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Maturation and ripening of three Berberis species from different altitudes
Scientia Horticulturae, 19 (1983) 91--95 Elsevier Scientific Publishing Company, Amsterdam -- Printed in The Netherlands
91
MATURATION AND RIPENING OF THREE BERBERIS SPECIES FROM DIFFERENT ALTITUDES
PRAKASH CHANDRA
and N.P. T O D A R I A
High Altitude Plant Physiology Research Centre, and Department of Botany, Garhwal University,Srinagar (Garhwal)-246174, U.P. (India) (Accepted for publication4 May 1982)
ABSTRACT Chandra, P. and Todaria, N.P., 1983. Maturation and ripening of three Berberis species from different altitudes. Scientia Hortic., 19: 91--95. Changes in chlorophyll, carotenoids, anthocyanins, soluble sugars, phenolics and berberine contents in the fruit pulp of one high-altitude, one middle-altitude and one low-altitude species of Berberis were studied during their maturation and ripening. Maturation and ripening of Berberis fruit was accompanied by an increase in soluble sugar and anthocyanin and a decrease in chlorophyll and berberine contents. The changes in the carotenoids and phenolics varied with species, and did not seem to be directly related to maturation and ripening of the fruits. The fruits of the low altitude species, which were more palatable, had high edibility, contained more sugar and a lower concentration of phenolics than the other species.
INTRODUCTION
There are many seasonal fruit-bearing wild plant species in the Indian mountains. Their fruits are eaten by the local inhabitants. However, the physiology and biochemistry of these fruits is little understood. Since they are natural components of the mountain ecosystem and fruit profusely even under harsh conditions, their breeding and cultivation could have a strong impact on the economy and nutrition ecology of the mountain regions. The genus Berberis (Berberidaceae) has nearly 150 species world-wide (Pomilio, 1973} of which 14 have been reported from the Indian Himalayas (Osmaston, 1927). With such a diversity of genetic material available, members of the genus could be suitable for domestication. The present study was therefore undertaken to investigate the physiology and biochemistry of fruits of 3 species of this genus from different altitudes in The Garhwal Himalayas.
0304-4238/83/0000--0000/$03.00
© 1983 Elsevier Scientific Publishing Company
92 MATERIALS AND METHODS
Twenty to 25 fruits from 4 bushes each of Berberis lycium Royle, B. chitria Lindl and B. vulgaris Linn, from altitudes of 1650, 2500 and 3000 m, respectively, were collected at regular intervals from the date of apparent fruit-set (fall of petals) until their full ripening on the plant. The fruits were brought to the laboratory on the day of sampling and immediately analysed. Chlorophylls and carotenoids in fresh fruit pulp were analyzed by a spectrophotometric method (Holm, 1954) using an ECIL spectrophotometer, model GS 865 B. For analysis of anthocyanin content, 1 g fresh fruit pulp was extracted in 1% HC1 in methanol (v/v) for 4 days at 10°C in the dark. The extract was clarified by filtration, and the absorbance of the filtrate was measured at 530 and 657 nm. The anthocyanin content of the extract was calculated following the method described by Mancinelli et al. (1976). Total phenolics and berberine in dried pulp were analyzed following the methods described by Swain and Hills (1959) and Reichter (1914), respectively. The method described by Dubois et al. (1951) was followed for determining the total soluble sugar content of dried pulp. All assays were replicated twice at each sampling. 070
la ~Jyc 060
0.24
~050
0 2O
]a lye
¢-
\040
~= 0 1 6
~
0 30
~ 012
O20
~ 0 08
ma
k.)
chick
--
ma
0 04
chit
Q.IO
~ 15
30
45
60
Days after fruit setting
75
90
105
0
15
30
45
Days after
60
75
ha
90
105
fruit setting
Fig. 1. Changes in chlorophyll content (mg/g fresh weight) during maturation and ripening of 3 Berberis species. ~--~, l o w altitude Berberis lycium = 1.a. lyc. , middle altitude Berberis chitria = m.a. chit. o--~, high altitude Berberis vulgaris = h.a. vulg. Fig. 2. Changes in carotenoids content (mg/g fresh weight) during maturation and ripening of 3 Berberis species. S y m b o l s as Fig. 1.
