Sugar Accumulation Difference between the Various Sections during

Pineapple Development
M.A. Dou, Y.L. Yao, L.Q. Du, J.G. Li
G.M. Sun and X.M. Zhanga College of Horticulture
South Subtropical Crop Research Institute South China
Chinese Academy of Tropical Agricultural University Guangzhou
Agricultural Science Guangdong 510642
Zhanjiang, Guangdong 524091 China
China

Keywords: pineapple (Ananas comosus), various sections, sucrose, sucrose phosphate

synthase, sucrose synthase

Abstract
Sugar accumulation of ‘Comte de Paris’ pineapple fruits was measured from
20 days after anthesis (DAA) to maturity at 80 DAA. The sugar content was highest
in the basal section of ripe fruit, followed by the medial section, apical section and
fruit core. Changes in activity of acid and neutral invertases (AI and NI respectively)
and sucrose phosphate synthase (SPS) corresponded to changes in sugar
concentration gradients within various sections during fruit maturation. The higher
activity of invertase in the basal section of the fruit likely was associated with rapid
cleavage of sucrose. High invertase activity would also help to form a sucrose
gradient between the leaves and the fruit, which facilitated sucrose transport to the
fruit. Higher SPS activity in the apical section could promote the synthesis of
sucrose, which indicated that the difference between activity of invertase and SPS
resulted in the difference of sugar accumulation among the various sections. Our
results indicated that the SPS and invertase activity was a gradient distribution
among the sections at the mature stage, which was similar with the sugar
concentration, which showed the activities of SS and SPS had a great impact on the
sugar accumulation in the medial and the basal sections of fruit during the period of
70 days after anthesis (DAA). Rapid sucrose accumulation of different sections was
related with the increase of SPS activity and the reduction of NI activity from 70 to
80 d. The activity of SS regulated the sucrose accumulation in the fruit apical and
core section in the period of 70 DAA, However, the activities of AI, NI and SPS
regulated the sucrose accumulation of the fruit core after 70 DAA, whereas the
activities of NI and SPS regulated the sucrose accumulation in the apical, medial and
basal sections of the fruit.

INTRODUCTION
The pineapple sugar content and composition greatly affects fruit flavor, color and
nutrient components. Sucrose, glucose and fructose are the main fruit soluble sugars,
which are not only the basic materials for the synthesis of carotene, vitamins, pigment and
aroma, but also substrates for metabolism and energy supply. Sugars also function as a
signal factor in cell signal transduction. Many studies show that the sucrose metabolic
enzymes are important in fruit sugar metabolism in strawberry (Xie et al., 2007), mango
(Hubbard et al., 1991), peach (Moriguchi, 1992) and citrus (Komatsu et al., 1999).
Pineapple is one of the four most important fruits in tropical and subtropical areas,
and gradually has become an important fruit in the world. Sugar content is an important
component of pineapple quality (Py et al., 1987). Even though the importance of sugar
content in pineapple quality is recognized, our knowledge about sugar metabolism in
pineapple is limited (Chen and Paull, 2000a,b), especially sugar accumulation difference
among fruit sections. Because the pineapple fruit is a syncarp flowering, development and
a
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Proc. 7th International Pineapple Symposium 141

Eds.: H. Abdullah et al.
Acta Hort. 902, ISHS 2011
maturation of the pineapple fruit occurs acropetally. Thus, flowers at the base of the
inflorescence open first followed progressively by others until the apex is reached. And
the development period of the base of the inflorescence is longer than that of the apical
(Okimoto, 1948). Up to now no reports were found on the sugar content difference
between the various parts of the fruit during the 80 days of maturation. Therefore, our
experiment used ‘Comte de Paris’ as a material to study the change of the sugar content
and the sucrose metabolic enzymes to arrive at a better understanding of the sugar
differences in the sections of the pineapple fruit and the main regulation factors.

MATERIAL AND METHODS

Plant Material
Ananas comosus ‘Comte de Paris’ for this study was cultivated in the pineapple
plantation of the South Subtropical Crops Research Institute. In April of 2006 150 well-
developed fruits were selected near the time anthesis was completed full florescence
period. From 20 d after anthesis until the fruit was fully mature as indicated by yellow
coloration of two to three “rows” of fruitlets at the base of the fruit, 5 fruits were
randomly sampled every 10 days. The fruits were cut into apical, medial, basal and core
fractions (Fig. 1), immediately frozen in liquid nitrogen and then transferred into a -80°C
refrigerator for later analysis. Each treatment consisted of 30 fruits with five replicates.

