Complete nucleotide sequence of a viroid isolated from Etrog citron, a new member of hop stunt viroid group

The c o m p l e t e n u c l e o t i d e s e q u e n c e of a viro i d i s o l a t e d from c i t r u s ( C i t r u s m e d i c a L. U S D C S 6 0 1 3 ) , d e s i g n a t e d as ci t r u s isolate of hop s t u n t v i r o i d ( H S V d t ) ( 1 ) , was e s t a b l i s h e d . T he sin g l e s t r a n d e d c i r c u l a r m o l e c u l e s of c i t r u s v i r o i d RNA c o n s i s t s of 3 02 n u c l e o t i d e s , w h i c h can form a r o d l 1 k e s t r u c t u r e with e x t e n s i v e b a s e p a 1 r 1 n g c h a r a c t e r i s t i c to the o t h e r v i r o i d s . H e t e r o g e n e i t y was found in the s e q u e n c e No. 2 0 7 2 0 9 . T he seq u e n c e w a s GAG whe n d e t e r m i n e d from c D N A s by re v e r s e t r a n s c r i p t a s e d i d e o x y m e t h o d using v i r o i d RNA and ol i g o n u c l e o t i d e p r i m e r s (Fig.l v a r i a n t 1, m a j o r s e q u e n c e ) . But 1t was CGA wh e n d e t e r m i n e d f r o m t w o cDNA c l o n e s by Ml 3 d i d e o x y method (Fig.l v a r i a n t 2, m i n o r s e q u e n c e ) . C o m p a r a t i v e a n a l y s i s of t h e s e q u e n c e of t h e four HSV I s o l a t e s i n d i c a t e s t h a t H S V c i t r u s 1s more c l o s e l y r e l a t e d to HSV-c Isolated from c u c u m b e r in The N e t h e r l a n d s ( 2 , 3 ) ( s e q u e n c e h o m o l o g y 9 9 Z ) than H S V h o p and H S V g r a p e v i n e ( 4 , 5 ) from hop and gr a p e v i n e 1n Jap a n , r e s p e c t i v e l y ( s e q u e n c e h o m o l o g y 9 6 Z ) .