93 RESULTS
Young fruits of the low-altitude (1.a.) B. lycium contained high concentrations of chlorophyll (Fig. 1) and carotenoids (Fig. 2), which decreased with maturation, although the fall was sharper in chlorophyll than in carotenoid content. In the middle-altitude (m.a.) B. chitria and high,altitude (h.a.) B. vulgaris, chlorophyll content was relatively low and showed a slight decrease with maturation (Fig. 1), but the carotenoid content gradually increased with maturation. The increase was more in the m.a.B, chitria than in the h.a.B. vulgaris (Fig. 2). The anthocyanin content was initially low and increased sharply after about 45 days in all the 3 species (Fig. 3). The total soluble sugar content (Fig. 4) was very low and altered little during maturation. Total phenolic content was high in the initial stages in all 3 species and decreased with maturation (Fig. 5). The rate of decrease was greatest in the 1.a.B. lycium and smallest in the h.a.B, vulgaris. The berberine contents {Fig. 6) were initially low, increased to a very high level during maturation and
la
10
06t
chit
24
20
%
lyc.
~)~045f
ha. ma
o
S E
e,
12
D 4
O05F
I Days
after
fruit
setting
I
i
Days
i
after
i
fruit
i
i
I
setting
Fig. 3. Changes in anthocyanin content (A530-0.33 A657) during maturation and ripening of Berberis species. Symbols as Fig. 1. Fig. 4. Changes in sugar content (mg/g dry weight) during maturation and ripening of 3 Berberis species. Symbols as Fig. 1.
94 16
lyc 14
012
~ 12
h"--~
h
Of O
$
4
00,~
\
002
0
"4
i
I
I
"I
15
30
45
60
"
I
I
75
90
Days after fr'ult setting
2
I
105
O
I
15
l
30
4
;
~
0
I
75
i0
I
105
Daysafter" fruit setting
Fig. 5. Changes in total phenolics (mg/g dry weight) during maturation and ripening of 3 Berberis species. Symbols as Fig. 1. Fig. 6. Changes in berberine content (mg/g dry weight) during maturation and ripening of 3 Berberis species. Symbols as Fig. 1.
decreased sharply during ripening in the 1.a.B. lycium. In the other 2 species, the berberine content remained high for a b o u t 75 days and decreased sharply with ripening. DISCUSSION The loss in chlorophyll content, associated with the maturation of many fruits {Workman, 1963; Rhodes and Wooltorton, 1967; L o o n e y and Patterson, 1967), appears also to be a characteristic feature in the fruits of Berberis. H o w ever, rapid destruction of chlorophyll in the 1.a.B. lycium as compared to the h.a.B, vulgaris might have been due to a faster rate of ripening through higher light intensity and higher temperature at lower elevations. Increase in carotenoid content during the ripening of fruits has been reported in many fruits (Workman, 1963; Francis et al., 1955). The dependence of such changes is evident in the findings reported here. In middle-altitude species carotenoid content increased with ripening, whereas in low-altitude species there was a sharp decline. In these fruits, accumulation of anthocyanin seems to be a characteristic feature of fruit maturation. The increase in anthocyanin content in all the species could support the earlier findings that sugar flux is essential for anthocyanin accumulation (Pirie and Mullins, 1976), although our data are insufficient.
95 T h e changes in p h e n o l i c s d u r i n g f r u i t m a t u r a t i o n are indicative o f the f a c t t h a t , w i t h m a t u r i t y , t h e p h e n o l i c s c o n t e n t decreases m o r e r a p i d l y in t h e 1. a. B. lycium t h a n in t h e h . a . B , vulgaris. This d i f f e r e n c e does n o t s e e m t o be r e l a t e d t o t h e f r u i t m a t u r a t i o n process, b u t r a t h e r t o the geographical distrib u t i o n o f t h e species. Since, at 75 days, the f r u i t o f even t h e h . a . B , vulgaris w e r e a l r e a d y ripe, t h e p h e n o l i c s c o n t e n t r e m a i n e d high. R a p i d fall in t h e b e r b e r i n e c o n t e n t w i t h t h e o n s e t o f m a t u r a t i o n was f o u n d in all t h e 3 species, indicating its close a s s o c i a t i o n w i t h t h e fruit-ripening process. A h o r t i c u l t u r a l e v a l u a t i o n o f t h e 3 species is in progress. ACKNOWLEDGEMENTS T h e a u t h o r s wish t o express t h e i r d e e p g r a t i t u d e to Dr. A.N. P u r o h i t , D i r e c t o r , High A l t i t u d e Plant P h y s i o l o g y R e s e a r c h Centre, G a r h w a l University, Srinagar ( G a r h w a l ) , f o r supervision a n d p r o v i d i n g necessary facilities f o r this w o r k . P. C h a n d r a is also grateful t o t h e University G r a n t s C o m m i s s i o n . New-Delhi, f o r financial assistance.
