Research NoteTen cultivars representing Riesling-type vines and 12 cultivars of the Burgundy family were analyzed by RAPD-PCR. Employing computer programs, similarity degrees were calculated from the RAPD patterns obtained from a set of 20 dekamer primers and used to perform cluster analysis. The Riesling-type cultivars could be differentiated from each other and from the progenitor of a cross Pinot noir x Chambourcin ("Spezial" in Fig.l). With the exception of Aubin blanc, Aubin vert and Elbling white, the Riesling-type cultivars did not show any close relationship. From the Burgundies, Pinot noir, Pinot meunier, Pinot blanc and Pinot gris could not be differentiated. Neither could Chardonnay blanc be differentiated from Chardonnay musque, nor Aligoté from Aligoté vert. Discrimination between the other cultivars was possible. Two Morillon samples from different origins were analyzed and showed clear differences in their RAPD patterns. Furthermore, they seemed to be more similar to Riesling white than to the Burgundies

Tschammer, J.

Zyprian, Eva

English

JKI Open Journal Systems (Julius Kühn-Institut)

Vitis 33, 249-250 (1994) Research Note Molecular characterization of grapevine cultivars of Riesling-type and of closely related Burgundies J. TscHAMMER and EvA ZYPR!AN S u m m a r y : Ten cultivars representing Riesling-type vines and 12 cultivars of the Burgundy family were analyzed by RAPD-PCR. Employing computer programs, similarity degrees were calculated from the RAPD patterns obtained from a set of 20 dekamer primers and used to perform cluster analysis. The Riesling-type cultivars could be differentiated from each other and from the progenitor of a cross Pinot noir x Chambourcin ("Spezial" in Fig.l). With the exception of Aubin blanc, Aubin vert and Elbling white, the Riesling-type cultivars did not show any close relationship. From the Burgundies, Pinot noir, Pinot meunier, Pinot blanc and Pinot gris could not be differentiated. Neither could Chardonnay blanc be differentiated from Chardonnay musque, nor Aligote from Aligote vert. Discrimina-tion between the other cultivars was possible. Two Morillon sam-ples from different origins were analyzed and showed clear dif-ferences in their RAPD patterns. Furthermore, they seemed to be more similar to Riesling white than to the Burgundies. K e y w o r d s : DNA, RAPD-PCR, Burgundy cultivar, Riesling cultivar, identification. Introduction: Today, the number of different grape-vine cultivars has been estimated tobe in the range of 5000 to 15000. The origin of many of them and the relationship between some of the cultivars is uncertain. Distinction of closely related cultivars by ampelography often is diffi-cult. Genomic analysis can help to solve these problems. RAPD (random amplified polymorphic DNA) analysis e.g. is one technique recently used with success to differenti-ate grapevine cultivars. It is rather cheap and easily per-formed, but nevertheless provides valuable information. Here we employed this method to investigate the distin-guishability of several grapevine cultivars and their rela-tionship . Materials and methods: DNA was prepared from young, expanding leaves using the protocol described by THOMAS et al. (1993). RAPD analysiswas performed with 20 dekamer primers (FO 1 to F20, Operon Technologies , Alameda, CA, USA) using the protocol described earlier (BüscHER et al. 1993; 1994). Amplification products were resolved by electrophoresis on 1.5% agarose gels made in 0.5 x TBE (44.5 mM Tris, 44.5 mM borate, 1 mM EDTA) . Similarity degree values were calculated from the RAPD patterns with the help of the computer program "Rapid" developed by R.BLATCH at the institute. Cluster analysis based on these similarity indices was performed with a self-made computer program. This program uses the algo-rithm described by NöBAUER and TrMrSCHL (1979). Tbe re-sults are represented as dendrograms showing the simi-larities between the cultivars analyzed. Results and discussion: The cultivars of the Riesling-type grapevines can be clearly distinguished by their RAPD patterns (Fig. l).The cultivars Aubin vert, Aubin blanc and q,o