u p a n d s u b s e q u e n to z o n o l y s i s. T h e e p o x i d e g r o u p w a s r e m o v e d , w i t h o u t a l t e r a t i o n o f t h e d o u b l eb o n d s , b y t h e f o l lo w i n g s e q u e n c e o f r e a c t io n s : 1 ) t r e a t m e n t o f I I I w i t h L A Hi n d r y E t 2 0 t o p r o d u c e a s e c o n d a r y a l c o h o l , 2 ) r e a c t i o n o f t hi s a l c o h o l w i t hf re s h ly p r e p a r e d T P P D B in b e n z e n e t o p r o d u c e th e c o r r e s p o n d i n g b r o m i d e ,a n d 3 ) t r e a t m e n t w i th L A H in d r y E t 2 0 to r e m o v e th e b r o m i d e g r o u p . T h ep r o d u c t w a s c o l l e c t e d f r o m H i - E f t , 1 90 ~ , a t 3 . 2 - 4 .2 5 m i n w i t h n - h e n e i c o s a n ea t 2 .9 m i n ). T h i s m a t e r i a l h a d a n e q u i v a l e n t c a r b o n n u m b e r o f 2 0.9 o n

O V - 1 0 1 , 2 1 0 ~ . T h i s p u r i fi e d h y d r o c a r b o n w a s o z o n i z e d i n C S 2 t o p r o d u c e a st h e m a j o r p r o d u c t a m a t e r i a l w i t h re t e n t i o n t i m e s o n O V - 1 0 1 , 1 7 0 ~ 6 .5 m i n ) ,a n d o n X F - 1150 , 1 7 0 ~ 4 .2 5 m i n ) , s i m i l a r t o t h o s e o f s y n t h e t ic n - p e n t a d e c a n a l6 .4 m i n a n d 4 .3 5 r a in o n t h e t w o c o l u m n s , r e s p e c ti v e l y ).

T h e p o s i t i o n o f th e o t h e r d o u b l e b o n d w a s d e t e r m i n e d b y o z o n o ly s is o fu n t r e a t e d I I I in C S 2, w h i c h y i el d e d a p r o d u c t w i t h a s i m i l a r r e te n t i o n t i m e o nX F - 11 5 0 4 .4 m i n a t 50 ~ a s p r o p i o n a l d e h y d e .

A n F T - I R o f I I I s h o w e d t h a t t he t w o d o u b l e b o n d s a r e o f t h e Zc o n f i g u r a t i o n , s in c e t h e r e w a s n o p e a k i n t h e 9 6 0 - 9 8 0 c m -~ r e g i o n F i g u r e 2 ) .

0g a a a L ' k f f

d I~ ~ : ~ 'e f - - f g

(1

- T- -6

1 00 M H z

L I I I

5 4 3 2 PPM

g

I

1

FIG. 3 . FT -P M R o f I I1C6D6), with chem ical shift assignmen ts.


9/14

P H E R O M O N E O F T H E S A L T M A R S H C A T E R P I L L A R 66

A [ 1 H ] N M R s p e c t r u m F T ) o f I I I is s h o w n i n F i g u r e 3 w i t h t h e c h e m i c a ls h i ft a s s i g n m e n t s , w h i c h a r e a l l c o n s i s t e n t w i t h t h e n o n c o n j u g a t e d d i e n e -e p o x i d e s t r u c t u r e a s s i g n e d t o I I I .

A l l t h e a b o v e d a t a l e a d t o t h e u n e q u i v o c a l s t r u c t u r e a s s i g n m e n t o fZ , Z ) - 3 , 6 - c i s - 9 , 1 0 - e p o x y h e n e i c o s a d i e n ef o r I I I , w h i c h c a n e x i s t i n t w o

e n a n t i o m e r i c f o r m s , 9 S , 1 0 R a n d 9 R , 10 S . Te s t s h a v e , a s y e t, n o t e s t a b l is h e dt h e e n a n t i o m e r i c c o m p o s i t io n o f II I p r o d u c e d b y f e m a l eE. ac r ea . I t a lso isp o s s i b le t h a t m i n o r q u a n t i t i e s o f t h e v a r i o u s g e o m e t r i c is o m e r s o f I, II , a n d I I Ia r e p r e s e n t i n th e g l a n d e x t r a c t .