REFERENCES Dubois, M., Gilles, K., Hamilton, J.K., Rebers, P.A. and Smith, F., 1951. In: K.Plaech and M.V. Tracey (Editors), Modern Methods of Plant Analysis. Vol. II, 1955, SpringerVerlag, Berlin, p. 22. Francis, F.J., Harney, P.H. and Bulstrode, P.C., 1955. Color and pigment changes in the flesh of McIntosh apples after removal from storage. Proc. Am. Soc. Hortic. Sci., 65: 211--213. Holm, G., 1954. Chlorophyll mutations in barley. Acta Agric. Scand., 4: 457--471. Looney, N.E. and Patterson, M.E., 1967. Chlorophyllase activity in apples and bananas during the climateric phase. Nature (London), 214: 1245--1246. Mancinelli, A.L., Yang, C.H., Rabino, I. and Kuzmanoff, K.M., 1976. Photocontrol of anthocyanin synthesis. V. Further evidence against the involvement of photosynthesis in high irradiance reaction anthocyanin of young seedlings. Plant. Physiol., 58 : 214--217. Osmaston, A.E., 1927. A Forest Flora for Kumaon. Govt. Press, United Provinces, Allahabad, pp. 17--22. Pirie, A. and Mullins, M.G., 1976. Changes in anthocyanin and phenolics content of grapevine leaf and fruit tissue treated with sucrose, nitrate and abscisic acid. Plant Physiol., 58 : 468--472. Pomilio, A.B., 1973. Anthochyanins in fruits of Berberis buxifolia. Phytochemistry, 12: 218--220. Reichter, V.E., 1914. Arch. Pharmacol., 2:371--383 (In: K. Paedh and M.V. Traeey, (Editors), Modern Methods of Plant Analysis. Vol. IV, 1955, Springer-Verlag, Berlin, p. 390 Rhodes, M.J.C. and Wooltorton, L.S.C., 1967. The respiratory climateric in apple fruits. The action of hydrolytic enzymes in peel tissue during climateric period in fruit detached from the tree. Phytochemistry. 6 : 1--12. Workman, M., 1963. Color and pigment changes in Golden Delicious and Grimes Golden apples. Proc. Am. Soc. Hortic. Sci., 83: 149--161. Swain, T. and Hills, N.E., 1959. The phenolic constituents of Prunus domestica. I. The quantitative analysis of phenolic constituents. J. Sci. Food Agric., 10: 63--67.
91
MATURATION AND RIPENING OF THREE BERBERIS SPECIES FROM DIFFERENT ALTITUDES
PRAKASH CHANDRA
and N.P. T O D A R I A
High Altitude Plant Physiology Research Centre, and Department of Botany, Garhwal University,Srinagar (Garhwal)-246174, U.P. (India) (Accepted for publication4 May 1982)
ABSTRACT Chandra, P. and Todaria, N.P., 1983. Maturation and ripening of three Berberis species from different altitudes. Scientia Hortic., 19: 91--95. Changes in chlorophyll, carotenoids, anthocyanins, soluble sugars, phenolics and berberine contents in the fruit pulp of one high-altitude, one middle-altitude and one low-altitude species of Berberis were studied during their maturation and ripening. Maturation and ripening of Berberis fruit was accompanied by an increase in soluble sugar and anthocyanin and a decrease in chlorophyll and berberine contents. The changes in the carotenoids and phenolics varied with species, and did not seem to be directly related to maturation and ripening of the fruits. The fruits of the low altitude species, which were more palatable, had high edibility, contained more sugar and a lower concentration of phenolics than the other species.
INTRODUCTION
There are many seasonal fruit-bearing wild plant species in the Indian mountains. Their fruits are eaten by the local inhabitants. However, the physiology and biochemistry of these fruits is little understood. Since they are natural components of the mountain ecosystem and fruit profusely even under harsh conditions, their breeding and cultivation could have a strong impact on the economy and nutrition ecology of the mountain regions. The genus Berberis (Berberidaceae) has nearly 150 species world-wide (Pomilio, 1973} of which 14 have been reported from the Indian Himalayas (Osmaston, 1927). With such a diversity of genetic material available, members of the genus could be suitable for domestication. The present study was therefore undertaken to investigate the physiology and biochemistry of fruits of 3 species of this genus from different altitudes in The Garhwal Himalayas.