Determinations of Fructose, Glucose and Sucrose Content

Frozen fruit fresh tissue (1 g) was homogenized with 5 ml of deionized water,
incubated at 80 C in a water bath for 15 min, cooled and centrifuged at 10,000 g for
15 min. The residue was re-dissolved with 4 ml deionized water and re-centrifuged at
10,000 g for 15 min, and the two clear supernatant fractions were combined and made to
10 ml volume with distilled water. Individual sugars were quantified by injecting a 10 μl
aliquot, filtered through a 0.45 μm filter, into a high performance liquid chromatograph
(HPLC) (Perkin Elmer Corp., USA) equipped with an automatic pump, an automatic
injector, an analysis column (Series 200, 250×4.6 nm, 5 μm), a differential refractometer
(PE200), and a reporting integrator. The mobile phase was degassed CH3CN:H2O (70:30,
V/V) at a flow rate of 1.0 ml·min-1 and 35 C. Peak height measurements were used to
quantify the soluble sugars by comparing them to the peak height of a standard solution.

Analysis of Enzyme Activities

The activities of the enzymes were analyzed according to the methods of Eduardo
(2001) and Wang et al. (2003), (the related enzymes activity were measured with some
development).

Statistical Analysis
Excel 2003 was used for data collation and mapping, the variance analysis was
done by DPS v3.01 software and the correlation analysis by SAS 9.0.

RESULTS

Change of Sugar Content in the Different Fruit Sections during Development

In general, the sucrose content rapidly increased from 20 or 30 d, but, depending
on the fruit section, dropped slightly between day 40 and day 60. Then, sucrose continued
to accumulate and reached the maximum at the mature stage (Fig. 2A). In the core
section, it rose from 9.43 mg·g-1 FW at 20 d to 34.96 mg·g-1 FW at 40 d. Then, it declined
by 19.7% from 50 to 60 d. After that, it sharply increased and reached a peak at the
mature stage. A comparison of the data for the four parts shows that before day 50,
sucrose accumulated most rapidly and in the greatest amount in the core, while in the
basal fraction rapid accumulation of large amounts of sucrose occurred in the late stage.
At the mature stage, the order of sucrose content was from the basal, medial, core to the

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apical.
Fructose decreased slightly from day 20 to day 40, increased relatively rapidly
from day 40 to day 70 and then declined between day 70 and day 80 in fruit tissue while
levels decreased more or less continuously until day 80 in core tissue (Fig. 2B). With
relatively minor variations, changes in glucose content of tissues were similar to those for
fructose (Fig. 2C).
Changes in the ratio hexose, principally the sum of fructose and glucose, to
sucrose, declined more or less consistently over time in core tissue and fruit basal tissue
(Fig. 3). In medial and apical tissue the ratio increased between day 20 and 30 and then
declined until day 50, increased somewhat at day 60, after which the ratio declined.

Activity of Sucrose Phosphate Synthase (SPS)

The activity of SPS went up slightly between day 20 and day 30, then dropped
slightly, finally sharply increased and reached the maximum in all the sections (Fig. 4).
But the big increment occurred at different periods among the sections, from day 40 to 50
in the medial and basal, after the 70 d in the apical and core. The basal section had the
highest SPS activity, followed by the medial, finally the core and the apical. But there was
no significant difference between the apical and the core.

Activity of Sucrose Synthase (SS)

Sucrose synthase synthetic activity in the base of the fruit was similar to that in the
core (Fig. 5A). The apical and medial section also had a similar change pattern. In the
early stage (before 40 d for the apical section and 50 d for the medial one), there was no
significant change in SS activity after which it quickly rose and reached the maximum at
70 d and finally dropped relatively rapidly. A comparison of the data for the four sections
shows that before the peak, the basal section had highest SS activity, then the core. But
the results for the apical and medial sections was complex. The significant difference
among each part was 10 days before the peak, the order of SS activity was the basal, core,
apical and medial. However, it was the smallest at 10 d after the peak.
The SS cleavage activity in the apical section was consistent with that in the core
(Fig. 5B). In the basal section, cleavage activity increased to a maximum (15.8 μmo1
h-1 g-1 FW) from day 20 to day 70 and then dropped rapidly. In the medial section, change
followed a wave-like pattern. However, at the peak, the difference was relatively small
among the sections. At the mature stage (80 d), SS cleavage activity in the basal section
was significantly different from the medial, apical and core. In addition, no matter which
sections and stages, the synthesis activity of SS was far higher than the cleavage activity.
Activity of the Neutral Invertase (NI)
During the entire period of fruit development, the NI activity among the sections
was different (Fig. 6). Basically, all sections reached a maximum at 40 d, the order of NI
activity decreased in the order basal, core, medial and apical and differences among the
sections were significant. At the maturity, the basal had highest activity, then the medial,
finally the apical and the core, which were not different.

