,.
R A P I D METHOD FOR IDENTIFYING THEBAINE-RICH PAPAVER B~~CTEATUM /
J.W. F a i r b a i r n and E.M. U n i v e r s i t y of London
,
Williamson, The School of Pharmacy, 29/39, Brunswick Square, London WCIN
p . b r a c t e a t u m (Chromosome number 2 2 = 1 4 ) i s a p o t e n t i a l l y Ymportant s o u r c e of o p i a t e s ( F a i r b a i r n , l 9 7 6 ) b u t t h e r e has been considerable d i f f i c u l t y i n d i s t i n g u i s h i n g t h i s thebaine-rich s p e c i e s from t h e c l o s e l y a l l i e d p. o r i e n t a l e ( 2 2 = 28) and g. s e u d o - o r i e n t a l e ( 2 2 = 42) and t h e i r numerous c u l t i v a t e d h y b r i d s , Ztc. a s b o t a n i c a l c h a r a c t e r s show c o n s i d e r a b l e o v e r l a p . Q u a n t i t a t i v e a n a l y s i s of t h e l a t e x f o r t h e b a i n e i s a n o b v i o u s pragmatic t e s t , b u t i t may t a k e a l m o s t 2 y e a r s from seed sowing t o f o r m a t i o n W e have t h e r e f o r e d e v i s e d a method based of c a p s u l e s f o r t e s t i n g . on measuring s t o m a t a l l e n g t h a few weeks a f t e r s e e d g e r m i n a t i o n . G o l d b l a t t (1974) used s t o m a t a l l e n g t h s based on v e g e t a t i v e l e a v e s , b u t t h e mean l e n g t h s overlapped b a d l y and w e found t h e y v a r i e d w i t h age of t h e l e a f . Our method u s e s c o t y l e d o n s a t a s p e c i f i e d physiol o g i c a l a g e , a f u r t h e r advantage b e i n g t h a t no t r i c h o m e s a r e Seeds of 7 c o l l e c t i o n s of a u t h e n t i c a t e d p. present t o interfere. b r a c t e a t u m , 4 of 2. o r i e n t a l e and 4 of P. p s e u d o - o r i e n t a l e were germinated and a t t h e s t a g e when t h e second p a i r of t r u e l e a v e s were j u s t a p p e a r i n g c e l l u l o s e a c e t a t e i m p r e s s i o n s w e r e made of t h e c o t y l e d o n s and s t o m a t a l l e n g t h s measured. About 5 p l a n t s from each c o l l e c t i o n w e r e used and 20 stomata measured from each. Chromosome c o u n t s from r o o t m a t e r i a l were a l s o made by M i s s C. Brighton, J o d r e l l L a b o r a t o r y , K e w , and t h e a l k a l o i d a l spectrum of t h e s e e d l i n g s determined by t . 1 . c .
Over 1 2 0 0 s t o m a t a l measurements were made and t h e d a t a f e d i n t o a computer. S t a t i s t i c a l a n a l y s i s showed t h a t t h r e e homogeneous p o p u l a t i o n s , which w e r e s i g n i f i c a n t l y d i s t i i i c t , e x i s t e d and corresporided t o t h e t h r e e s p e c i e s . The mean f o r p. b r a c t e a t u m (614 d e t . ) was 58 pm s . d . 6.89; p. o r i e n t a l e (300 d e t . ) was 74 pm s . d . 9.86 and p. p s e u d o - o r i e n t a l e (230 d e t . ) was 84 pm s . d . 11.40. I f t h e r e f o r e 25 d e t e r m i n a t i o n s of s t o m a t a l l e n g t h s made from 5 d i f f e r e n t s e e d l i n g s a r e made, t h e means would f a l l i n t o t h e following ranges ( P = 0 . 9 5 ) ; P . b r a c t e a t u m ( 5 5 . 5 t o 6 0 . 5 ) ; p. o r i e n t a l e ( 7 0 t o 78) and P. p s e u d o - o r i e n t a l e (79.5 t o 8 8 . 5 ) . The method t h e r e f o r e c l e a r l y d i s t i n g u i s h e s p. b r a c t e a t u m s e e d l i n g s from t h o s e of t h e o t h e r two s p e c i e s . However t h e r e i s a s l i g h t chance of o v e r l a p of r e s u l t s between t h e l a t t e r two and t h i s was a l s o s e e n w i t h t h e chromosome c o u n t s as t h e y corresponded t o s t o m a t a l l e n g t h s f o r a l l v a r i e t i e s of p. b r a c t e a t u m b u t d i d not always s o c o r r e s p o n d w i t h p. o r i e n t a l e and p. p s e u d o - o r i e n t a l e . The a l k a l o i d a l p a t t e r n of t h e s e e d l i n g s was n o t a r e l i a b l e i n d i c a t o r of species a s numerous a l k a l o i d s p o t s appeared w i t h o u t anyrecognisable p a t t e r n . F a i r b a i r n , J . W . (1976). P l a n t a Medica, 30, 26-31. G o l d b l a t t , P . (1974). Ann. M i s s o u r i Bot. Gard., 61 , 264-296.