q.1 q.2 q,3 q.4 q,s q.s q,? q.s q,9 1,,0 ~ , similarity degree value Räuschling weiß 2 Räuschling weiß L----+- Arvine petite r---+ Gauais blanc IL---'- Arbois Aubin vert 2 Aubin vert 1 Aubin vert 3 Elbling weiß Au bin blanc Au bin blanc 1 1c_ ___ ....._ Reze L-___ ..i... Humagne L-----"- Amigne L-____ .;... Riesling weiß -'------'- Spezial Fig. 1: Dendrogram of the Riesling-type grapevines. The simi-larity degree values were calculated using the bands generated by all20 primers (FO 1 to F20) and and with the help of the "Rapid" program. Cluster analysis was done with a self-made program (see Materials and methods for details). Fig. 2: Ethidiumbromide-stained 1.5 % agarase gel showing re-solved amplification products obtained with primer F04 (Operon Technologies) from some cu ltivars of the Burgundy group: a) A. DNA cut with Hind/ll and EcoRI as size standard; band c) Aligote; d and e) Aligote vert; fand g) Auxerrois; h and i) Affen-thaler ; j and k) Chardonnay blanc ; l) Chardonnay musque ; m) pBR322 cut with Hinfl as size marker. Duplicate assays were prepared independently from two individual plants. Bundesanstalt für Züchtungsforschung an Kulturpflanzen, Inst. für Rebenzüchtung Geilwellerhof, D-76833 Siebeldingen, Germany. 2 5 0  J .  T s c H A M M E R  a n d  E v A  Z Y P R I A N  E l b l i n g  w h i t e  a p p e a r  r a t h e r  c l o s e l y  r e l a t e d ,  i n  a g r e e m e n t  w i t h  t h e i r  m o r p h o l o g i c a l  s i r n i l a r i t y  ( G A L E T  1 9 9 0 ) .  G o u a i s  b l a n c  i s  c l e a r l y  d i s t i n g u i s h a b l e  f r o m  E l b l i n g  w h i t e ,  a  r e -s u l t  i n  c o n t r a s t  t o  G o E T H E  ( 1 8 8 7 ) .  T h e  o t h e r  r e p r e s e n t a -t i v e s  o f  t h e  R i e s l i n g - t y p e  v i n e s  d o  n o t  s e e m  t o  b e  c l o s e l y  r e l a t e d  t o  e a c h  o t h e r .  T h e  p r o g e n i t o r  o f  a  c r o s s  P i n o t  n o i r  x  C h a m b o u r c i n  ( " S p e z i a l " )  i s  w e i l  s e p a r a t e d  f r o m  t h e  R i e s -l i n g - t y p e  v i n e s  a n d  s u r e l y  n o t  c l o s e l y  r e l a t e d  t o  R i e s l i n g .  q . o  g.  1  q. 2  q , 3  q. 4  q. 5  q . 6  q. 7  q. s  q . 9  1 , . o  )  .  .  .  ;  S 1 m 1 l a n t y  d e g r e e  v a l u e  ~----'- M o r i l l o n  N W  j  M o r i l l o n  Ö s t e r r e i c h  L _  _ _ _  _ _ , _  R i e s l i n g  w e i ß  C h a r d o n n a y  m u s q u e  C h a r d o n n a y  b l a n c  2  C h a r d o n n a y  b l a n c  1  A u x e r r o i s  A l i g o t e  1  A l i g o t e  v e r t  A l i g o t e  2  A f f e n t h a l e r  P i n o t  b l a n c  P i n o t g r i s  2  P i n o t  m e u n i e r  P i n o t  g r i s  1  P i n o t  n o i r  S a i n t  L a u r e n t  1  S a i n t  L a u r e n t  2  F i g .  3 :  D e n d r o g r a m  o f  t h e  B u r g u n d y  g r o u p .  C a l c u l a t i o n  o f  s i m i -l a r i t y  d e g r e e s  a n d  d u s t e r  a n a l y s i s  w a s  p e r f o r m e d  a s  d e s c r i b e d  i n  F i g .  1 .  Q u i t e  i n  c o n t r a s t  t o  t h e  R i e s l i n g - t y p e  c u l t i v a r s ,  t h e  B u r -g u n d y  g r o u p  i s  m o r e h o m o g e n e a u s  ( F i g s .  2  a n d  3 ) .  l t  w a s  n o t  p o s s i b l e  t o  d i f f e r e n t i a t e  b e t w e e n  t h e  P i n o t  ( B u r g u n d y )  v a r i e t i e s  ( P i n o t  n o i r ,  b l a n c ,  m e u n i e r ,  g r i s )  t h e m s e l v e s ,  d u e  t o  t h e i r  c l o s e  r e l a t i o n s h i p .  T h e s e  B u r g u n d i e s  p r o b a b l y  d i f -f e r  o n l y  i n  s i n g l e  b a s e  c h a n g e s .  T h u s  i t  s e e m s  v e r y  d i f f i -c u l t ,  h o w e v e r  n o t  i m p o s s i b l e  t o  f i n d  a n  a p p r o p r i a t e  p r i m e r  a b l e  t o  d i s c r i r n i n a t e  t h e m .  N e i t h e r  C h a r d o n n a y  b l a n c  c o u l d  b e  d i f f e r e n t i a t e d  f r o m  C h a r d o n n a y  m u s q u e ,  n o r  A l i g o t e  f r o m  A l i g o t e  v e r t  f o r  s i r n i l a r  r e a s o n s  ( g e n e t i c  a l t e r a t i o n  b y  p o i n t  m u t a t i o n s  o n l y ) .  