Planta
Planta 140, 151 153 (1978)
9 by Springer-Verlag 1978
Dihydrozeatin Riboside, a Minor Cytokinin from the Leaves of Phaseolus vulgaris L. T.L. W a n g * a n d R. H o r g a n Department of Botany and Microbiology, University College of Wales, Penglais, Aberystwyth, Dyfed SY23 3DA, U.K.
Abstract. D i h y d r o z e a t i n r i b o s i d e has been identified in the leaves o f d e c a p i t a t e d b e a n p l a n t s by S e p h a d e x L H 2 0 c h r o m a t o g r a p h y a n d c o m b i n e d gas c h r o m a t o g r a p h y - m a s s s p e c t r o m e t r y . The r e l a t i o n s h i p b e t w e e n the c y t o k i n i n s i s o l a t e d a n d identified f r o m this system a n d those p r e v i o u s l y r e p o r t e d in P h a s e o l u s is discussed.
Key words: C y t o k i n i n - Leaves - P h a s e o l u s .
to a ZR marker was purified by successive chromatography on paper in Solvents B, D and E before analysis by GLC and GCMS. Chromatography
Column, paper and gas-liquid chromatography were carried out essentially as described previously (Wang et al., 1977) with the following differences: one tenth of each zone from the chromatogram developed in Solvent E was used for bioassay; GLC was performed isothermally at 280~ C. Solvents
Introduction D i h y d r o z e a t i n - O - f i - D - g l u c o s i d e has recently been identified f r o m the leaves o f P h a s e o l u s v u l g a r i s L. ( W a n g et al., 1977). I n the following work, the presence o f d i h y d r o z e a t i n riboside, as a m i n o r c o m p o n e n t of the c y t o k i n i n c o m p l e m e n t in these leaves, is reported.
A: 35% ethanol (v/v); B: sec-butanol: 25% ammonia (4: 1, v/v); C: 80% ethanol (v/v); D: n-butanol:acetic acid:water (12:3:5, v/v) ; E : water. Bioassay
The assay used throughout was the tobacco stem pith callus assay (Murashige and Skoog, 1962).
Results Materials and Methods Plant Material and Cytokinin Extraction
Plants of Phaseolus vulgaris L. var. "Canadian Wonder" were grown and extracted as previously described (Wang et al., 1977). After extraction in aqueous methanol, the sample was subjected to ion-exchange and partitioned against water-saturated n-butanol. Following chromatography of the combined butanol phases on Sephadex LH20 in Solvent A, the active fraction (15) corresponding Present address." Botany Department, University of Leicester, Leicester, LE1 7RH, U.K. *
Abbreviations : DHZ = dihydrozeatin ; DHZOG = dihydrozeatin-O-
fi-D-glucoside; DHZR=dihydrozeatin riboside: GCMS=combined gas chromatography-mass spectrometry; GLC=gas-liquid chromatography; TIC=total ion current; TMS=trimethylsilyl; Z=zeatin; ZR=zeatin riboside
In a d d i t i o n to the m a j o r p e a k o f c y t o k i n i n activity detected f r o m S e p h a d e x L H 2 0 c h r o m a t o g r a p h y o f an extract o f b e a n leaves which has p r e v i o u s l y been identified as 6-(4-O-fi-D-glucosyl-3-methylbutyla m i n o ) p u r i n e ( W a n g et al., 1977), a small p e a k o f activity was o b s e r v e d which c o - c h r o m a t o g r a p h e d with a zeatin r i b o s i d e m a r k e r . This p e a k was c h r o m a t o g r a p h e d on p a p e r in Solvent B a n d the b i o l o g i c a l l y active r e g i o n (centered on R f 0.65) eluted with Solvent C. T h e eluate was t a k e n to dryness a n d c h r o m a t o g r a p h e d on p a p e r in Solvent D. A single U V abs o r b i n g s p o t ( c o n t a i n i n g all the b i o l o g i c a l activity) was n o t e d at R f 0.72. H o w e v e r , after elution o f this s p o t with Solvent C, the U V a b s o r p t i o n s p e c t r u m revealed the presence o f n o n - c y t o k i n i n substances.
0032-0935/78/0140/0151/$01.00
152
T.L. Wang and R. Horgan: Dihydrozeatin Riboside from Leaves KI NETIN CONC.