E A G R e s p o n s e s o f M a l e A n t e n n a e to § - ) - D i s p a r lu r e s .Ty p i c a lE A G r e s p o n s e s g i v en b y a m a l eE. ae r ea a n t e n n a e t o + ) - d i sp a r l u re , - ) -d i s p a r l u r e a n d I I I a r e p r e s e n t e d i n F i g u r e 4 . S t r u c t u r e s o f t h e s e c o m p o u n d sa r e g i v e n i n F i g u r e 5 . T h e t w o d i s p a r l u r e s a m p l e s w e r e e v a p o r a t e d i n t o t h es t r e a m o f a i r g o i n g o v e r th e t e s t a n t e n n a f r o m f i l te r p a p e r c a r t r i d g e s h a v i n g100 g o f e a c h m a t e r ia l ; t h e s a m p l e o f I I I w a s f r o m a c a p i l la r y tu b e G Cc o l l e c te d ) . T h e h i g h e s t r e s p o n s e w a s t o I I I 1 . 8 - 2 . 9 m V ) , fo l l o w e d b y t h a t t o- ) - d i s p a r l u r e 1 . 0 - 1 .4 m V ) , a n d t h e n b y a d i s ti n c t iv e l y l o w e r re s p o n s e t o th e+ ) - d i s p a r l u r e 0 . 2 - 0 . 6 m V ) . I n a d d i t i o n , t h e r e s p o n s e t o - ) - d i s p a r l u r e

s h o w e d t h e s a m e s l o w r e c o v e r y a s th e r e s p o n s e o f I I I , in d i c a t i v e o f a b i n d i n gi n t e r a c t i o n w i t h t h e a c c e p t o r s i te .

O b s e r v a t i o n s o f M a l e F l i g h ts in a W i n d Tu n n e l .Ta b l e 1 p r e s e n t s t h er e s u lt s o b t a i n e d w h e n m a l eE. ae r e a m o t h s w e r e e x p o s e d i n a f li g h t t u n n e l t o I ,I I , II I , a n d v a r i o u s c o m b i n a t i o n s o f t h e s e m a t e r i a ls ; t hi s in c l u d e s t h e s a m p l e si s o l a t e d f r o m f e m a l eE. a c r ea as we l l a s syn the t i c s am ples o f t he t h r eec o m p o u n d s I I I w a s r a c em i c ) .

Male E. c r e

EAG responses to Disparlure enantiomers

and to female-produced epoxide 1TO

- - 4 + (_ )- 4- 4- b~ . . . . ~ L ~ t

r

, l l . . . .

/1I

FIG. 4. E lec t roa n t e nnog ra mres ponse so fm a l e E . a c r eaan t enna to l I I , + ) -d i s pa r l u r e ,and -)-dispar lure .


10/14

664 H I L L A N D R O E L O F S

0/ \

E s t i g m e n e a c r e a

U t e t h e i s a o r n a t r i x

0

O r g y i a p s e u d o t s u g a t a

F IG . 5. C o m p a r i s o n o f II I w i th o t h e r fe m a l e - p r o d u c e d s ex p h e r o m o n e c o m p o n e n t sk n o w n fo r L e p i d o p t e r a .

I t is c l e ar t h a t m a l e s w i l l n o t f l y t o I I I a l o n e , b u t t h a t I I I in c o m b i n a t i o n