A u x e r r o i s  a p p e a r e d  c l o s e l y  r e l a t e d  t o  C h a r d o n n a y ,  b u t  w a s  c l e a r l y  d i f f e r e n t .  C h a r d o n n a y  c o u l d  w e l l  b e  d i f f e r e n t i a t e d  f r o m  t h e  P i n o t s  a s  w e i l  a s  S a i n t  L a u r e n t ,  A u x e r r o i s  a n d  M o r i l l o n .  T h i s  i n d i c a t e s ,  t h a t  t h e s e  c u l t i v a r s  a r e n o t  a s  c l o s e l y  r e l a t e d  a s  t h e  P i n o t s  t h e m s e l v e s .  T h i s  r e s u l t  c o n t r a s t s  Ü A L E T  ( 1 9 9 0 ) ,  M Ü L L E R  ( 1 9 3 0 )  a n d  H r L L E B R A N D T  e t  a l .  ( 1 9 9 3 ) .  E s p e c i a l l y  M o r i l l o n  e x h i b i t e d  s i g n i f i c a n t  d i f f e r e n c e s  i n  R A P D  p a t t e r n s  t o  t h e  o t h e r  B u r -g u n d y  c u l t i v a r s .  T h e  s i m i l a r i t y  o f  M o r i l l o n  a n d  R i e s l i n g  s e e m s  t o  b e  h i g h e r .  M o r i l l o n  t h e r e f o r e  i s  c o n s i d e r e d  n o t  c l o s e l y  r e l a t e d  t o  t h e  P i n o t  c u l t i v a r s .  I n  a d d i t i o n ,  M o r i l l o n  p l a n t s  f r o m  d i f f e r e n t  l o c a t i o n s  ( N e u s t a d t / W e i n s t r a ß e ,  G e r -m a n y  a n d  K l o s t e r n e u b u r g /  A u s t r i a )  e x h i b i t e d  d i f f e r e n t  R A P D  p a t t e r n s .  T h e  n a m e  M o r i l l o n  t h e r e f o r e  r e f e r s  t o  d i f -f e r e n t  c u l t i v a r s .  A f f e n t h a l e r  a p p e a r e d  t o  b e  s i m i l a r  t o  t h e  P i n o t  c u l t i v a r s ,  a l t h o u g h  i t  h a s  b e e n  c o n s i d e r e d  n o t  r e l a t e d  t o  t h e  P i n o t s  ( H r L L E B R A N D  e t  a l .  1 9 9 3 ) .  B ü s c H E R ,  N . ;  Z Y P R I A N ,  E . ;  B L A I C H ,  R . ;  1 9 9 3 :  I d e n t i f i c a t i o n  o f  g r a p e v i n e  c u l t i v a r s  b y  D N A  a n a l y s e s :  P i t f a l l s  o f  r a n d o m  a m p l i f i e d  p o l y m o r -p h i c  D N A  t e c h n i q u e s .  V i t i s  3 2 ,  1 8 7 - 1 8 8 .  - - ; - - ;  B A C H M A N N ,  Ü . ;  B L AI C H ,  R . ;  1 9 9 4 :  Ü n  t h e  o r i g i n  o f  t h e  g r a p e v i n e  v a r i e t y  M ü l l e r - T h u r g a u  a s  i n v e s t i g a t e d  b y  t h e  i n h e r i t a n c e  o f  r a n d o m  a m p l i f i e d  p o l y m o r p h i c  D N A  ( R A P D ) .  V i t i s  3 3 ,  1 5 - 1 7 .  G AL E T ,  P . ;  1 9 9 0 :  C e p a g e s  e t  V i g n o b l e s  d e  F r a n c e .  T o m e  1 1 :  L ' A m p e -l o g r a p h i e  Fran~aise , 2 e m e  E d i t i o n .  l m p r i m e r i e  C h a r l e s  D e h a n ,  M o n t p e l l i e r .  G o E T H E ,  H . ;  1 8 8 7 :  H a n d b u c h  d e r  A m p e l o g r a p h i e ,  2 .  A u f l .  V e r l a g  P a u l  P a r e y ,  B e r l i n .  H I L L E B R A N D,  W . ;  L o r r ,  H . ;  P F A F F ,  F . ;  1 9 9 3 :  T a s c h e n b u c h  d e r  R e b s o r t e n ,  I  0 .  A u f ! .  F a c h v e r l a g  D r .  F r a u n d  G m b H ,  M a i n z .  M ü L L E R ,  K .  ( E d . ) ;  1 9 3 0 :  W e i n b a u  L e x i k o n . V e r l a g s b u c h h a n d l u n g  P a u l  P a r e y ,  B e r l i n .  N ö B A U E R ,  W . ;  T I MIS C H L ,  W . ;  1 9 7 9 :  M a t h e m a t i s c h e  M o d e l l e  i n  d e r  B i o l o g i e .  V i e w e g  V e r l a g ,  B r a u n s c h w e i g ,  W i e s b a d e n .  T H O M A S ,  M .  R . ;  M A T S U M O T O ,  S . ;  C AIN ,  P . ;  S c o r r ,  N .  S . ;  1 9 9 3 :  R e p e t i t i v e  D N A  o f  g r a p e v i n e :  c l a s s e s  p r e s e n t  a n d  s e q u e n c e s  s u i t a b l e  f o r  c u l t i v a r  i d e n t i f i c a t i o n .  T h e o r .  A p p l .  G e n e t .  8 6 ,  1 7 3 - 1 8 0 .  

Molecular characterization of grapevine cultivars of Riesling-type and of closely related Burgundies

https://ojs.openagrar.de/index.php/VITIS/article/download/5158/4946

Molecular characterization of grapevine cultivars of Riesling-type and of closely related Burgundies

Abstract

Similar works

Full text

Available Versions

JKI Open Journal Systems (Julius Kühn-Institut)