0304-4238/83/0000--0000/$03.00
© 1983 Elsevier Scientific Publishing Company
92 MATERIALS AND METHODS
Twenty to 25 fruits from 4 bushes each of Berberis lycium Royle, B. chitria Lindl and B. vulgaris Linn, from altitudes of 1650, 2500 and 3000 m, respectively, were collected at regular intervals from the date of apparent fruit-set (fall of petals) until their full ripening on the plant. The fruits were brought to the laboratory on the day of sampling and immediately analysed. Chlorophylls and carotenoids in fresh fruit pulp were analyzed by a spectrophotometric method (Holm, 1954) using an ECIL spectrophotometer, model GS 865 B. For analysis of anthocyanin content, 1 g fresh fruit pulp was extracted in 1% HC1 in methanol (v/v) for 4 days at 10°C in the dark. The extract was clarified by filtration, and the absorbance of the filtrate was measured at 530 and 657 nm. The anthocyanin content of the extract was calculated following the method described by Mancinelli et al. (1976). Total phenolics and berberine in dried pulp were analyzed following the methods described by Swain and Hills (1959) and Reichter (1914), respectively. The method described by Dubois et al. (1951) was followed for determining the total soluble sugar content of dried pulp. All assays were replicated twice at each sampling. 070
la ~Jyc 060
0.24
~050
0 2O
]a lye
¢-
\040
~= 0 1 6
~
0 30
~ 012
O20
~ 0 08
ma
k.)
chick
--
ma
0 04
chit
Q.IO
~ 15
30
45
60
Days after fruit setting
75
90
105
0
15
30
45
Days after
60
75
ha
90
105
fruit setting
Fig. 1. Changes in chlorophyll content (mg/g fresh weight) during maturation and ripening of 3 Berberis species. ~--~, l o w altitude Berberis lycium = 1.a. lyc. , middle altitude Berberis chitria = m.a. chit. o--~, high altitude Berberis vulgaris = h.a. vulg. Fig. 2. Changes in carotenoids content (mg/g fresh weight) during maturation and ripening of 3 Berberis species. S y m b o l s as Fig. 1.
93 RESULTS
Young fruits of the low-altitude (1.a.) B. lycium contained high concentrations of chlorophyll (Fig. 1) and carotenoids (Fig. 2), which decreased with maturation, although the fall was sharper in chlorophyll than in carotenoid content. In the middle-altitude (m.a.) B. chitria and high,altitude (h.a.) B. vulgaris, chlorophyll content was relatively low and showed a slight decrease with maturation (Fig. 1), but the carotenoid content gradually increased with maturation. The increase was more in the m.a.B, chitria than in the h.a.B. vulgaris (Fig. 2). The anthocyanin content was initially low and increased sharply after about 45 days in all the 3 species (Fig. 3). The total soluble sugar content (Fig. 4) was very low and altered little during maturation. Total phenolic content was high in the initial stages in all 3 species and decreased with maturation (Fig. 5). The rate of decrease was greatest in the 1.a.B. lycium and smallest in the h.a.B, vulgaris. The berberine contents {Fig. 6) were initially low, increased to a very high level during maturation and
la
10
06t
chit
24
20
%
lyc.
~)~045f
ha. ma
o
S E
e,
12
D 4
O05F
I Days
after
fruit
setting
I
i
Days
i
after
i
fruit
i
i
I
setting
Fig. 3. Changes in anthocyanin content (A530-0.33 A657) during maturation and ripening of Berberis species. Symbols as Fig. 1. Fig. 4. Changes in sugar content (mg/g dry weight) during maturation and ripening of 3 Berberis species. Symbols as Fig. 1.
94 16
lyc 14
012
~ 12
h"--~
h
Of O
$
4
00,~
\
002
0
"4
i
I
I
"I
15
30
45
60
"
I
I
75
90
Days after fr'ult setting
2
I
105
O
I
15
l
30
4
;
~
0
I
75
i0
I
105
Daysafter" fruit setting
Fig. 5. Changes in total phenolics (mg/g dry weight) during maturation and ripening of 3 Berberis species. Symbols as Fig. 1. Fig. 6. Changes in berberine content (mg/g dry weight) during maturation and ripening of 3 Berberis species. Symbols as Fig. 1.
decreased sharply during ripening in the 1.a.B. lycium. In the other 2 species, the berberine content remained high for a b o u t 75 days and decreased sharply with ripening. DISCUSSION The loss in chlorophyll content, associated with the maturation of many fruits {Workman, 1963; Rhodes and Wooltorton, 1967; L o o n e y and Patterson, 1967), appears also to be a characteristic feature in the fruits of Berberis. H o w ever, rapid destruction of chlorophyll in the 1.a.B. lycium as compared to the h.a.B, vulgaris might have been due to a faster rate of ripening through higher light intensity and higher temperature at lower elevations. Increase in carotenoid content during the ripening of fruits has been reported in many fruits (Workman, 1963; Francis et al., 1955). The dependence of such changes is evident in the findings reported here. In middle-altitude species carotenoid content increased with ripening, whereas in low-altitude species there was a sharp decline. In these fruits, accumulation of anthocyanin seems to be a characteristic feature of fruit maturation. The increase in anthocyanin content in all the species could support the earlier findings that sugar flux is essential for anthocyanin accumulation (Pirie and Mullins, 1976), although our data are insufficient.