Activity of the Acid Invertase (AI)

Acid invertase activity (Fig. 6) during the whole period of fruit development
basically increased then declined. At 40 d, the apical, basal and core showed a maximum
of AI activity, however the peak of the medial was day 50. After the peak, the apical,
basal and medial dropped to the lowest levels and then increased somewhat at the mature
stage. AI activity in the core reached a peak at day 50. At the mature stage, AI activity in
all sections was maintained at a relatively low level. However, the basal had significantly
higher activity than the other sections.

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DISCUSSION

Sugar Accumulation Difference between Sections

Sugar content was an important index of the fruit quality and directly affected the
fruit flavor. The main soluble sugars in the pineapple fruit included sucrose, fructose and
glucose and these sugars have different accumulation patterns between the fruits, cultivars
and the fruit sections. Zhang et al. (2003) reported that the sucrose accumulation mode
was different between the parts of the muskmelon. The highest accumulation rate was at
the fruit navel, but the lowest at the pedicle flesh. In the whole development, there was a
sucrose content gradient from the navel, central and the pedicle. The research on the
strawberry indicated that at the beginning period, the sucrose, fructose and glucose
content were basically close among the parts. The sugar accumulation started from the
20th day, the top and central part had the highest increment rate. At the mature stage, the
three sugar contents had a little reduction at the basal. The top had highest sugar content,
followed by the central and basal part, which was the result of the high invertase activity
at the top part and the hexose metabolic enzymes activity at the basal part (Xie et al.,
2007). Our data showed that in ‘Comte de Paris’ pineapple fruit development, the pattern
of sucrose, fructose and glucose in the sections was different. At the mature stage, the
sugar content was significantly different among the sections (Fig. 2). The basal and
medial sections had higher sugar contents than the apical and core tissues, which was
consistent with the results of Siders and Krauss (1933) but different from the muskmelon
(Zhang et al., 2007) and strawberry (Xie et al., 2007). The difference between the syncarp
fruit and single fruit resulted in the different sugar accumulation and metabolic model
between the pineapple and the tomato (Qi et al., 2006), citrus (Zhao et al., 2001) and
muskmelon (Zhang et al., 2003). The pineapple fruit is a syncarp formed by the fusion of
individual fruits produced by each flower and the basal segment is more mature than the
middle portion, and this in turn is more mature than the top segment (Miller and Hall,
1953). Therefore, the basal segment accumulated more sugar than the medial and apical
segment.
Our results indicated that the sugar composition was different between the sections
and the different parts accumulated different kinds of sugar, which was consistent with
the result on tomato (Qi et al., 2006), strawberry (Xie et al., 2007), citrus (Zhao et al.,
2001) and muskmelon (Zhang et al., 2003).

Sugar Accumulation Difference at the Sections Related to the Sucrose Metabolic

Enzymes
In the pineapple fruit, the sucrose was the main transport form of photosynthetic
products. The sucrose input rate was regulated by the sucrose concentration gradient
between the source and the sink. The sucrose metabolic enzymes played an important role
in the sucrose concentration and sucrose input. The acid and neutral invertases, which are
able to decompose sucrose, had low activities and a gradient distribution among the
sections at the mature stage which was similar to data for sugar concentration. At the
same time, the basal and the medial section had higher AI and NI than the apical (Fig. 6).
It suggested that there was the sucrose gradient between the apoplast and the phloem at
the basal by the high invertase activity, which was conducive to the transport of
photosynthetic product into the fruit, and resulted in the sugar accumulation. The study on
strawberry also indicated that the invertase regulation was the main factor for the sugar
concentration gradient (Xie et al., 2007). The invertase played a role in the sink strength
regulation, kept the concentration gradient between the sink cell and the phloem, and
promoted the sugar input from the source. Plants that mainly accumulate sucrose, such as
golden pear (Li et al., 2007), strawberry (Hubbard et al., 1991), navel orange (Liu et al.,
2003), litchi (Wang et al., 2003), bayberry (Xie et al., 2005), had relatively low acid
invertase activity during the fruit development. The decline and disappearance of acid
invertase was a prerequisite for sucrose accumulation of these plants. However, whether
the hexose metabolic enzymes regulated the sugar content among sections, we will wait