w i t h e i t h e r I o r I I w i l l c a u s e t h e m a l e s t o f ly u p t o t h e s o u r c e . T h e y w i l l a l s oh o v e r n e a r it , e x t e n d t h e i r c l a s p e r s w h i l e w a l k i n g o n t h e s o u r c e w i t h v i g o r o u sw i n g f a n n i n g , a n d e v e n a t t e m p t t o m a t e w i t h o t h e r i n d i v i d u a l s e it h e r a t th es o u r c e o r a t t h e s i te o f f l i g h t i n i t i a t i o n . A f t e r f l y i n g t o t h e s o u r c e , i n d i v i d u a l sc o u l d b e p l a c e d d o w n w i n d r e p e a t e d ly a n d a g a i n w o u l d f ly t o t h e so u r ee . T h e yc o u l d a l s o b e k e p t i n f l ig h t fo r se v e r a l m i n u t e s (u p t o 1 0 - 1 5 m i n ) b y m o v i n gt h e s t r i p e d f l o o r u n d e r n e a t h t h e m b a c k w a r d s ( M i l l e r a n d R o e l o f s , 1 97 8) . T h ep e r i o d o f th i s f l ig h t a c t i v i t y s t a r t e d a b o u t 15 m i n a f t e r i n i t i a t i o n o f th ep h o t o p h a s e a n d l a s t e d a b o u t 1 h r .

TABL E 1. RESPONSESOF MALEE . a c r e a MO THS IN A FLIGHT TUNNEL TO I , I I , I11, ANDCOMBINATIONS THEREOF

No. of male respondingb(% of total)

Average flightNo. of Flying t ime to

Samplea observat ions Fa nn ing at source source c

E . a c r e a effluvium 3 3 (100)a 3 (100)aE . a c r e a tip extr act 2 2 (100)a 2 (100)a1 4 2 5 0 ) b 0 0 ) bI 2 O ( O ) b O ( O ) bIH ( ) 10 8 (8 0 ) b 0 ( 0 ) bH I ( ) + 1 14 13 ( 93)ab 8 ( 57)aII1 ( ) + II 20 20 (100)a 14 ( 70)aI l i ( ) + I + II 13 13 (100)a 10( 77)a111( ) + Z 9 - 1 8 : A L D 5 1 ( 2 0 ) b 0 ( 0 ) bIII + I + I1 6 6 (100)a 5 ( 83)a

226 sec659 sec204 sec

300 sec

aSamples w ere synth etic unless indica ted to be from females. Sa mp les from m oths were purifiedby GC collection. Z 9- 18 :A L D is (Z)-9-octadecenal . Synthet ic I II was racemic.

b - 9 . . . . .

Figures fol lowed by the sam e let ter in each co lum n are not s ignif icant ly different, according tothe m ethod of R yan (1960), a t the 5% experimental error rate.

CTo tal nu m be rs of flights tim ed w ere 7, 9, 10, an d 1, respectively (for sam ples 6, 7, 8, an d 10).


11/14

PHEROMONE OF THE S LTM RSH C TERP ILL R 66

Observation of Calling E . a c r e a Females and Location of Female SexPheromone Gland. F e m a l e s w e r e k e p t r o u t i n e l y o n a 1 6 : 8 l i g h t - d a r k c y c le .U n d e r t h e s e c o n d i t i o n s , f e m a l e s w e r e s e e n c a l li n g j u s t b e f o r e s c o t o p h a s e a n da l s o j u s t a f t e r s c o t o p h a s e . C a l l in g c o u l d b e i n i ti a t e d i n t h e fe m a l e s b y r e m o v a lo f t h e f e m a l e s f r o m s c o t o p h a s e a t a n y t i m e d u r i n g t h e l a s t 5 h r o f s c o t o p h a s e ;w h e n t h is w a s d o n e , t h e f e m a l e s w o u l d c a l l s t a r t in g a b o u t 15 m i n a f t e ri n i ti a t io n o f t h e p h o t o p h a s e .

Th e ca l l i ng s t ance is one i n wh ich the f em a le e l eva t e s he r w ings s l i gh tly,s o m e t i m e s f l u t t e r s t h e m w h i l e w a l k i n g a r o u n d , a n d h e r a b d o m i n a l t i p i sp u s h e d o u t s l ig h t l y a n d r e t r a c t e d in a p u l s i n g m o t i o n . T h e r a t e o f p u l s i n g a tr o o m t e m p e r a t u r e w a s ti m e d a t a b o u t 8 0 p u ls e s p e r m i n t h r e e i n d i v id u a l sw e r e o b s e r v e d ) .