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Planta 140, 151 153 (1978)
9 by Springer-Verlag 1978
Dihydrozeatin Riboside, a Minor Cytokinin from the Leaves of Phaseolus vulgaris L. T.L. W a n g * a n d R. H o r g a n Department of Botany and Microbiology, University College of Wales, Penglais, Aberystwyth, Dyfed SY23 3DA, U.K.
Abstract. D i h y d r o z e a t i n r i b o s i d e has been identified in the leaves o f d e c a p i t a t e d b e a n p l a n t s by S e p h a d e x L H 2 0 c h r o m a t o g r a p h y a n d c o m b i n e d gas c h r o m a t o g r a p h y - m a s s s p e c t r o m e t r y . The r e l a t i o n s h i p b e t w e e n the c y t o k i n i n s i s o l a t e d a n d identified f r o m this system a n d those p r e v i o u s l y r e p o r t e d in P h a s e o l u s is discussed.
Key words: C y t o k i n i n - Leaves - P h a s e o l u s .
to a ZR marker was purified by successive chromatography on paper in Solvents B, D and E before analysis by GLC and GCMS. Chromatography
Column, paper and gas-liquid chromatography were carried out essentially as described previously (Wang et al., 1977) with the following differences: one tenth of each zone from the chromatogram developed in Solvent E was used for bioassay; GLC was performed isothermally at 280~ C. Solvents
Introduction D i h y d r o z e a t i n - O - f i - D - g l u c o s i d e has recently been identified f r o m the leaves o f P h a s e o l u s v u l g a r i s L. ( W a n g et al., 1977). I n the following work, the presence o f d i h y d r o z e a t i n riboside, as a m i n o r c o m p o n e n t of the c y t o k i n i n c o m p l e m e n t in these leaves, is reported.
A: 35% ethanol (v/v); B: sec-butanol: 25% ammonia (4: 1, v/v); C: 80% ethanol (v/v); D: n-butanol:acetic acid:water (12:3:5, v/v) ; E : water. Bioassay
The assay used throughout was the tobacco stem pith callus assay (Murashige and Skoog, 1962).
Results Materials and Methods Plant Material and Cytokinin Extraction
Plants of Phaseolus vulgaris L. var. "Canadian Wonder" were grown and extracted as previously described (Wang et al., 1977). After extraction in aqueous methanol, the sample was subjected to ion-exchange and partitioned against water-saturated n-butanol. Following chromatography of the combined butanol phases on Sephadex LH20 in Solvent A, the active fraction (15) corresponding Present address." Botany Department, University of Leicester, Leicester, LE1 7RH, U.K. *
Abbreviations : DHZ = dihydrozeatin ; DHZOG = dihydrozeatin-O-
fi-D-glucoside; DHZR=dihydrozeatin riboside: GCMS=combined gas chromatography-mass spectrometry; GLC=gas-liquid chromatography; TIC=total ion current; TMS=trimethylsilyl; Z=zeatin; ZR=zeatin riboside
In a d d i t i o n to the m a j o r p e a k o f c y t o k i n i n activity detected f r o m S e p h a d e x L H 2 0 c h r o m a t o g r a p h y o f an extract o f b e a n leaves which has p r e v i o u s l y been identified as 6-(4-O-fi-D-glucosyl-3-methylbutyla m i n o ) p u r i n e ( W a n g et al., 1977), a small p e a k o f activity was o b s e r v e d which c o - c h r o m a t o g r a p h e d with a zeatin r i b o s i d e m a r k e r . This p e a k was c h r o m a t o g r a p h e d on p a p e r in Solvent B a n d the b i o l o g i c a l l y active r e g i o n (centered on R f 0.65) eluted with Solvent C. T h e eluate was t a k e n to dryness a n d c h r o m a t o g r a p h e d on p a p e r in Solvent D. A single U V abs o r b i n g s p o t ( c o n t a i n i n g all the b i o l o g i c a l activity) was n o t e d at R f 0.72. H o w e v e r , after elution o f this s p o t with Solvent C, the U V a b s o r p t i o n s p e c t r u m revealed the presence o f n o n - c y t o k i n i n substances.
0032-0935/78/0140/0151/$01.00
152
T.L. Wang and R. Horgan: Dihydrozeatin Riboside from Leaves KI NETIN CONC.
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