95 T h e changes in p h e n o l i c s d u r i n g f r u i t m a t u r a t i o n are indicative o f the f a c t t h a t , w i t h m a t u r i t y , t h e p h e n o l i c s c o n t e n t decreases m o r e r a p i d l y in t h e 1. a. B. lycium t h a n in t h e h . a . B , vulgaris. This d i f f e r e n c e does n o t s e e m t o be r e l a t e d t o t h e f r u i t m a t u r a t i o n process, b u t r a t h e r t o the geographical distrib u t i o n o f t h e species. Since, at 75 days, the f r u i t o f even t h e h . a . B , vulgaris w e r e a l r e a d y ripe, t h e p h e n o l i c s c o n t e n t r e m a i n e d high. R a p i d fall in t h e b e r b e r i n e c o n t e n t w i t h t h e o n s e t o f m a t u r a t i o n was f o u n d in all t h e 3 species, indicating its close a s s o c i a t i o n w i t h t h e fruit-ripening process. A h o r t i c u l t u r a l e v a l u a t i o n o f t h e 3 species is in progress. ACKNOWLEDGEMENTS T h e a u t h o r s wish t o express t h e i r d e e p g r a t i t u d e to Dr. A.N. P u r o h i t , D i r e c t o r , High A l t i t u d e Plant P h y s i o l o g y R e s e a r c h Centre, G a r h w a l University, Srinagar ( G a r h w a l ) , f o r supervision a n d p r o v i d i n g necessary facilities f o r this w o r k . P. C h a n d r a is also grateful t o t h e University G r a n t s C o m m i s s i o n . New-Delhi, f o r financial assistance.
REFERENCES Dubois, M., Gilles, K., Hamilton, J.K., Rebers, P.A. and Smith, F., 1951. In: K.Plaech and M.V. Tracey (Editors), Modern Methods of Plant Analysis. Vol. II, 1955, SpringerVerlag, Berlin, p. 22. Francis, F.J., Harney, P.H. and Bulstrode, P.C., 1955. Color and pigment changes in the flesh of McIntosh apples after removal from storage. Proc. Am. Soc. Hortic. Sci., 65: 211--213. Holm, G., 1954. Chlorophyll mutations in barley. Acta Agric. Scand., 4: 457--471. Looney, N.E. and Patterson, M.E., 1967. Chlorophyllase activity in apples and bananas during the climateric phase. Nature (London), 214: 1245--1246. Mancinelli, A.L., Yang, C.H., Rabino, I. and Kuzmanoff, K.M., 1976. Photocontrol of anthocyanin synthesis. V. Further evidence against the involvement of photosynthesis in high irradiance reaction anthocyanin of young seedlings. Plant. Physiol., 58 : 214--217. Osmaston, A.E., 1927. A Forest Flora for Kumaon. Govt. Press, United Provinces, Allahabad, pp. 17--22. Pirie, A. and Mullins, M.G., 1976. Changes in anthocyanin and phenolics content of grapevine leaf and fruit tissue treated with sucrose, nitrate and abscisic acid. Plant Physiol., 58 : 468--472. Pomilio, A.B., 1973. Anthochyanins in fruits of Berberis buxifolia. Phytochemistry, 12: 218--220. Reichter, V.E., 1914. Arch. Pharmacol., 2:371--383 (In: K. Paedh and M.V. Traeey, (Editors), Modern Methods of Plant Analysis. Vol. IV, 1955, Springer-Verlag, Berlin, p. 390 Rhodes, M.J.C. and Wooltorton, L.S.C., 1967. The respiratory climateric in apple fruits. The action of hydrolytic enzymes in peel tissue during climateric period in fruit detached from the tree. Phytochemistry. 6 : 1--12. Workman, M., 1963. Color and pigment changes in Golden Delicious and Grimes Golden apples. Proc. Am. Soc. Hortic. Sci., 83: 149--161. Swain, T. and Hills, N.E., 1959. The phenolic constituents of Prunus domestica. I. The quantitative analysis of phenolic constituents. J. Sci. Food Agric., 10: 63--67.