Va r i o u s c r u d e d i s s e c ti o n s o f f e m a l e a b d o m i n a l t ip s w e r e c a r r i e d o u t a n de a c h p o r t i o n w a s e x t ra c t e d a n d a n a l yz e d f o r t he p h e r o m o n e c o m p o n e n t s .O n l y a p a i r o f t u b e s , e x i st in g d o r s a l l y a t t h e b a s e o f t h e p e n u l t i m a t ea b d o m i n a l s e g m e n t , w e r e f o u n d u s in g G C t r a ci n g s) to c o n t a i n a n y a p p r e c i -a b le a m o u n t s o f t h e s ex p h e r o m o n e c o m p o n e n t s , I , II , a n d I II . T h e se tu b e s a r ev i si b le u n d e r t h e c u t i c u l a r a b d o m i n a l c o v e r i n g u n d e r s l ig h t m a g n i f i c a t i o n2 5 • E a c h is a b o u t 3 m m i n l e n gt h a n d a b o u t 0.2 m m O D , t r a n s lu c e n t , a n d

a l m o s t c o l o r le s s l ig h t c r e a m c o l o r ) . T h e y a p p e a r t o be si m i l a r t o t h e t u b u l a r

g l a n d s o f th e g e o m e t r i d m o t h ,Rheumaptera hastata L.) Werner, 1977) ,e x c e p t t h e p a i r o f tu b e s i nE. acrea e x i s t s e p a r a t e l y, w h e r e a s t h e t u b e s o f R .hastata j o i n a n d h a v e a c o m m o n f u n n e l - s h a p e d o p e n i n g .

D I S C U S S I O N

T h e th r e e c o m p o u n d s I , I1 , a n d I I I; F i g u r e 1) i d e n t i f ie d as s e xp h e r o m o n e c o m p o n e n t s f o rE. acrea a n d Hyphantria cunea D r u r y ) H i l l

e t a l. , 1 98 1) h a v e n o t b e e n r e p o r t e d p r e v i o u s l y as c o m p o n e n t s o f o t h e r s e xp h e r o m o n e s y st em s . C o m p o u n d s I a n d I I a r e th e a l d eh y d e s c o r r e s p o n d i n g tot h e w i d e l y d i s t r i b u t e d p o l y u n s a t u r a t e d f a t t y a c id s l i n o l e ic a n d l i n o le n i c a c i d s,w h i c h a r e k n o w n t o b e e ss e n t ia l f a t t y a c i d s f o r in s e c ts D o w n e r, 1 97 8). T h ee p o x i d e c o m p o n e n t , I I I , a l t h o u g h h i t h e r t o u n k n o w n , i s s i m i l a r t o t h ec o m p o u n d s s h o w n i n F i g u re 5 , w h i c h ha v e b e e n re p o r t e d a s s ex p h e r o m o n ec o m p o n e n t s f o r o t h e r l e p i d o p t e r a : o n e i s f r o m a n o t h e r a r c t i i d ,Utetheisaornatrix bella L . ) C o n n e r e t a l ., 1 98 0) ; t h e o t h e r tw o a r e f r o m l y m a n t r i i d s ,t h e D o u g l a s - f i r t u s s o c k m o t h ,Orgyiapseudotsugata M c D u n n o u g h ) S m i t h

e t a l ., 1975) and the gypsy m o th ,Lymantria dispar L.) Bier l e t a l . , 1970). Th et o p t w o p h e r o m o n e s i n F i g ur e 5 a n d c o m p o u n d I II a r e all u n b r a n c h e d2 1 - c a r b o n c h a i n s t r u c t u r e s w i t h a Z d o u b l e b o n d a t t h e 6 p o s i t i o n . T h e U .ornatrix c o m p o n e n t a l so h as a Z ) - 3 d o u b l e b o n d i n c o m m o n w i th I II , a n d itso t h e r d o u b l e b o n d , a t p o s i t io n 9 , o c c u r s a t th e s a m e l o c us o n t h e c a r b o n c h a i n