R A P I D METHOD FOR IDENTIFYING THEBAINE-RICH PAPAVER B~~CTEATUM /
J.W. F a i r b a i r n and E.M. U n i v e r s i t y of London
,
Williamson, The School of Pharmacy, 29/39, Brunswick Square, London WCIN
p . b r a c t e a t u m (Chromosome number 2 2 = 1 4 ) i s a p o t e n t i a l l y Ymportant s o u r c e of o p i a t e s ( F a i r b a i r n , l 9 7 6 ) b u t t h e r e has been considerable d i f f i c u l t y i n d i s t i n g u i s h i n g t h i s thebaine-rich s p e c i e s from t h e c l o s e l y a l l i e d p. o r i e n t a l e ( 2 2 = 28) and g. s e u d o - o r i e n t a l e ( 2 2 = 42) and t h e i r numerous c u l t i v a t e d h y b r i d s , Ztc. a s b o t a n i c a l c h a r a c t e r s show c o n s i d e r a b l e o v e r l a p . Q u a n t i t a t i v e a n a l y s i s of t h e l a t e x f o r t h e b a i n e i s a n o b v i o u s pragmatic t e s t , b u t i t may t a k e a l m o s t 2 y e a r s from seed sowing t o f o r m a t i o n W e have t h e r e f o r e d e v i s e d a method based of c a p s u l e s f o r t e s t i n g . on measuring s t o m a t a l l e n g t h a few weeks a f t e r s e e d g e r m i n a t i o n . G o l d b l a t t (1974) used s t o m a t a l l e n g t h s based on v e g e t a t i v e l e a v e s , b u t t h e mean l e n g t h s overlapped b a d l y and w e found t h e y v a r i e d w i t h age of t h e l e a f . Our method u s e s c o t y l e d o n s a t a s p e c i f i e d physiol o g i c a l a g e , a f u r t h e r advantage b e i n g t h a t no t r i c h o m e s a r e Seeds of 7 c o l l e c t i o n s of a u t h e n t i c a t e d p. present t o interfere. b r a c t e a t u m , 4 of 2. o r i e n t a l e and 4 of P. p s e u d o - o r i e n t a l e were germinated and a t t h e s t a g e when t h e second p a i r of t r u e l e a v e s were j u s t a p p e a r i n g c e l l u l o s e a c e t a t e i m p r e s s i o n s w e r e made of t h e c o t y l e d o n s and s t o m a t a l l e n g t h s measured. About 5 p l a n t s from each c o l l e c t i o n w e r e used and 20 stomata measured from each. Chromosome c o u n t s from r o o t m a t e r i a l were a l s o made by M i s s C. Brighton, J o d r e l l L a b o r a t o r y , K e w , and t h e a l k a l o i d a l spectrum of t h e s e e d l i n g s determined by t . 1 . c .
Over 1 2 0 0 s t o m a t a l measurements were made and t h e d a t a f e d i n t o a computer. S t a t i s t i c a l a n a l y s i s showed t h a t t h r e e homogeneous p o p u l a t i o n s , which w e r e s i g n i f i c a n t l y d i s t i i i c t , e x i s t e d and corresporided t o t h e t h r e e s p e c i e s . The mean f o r p. b r a c t e a t u m (614 d e t . ) was 58 pm s . d . 6.89; p. o r i e n t a l e (300 d e t . ) was 74 pm s . d . 9.86 and p. p s e u d o - o r i e n t a l e (230 d e t . ) was 84 pm s . d . 11.40. I f t h e r e f o r e 25 d e t e r m i n a t i o n s of s t o m a t a l l e n g t h s made from 5 d i f f e r e n t s e e d l i n g s a r e made, t h e means would f a l l i n t o t h e following ranges ( P = 0 . 9 5 ) ; P . b r a c t e a t u m ( 5 5 . 5 t o 6 0 . 5 ) ; p. o r i e n t a l e ( 7 0 t o 78) and P. p s e u d o - o r i e n t a l e (79.5 t o 8 8 . 5 ) . The method t h e r e f o r e c l e a r l y d i s t i n g u i s h e s p. b r a c t e a t u m s e e d l i n g s from t h o s e of t h e o t h e r two s p e c i e s . However t h e r e i s a s l i g h t chance of o v e r l a p of r e s u l t s between t h e l a t t e r two and t h i s was a l s o s e e n w i t h t h e chromosome c o u n t s as t h e y corresponded t o s t o m a t a l l e n g t h s f o r a l l v a r i e t i e s of p. b r a c t e a t u m b u t d i d not always s o c o r r e s p o n d w i t h p. o r i e n t a l e and p. p s e u d o - o r i e n t a l e . The a l k a l o i d a l p a t t e r n of t h e s e e d l i n g s was n o t a r e l i a b l e i n d i c a t o r of species a s numerous a l k a l o i d s p o t s appeared w i t h o u t anyrecognisable p a t t e r n . F a i r b a i r n , J . W . (1976). P l a n t a Medica, 30, 26-31. G o l d b l a t t , P . (1974). Ann. M i s s o u r i Bot. Gard., 61 , 264-296.