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for further research.
SPS activity in pineapple was also related to fruit sugar accumulation. During fruit
development, the change of SPS was basically accompanied with the sucrose
accumulation, it was gradually increasing. The high SPS activity benefitted sugar
accumulation, which affected the different sugar accumulation among the sections.
Similar findings reported that the sucrose accumulation paralleled the SPS activity of
Citrus unshiu (Komatsu et al., 1999), and pear (Moriguchi et al., 1992). In the mature
stage, the SS in pineapple fruit was mainly synthetic activity, but differences among the
sections were not significant, which indicated that the SS was not closely related to the
sugar content among the sections.
Thus, the change of invertase and sucrose phosphate synthase was the main reason
for the sugar accumulation difference among the sections. Therefore decreasing invertase
activity and increasing sucrose phosphate synthase activity were the main intrinsic
regulation factors of pineapple fruit sucrose accumulation in the different parts.

ACKNOWLEDGEMENTS
Financial support was provided by the Guangdong Natural Science Fund of China,
the Hainan Natural Science Fund of China (808182), Special Fund for Agro-scientific
Research in the Public Interest (3-41), Fund on Basic Scientific Research Project of
Nonprofit Central Research Institutions (200701).

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Figures

Fig. 1. Sampling sections in different fruit tissues.

146
 70.00 40.00
apical section
A a 35.00
60.00 b

F ru cto s e C o n te n t （ m g .g- 1 F W ）
Medial section
c 30.00 B
S u cro s e C o n ten t （ m g .g- 1 F W ）

50.00 Basal section

25.00 a
Core section d
40.00
20.00 b
30.00 c
15.00
20.00
10.00
d
10.00 5.00

0.00 0.00
20 30 40 50 60 70 80 20 30 40 50 60 70 80

45.00

40.00
G lu co s e C o n ten t （ m g .g- 1 F W ）

35.00
C
30.00

25.00 a

20.00 b
c
15.00

10.00 d

5.00

0.00
20 30 40 50 60 70 80

Days after anthesis(d)

Fig. 2. Changes in sucrose (A), fructose (B) and glucose (C) content in different sections
of ‘Comte de Paris’ pineapple fruit during the period from 20 to 80 d after anthesis.

147
 4.5 apical section

4 M edial section

3.5 Basal section

Hexose to sucrose ratio

Core section
3

2.5

1.5

0.5

0
20 30 40 50 60 70 80
Days after anthesis(d)

Fig. 3. Changes of the ratios of hexose to sucrose in various sections of ‘Comte de Paris’
pineapple fruit during the period from 20 to 80 d after anthesis.

25

a
Apical section
20
SPS activity(μmol·g-1h-1FW)

Medial section b

Basal section c
15
Core section

10

0
20 30 40 50 60 70 80
Days after anthesis(d)

Fig. 4. Changes in sucrose phosphate synthase (SPS) activity in various sections of

‘Comte de Paris’ pineapple fruit during the period from 20 to 80 d after anthesis.

148
 100 100
A
90 90 B
Sucrose synthase activity （μmol·g-1 h -

Apical section
80 Medial section 80
70 Basal section 70
60 Core section 60
FW）

50 50
a
1

40 40
30 30
20 20
a
10 10 b
0 c
0
20 30 40 50 60 70 80 20 30 40 50 60 70 80
Days after anthesis（d） Days after anthesis(d)

Fig. 5. Changes in sucrose synthase synthetic activity (A) and cleavage activity (B) in
various sections of ‘Comte de Paris’ pineapple fruit during the period from 20 to
80 d after anthesis.

100 140
Apical section Apical section
90 Medial section Medial section
120
80 Basal section Basal section
N I a c tiv ity (μ m o l·g - 1 h - 1 F W )

A I activ ity (μ m o l· g - 1 h - 1 )

70 Core section 100 Core section

60
80
50

40 60

30 40
a
20 a
b
20
10 c b
c
0 0
20 30 40 50 60 70 80 20 30 40 50 60 70 80
Days after anthesis(d) Days after anthesis(d)

Fig. 6. Changes in neutral (NI) and acid invertase (AI) activity in various sections of
‘Comte de Paris’ pineapple fruit during the period from 20 to 80 d after anthesis.

149
150