12/14

HILL ANDROELOFS

a n d h a s th e s a m e Z c o n f i g u r a t i o n a s t h e e p o x y g r o u p o f II I. T h e p o i n t s o fs i m i l a r it y b e t w e e n I I I a n d t h e O .p s e u d o t s u g a t ap h e r o m o n e c o m p o n e n t a ref e w e r. A l t h o u g h t h e o x y g e n f u n c t i o n a l it i e s o f t h e s e tw o c o m p o u n d s a r e n o t ine x a c t l y t h e s a m e p l a c e a l o n g t h e c a r b o n c h a i n , t h e y d o o c c u r a t v i c i n a lp o s i ti o n s . T h e o n l y o t h e r e p o x i d e s o fa r r e p o r t e d a s a n i ns e c t s e x p h e r o m o n e ,c i s -7 ,8 - epoxy-2 -me thy toc t adecane(d i spa r u re ) , i s t ha t f romL. dispar.Fie ldda t a (M i l l e r e t a l ., 1977; Ca rde ' e t a l ., 1977) s t rong ly sup po r t a s s ignm en t o f t he7 R , 8 Ss t r u c t u re [ ( + ) - d is p a r l u re ] a s th a t o f th e n a t u r a l p h e r o m o n e . N o o t h e rs e x p h e r o m o n e c o m p o n e n t s f o r l y m a n t r i i d s h a v e b e e n r e p o r t e d , a l t h o u g hm a l e s o f o n e o t h e r s p e c ie s in t h is f a m i l y, th e n u n m o t h ,Ly m a n t r i a m o n a c h a(L . ) , ha s been cap tu red in t r aps ba i t ed w i th d i spa r lu re .

T h e o n l y o t h e r k n o w n a r c t i i d s e x p h e r o m o n e s t r u c t u r e i s 2 - m e t h y l -h e p t a d e c a n e , w h i ch h a s b e e n r e p o r t e d f o r t h e b a n d e d w o o l l y b e a r m o t h ,l s iaisabel la ( J. E . S m i t h ; f o r m e r l yPyrrarc t ia i sabel la ,a n d f o r a n u m b e r o f t ig e rm o t h s i n th eH o l o m e l i n ag e n u s ( R o e l o f s a n d C a rd e ', 1 97 1). T h i s h y d r o c a r b o nh a s t h e 2 - m e t h y l b r a n c h f e a t u r e i n c o m m o n w i t h th e d i s p a r l u r e s t r u c tu r e .

M a l e E. acrea a n t e n n a e c a n d i s c r im i n a t e b e t w e e n t h e l e v o - a n dd e x t r o r o t a t o r y i s om e r s o fcis-disparlure,w i t h t h e ( - ) - i s o m e r c o n s i s t e n t l yp r o d u c i n g E A G r e s p o n se s t h a t a r e 2 - 5 t i m e s g r e a t e r in a m p l i t u d e t h a n t h o sep r o d u c e d w i th t h e ( + ) - i s o m e r. T h e a c c e p t o r si te is , p r e s u m a b l y, t h a t f o r I I I, s o

th is acc ep to r s i te appe a r s t o be ch i r a l. I f t he s t ruc tu re s o f t he two i some rs o f II1a n d t h e t w o e n a n t i o m e r s o fcis-dispar lurec a n b e c o r r e l a t e d a s s h o w n i nF i g u r e 6 , t h e n a p r e d i c t i o n o f th e a b s o l u t e c o n f i g u r a t i o n o f I II s e e m s p o s s ib l e .T h e c o r r e s p o n d e n c e o f t h e tw o l o n g e r - c h a i n p o r t i o n s o f e a c h c o m p o u n d ,s h o w n i n F i g u r e 6 , t o th e r i g h t o f e a c h e p o x i d e g r o u p , is m a d e o n t h ea s s u m p t i o n t h a t t h e 1 0 - c ar b o n c h a in p o r t i o n o f t h ecis-dispar lurem olecu le ist o o b u l k y t o b e a c c o m o d a t e d a t t h e l o c u s o n t h e a c c e p t o r s ite t h a t o r d i n a r i l y

7R,8S +)

7S,8R -)

FIG. 6. Stereochemical relationships between 11I and the dispa rlure enan tiom ers.


13/14

PHEROMONE OF THE SALTMARSH CATERPILLAR 667

f it s the 8-carbo n unsa tu ra te d por t i on of I l l . Based on these assumpt ions , a9S, 10R con figu rat i on at the epoxide gr oup of III is predicted. Addit io nally,because of the marked difference in the EAG responses to (+) and (-)disparlures, it seems reasonable to expect that III produc ed by female E . a c r e awill be found to be exclusively or pre dom ina ntl y the 9S,10R e nan tiom er.

Wind tunnel observat ions of male E . a c r e a f l ights have demonstratedthat upwi nd f l ight by males is ini t iated and sustained in the presence of III inco mb in at io n with either I or II and t hat racemic, synth etic III is effective.Since al l three components are emit ted by the females, but only two of thethree appear to be requi red for sus ta ined upwind anemot axis , the funct ion alreason for emission of both aldehydes by the female is not clear at present.

At this stage it is evident that all three compounds, I, II, and III, areemitted by female E . a c r e a and that all three can be perceived by the males andmediate the specif ic beha vior of upwi nd f l ight (amen otaxi s) by them. Fur the rtest ing of the biological s ignif icance of these co mpo nen ts, incl udin g f ieldtra ppin g tests and invest ig at ion of the biosynthesis of these co mp ou nd s, wil lbe carried out.

A c k n o w l e d g m e n t s - - W eare very grateful to the following for providing specializedservices: Dr. J.H. Tumlinson and R.R. Heath for the FT-1R spectrum of III; Dr. J. Kochansky forthe FT-IR spectrum of II; Dr. R.M. Silverstein for the 100-MHz FT-NMR of III; Dr. T. Wachsand D. Angell for the mass spectra. We are grateful also to F. Wadhams and K. Poole formaintaining the insect cultures and for preparation of most of the female tip extracts, to Drs. R.T.Carde'and T.C. Baker for some of the early EAG determinations, to Dr. J. Barnard for thestatistical analyses, to C. Anway for assistance with the flight tunnel bioassays, and to Drs. P.J.Chapman and S.E. Lienk for providing the gravid E . ac r e a female from the blacklight trap.

We are also extremely grateful to Dr. B. Kovalev for providing the synthetic sample of III,and to Dr. K. Mori for the enantiomers of c is -d ispar lure .

REFERENCES

BEROZA M., and BIERL B.A. 1967. Rapid determination of olefin position in organic compoundsin microgram range by ozonolysis and gas chromatography. A n a l . C h e m .39:1131-1135.

BIERL B.A., BEROZA M. and COLLIER C.W. 1970. Potent sex attractant for the gypsy moth: Itsisolation, identification and synthesis. S c i e n c e 170:87-89.

CAPELLA P. and ZORZUT, C.M. 1968. Determination of double bond position in monounsatu-rated fatty acid esters by mass spectrometry of their trimethylsilyloxy derivatives. An a l .C h e m . 40:1458-1463.

CAROL R.T., DOANE, C.C., BAKER, T.C., IWAKI, S., and MARUMO S. 1977. Attractancy ofoptically active pheromone for male gypsy moths. E n v i r o n . E n t o m o l .6:768-772.

CONNER, W.E., EISNER T. VANDER MEER R.K., GUERRERO, A., GHIRINGELLI, D., andMEINWALD J. 1980. Sex attractant of an arctiid moth U t e t h e s i a o r n a t r i x ) : A pulsedchemical signal. Behav. Eco l . Soc iob ioL7:55-63.

COREY E.J., and SUGGS, J.W. 1975. Pyridinium chlorochromate. An efficient reagent foroxidation of primary and secondary alcohols to carbonyl compounds. Te t r a h e d r o n L e t t .31 :2647-2650.


14/14

668 HILL AND ROELOFS

DOWNER, R.G.H. 1978. Chapter 2, Functional role of lipids in insects, pp. 58-92, in M. Rockstein(ed0. Biochemistry of Insects. Academic Press, New York.

EGLINTON~ G., HUNNEMAN, D.H., and McCORMiCK, A. 1968. Gas chromatographic-mass

spectrometri c studies o f long-chain hydroxy acids. 11I. The mass spectra of the methyl esterstrimethylsily ethers of aliphatic hydroxy acids. A facile method of double bond location.Org . Mass . Spec t rom.1:593-61 I.

FARI(AS~ S.R., SrIOREY, H.H., and GASTON, L.K. 1974. Sex pheromones of Lepidoptera:Influence of pheromone concentration and visual cues on aerial odor-trail following bymales of Pec t inophora gossyp ie tl a . Ann . En tomol . Soc . A m .67:633-638.

GREEN, N., JACOBSON, M, HENNEBERRY, T.J., and KISHABA A.N. 1967. Insect sex att ractants. VI.7-Dodecen-l -ol acetates and congeners. J. M e d . C h e m . 10:533-535.

HILL A.S.~ CARDE~ R.T. BODE W.M. and ROELOES W.L. 1977. Sex pheromone components ofthe variegated leafroller moth, Pla tyno ta f l avedana . J . Chem. Ecok3:371-378.

HILL, A.S., KOVALEV, B.G., NIKOLAEVA, L.N., and ROELOES, W.L. 1981. Sex pheromone of thefail webworm moth, Hypha n t r i a cunea . J. Chem. Eco l .6:Submitted.

KENNEDY, J.S. 1977. Behaviorally discriminating assays of att rac tants and repellents, pp. 215-229, in H.H. Sborey, J.J. McKelvey, Jr., (eds.). Chemical Control of Insect Behavior--Theory and Appl icat ion. Wiley-Interscience, New York.

METCALF, C.L., FLINT, W.P., and METCALE, R.L. 1962. Destructive and Useful Insects, 4th ed.,pp. 692-693. McGraw-Hill Book Co., New York.

MILLER, J. R., and ROELOFS, W.L. 1978. Sustained-flight tunnel for measuring insect responses towind-borne sex pheromones. J. Chem. Ecol .4:187-198.

MILLER, J.R. , MORI, K., and ROELOFS, W.L. 1977. Gypsy moth field trapp ing and electro-antennogram studies with pheromone enantiomers. J. Insec t Phys io l .23:1449-1453.

MORI, K., TAKIGAWA, T., and MATSUI, M. 1979. Stereoselective synthesis of both enantiomers ofdisparlure, the pheromone of the gypsy moth. Te t rahed ron 35:833-837.

ROELOFS, W.L. 1977. The scope and limitat ions of the elect roantennogram technique inidentifying pheromone components , pp. 147-165 in McFarlane (ed.). Crop ProtectionAgents, Academic Press, London.

ROELOFS, W.L., and CARDE7 R.T. 1971. Hydrocarbon sex pheromone in tiger moths (Arctiidae).Sc ience 171 : 684-686.

RYAN, T.A. 1960. Significance tests for multiple comparisons of proport ion , variances and otherstatistics. Psychol . Bul l .59:318-328.

SHOREY, H.H. , and HALE, R.L. 1965. Mass-rear ing of the larvae o f nine noctuid species on asimple artificial medium. J. E c o n . E n t o m o l .58:522-524.

SMITH R.G. DATERMAN G.E., and DAVES G.D., Jr. 1975. Douglas-fir tussock moth: Sexpheromone identificat ion and synthesis. Sc ience 188: 63-64.

SONNET, P.E., and OLIVER, J.E. 1976. Olefin inversion. I. Reaction o f aliphatic epoxides withtriphenylphosphine dihalides. J. Org. Chem.41:3279-3283.

SWEELEY C.C. BENTLEY R. MAKITA M. and WELLS, W.W. 1963. Gas-l iquid chr omat ographyof trimethylsilyl derivatives of sugars and related substances. J . Am. Chem. Soc .85:2497-2507.

WARTHEN J.D., Jr., and JACOBSON M. 1973. Insect sex attractants. XIV. Al l t r ans a lkeno lacetates, via sodium-l iquid ammonia reduction. Syn thes i s 10:616-617.

WERNER, R.A. 1977. Morphology and histo logy of the sex pheromone gland of a geometrid ,

R h e u m a p t e r a h a s t a t a . A n n . E n t o m o l . S o c . A m .701264-266.

hill1981-sex pheromone of the saltmarsh

Documents